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Functional Analysis of VvARF18 Gene in Red Globe Grape YUAN Miao, ZHOU Juan, DANG ShiZhuo, TANG XueShen, ZHANG YaHong Scientia Agricultura Sinica    2024, 57 (7): 1363-1376.   DOI: 10.3864/j.issn.0578-1752.2024.07.012 Abstract （30）   HTML （4）    PDF （3353KB）（33）       Knowledge map    Save 【Objective】Auxin response factor (ARF) is a significant regulatory factor in the auxin signaling pathway and plays an important role in plant growth and development as well as various physiological processes. Analysis of the Red Globe grape VvARF18 promoter, heterologous expression, endogenous hormone content and its expression in response to hormones was made in order to explore the mechanism of VvARF18 gene in the auxin (IAA) signaling pathway and flower bud differentiation process in Red Globe grapes. 【Method】The VvARF18 gene sequence was obtained by homologous cloning by using facility Red Globe grape flower buds as experimental materials. The cis-acting elements of the promoter were analyzed using the online database PLACE. The plant overexpression vector pC2300-VvARF18 was constructed based on the pCAMBIAI2300 plant expression vector by double enzyme digestion and homologous recombination method. The recombinant vector pC2300-VvARF18 was transformed into Agrobacterium tumefaciens strain GV3101 by using electrical shock method. The tobacco leaves were used as explants and transferred into tobacco by Agrobacterium-mediated callus transformation method, and positive transgenic seedlings were obtained by PCR. The quantitative real-time PCR (qRT-PCR) was used to analyze the expression level of VvARF18 transgenic tobacco lines, and the transgenic lines with high expression level were screened and cultured to T3 generation, and treated with IAA and GA3 to analyze the expression level of VvARF18. The content of IAA, GA, ABA and CTK in flower buds and leaves of transgenic tobacco were determined by enzyme-linked immunosorbent assay. 【Result】VvARF18 of Red Globe grape located on chromosome 13, and contained 3 exons and 2 introns. There are multiple cis-acting elements in the VvARF18 promoter region that respond to light, plant hormones, and stress. The phenotypic analysis found that the process of flower bud differentiation was faster in transgenic tobacco than in wild-type tobacco. The qRT-PCR results showed that the expression of VvARF18 showed an increasing and then decreasing trend during the four periods of flower bud development in transgenic tobacco, and the highest expression level was reached in the S3 stage. The results of IAA, CTK, GA and ABA determination in flower buds and leaves of transgenic tobacco plants showed that the content of four plant hormones in flower buds and leaves of transgenic tobacco plants were higher than those of wild-type plants. The change trend of GA/IAA during the four periods of transgenic tobacco flower bud development were consistent with the expression trend of VvARF18. The expression level of VvARF18 in transgenic tobacco plants treated with IAA and GA3 decreased with the increase of IAA treatment concentration and also decreased with the extension of GA3 treatment time. 【Conclusion】Grape VvARF18 negatively regulated auxin to participate in the process of plant flower bud differentiation, which could interact with key factors in the gibberellin signaling pathway to synergistically regulate hormone levels in plant flower buds and had a facilitative effect on plant flower bud differentiation. Table and Figures | Reference | Related Articles | Metrics

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Effect of Methyl Jasmonate on Energy Metabolism and Membrane Lipid Metabolism During Resistance to Botryosphaeria dothidea in Kiwifruit XIAO LiuHua, KANG NaiHui, LI ShuCheng, ZHENG ZhiYuan, LUO RaoRao, CHEN JinYin, CHEN Ming, XIANG MiaoLian Scientia Agricultura Sinica    2024, 57 (7): 1377-1393.   DOI: 10.3864/j.issn.0578-1752.2024.07.013 Abstract （32）   HTML （3）    PDF （4553KB）（27）       Knowledge map    Save 【Objective】Soft rot caused by Botryosphaeria dothidea is one of important diseases during kiwifruit storage, which causes serious economic losses for kiwifruit industry. Methyl jasmonate (MeJA) is a kind of biological signal molecule widely existing in plants, which had been found that it could effectively induce kiwifruit’s resistance to B. dothidea in previous studies. In order to further investigate the mechanism of MeJA-mediated resistance to disease in postharvest kiwifruit, the effects of MeJA on energy metabolism and membrane lipid metabolism were analyzed in this study. 【Method】 Hongyang kiwifruit (Actinidia chinensis cv. Hongyang) were used as experiment material and divided into three groups as follows: inoculation group, which the fruits were inoculated with B. dothidea without MeJA treatment; MeJA+inoculation group, which the fruits were fumigated with 0.1 mmol·L-1 MeJA for 24 h then inoculated with B. dothidea; control group, the fruits without MeJA treatment or inoculation with B. dothidea. All fruits were stored in an incubator ((20±1) ℃, 90%-95% relative humidity) for 8 d. The reactive oxygen species (ROS) accumulation was analyzed by 2, 7-dichlorodihydrofluorescein acetoacetate (H2DCFDA) method, malondialdehyde (MDA) content and relative conductivity were also measured. Meanwhile, phospholipase D (PLD), lipase (LPS), lipoxygenase (LOX), succinate dehydrogenase (SDH), malate dehydrogenase (MDH), H+-ATPase, Ca2+-ATPase and cytochrome C oxidase (CCO) activities and the related genes expression were analyzed. The content of Adenosine triphosphate (ATP), Adenosine diphosphate (ADP), Adenosine monophosphate (AMP), oxaloacetic acid and fenugreek acid were determined by high performance liquid chromatography (HPLC). Then the correlation between membrane lipid metabolism and energy metabolism of kiwifruit in inoculation group and MeJA+ inoculation group was analyzed. 【Result】Compared with the inoculated group, MeJA treatment could inhibit the excessive accumulation of ROS in inoculated B. dothidea fruits, slowed down the degree of membrane lipid peroxidation, and effectively decreased PLD, LPS, and LOX activities and the expression levels of AcLOXs, AcPLD and AcLPS. However, MeJA increased the activities of SDH, MDH, H+-ATPase, Ca2+-ATPase and CCO and the transcription levels of AcSDH, AcMDH, AcH+-ATPase, AcCa2+-ATPase and AcCCO, promoted the production of ATP, ADP, oxaloacetic acid and fumaric acid, delayed the decline of fruit energy charge then ensured the energy required for fruit resistance to pathogens. In addition, correlation analysis showed that kiwifruit membrane lipid metabolism was positively correlated with ROS accumulation, but negatively correlated with energy charge. 【Conclusion】The above results indicated that MeJA-induced resistance to B. dothidea in kiwifruit was related to its regulation of fruit energy levels and mitigation of ROS-involved membrane lipid metabolism. Table and Figures | Reference | Related Articles | Metrics

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Research on Leaf Nutritional Diagnosis of Liaoning 1 Walnut in Taihang Mountain Area of Hebei Province AN XiuHong, SUN Yan, WANG Fang, FENG QiKe, WANG Ning, LI JinJin, ZHANG JunPei, WANG HongXia Scientia Agricultura Sinica    2024, 57 (6): 1153-1166.   DOI: 10.3864/j.issn.0578-1752.2024.06.010 Abstract （94）   HTML （7）    PDF （689KB）（70）       Knowledge map    Save 【Objective】 The mineral element content in leaves of Liaoning 1 walnut in Taihang Mountain area of Hebei Province were determined and the nutritional status of leaves were diagnosed, so as to provide the basis for scientific fertilization for Liaoning 1. 【Method】 The content of mineral elements in Liaoning 1 leaves at different developmental stages were measured to determine the sampling time of leaf nutritional diagnosis. Component nutritional diagnosis (CND) method was used to establish the standard values of element contents for the nutritional diagnosis of Liaoning 1. The diagnosis and recommendation integrated system (L-DRIS), CND, and deviation from optimum percentage (DOP) methods were used to diagnose the leaf nutritional status in low-yielding orchards to provide fertilization strategies. 【Result】 The suitable period for leaf nutritional diagnosis of Liaoning 1 in Taihang Mountains of Hebei province from August to September. Correlation analysis showed that the mineral elements in Liaoning 1 leaves from Taihang Mountain area mainly had synergistic effects, and there was strong antagonistic interaction between K and Mg. Based on the CND analysis, the yield of 3.64 t∙hm-2 was selected as the yield cutoff value of high-yielding orchards. The optimal values of mineral element content determined by high-yielding orchards were N (13.04-17.04) g∙kg-1, P (0.67-1.23) g∙kg-1, K (5.94-10.64) g∙kg-1, Ca (15.64-22.16) g∙kg-1, Mg (3.80-6.62) g∙kg-1, Cu (5.14-8.48) mg∙kg-1, Fe (437.38-794.58) mg∙kg-1, Mn (126.94-172.02) mg∙kg-1, and Zn (14.59-34.25) mg∙kg-1. There were differences in the abundance and deficiency of mineral elements in Liaoning 1 leaves of low-yielding orchards obtained through L-DRIS, CND, and DOP methods, and the diagnosis results of L-DRIS and CND methods were more consistent. The fertilization order of low-yielding orchards in Taihang Mountain of Hebei province was K>P>N>Zn>Mn>Ca>Fe>Mg>Cu. 【Conclusion】 It was suggested that K should be the main fertilizer, combined with N and P, while excessive applications of Mg and Cu should be avoided in low-yielding orchards. Table and Figures | Reference | Related Articles | Metrics

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Effects of Different Ratios of Green-Blue Light on Basil Growth and Its Energy Use Efficiency YANG Yang, JIA MengHan, CHEN Can, ZHANG YiHan, TONG YuXin Scientia Agricultura Sinica    2024, 57 (6): 1167-1179.   DOI: 10.3864/j.issn.0578-1752.2024.06.011 Abstract （70）   HTML （8）    PDF （587KB）（52）       Knowledge map    Save 【Objective】 This study aimed to investigate the effects of different ratios of green-blue light (G/B) with constant red light (R, 660 nm) on the growth, stomatal characteristics, photosynthetic capacity, and energy utilization efficiency of basil plants (Ocimum basilicum Linn.). The ultimate goal of this study was to provide the technical support for efficient and energy-saving production of basil plants in plant factory. 【Method】 Basil seedlings were subjected to seven different light quality treatments of RB (R:G:B=8:0:8, set as control), RG1B7 (R:G:B=8:1:7), RG1B3 (R:G:B=8:2:6), RG1B1 (R:G:B=8:4:4), RG3B1 (R:G:B=8:6:2), RG7B1 (R:G:B=8:7:1), and RG (R:G:B=8:8:0) for a duration of 19 days after transplanting. The total light intensity was 160 μmol∙m-2∙s-1, with red light accounting for 50% of the total light intensity. The photo/dark period was 16/8 hours per day. The air temperature during the light/dark period was set at 25 ℃/23 ℃, and the relative humidity was set at 65%. Additionally, the CO2 concentration during the light period was 1 000 μmol∙mol-1. The photosynthesis parameters of basil plants were determined after 13 days of experimental treatment, while the growth parameters, stomatal characteristics, and energy use efficiency of basil plants were determined after 19 days of experimental treatment. 【Result】 Except for treatment RG1B3, the addition of green light significantly enhanced height, leaf area, and dry and fresh weight of basil plants. The shoot dry and fresh weights of basil plants under different G/B treatments showed an increase of 19%-51% and 26%-64%, respectively, compared with those under the control. However, no significant differences in leaf area and dry/fresh weights were observed among the G/B treatments, except those under treatment RG. The treatment with low G/B (RG1B7) only caused a reduction in the stomatal density on the abaxial surface of basil lower leaves. As G/B increased, the stomatal density decreased on the adaxial surface of the lower leaves and both surfaces of the upper leaves. In general, the leaf nitrogen content decreased with an increase in the G/B. This decrease in leaf nitrogen content and stomatal density resulted in a decline in CO2 assimilation capacity and light utilization capacity of basil plants. The reduction in these capacities generally increased with the increasing G/B. Due to the lower light-emitting efficiency of green LEDs compared with blue light, the total power consumption increased with an increase in the G/B. Only the treatment RG1B7 significantly improved electric use efficiency by 25% than that under the control. With the exception of RG1B3, all other G/B treatments improved light utilization efficiency by 30%-57% than that under the control, and no significant differences were found among different G/B treatments, except for RG. RG3B1 and RG improved water use efficiency of both upper and lower basil leaves, with RG3B1 showing a 58% and 74% increase, and RG showing a 67% and 90% increase, respectively compared with that under the control. 【Conclusion】 By comprehensively analyzing, basil plants growth and energy utilization efficiency, the RG1B7 could be considered as a more suitable light quality combination for basil plants production in plant factories. The findings of this study could serve as a technical support for understanding the impact of different G/B ratios on basil growth, photosynthetic characteristics, and energy utilization efficiency. This research provided the valuable insights for developing optimized light formulations that enable efficient and energy-saving production of basil plants in plant factory environments. Table and Figures | Reference | Related Articles | Metrics

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Light Interception Rate and Canopy Structure Optimization of Korla Fragrant Pear YAN Pan, WANG ZhenDong, DENG YongHui, CHEN QiLing, ZHENG QiangQing Scientia Agricultura Sinica    2024, 57 (5): 965-979.   DOI: 10.3864/j.issn.0578-1752.2024.05.011 Abstract （108）   HTML （9）    PDF （5432KB）（62）       Knowledge map    Save 【Objective】 This study investigated the canopy structure of five tree shapes of Korla fragrant pears, analyzed the differences in light distribution within different tree shapes, established a daily change model of light interception, calculated the light interception rate, clarified the regulation methods and target parameters of the canopy structure, to provide a reference basis for cultivating high-light-efficiency tree shapes. 【Method】 Korla fragrant pear trees with different shapes were chosen as the test materials, the canopy structural parameters and photosynthetic-effective-radiation (PAR) passing through different tree shapes were measured. Based on a quadrant approach, a three-dimensional space was constructed, the light distribution maps were drawn, an evaluation index system of light interception was established, and the light interception amount and light interception rate were calculated. Through correlation analysis and principal component analysis, the main canopy structure and regulation methods that affect light interception were identified. 【Result】 (1) The ground diameter and average branch diameter of large cylindrical-shaped trees were larger, with main branches with imbalanced branch-to-trunk ratios. The number of branches and branch diameter through the canopy of the cylindrical-shaped trees were more uniform. The number of branches, total length of branches, and average branch length of the narrow cylindrical-shaped trees were significantly lower than those of the cylindrical-shaped trees. The height of trees with the short cylindrical-shaped trees was significantly lower and other parameters were similar to the narrow cylindrical-shaped trees. The ground diameter, average branch length, average branch diameter, average branch angle, and average distance of branches of slender- spindle-shaped trees were significantly lower than those of cylindrical-shaped trees, but the proportion of short branches was significantly larger. (2) The distance from the trunk and height were the main factors which affected the canopy light interception (LI), and the average light interception(ALI) at a distance of 100 cm from the trunk increased significantly, reached 572 μmol·m-2·s-1, which was approximately twice that of the inner chamber. The lighting conditions below 220 cm were poor. The south, north, southeast, and southwest sides were high-light areas, while the west, east, northeast, and northwest sides were low-light areas. The daily variation in ALI can be roughly divided into five periods. (3)As the tree shape became narrower and shorter, the ALI significantly increased in the inner chamber, in each or partial layer, in all or partial directions, and in partial periods. (4) The single-day cumulative light interception (CLI) of slender-spindle-shaped trees was 22.2 mol·m-2, the group CLI was 3 712 mol/667 m2, and the light interception rate (LIR) was 35.6%, which was significantly higher than that of other tree shapes. The proportion of low-light area (PLL) was 50.9%, which was significantly lower than that of other tree shapes. (5) Five canopy structural parameters were significantly positively correlated with the LIR while fifteen canopy structural parameters were significantly negatively correlated. 【Conclusion】 The proportion of short branches is the most important parameter affecting the light interception in Korla fragrant pear trees, while the length of branches is the most important parameter affecting the proportion of low-light area. Control the tree height and canopy width can improve the light interception rate and the uniformity of light distribution. The slender-spindle-shaped canopy has a more uniform light distribution and the highest light interception rate. It can maintain a larger number of branches and a larger proportion of short branches with reduced average branch length to improve the lighting level of the canopy. Table and Figures | Reference | Related Articles | Metrics

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Isolation and Functional Verification of Genes Mediating Mineral Element Stress Tolerance in Lotus HU HengLiang, GU TianYu, CHEN SiYing, WANG Yao, PENG JiaShi Scientia Agricultura Sinica    2024, 57 (5): 980-988.   DOI: 10.3864/j.issn.0578-1752.2024.05.012 Abstract （53）   HTML （5）    PDF （2538KB）（43）       Knowledge map    Save 【Objective】 Lotus (Nelumbo nucifera) is a traditional edible and medicinal homologous crop in China, yet its abundant genetic resources lack functional characterization. This research aimed to functionally identify genes involving in the accumulation/tolerance of trace mineral nutrients and heavy metal elements in lotus, thereby accumulated gene resources that facilitated increased nutrient efficiency and stress tolerance, so as to provide a theoretical basis for the genetic improvement of lotus. 【Method】The representative Xianglian lotus variety, Cunsanlian was used as the experimental material. Initially, a yeast expression cDNA library of lotus was constructed and transformed into Saccharomyces cerevisiae. It was then screened on plates containing excessive cadmium (Cd), manganese (Mn), zinc (Zn), copper (Cu), iron (Fe), and aluminum (Al) stress to isolate target genes mediating stress tolerance in positive yeast clones. Finally, the functional verification of the selected tolerance genes was conducted through combining bioinformatics analysis and yeast complementation verification. 【Result】A yeast cDNA library of lotus was constructed with a capacity exceeding 106 yeast monoclonal clones, a recombination rate of 100%, and an average insertion fragment size greater than 1 000 bp. Following the screening on stress plates, 13 genes for Cd tolerance, 4 for Mn tolerance, 4 for Zn tolerance, 3 for Cu tolerance, 7 for Fe tolerance, and 1 for Al tolerance were identified. Among these, 3 genes were able to mediate tolerance to both Fe and Mn. These genes were distributed on all lotus chromosomes except chromosome 6.【Conclusion】A high-quality yeast expression cDNA library of lotus was constructed, and screened 29 genes mediating resistance to excess trace mineral nutrients or heavy metal elements, accumulating gene resources for lotus genetic improvement for enhanced nutrient efficiency and the prevention of heavy metal accumulation. Table and Figures | Reference | Related Articles | Metrics

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Observation of Flower Bud Differentiation Process and Fitting of Flower Growth Model of Passion Fruit TIAN QingLan, ZHOU JunNiu, WU YanYan, LIU JieYun, HUANG WeiHua, ZHANG YingJun, XIE WenLian, WEI GuangTan, MOU HaiFei Scientia Agricultura Sinica    2024, 57 (4): 765-778.   DOI: 10.3864/j.issn.0578-1752.2024.04.011 Abstract （134）   HTML （10）    PDF （6115KB）（82）       Knowledge map    Save 【Objective】 This study aimed to elucidate the flower bud differentiation process in the primary passion fruit (Passiflora edulis) varieties within Southern China’s ecological region. Additionally, it sought to establish a flower growth model depicting the morphological differentiation of passion fruit flower buds, so as to serve as a reference for promoting and retenting passion fruit flowers.【Method】Tainong No. 1 (TN) as the predominant cultivar of the purple fruit variety, and Zhuangmi 05 (also known as Zhuangxiangmibao, MB) as the main cultivar of the yellow fruit variety in Guangxi, China, were utilized as experimental materials. The scanning electron microscopy and paraffin section dissection were employed to observe flower bud differentiation process in these varieties. The differentiation process of nodes below the shoot apical meristem, and measured the first bract length of flower buds, leaf length, and tendril length at each node were observed too. The 7th node below the shoot apical meristem was marked as the initial marker node, and this marked the onset of the first bract differentiation phase of the flower bud. The flower bud differentiation from 0 to 16 days after marking were investigated, as well as the dynamic changes in five flower morphological indicators, such as the length of the first bract and bud from marking to flowering were tracked, for facilitating the construction of a flower growth model.【Result】The primary process of flower bud differentiation in the tested passion fruit varieties encompassed first bract formation stage, additional bract formation stage, sepal formation stage, petal formation stage, stamen formation stage, pistil formation stage, corona formation stage, etc. The duration from first bract formation to pistil formation spanned 10-12 days, with TN exhibiting 1-2 day advancement over MB. Pistil formation emerged as a vital marker for successful passion fruit flower bud development, characterized by a 3-4 mm first bract length. The duration from first bract formation to flowering ranged between 36-44 days, with TN progressing 3-4 days earlier than MB. Tendril differentiation was visible at the 4th-5th nodes below the shoot apical meristem, flower primordium differentiation at the 6th node, appearing alongside the tendril, first bract differentiation at the 7th node, vegetative meristem appearing at the 9th node, which were positioned separately on the inner side of the tendril and flower bud, axillary vegetative shoot formation at the 10th-11th nodes, and pistil primordium differentiation at the 14th-15th nodes. The non-linear regression fitting of a Logistic model in Origin software was performed for the first bract length, width, pedicle length, flower bud length, length of flower bud and pedicle along with days after making for both MB and TN, and the results showed the determination coefficient R2 ranged from 0.9524 to 0.9988, and the normalized root mean square error (nRMSE) was between 8.54% to 19.62%, indicating a good fit of the model equation. Based on the model parameters and actual observations, the length of bract entered a rapid growth period 11-12 days after marking (i.e. after the first bract formation stage) and stabilized 24-26 days after marking. Subsequently, the pedicels and sepals grew rapidly, with the sepals surpassing the bracts length, and the total length of the flower bud and pedicel entered a rapid growth phase 24-25 days after marking, blooming after reaching maximum growth at 41-42 days after marking.【Conclusion】The flower buds of passion fruit were closely associated with tendrils, and independent of nutritional buds. The flower bud differentiation and morphological differentiation of passion fruit could be divided into three stages: the first bract formation period to the pistil formation period, lasting 10-12 days; the bract growth period, lasting 12-14 days; and the flower buds and pedicels growth period, lasting 15-17 days. In actual production of passion fruit, the flowering time could be predicted using the flower growth model, and the flower bud differentiation process could be assessed using morphological indicators, such as bract length, which provided a reference basis for promoting flowering and promoting flowers. Table and Figures | Reference | Related Articles | Metrics

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Effects of Tomato Grafting and Nitrogen Fertilization on Fertilizer Nitrogen Fate and Nitrogen Balance SUN ZhaoAn, ZHANG YiWen, JIANG LiHua, LI ZhaoJun, GUO Xin, CAO Hui, MENG FanQiao Scientia Agricultura Sinica    2024, 57 (4): 755-764.   DOI: 10.3864/j.issn.0578-1752.2024.04.010 Abstract （93）   HTML （5）    PDF （1022KB）（48）       Knowledge map    Save 【Objective】 By quantifying the amounts of soil-derived nitrogen (N) uptake by tomato aboveground and residual fertilizer N in soil, this study evaluated the impacts of tomato grafting and N fertilization on the fertilizer N fate, net residual fertilizer N, and N balance in soil-tomato production.【Method】A pot experiment with 15N-labeled urea and the experiment were conducted. Tomato varieties were Qidali and 017, included grafting (grafting and self-rooted), and fertilization (control and N-fertilization) treatments. The 15N tracer was used to distinguish the contribution of fertilizer- and soil-derived N in plants and soils, the fertilizer N fate and the added-N interaction (ANI, i.e. the difference of soil-derived N uptake between N-fertilized and -unfertilized treatments), and the N balance in soil-tomato production was also evaluated.【Result】The contribution of fertilizer N to whole plant N uptake was 35.9%-38.8%, and the contribution to aboveground plant N uptake (35.9%-39.9%) was higher than that in root N uptake (31.6%-36.2%). The ANI exhibited positive values in most treatments, and there was no significant impacts from grafting. Under all the treatments, the average fertilizer N allocated to aboveground, soil and loss was 4.0﹕2.6﹕3.4, and the total recovery of fertilizer N (aboveground N uptake + soil residue) was about 70%. At the fertilizer N level of 250 kg·hm-2, the residual fertilizer N in the soil could not compensate for the soil-derived N allocation to tomato aboveground, and this might cause the depletion of soil N stock in the long run. 【Conclusion】The risk of fertilizer N loss was high if large rate of N fertilization was chosen compensate the consumption of soil native N. Compared with tomato 017 scion and self-rooted treatment, the combination of tomato Qidali scion and pumpkin rootstock increased the fertilizer N stay in soil and reduced the fertilizer N loss. Appropriate tomato scion and rootstock could be an feasible and effective measure to maintain the soil N fertility in tomato production. Table and Figures | Reference | Related Articles | Metrics

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Effects of Wheat and Common Vetch Cover Crops on Chinese Cabbage Seedling Growth and Soil Microbial Community Structure WANG QingHui, LI NaiHui, ZHANG YiPing, DI ChengQian, WU FengZhi Scientia Agricultura Sinica    2024, 57 (3): 555-569.   DOI: 10.3864/j.issn.0578-1752.2024.03.010 Abstract （86）   HTML （17）    PDF （1516KB）（90）       Knowledge map    Save 【Objective】 This study aimed to investigate the impact of cover crops (wheat and common vetch) on the growth of Chinese cabbage seedlings and the structure of the soil microbial community. The findings could provide the theoretical and technical support for using wheat and common vetch cover crops to alleviate continuous cropping obstacles in Chinese cabbage production. 【Method】In this experiment, there were four treatments: wheat cover crop treatment (W), common vetch cover crop treatment (P), mixture of wheat and common vetch treatment (WP), and no cover crop treatment (CK). The effects of different cover crop treatments on the growth of Chinese cabbage seedlings were studied. Moreover, the effects of wheat and common vetch cover crops on the microbial community of Chinese cabbage rhizosphere were also investigated through qPCR and Illumina MiSeq techniques. In addition, Spearman correlation analysis was conducted to identify the key soil microbial taxa related to Chinese cabbage growth. Then, the changes in soil chemical properties on soil microbial community structure were explored by environmental factor correlation analysis. 【Result】Compared with CK, the cover crop treatments had positive effects on Chinese cabbage growth and decreased soil electrical conductivity (EC) value. The mixed cover crop treatment significantly decreased soil available potassium content, whereas wheat cover crop treatment increased soil pH. The qPCR results showed that the abundance of soil bacterial community was not significantly affected by the cover crop treatment, but increased the abundance of soil fungal community. Both common vetch cover crop treatment and mixed wheat and common vetch cover crop treatment significantly reduced the abundances of Bacillus spp. and Pseudomonas spp. communities. The Illumina MiSeq analysis showed that the relative abundance of genus TM7a was significantly increased by cover treatments, while the relative abundances of Leptolyngbya_EcFYyyy-00, Lophotrichus, Acaulium, and Sodiomyces were decreased. The mixed cover crop treatment significantly increased the relative abundance of Sphingomonas and Massilia and significantly decreased the relative abundance of Fusarium. Spearman correlation analysis showed that Sphingomonas, TM7a, Massilia, and Gemmatimonas were positively correlated with growth. Leptolyngbya_EcFYyyy-00, Acaulium, Lophotrichus, Sodiomyces, and Fusarium were significantly negatively correlated with the growth of Chinese cabbage. Moreover, these cover crop treatments influenced bacterial and fungal diversity indices. The Shannon index and inverse Simpson index for soil bacterial community and Shannon index of soil fungal community significantly decreased in cover common vetch treatment. In contrast, the inverse Simpson index of soil fungal community was increased. The mixed cover crop treatment increased the fungal Shannon index, while the inverse Simpson index of soil fungal community decreased. Principal Coordinates Analysis (PCoA) showed significant differences in soil microbial community structure, with soil EC value as a major environmental factor affecting the structure.【Conclusion】The cover crop treatments exhibited growth-promoting effects on Chinese cabbage seedlings, and the best effect was found in the mixed cover crop treatment. The relative abundances of some Sphingomonas, TM7a, Massilia, other potential growth-promoting bacteria were increased in the cover crop treatments. The relative abundances of some potential plant pathogens Leptolyngbya_EcFYyyy-00 and Fusarium were decreased and the relative abundance of potential biocontrol agent Chaetomium was increased in the mixed cover crop treatment. Table and Figures | Reference | Related Articles | Metrics

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Physiological Response of Potted Tomatoes to NaCl and Na2SO4 Brackish Water Irrigation PEI ShuYao, CAO HongXia, ZHANG ZeYu, ZHAO FangYang, LI ZhiJun Scientia Agricultura Sinica    2024, 57 (3): 570-583.   DOI: 10.3864/j.issn.0578-1752.2024.03.011 Abstract （84）   HTML （4）    PDF （702KB）（55）       Knowledge map    Save 【Objective】Brackish water irrigation is one of the important means to increase irrigation water sources and to alleviate the shortage of agricultural water in arid areas. However, unreasonable irrigation water quality can severely limit the physiological activity and growth of plants. Carrying out research on the effects of different salt types of brackish water irrigation on the physiological changes of tomato leaves is conducive to reveal the mechanisms of salt tolerance to different types of salt in the salt-sensitive crop tomato at the physiological level, which is of great significance to agricultural production as well as to the use of brackish water for water conservation and salt control. 【Method】In this study, tomato was used as an object of study in a brackish water irrigation pot experiment, and the two factors of irrigation water salt type (NaCl (T1) and Na2SO4 (T2)) and salinity (0, 1.5 (S1), 3.0(S2), 4.5 (S3) and 6.0 (S4) dS·m-1) were set to analyze the changes of physiological indexes, such as leaf gas exchange parameters, osmotic and antioxidant physiological regulation, and ionic balance, in tomato plants subjected to different types and degrees of stress at different reproductive periods. The reasons for the differences in the degree of decline in photosynthetic capacity of tomato under NaCl and Na2SO4 stress were explored too. 【Result】Brackish water irrigation caused significant decreases in leaf net photosynthetic rate (Pn), stomatal conductance (Gs), and transpiration rate (Tr) in the late stage of fertility (mature picking stage) and high salinity (S4) treatments compared with CK, and the contents of proline (pro), soluble sugar (SS), malondialdehyde (MDA), and leaf Na+ increased continuously during the salt stress and the progression of the fertility period (P<0.05). The superoxide dismutase activity (SOD) showed a trend of increasing and then decreasing with increasing salinity in the late reproductive stage. The highest decreases in Pn, Gs, and Tr could be up to 44.13%, 64.53%, and 33.75%, respectively, whereas the increases in pro, SS, MDA, and Na+ could be up to 2.31, 0.77, 0.55, and 5.81 times higher than that of CK, respectively, all of which achieved under T1 stress. The correlations of SS, SOD and K+/Na+ with Pn were significantly changed under the two salt stresses, in which the slopes of the regression lines of SS and Pn were significantly higher under T1 treatment than T2 (P<0.05), the slopes of the regression lines of SOD and Pn were significantly lower under T1 treatment than T2 (P<0.05), and the regression curves of K+/Na+ and Pn showed that the T1 curves were relatively leftward. The results of principal component analysis showed that, under T1 treatment, SOD activity value was higher, which had an important role in Pn stabilization, but it was suppressed in the late reproductive stage, and could only enhance the water use efficiency to a certain extent; under T2 treatment, the physiological indexes were less stressed, in which the SS accumulation was related to the photosynthetic products, which could promote the biomass accumulation. 【Conclusion】 Salt stress led to excessive Na+ absorption by leaves, causing a decrease in net photosynthetic rate, stomatal conductance and transpiration rate of tomato, and the accumulation of malondialdehyde in leaves, while leaves increased superoxide dismutase activity as well as proline and soluble sugar content to cope with the stress. Under the same irrigation salinity, the net photosynthetic rate and stomatal conductance of tomato leaves were more affected by NaCl stress, soluble sugars maintained the stability of net photosynthetic rate under Na2SO4 stress, superoxide dismutase was able to protect the photosynthetic system under NaCl stress, and the NaCl treatment was required to maintain a higher leaf K+/Na+ level when the net photosynthetic rate was the same. Tomato leaves under Na2SO4 stress were less affected by stress, whereas tomato under NaCl stress had higher water use efficiency at the same salinity. The recommended salinity for irrigation of brackish water containing mainly NaCl was less than 3 dS·m-1, and the salinity for irrigation of brackish water containing mainly Na2SO4 was not more than 4.5 dS·m-1. Table and Figures | Reference | Related Articles | Metrics

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Identification of Salt Resistance Functional of Grape Transcription Factor VvERF2 DAI YingZi, GUO HongYang, YANG ZhiFeng, WANG XianPu, XU LiLi Scientia Agricultura Sinica    2024, 57 (2): 336-348.   DOI: 10.3864/j.issn.0578-1752.2024.02.009 Abstract （205）   HTML （13）    PDF （2770KB）（139）       Knowledge map    Save 【Objective】 In order to give references for future study on the mechanism of the AP2/ERF superfamily on grapes, the protein bioinformatics analysis of grape transcription factor VvERF2 was performed. Additionally, the procedures of gene cloning and homologous genetic transformation were employed for exploring the function of VvERF2 under salt stress in grape callus. 【Method】 For the bioinformatics analysis of the VvERF2 protein, the NCBI Blast database (https://blast.ncbi.nlm.nih.gov/Blast.cgi) and additional online resources were utilized. The Thompson seedless (Vitis vinifera L.) callus was used as the material, and the grape homologous genetic transformation system of VvERF2 were constructed. The transgenic callus phenotype was determined by growth volume, total sugar, total acid, and other factors. Free proline, antioxidant enzyme activity, and other indices were used to assess the salt tolerance of transgenic callus. 【Result】 Based on the bioinformatical analysis of VvERF2 and the 7 most homologous orthologous protein sequences, the VvERF2 gene encoded 240 amino acids, which were quite similar to those of tomatoes and figs, with protein homology percentages of 78% and 67%, respectively. The amino acid residues in eight species varied from 240 to 348, their molecular weights from 26.43 to 38.60 kDa, their theoretical isoelectric points from 5.54 to 8.68, and their index of fatty amino acids were all belonged to unstable proteins, which was higher than 66%. The physicochemical properties of amino acid sequences in different species were quite different. In addition, the promoter of VvERF2 gene had a variety of cis-acting element related to abscisic acid and other transcription factors, such as MYB. Particularly, VvERF2 expressed specificity in different tissues, with callus exhibiting the lowest level of expression. Following salt stress, however, VvERF2 gene expression increased to three times that of the control group. Transgenic results showed that after overexpression of VvERF2 gene in grape callus, growth amount, total acid, total phenol content and antioxidant activity of DPPH (1, 1-diphenyl-2-trinitrophenylhydrazine) were significantly increased. The content of total protein and free proline in transgenic callus were almost higher than those in wild-type callus treated with different concentrations of NaCl. 【Conclusion】 The overexpression of VvERF2 promoted callus growth and accumulation of secondary metabolites, such as phenolic substances, and improved salt tolerance of grape. Table and Figures | Reference | Related Articles | Metrics

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Effects of Soil Texture on Rhizosphere Microbial Carbon Source Utilization, Nematode Community and Fruit Sugar of Jackfruit SU LanXi, PU QiuJie, BAI TingYu, WU YueXian, WU Gang, TAN LeHe, HU YaLi Scientia Agricultura Sinica    2024, 57 (2): 349-362.   DOI: 10.3864/j.issn.0578-1752.2024.02.010 Abstract （108）   HTML （2）    PDF （1091KB）（61）       Knowledge map    Save 【Objective】 This study aimed to investigate the variations in soil microenvironments with different textures under identical fertilization pattern and their potential relationship with the key fruit quality (sugar content), so as to provide a theoretical foundation for regional orchard fertilization and enhancing yield and quality. 【Method】 Standardized jackfruit Malaysia No. 1 orchards, characterized by consistent fertilization practices and soil textures including loam, sandy loam and sand, was carefully selected for this study. Rhizosphere soil and mature fruit of jackfruit were collected during the tree growing, flower and fruit stages to investigate the variation of metabolic functions of soil microbial communities, nematode community structure, soil physicochemical properties and fruit sugars and potential relationships of them. 【Result】 The trend of carbon source utilization of soil microorganisms in sandy loam and sandy soil followed the pattern of tree growing stage > flower stage > fruit stage. The carbon source metabolic capacity of soil microorganisms in loamy soils was ranked as fruit stage > tree growing stage > flower stage. The main types of carbon sources utilized by soil microorganisms were carbohydrates, amino acids, carboxylic acids, and polymers. The diversity index of the soil microbial community and nematode abundance in each trophic group were exhibited consistently higher values in sandy soils compared with other soil types, with particularly elevated levels observed during the tree growing and fruit stages. The ecological index of nematode, soil pH and organic matter in loamy soils were generally higher than that in the other two soil types. The glucose content in fruit showed the following order: sandy loam soil> loam soil > sandy soil. The differences in other sugar components among different soil textures were not significant. The sampling stage had a significant direct effect on the diversity of soil nematode communities, and the soil texture had a significant indirect effect and total effect on the sugar content of jackfruit. 【Conclusion】 The combined application of organic and inorganic fertilizers had different effects on the microecological environment of soils with different textures. The soil texture exerted a direct or indirect influence on the composition and functioning of microbial and nematode communities in rhizosphere soil of jackfruit throughout the tree growing and fruit stages, thereby ultimately impacting both the soil nutrient transformation and fruit quality. Table and Figures | Reference | Related Articles | Metrics

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Effects of Inoculation with Indigenous and Exogenous Arbuscular Mycorrhizal Fungi on Drought Resistance of Pyrus betulaefolia and Its Adaptation Mechanism LI Han, JIANG ShangTao, PENG HaiYing, LI PeiGen, GU ChangYi, ZHANG JinLian, CHEN TingSu, XU YangChun, SHEN QiRong, DONG CaiXia Scientia Agricultura Sinica    2024, 57 (1): 159-172.   DOI: 10.3864/j.issn.0578-1752.2024.01.011 Abstract （157）   HTML （26）    PDF （3895KB）（115）       Knowledge map    Save 【Objective】Arbuscular mycorrhizal (AM) fungi play a significant role in enhancing plant growth and improving resistance to environmental stress. The aim of this study was to screen AM fungi that can withstand drought stress in pear seedlings, so as to establish a theoretical foundation and technical approach for pear mycorrhizal cultivation. 【Method】In this study, a pot experiment and high-throughput sequencing technology were employed to investigate the effects of single and mixed inoculation with indigenous AM fungi, including Claroideoglomus lamellosum (Cl), the exogenous fungus Funneliformis mosseae (Fm), Rhizophagus intraradices (Ri), and Acaulospora mellea (Am), on the growth of Pyrus betulaefolia seedlings under normal and drought conditions. The changes in the AM fungal community in the roots and rhizosphere soil of P. betulaefolia seedlings were analyzed under mixed inoculation (Mix) after 0, 3, and 6 weeks of drought treatment. 【Result】 Under normal and drought conditions, the single inoculation with Cl, Ri, and Mix significantly increased the plant height, stem diameter, leaf area, and relative water content of the leaves of pear seedlings, resulting in a dry weight increase of 35.26% to 52.20%. Additionally, the uptake of phosphorus, potassium, calcium, and magnesium in the aboveground part of the seedlings was enhanced, especially phosphorus uptake, with a mycorrhizal phosphorus uptake effect of up to 1.0. The exogenous fungus Am showed less effectiveness, while Fm inhibited the growth of P. betulaefolia seedlings under normal water supply conditions. Regression analysis indicated that the growth and element absorption effects of mycorrhizal fungi increased with the degree of infection. Under drought stress, AM fungal inoculation significantly reduced the MDA (malondialdehyde) content in the leaves of P. betulaefolia seedlings and increased the activity of antioxidant enzymes and the content of proline to varying degrees. Sequencing results demonstrated that the structure of the AM fungal community in the roots and rhizosphere soil of P. betulaefolia seedlings changed significantly under drought stress compared with normal water conditions. The exogenous fungus Ri dominated the community, followed by Cl and Am, while Fm was the least abundant. The abundance of Ri in the roots significantly increased with the degree of drought stress. 【Conclusion】In conclusion, the different AM fungi had varying effects on the growth of P. betulaefolia seedlings, with the indigenous strain Cl and the exogenous strain Ri showing strong growth-promoting effect and drought resistance. The increase in Ri abundance in the AM fungal community was an important adaptation mechanism for P. betulaefolia seedlings to withstand drought stress. Table and Figures | Reference | Related Articles | Metrics

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Comprehensive Evaluation and Selection of Hybrid Offsprings of Early Flowering Spray Outdoor Chrysanthemum SONG Xiang, WANG ZhongMan, ZHANG QiuLing, WEI YuanYuan, ZHAO XiaoGang, LIU Bo, DAI SiLan Scientia Agricultura Sinica    2024, 57 (1): 173-189.   DOI: 10.3864/j.issn.0578-1752.2024.01.012 Abstract （82）   HTML （12）    PDF （6764KB）（84）       Knowledge map    Save 【Objective】The aim of this study was to establish a comprehensive evaluation system for early flowering garden chrysanthemums, and to select plant lines with flowering periods from mid-September to mid-October, of which flower colors were close to pure red or yellow with novel flower type, round plant shape and excellent comprehensive performance to enrich ornamentals for early autumn festivals in Northen China, so as to provide breeding materials and selection methods for further breeding of early flowering garden chrysanthemum. 【Method】Using 3 early-flowering parents and 1 high double-flowering parent with bright color of small flowered chrysanthemum and 25 asexual hybrid progenies as test materials, 30 original traits were observed from 5 aspects, including flower color, flowering period, flower type, plant type, and leaf type. The selected 18 key traits were used as evaluation factors, and then a comprehensive evaluation system was established using Analytic Hierarchy Process and K-Means clustering analysis to conduct the comprehensive screening of hybrid lines. 【Result】The weight values of each trait of analytic hierarchy process were determined with the breeding objectives of early flowering, round flower type, and stable red and yellow flower color. Among them, the weight value of the initial flowering time (0.1817) was the largest, followed by the flower color stability (0.1695), the number of flowers per plant in the full flowering period (0.1383), the flowering duration (0.1301), the flower color suitability (0.0565), the flower uniformity in the full flowering period (0.0565 ), the inflorescence diameter (0.0539), and the number of ligulate flowers (0.0574). The 29 materials were divided into 3 grades by K-Means clustering analysis: 10 in grade I, 11 in grade II, and 8 in grade III, accounting for 34.38%, 37.93% and 27.59%, respectively. The traits of the hybrid progeny were widely separated, and the flower color, flower type, and plant type all showed transgressive traits, but the flowering period was still the earliest in the two early flowering parents, which was in line with the previous studies that the flowering period was a quantitative trait controlled by multiple genes. Therefore, the early flowering breeding still needs to screen parents and increase the resources of open-field small chrysanthemums that can be used in early autumn. 【Conclusion】 This evaluation method integrated the advantages of various evaluation systems, characterized the traits observed in the field with quantitative results, and conducted hierarchical analysis and evaluation guided by breeding objectives, which provided a quantitative method for comprehensive evaluation and screening of excellent varieties of early flowering garden chrysanthemum. The data analysis results indicated that the evaluation system established in this study could effectively evaluate and screen early flowering garden chrysanthemums, providing a reliable method for the breeding of new varieties of early flowering garden chrysanthemums, and helping to improve the efficiency of selecting excellent germplasm for future breeding project. Finally, 10 early flowering garden chrysanthemum strains with excellent ornamental traits were selected, which could be used as new varieties or strains and also as further breeding materials. Table and Figures | Reference | Related Articles | Metrics

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Cloning and Expression Analysis of PtMLP1 Promoter in Poncirus trifoliata YAO LiXiao, SU Juan, GUO XingRu, LI FengLong, HE YongRui, ZOU XiuPing, CHEN ShanChun Scientia Agricultura Sinica    2023, 56 (24): 4906-4915.   DOI: 10.3864/j.issn.0578-1752.2023.24.009 Abstract （102）   HTML （10）    PDF （869KB）（75）       Knowledge map    Save 【Objective】Genetic transformation plays a significant role in exploring gene function and improving traits in citrus. Tissue-specific promoters is a key to regulate the expression of transgenes in particular tissues. Here, expression characteristics of the PtMLP1 promoter, isolated from the root subtractive library of Poncirus trifoliata, was thoroughly examined, which could lay a foundation for the specific expression of exogenous genes in citrus root tissue. 【Method】The complete sequence of PtMLP1 gene was cloned by PCR using DNA as a template. The physiochemical attributes, secondary and tertiary structures of PtMLP1 protein were predicted by ExPASy, PSIPRED, and SWISS-MODEL tools. Cis-acting elements in PtMLP1 promoter were predicted by PlantCARE. The expression pattern of PtMLP1 in P. trifoliata trees of diverse ages was examined by employing real-time qPCR. Furthermore, to investigate the tissue-specific expression of the PtMLP1 promoter in citrus, a pBI121-ProPtMLP1::GUS plasmid, in which GUS expression was controlled by the PtMLP1 promoter, was constructed and then introduced into P. trifoliata through Agrobacterium-mediated hypocotyl transformation. 【Result】PtMLP1 consisted of two exons and one intron, which possessed a 471 bp open reading frame encoding a protein with 156 amino acid residues. This protein had a molecular weight of 17.63 kilodaltons with an isoelectric point of 5.49 and contained a Bet v I functional domain in its primary structure. Moreover, the secondary structure of PtMLP1 contained three α-helices and seven β-folds, while its tertiary structure had a conserved hydrophobic binding site and a cyclic domain, which was rich in glycine. The PtMLP1 promoter was 1 666 bp long. Multiple root-specific expression elements, phytohormone response elements (such as the TGACG motif, P-box, and ABRE), and the TATA box and CAAT box core elements were predicted in the promoter. Additionally, the 3-terminal untranslated region of PtMLP1 was predicted to contain a poly (A) signal AATAAA. Notably, the expression of PtMLP1 was significantly higher in the roots of 1-month, 6-month, and 20-year-old P. trifoliata, with fold changes of 46.34, 74.82, and 110.25, respectively, compared with those in leaves. GUS expression analysis of pBI121-ProPtMLP1::GUS transgenic plants showed that PtMLP1 promoter exhibited specific and high expression in roots, and its expression levels were 7.76 to 124.78 times of that in the leaves. 【Conclusion】The sequences of the PtMLP1 gene and its promoter were successfully obtained, and the promoter demonstrated the ability to drive specific expression of GUS gene in citrus roots. Table and Figures | Reference | Related Articles | Metrics

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Effects of Intercropping with Vulpia myuros in Tea Plantation on Soil and Tea Quality Components CHEN YiYong, LI JianLong, ZHOU Bo, WU XiaoMin, CUI YingYing, FENG ShaoMao, HU HaiTao, TANG JinChi Scientia Agricultura Sinica    2023, 56 (24): 4916-4929.   DOI: 10.3864/j.issn.0578-1752.2023.24.010 Abstract （145）   HTML （15）    PDF （1581KB）（109）       Knowledge map    Save 【Objective】 The objective of this study was to analyze the impact of intercropping Vulpia myuros on soil nutrient composition, soil microbial community structure, and tea quality components in tea plantations, so as to provide the data support for intercropping V. myuros as a means to enhance the ecological environment and quality of tea cultivation.【Method】The experimental materials consisted of soil and fresh leaves from a tea garden that had been intercropped with V. myuros for a duration of 2 years, and the control group was a clear-ploughed tea garden. The pH, organic matter, and mineral nutrients of the topsoil in the tea garden were measured. Additionally, the population structure of soil bacteria and fungi was analyzed using 16S and ITS high-throughput sequencing techniques. The quality components of tea were determined through Agilent-7890B gas chromatography.【Result】After intercropping V. myuros in a tea garden for 2 years, the soil pH increased by 0.29, and the soil organic matter content increased by 16.46 g∙kg-1. Additionally, the available phosphorus, available potassium, ammonium nitrogen, and nitrate nitrogen also increased to varying degrees in the tea garden soil planted with V. myuros. Notably, the available phosphorus was 5.88 times higher in the intercropped tea garden compared with the clear-cultivated tea garden. The total nitrogen content in the V. myuros plantation soil was higher than that in the clear-cultivated tea plantation, while the total phosphorus, potassium, and sodium contents were lower. Moreover, the tea garden soil planted with V. myuros had higher levels of available zinc, available iron, available copper, and cation exchange capacity. The intercropping of V. myuros in the tea garden also led to an increase in the number of bacteria and fungi in the soil. Furthermore, the relative abundance of Actinobacteria and Ascomycota associated with organic matter decomposition increased in the soil of the V. myuros tea plantation. A total of 259 metabolites were identified from the fresh leaves of the grass plantation and the clear cultivation garden. Among them, the content of 20 metabolites showed significant differences, and these different metabolites mainly included sugars, fatty acids, and catechins. The tea leaves of the V. myuros plantation had more than 2 times the contents of leucrose, methyl-β-D-glucopyranoside, lacttol alcohol, galactoglycerol, and α-lactose compared to the tea plantation. On the other hand, the content of (9Z)-octadecatrienoic acid and (9Z,12Z,15Z)-octadecatrienoic acid were significantly lower in the cultivated tea garden. Additionally, compared with the clear-ploughed tea garden the intercultivated tea plantation had significantly lower contents of (+)-galligallocatechin, galligallocatechin, and epicatechin metabolites. 【Conclusion】When V. myuros was interplanted with tea gardens, the acidity of the soil might be successfully alleviated, and the amount of organic matter and mineral nutrient components in the soil increased. Then, tea plants could more easily absorb and use soil nutrients when there were changes in the amount and community structure of soil bacteria and fungi. The structure of the microbial population and variations in soil nutrients had a significant impact on the quality of tea. Table and Figures | Reference | Related Articles | Metrics

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Analysis of Genetic Effect of Fruit Color Controlled by Epistatic Genes in Eggplant CHEN JinRong, LÜ ZiJian, FAN LiSha, YOU Qian, LI Tao, GONG Chao, SUN GuangWen, LI ZhiLiang, SUN BaoJuan Scientia Agricultura Sinica    2023, 56 (23): 4729-4741.   DOI: 10.3864/j.issn.0578-1752.2023.23.014 Abstract （193）   HTML （15）    PDF （3332KB）（162）       Knowledge map    Save 【Objective】Fruit color is an important trait that affects the commercial value of eggplant fruit. By analyzing the causes for the special segregation ratio of individual plants with purple red peel and with white peel in the F2 population constructed by crossing between two white-fruit parents, this paper could lay the foundation for elucidating the mechanism of epistatic gene interaction on regulating eggplant fruit coloration.【Method】The white-flower and white-peel female parent 19141 with mutation at the structural gene ANS involving in the anthocyanin biosynthesis pathway, white-flower and white-peel male parent 19142 with unknown mutation genes involving in the anthocyanin biosynthesis pathway, and their F1 population with purple red peel and F2 population with separate peel colors were used to explore the epistatic inheritance of eggplant fruit coloration. Genes and their mutation patterns were studied by cloning the known genes, D (SmMYB1) and Y (SmDFR) related to peel color in male parent 19142. Molecular basis of peel color-controlling epigenetic genes was analyzed by developing molecular markers based on genetic variations of peel color genes, analyzing the relationship between genotype and phenotype in E4450F2 population, and crossing with other eggplant parents without anthocyanin pigmentation in peel.【Result】The segregation ratio of plants with purple red fruit and white fruit in E4450 F2 progeny was consistent with the segregation ratio of 27:37 controlled by three pairs of epistatic genes, that is, mutations occurred at the P gene locus in 19141, with genotype DDppYY, and mutations occurred at both D and Y gene loci in 19142, with genotype ddPPyy. The results of cloning and sequencing showed that alternative splicing occurred in SmMYB1 of 19142, which led to the second exon skipping. In 19142, SNP (C→G) in the promoter region’s -326 bp upstream of the start codon resulted in the absence of a CAAT-box cis-acting element in SmDFR gene. An SNP, G to C, at the last base of the second exon, was annotated as splicing mutation, which might cause abnormal function of SmDFR gene in 19142, resulting in the eggplant peel’s inability to synthesize anthocyanin. Based on the genetic variation of SmMYB1, SmANS and SmDFR, the functional molecular markers were developed, and the progenies of E4450F2 were genotyped. The results showed that genotype and phenotype were completely consistent. D_P_Y_ corresponded to phenotypes of purple flower and purple red peel, ddP_Y_ corresponded to phenotype of purple flower and white peel, D_ppY_, D_P_yy, D_ppyy, ddppY_, ddP_yy and ddppyy genotypes corresponded to white flower and white peel phenotypes. When 19142 was crossed with white-peel inbred line 19147 (dtdtPPYY), and green-peel inbred line 19144 (DDPPyy), it was found that the fruit color of the two F1 progenies were white and green, respectively, and there was no anthocyanin pigmentation in the peel, which further proved that 19142 was a double mutant in SmMYB1 and SmDFR.【Conclusion】When two eggplant parents without anthocyanin pigmentation in the peel were crossed, the peel of the F1 generation had anthocyanidin pigmentation, and the segregation ratio of plants with anthocyanin pigmentation and non-anthocyanin pigmentation in F2 population was 27:37, it was because one of the parents had a mutation at a gene locus in the anthocyanin biosynthesis pathway, and the other parent had mutations at two other loci in the anthocyanin biosynthesis pathway. When two parents without anthocyanin pigmentation were crossed, the peel of F1 was able to synthesize anthocyanin and present purple red color, which was due to the simultaneous dominance of three epistatic gene loci D, P and Y, and anthocyanin biosynthesis was restored. Mutation of the structural gene of SmANS or SmDFR inhibited anthocyanin biosynthesis in all parts of the plant. The regulation of transcription factor mutation SmMYB1 was tissue specific, and its mutation inhibited anthocyanin biosynthesis in the peel, but did not inhabit anthocyanin biosynthesis in the flower. Table and Figures | Reference | Related Articles | Metrics

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Evaluation of Heat Tolerance of Herbaceous Peony and Screening of Its Identification Indices HOU ZhaoYu, GONG YiZhao, QIAN Yi, CHENG ZhuoYa, TAO Jun, ZHAO DaQiu Scientia Agricultura Sinica    2023, 56 (23): 4742-4756.   DOI: 10.3864/j.issn.0578-1752.2023.23.015 Abstract （138）   HTML （11）    PDF （2787KB）（171）       Knowledge map    Save 【Objective】 The multivariate statistical analysis method was used to evaluate the heat-tolerance of different herbaceous peony (Paeonia lactiflora Pall.) varieties, to screen the heat-tolerance identification indexes of peony, and finally to establish a more comprehensive and reliable heat-tolerance evaluation system of peony. 【Method】 In this study, 140 peony varieties were used as materials, and field experiments were used to measure 8 morphological and structural indexes, including heat damage index, plant height, crown width, and SPAD value; at the same time, 13 physiological indexes, such as malondialdehyde and relative electrical conductivity, were measured in August after high-temperature stress in summer. Correlation analysis, subordination function method, principal component analysis, cluster analysis and stepwise regression analysis were used to comprehensively evaluate the peony heat-tolerance and to screen the identification indexes of heat-tolerance. 【Result】 There were different degrees of variation among the 21 indicators, and the variation coefficient ranged from 6.66% to 78.02%. The variation coefficient was shown as follows: Catalase (CAT)>POD>Pn>qN>SOD>stomatal density>barrier tissue thickness/sponge tissue thickness>SSC>SPC>heat damage index>SPAD>Y(Ⅱ)>b>MDA>Y(NO)>crown width>leaf thickness>plant height>REC>Fv/Fm>Hue angle (H), among which, CATwas the largest coefficient of variation, and H was the smallest coefficient of variation; through the correlation analysis of each index, it was found that X1 (heat damage index) and X2 (plant height), X3 (crown width), X4 (SPAD), X7 (Fv/Fm), X12 (leaf thickness), X17 (SSC ) were extremely significantly negatively correlated, which were significantly negatively correlated with X6 (Pn), X8 [Y(Ⅱ)], X13 (stomatal density), X20 (CAT), while they were extremely significantly positively correlated with X5 (REC), X9 [Y(NO)], X16 (MDA) and X18 (SPC). There were different degrees of correlation among the indicators, which was relatively complicated; 21 indicators were extracted into 7 principal component factors through the principal component analysis method, and the contribution rates were 20.50%, 11.66%, 8.24%, 7.24%, 7.06%, 5.31%, and 4.85%, respectively, while the cumulative contribution rate reached 64.87%; the comprehensive score (W) of 140 peony varieties were calculated by the membership function analysis method. On this basis, cluster analysis was used to classify the peony cultivars into four types of heat resistance: “excellent” “good” “medium” and “poor”. The “excellent” type accounted for 14.3%, “good” type accounted for 26.4%, “medium” type accounted for 46.4%, and “poor” type accounted for 12.9%; the stepwise regression analysis was further used to establish the optimal linear regression equation: W=0.228-0.166X1+0.002X4+0.325X7-0.257X9+0.112X10+0.00028X13+0.002X17+0.00015X19+0.001X20, and 9 indicators were selected from 21 indicators (heat damage index), including X1(heat damage index), X4 (SPAD), X7 (Fv/Fm), X9 [Y(NO)], X10 (qN), X13 (pore density), X17 (SSC), X19 (SOD), and X20 (CAT), which were used as identification peony indicators of heat-resistance. 【Conclusion】 By using multivariate statistical analysis method to evaluate the heat resistance of peony, 140 peony varieties were divided into 4 categories (excellent, good, medium, and poor). 9 indexes including heat damage index and SPAD value were screened as identification indexes of heat-resistance of peony, to quickly evaluate the heat-resistant ability of peony, thereby significantly improving the efficiency of heat-resistant identification of peony. Table and Figures | Reference | Related Articles | Metrics

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Development and Transferability of EST-SSR Markers Based on Transcriptome Data from Asparagus officinalis YI ZeHui, ZHAO Jing, MAO LiPing Scientia Agricultura Sinica    2023, 56 (22): 4490-4505.   DOI: 10.3864/j.issn.0578-1752.2023.22.011 Abstract （115）   HTML （7）    PDF （2671KB）（137）       Knowledge map    Save 【Objective】The aim of this study was to clarify the distribution law of SSR loci across the transcriptome of Asparagus officinalis, to develop highly informative EST-SSR markers and to analyze their transferability, so as to provide the tools for phylogenetic analysis, functional gene mining and molecular marker-assisted breeding of asparagus plants.【Method】Based on the RNA-seq data of 15 asparagus roots obtained from the previous stage by our research group, MISA software was used to retrieve SSR loci, and Primer 3.0 software was employed to design primers in batches. Then, the ineffective primers were eliminated by performing batch e-PCR with TB-tools software and one-to-one e-PCR with the Primer-blast programme. The information of EST-SSR markers (such as gene id, physical location, and potential function) was obtained by comparison with the genome of asparagus officinalis. The DNA of 9 A. officinalis varieties, 7 A. setaceus varieties, 5 A. cochinchinensis varieties, and 3 A. umbellatus varieties were used as templates, and 50 pairs of randomly synthesized primers were used as primers to detect the effectiveness, polymorphism and transferability of the primers developed.【Result】A total of 36 590 simple sequence repeats (SSRs) loci distributed in 30 229 unigenes with a frequency of 4.78% and an average distance of 9.17 kb were identified based on data from 15 root transcriptomes of A. officinalis. The SSRs were unevenly distributed in the 10 chromosomes, with the highest number in chromosome 7 (4 642) and the highest density in chromosome 3 (37.86 SSRs/Mb). The SRRs were distributed from di- to hexa-, with tri- (46.92%) and AG/CT (16.58%) as the most abundant repeat type and predominant repeat motif, respectively. A total of 19 695 pairs of EST-SSR primers were successfully designed, and 15 147 pairs ineffective primers were eliminated by e-PCR. Among them, 3 085 pairs ineffective primers didn’t produce any amplification products, 10 102 pairs produced severely inconsistent amplification products in terms of fragment size, 1 289 pairs had unknown physical positions in the genome, 402 pairs gave other amplification products of similar size to the target fragments, and 269 pairs generated amplification products without SSRs. Based on 2 517 EST-SSR markers located in the gene region developed in this study, the chromosome density distribution map was constructed, with a total coverage length of 1 125.51 Mb and an average distance of 447.16 kb. The potential functions of these markers were involved in many aspects, such as yield, quality, stress resistance, and so on. All 50 pairs of randomly synthesized primers could amplify target bands clearly, of which 36 pairs were polymorphic, and the average polymorphic information content was 0.330. These markers could be used in three other species of Asparagus: the transferability to A. cochinchinensis, A. setaceus, and A. umbellatus were 100%, 92%, and 88%, respectively. Cluster analysis based on the EST-SSR alleles grouped the 24 accessions into four clusters that corresponded to the species of A. officinalis, A. setaceus, A. cochinchinensis, and A. umbellatus.【Conclusion】In this study, 2 517 highly informative EST-SSR markers of asparagus were successfully developed, and the effective amplification rate was 100%. The total coverage length of the physical map was 1 125.51 Mb, and the average distance was 447.16 kb, which could be used for phylogenetic analysis of asparagus and related species. Moreover, it provided a reference for the development of EST-SSR markers in other species. Table and Figures | Reference | Related Articles | Metrics

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Screening Regulatory Genes Related to Luffa Fruit Length and Diameter Development Based on Transcriptome and WGCNA CHEN MinDong, WANG Bin, LIU JianTing, LI YongPing, BAI ChangHui, YE XinRu, QIU BoYin, WEN QingFang, ZHU HaiSheng Scientia Agricultura Sinica    2023, 56 (22): 4506-4522.   DOI: 10.3864/j.issn.0578-1752.2023.22.012 Abstract （150）   HTML （18）    PDF （3099KB）（176）       Knowledge map    Save 【Objective】 The aim of this study was to identify the co expression modules of luffa fruit length and diameter development and to screen key regulatory genes, so as to provide the theoretical basis for subsequent research on the molecular mechanism of fruit shape control in luffa. 【Method】 The luffa fruits in 9 fruit development stages (2 days before anthesis, and 0, 2, 4, 6, 8, 10, 15, and 20 days after anthesis) were applied as research materials. The fruit length and diameter of each stage were measured. The WGCNA method was used to jointly analyze transcriptome and fruit length and diameter data, to identify co-expressed gene modules of fruit length and diameter development, and to screen out key regulatory genes.【Result】A total of 14 co expression modules were identified by WGCNA, among which two modules (Turquoise and Lightpink4) were significantly correlated with fruit length and diameter (absolute value of correlation coefficient=0.9); Turquoise module was significantly positively correlated, while Lightpink4 module was significantly negatively correlated. KEGG enrichment analysis found that the Turquoise module was significantly enriched in endocytosis and phenylpropanoid biosynthesis pathways, which were closely related to fruit enlargement and growth regulation, and could be used as a key gene module for studying fruit length and diameter in luffa. According to the connectivity and functional annotation of genes in Turquoise module, ten key regulatory genes were screened, including xyloglucan endotransglucosylase/hydrolase gene XTH23, actin-depolymerizing factor gene ADF2, chaperone protein gene DnaJ10, expansin gene (EXPA1, EXPA4 and EXLA5), kinesin gene kinesin-13A, auxin response genes SAUR21, and Aux/IAA11. The RT-qPCR results showed that the expression levels of ten regulatory genes significantly increased after the fruit entered the rapid growth period (8 day after anthesis), with an increase of 2-50 times approximately. Through constructing a gene interaction network, it was found that some candidate genes interacted with the WRKY, bHLH, and HSF transcription factor families.【Conclusion】The Turquoise module, an important co expression module of luffa fruit length and diameter was obtained, and ten potential candidate genes for luffa fruit shape control were screened. It was found that luffa fruit length and diameter development regulation mainly involved the processes of cell wall reconstruction, cell development and differentiation, and auxin regulation. Table and Figures | Reference | Related Articles | Metrics

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Sequencing and Functional Analysis of Tomato circRNA During Flowering Stage YIN ZiHe, YANG ChengCheng, ZHAO YuHui, ZHAO Li, LÜ XiuRong, YANG ZhenChao, WU YongJun Scientia Agricultura Sinica    2023, 56 (21): 4288-4303.   DOI: 10.3864/j.issn.0578-1752.2023.21.012 Abstract （111）   HTML （9）    PDF （4532KB）（231）       Knowledge map    Save 【Background】As one of the most important periods in plant growth and development, the flowering period directly affects fruit ripening and seed development. circRNAs are a class of covalent closed-loop RNA molecules that are ubiquitous in eukaryotic cells and play an important role in the regulation of tomato development and stress response. However, the current circRNA studies on tomato mainly focus on fruit and leaves, and there is a lack of systematic studies on tomato circRNA at flowering stage.【Objective】Identification and analysis of circRNs in flowering tomato could be of great significance for the functional study of miRNA and circRNA in tomato, and also layed a foundation for the study of tomato growth, development and stress response mechanism.【Method】circRNA sequencing was performed on 3 tissue samples of flowers, roots and leaves of flowering tomato plants, with 3 replicates for each sample. circRNAs were identified and their basic characteristics were analyzed. The cycle-forming ability of tissue-specific circRNAs was screened, and the host genes of identified circRNAs were analyzed by GO analysis and KEGG analysis. The mode and site of action of circRNAs were predicted and analyzed by bioinformatics methods to construct a potential circRNA-miRNA-mRNA interaction regulatory network in response to tomato growth and development.【Result】A total of 532 circRNAs were obtained by high-throughput sequencing, 83% of which were exon types. The distribution of circRNA in each chromosome of flowering tomato was uneven, among which chromosome 1 produced the most circRNAs and chromosome 5 produced the least. circRNAs differentially expressed in flower, leaf and root tissues of flowering tomato showed that 79 circRNAs were differentially expressed in flower and leaf, 133 circRNAs were differentially expressed in flower and root, and 132 circRNAs were differentially expressed in leaf and root tissues. Among them, 14 circRNAs were differentially expressed in flower, leaf and root tissues. The cyclization ability of 8 circRNAs randomly selected from 14 differentially expressed circRNAs was tested, and the results showed that all 8 circRNAs had cyclization ability. GO analysis and KEGG analysis showed that circRNAs in flowering tomato were mainly related to the binding of nucleic acids, proteins and other small molecules, as well as the synthesis and metabolism of various biological macromolecules. Finally, the tomato circRNA-miRNA-mRNA interaction regulatory network composed of 14 circRNAs, 10 miRNAs and 136 mRNAs was constructed.【Conclusion】A total of 342 tissue-specific circRNAs were identified, among which 14 were significantly expressed, and 8 were successfully identified. A circRNA-miRNA-mRNA interaction regulatory network was constructed for tomato at flowering stage. This study laid a foundation for the subsequent research of circRNA in flowering period. Table and Figures | Reference | Related Articles | Metrics

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Identification of the Transcription Factor WRKY75 of CmPR4A in Citron C-05 and Its Function Analysis in Resistance to Citrus Canker Disease YAN PeiHan, LUO JianMing, HAO ChenXing, SUN ZiQing, YE RongChun, LI Yi, LIU Lian, SHENG Ling, MA XianFeng, DENG ZiNiu Scientia Agricultura Sinica    2023, 56 (21): 4304-4317.   DOI: 10.3864/j.issn.0578-1752.2023.21.013 Abstract （168）   HTML （24）    PDF （2450KB）（211）       Knowledge map    Save 【Background】Citrus canker is one of the serious citrus diseases caused by Xanthomonas citri subsp. citri (Xcc). There is currently no radical cure method for it, and few of the existing cultivars have sufficient resistance to citrus canker. Therefore, the breeding for resistant varieties is crucial for the radical cure of the disease, and the identification of resistant genes is beneficial to disease-resistant cultivar breeding.【Objective】The aim of this study was to use the resistance related gene CmPR4A to screen its upstream transcription factors, and to explore the role of transcription factors in resistance to Xcc, which could provide genetic information for the breeding of citrus disease resistant varieties.【Method】Based on the transcriptome results of Citron C-05 (resistant) and Bingtang Sweet orange (susceptible) after inoculated with Xcc, and combined with the results of qRT-PCR, PR4A was differentially expressed in resistant and susceptible genotypes. Differential analysis on PR4A promoter sequence of Citron C-05 and Bingtang Sweet orange was performed using PlantCARE. Yeast one hybrid was used to screen the upstream transcription factors of PR4A. Further interaction between CmPR4A and candidate transcription factors was verified by yeast gyration test and dual-Luciferase. The expression of candidate transcription factors was detected among 8 resistant and susceptible citrus genotypes after inoculation with Xcc at 0, 2, 4, 6, and 8 days to verify their relationship with disease resistance. By transient overexpression of candidate transcription factors in Citron C-05 and Bingtang sweet orange leaves, the expression of transcription factors and PR4A were analyzed using qRT-PCR. Xcc bacterial quantification and symptom observation were executed in transgenic leaves after 24 h inoculated with Xcc.【Result】The expression of PR4A was significantly higher in resistant Citron C-05 than that in the susceptible Bingtang sweet orange at 4, 6, and 8 dpi after inoculated with Xcc. There was a difference in the cis acting element W-box in PR4A promoter between Citron C-05 and Bingtang sweet orange at -236 bp location. Therefore, the CmPR4A promoter was truncated and the bait vector was constructed. Yeast one hybrid screening was conducted using Citron C-05 yeast library induced by Xcc, resulting in CmWRKY75 could interact with proCmPR4A-2. Further dual Luciferase reporting system also confirmed that the interaction between CmWRKY75 and CmPR4A, and CmWRKY75 was positive regulating the expression of CmPR4A. Additionally, the expression of WRKY75 was significantly upregulated in resistant genotypes Citron C-05, American citron and Aiguo citron after inoculation with Xcc, while it was only slight upregulation in susceptible genotypes Bingtang Sweet orange, Shatian Yu pummelo, lemon, Nanchuan and Danna citron. Transient overexpression WRKY75 in Citron C-05 and Bingtang Sweet orange leaves revealed a significant upregulation expression of PR4A at 4 dpi of Xcc and enhanced leaf resistance to Xcc.【Conclusion】CmWRKY75 could bind to the W-box in CmPR4A promoter and positively regulate the expression of CmPR4A, resulting in enhancing leaf resistance to Xcc. Moreover, the expression of WRKY75 was induced by Xcc and showed significant upregulation in disease-resistant genotypes, which was consistent with the expression pattern of PR4A. These results indicated that the differential expression of WRKY75 in different disease-resistant and susceptible citrus genotypes influenced the expression of PR4A, which made it play a role in the resistance of Citron C-05 to canker disease. Table and Figures | Reference | Related Articles | Metrics

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Functional Identification of Peach Gene PpSAUR73 YANG Li, CAO HongBo, ZHANG XueYing, ZHAI HanHan, LI XinMiao, PENG JiaWei, TIAN Yi, CHEN HaiJiang Scientia Agricultura Sinica    2023, 56 (20): 4072-4086.   DOI: 10.3864/j.issn.0578-1752.2023.20.012 Abstract （107）   HTML （6）    PDF （4518KB）（218）       Knowledge map    Save 【Objective】 The object of this study was to isolate a peach potential-related gene PpSAUR73, to analyze its expression response to hormones, and to identify its role in regulating seedling growth in transgenic Arabidopsis, so as to provide the molecular basis for the regulation of tree potential. 【Method】 Using Zhongyou Pan 9 as the material for hormone treatment, the real-time fluorescence quantitative analysis was used to analyze the dynamic response of PpSAUR73 within 24 hours. PpSAUR73 was cloned from the peach variety Jiuyan. PpSAUR73 overexpression vector was constructed and transformed into Arabidopsis. Phenotypic observation of genetical modified Arabidopsis was carried out, and the germination rate statistics of both genetically modified and wild-type Arabidopsis sown simultaneously were performed too. The root length and hypocotyl of 7-day growing Arabidopsis with consistent germination were measured, and Arabidopsis with consistent germination was treated with different concentration hormone. Transcriptome sequencing was performed using 7-day-old seedlings, and the differentially expressed genes were analyzed by functional analysis, KEGG pathway enrichment analysis, and regulatory genes analysis, respectively. 【Result】 PpSAUR73 could respond quickly to hormone treatments. The overexpression of PpSAUR73 could affect the germination of Arabidopsis seeds. The hypocotyl and root length of seedlings were longer than those of wild type. In addition, the rosette of transgenic Arabidopsis was larger, and the overall growth potential was larger than wild type. The transgenic Arabidopsis showed decreased sensitivity to auxin. The transcriptome analysis of overexpressing PpSAUR73 showed that there were 128 differentially expressed genes in both control groups, including 84 up-regulated genes and 44 down-regulated genes, and 20 differentially expressed genes were described. The GO function significant enrichment analysis of the differentially expressed genes generated by overexpression of PpSAUR73 showed that the differentially expressed genes were the most abundant in cell components, located in cytoplasm, cell membrane, organelle and extracellular regions. KEGG pathway enrichment analysis on differentially expressed genes were conducted, and the results showed that the differentially expressed genes in pairwise comparisons CK vs SAUR73-1 and CK vs SAUR73-14 were mainly enriched in phenylalanine biosynthesis pathway, plant hormone signal transduction pathway, starch sucrose metabolic pathway and other metabolic pathways. In the phenylalanine biosynthesis pathway, PpSAUR73 could regulate the upregulation of peroxidase encoding genes AT1G05260, AT3G01190, AT3G32980 and AT5G15180. Peroxidases were associated with lignin synthesis, and lignin content was significantly correlated with plant growth, suggesting that overexpression of PpSAUR73 might be involved in regulating lignin synthesis in Arabidopsis and thus growth. In plant hormone signal transduction pathway, the expression of some auxin responsive genes of AtSAUR41, AtSAUR71, AtSAUR51, AtSAUR72 and AtSAUR1 in auxin signal transduction pathway was down-regulated, the expression of phosphatase protein AtPP2CA in abscisic acid signal transduction pathway was up-regulated, and the expression of abscisic acid signal pathway gene AtPYL5 was down-regulated. PpSAUR73 could regulate the growth of Arabidopsis and participate in multiple hormone signal transduction pathways. 【Conclusion】 This study found that PpSAUR73 could quickly respond to hormones and regulate the growth in transgenic Arabidopsis. The differentially expressed genes caused by overexpressed genes caused by overexpression of PpSAUR73 were mainly enriched in metabolic pathways, such as phenylalanine biosynthesis pathway, plant hormone signaling pathway, and starch sucrose metabolism pathway. PpSAUR73 also played an important role in IAA and ABA signal transduction pathways, it was speculated that it played an important role in the growth and development of peach trees. Table and Figures | Reference | Related Articles | Metrics

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A Transcriptome Analysis Identifies Candidate Genes Related to Fruit Cracking in Pomelo Fruits LU YanQing, LIN YanJin, WANG XianDa, LU XinKun Scientia Agricultura Sinica    2023, 56 (20): 4087-4101.   DOI: 10.3864/j.issn.0578-1752.2023.20.013 Abstract （114）   HTML （10）    PDF （1114KB）（209）       Knowledge map    Save 【Objective】Fruit cracking is a universal physiological disorder that occurs during growth in citrus fruits. However, the molecular mechanisms that regulate cracking in citrus fruits remain unclear. The aim of this study was to screen genes that were related to resistance to fruit cracking. 【Method】 Normal fruits from a pomelo (Citrus grandis (L). Osbeck) cultivar (Duxin 1) resistant to cracking, as well as normal and cracked fruits from Duwei, a cultivar sensitive to cracking, were collected on August 3, 2021 and August 20, 2021, respectively. The pericarp surrounding blossom ends of the fruits (the blossom end was considered the center, approximate 30 millimeters radius) were sampled for RNA-seq. 【Result】 The differentially expressed genes (DEGs) in each stage were screened based on the comparisons of a transcriptome between cracked fruits from the cracking-sensitive cultivar and normal fruits from both cultivars. In the stage A, 1 660 DEGs were obtained, and 104 DEGs were common between the comparison. A total of 1 972 DEGs were screened in stage B, and 82 were common in the comparison. All the DEGs screened at both stages were used for a Gene Ontology enrichment analysis. In the classification of biological process, the major common sub-classifications, including ‘metabolic process’, ‘cellular process’, ‘single-organism process’, ‘biological regulation’, ‘response to stimulus’, and ‘signaling’ were identified in both stages. All the screened DEGs were also analyzed using Kyoto Encyclopedia of Genes and Genomes enrichment. Many genes were enriched in several metabolic pathways, including ‘carbon metabolism’, ‘MAPK signaling pathway-plant’, ‘plant hormone signal transduction’ and ‘protein processing in endoplasmic reticulum’. In addition, these pathways were identified in both stages. Several genes related to resistance to fruit cracking were identified in this study. The levels of transcription of Expansin-A1 were significantly higher in the pericarp of normal fruits from the two cultivars than that in the pericarp of cracked fruits from the sensitive cultivar. Calcineurin B-like protein gene was highly expressed in the pericarp of normal fruits from both cultivars when compared with the pericarp of cracked fruits from the sensitive cultivar. However, this difference disappeared at the stage B. The genes for heat stress transcription factor, serine/threonine-protein kinase, auxin-responsive protein, and dehydration-responsive element-binding protein were upregulated in the pericarp of cracked fruits from the sensitive cultivar compared with the pericarp of normal fruits from the two cultivars in both stages. 【Conclusion】These findings suggested that the genes related to strength of pericarp, water movement, and responsing to high temperature and water deficiency stresses were critical to regulating resistance to fruit cracking. Table and Figures | Reference | Related Articles | Metrics

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Nutritional Effects of Liquid Digestate on Tomatoes Grown in Facility Substrates TENG YunFei, SHANG Bin, TAO XiuPing Scientia Agricultura Sinica    2023, 56 (19): 3869-3878.   DOI: 10.3864/j.issn.0578-1752.2023.19.013 Abstract （110）   HTML （7）    PDF （457KB）（138）       Knowledge map    Save 【Objective】 The nutritional effects of liquid digestate on tomatoes in facility soilless cultivation were investigated, so as to provide scientific basis for the substrate cultivation application of biogas slurry. 【Method】 Here, tomato was used as subject, and the dual inputs of fertilizers (liquid digestate (D) and mineral fertilizer (M)) and soilless substrates (peat substrate (P) and cinder substrate (C)) consisted of four treatments. The parameters of tomato growth, photosynthesis and nutrients uptake were recorded during the experimental period, as well as the properties of substrates. 【Result】The tomato dry biomass under DP treatment was higher than that under MP treatment by 12.6%, and DC treatment was higher than MC treatment by 70.9%. However, the dry biomass under DP and MP treatments were significantly higher than that under DC and MC treatments (P<0.05). The Cond, Ci and Tr of tomato leaf under DP treatment was significantly higher than that under other treatments (P<0.05), and the greatest leaf photosynthetic rate was also observed under DP treatment of (18.17±0.47) µmol·m-2·s-1. Liquid digestate significantly increased the dry biomass and photosynthesis of tomato plants. The total N and total P content of plant under DP treatment were higher than that under MP treatment by 46.9% and 19.7%, respectively, and DC treatment were 1.38 and 2.45 times higher than MC treatment, respectively. Moreover, liquid digestate treatments significantly increased the pH value, organic matter and available phosphate content of substrates compared with chemical fertilizer treatments (P<0.05). The highest fruit yield was obtained under DP treatment of (6.0±0.4) kg·m-2, and the yields between DP and MP treatments had no significant difference. The liquid digestate treatments could significantly increase the soluble sugar content and vitamin C content in tomato fruits (P<0.05), and improve fruit quality. Meanwhile, the results of principal component analysis showed that DP treatment had the best overall performance, followed by MP and DC treatments, while MC treatment was the worst. 【Conclusion】The application of liquid digestate into peat substrate increased the photosynthetic efficiency of leaves and the absorption of nitrogen and phosphorus of tomato plants, as well as the properties of substrate and the yield and quality of fruits. Therefore, the combination was recommended for use in facility tomato cultivation. Table and Figures | Reference | Related Articles | Metrics

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Differences and Genesis of Grape Phenolic Compounds Among Different Altitudes in Yunnan Shangri-la ZHANG KeNan, YIN HaiNing, WANG JiaKui, CAO JianHong, XI ZhuMei Scientia Agricultura Sinica    2023, 56 (19): 3879-3893.   DOI: 10.3864/j.issn.0578-1752.2023.19.014 Abstract （199）   HTML （12）    PDF （2810KB）（444）       Knowledge map    Save 【Objective】Phenolic compounds are important secondary metabolites of wine grape, which have an important influence on the quality of grape and wine. In this study, the differences and genesis of phenolic substances in grape skins among different altitudes were studied combined with soil and climate factors, so as to provide a theoretical basis for the planting management of wine grapes at high altitudes region. 【Method】In the present research, Merlot wine grape was used as the test material. For three consecutive years (2020, 2021 and 2022), the differences of total phenols, flavonoids, tannins, total anthocyanins, the content of individual and non-individual anthocyanin components in grape skins at two altitudes (2 181, 2 300 m) at maturity stage were analyzed. Meanwhile, climate factors such as light, temperature and humidity at different altitudes were monitored during grape development, and the influences of climatic factors on phenolic substances of grape skins were analyzed. 【Result】There were no significant differences in the main mineral nutrients of the soils of the two altitude vineyards, and some differences in climatic factors, such as light, UV intensity, temperature and humidity. Altitude had a significant effect on the content of phenolic substances in grape skins. In the years of 2020-2022, the higher altitude was conducive to the accumulation of phenolic substances in grape skins. the content of total phenols, total tannin, total anthocyanins, most of the individual anthocyanins and the quercetin in berry skins were higher at the altitude of 2 300 m; compared with that at 2 181 m, the content of total tannin in grape skins at 2 300 m increased by 56.27%-174.49%. The flavonoid content at 2 181 m altitude were significantly higher than that at 2 300 m, with an increase of 32.25% to 79.48%. OPLS-DA analysis showed that, the main different compounds of phenolic compounds between the two altitudes were total tannin (TTC), total flavonoids (TFo), malvidin-3-glucoside (Mv), malvidin-3-acetly-glucoside (Mv-Ace), cyanidin-3- glucoside (Cy), and peonidin-3-glucoside (Pn). Grey correlation analysis showed that day-night temperature difference in grape growing season had a great effect on the content of total phenols and total flavonoids in grape skins. The content of total anthocyanins, individual anthocyanins and quercetin in skins were significantly affected by light and ultraviolet intensity. The content of three anthocyanins (Pt, Pn-Ace and Pn-Cou) and quercetin were mainly affected by the light intensity during grape veraison (July).【Conclusion】The climatic conditions of different altitudes, especially day-night temperature difference, light and ultraviolet intensity were the main factors causing the differences of phenolic content. The larger day-night temperature difference, stronger light and ultraviolet intensity at higher altitude were conducive to the accumulation of phenolic substances in grape skins. Table and Figures | Reference | Related Articles | Metrics

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Identification and Cloning of Ferritin Family Genes in Grape and Response to Compound Amino Acid-Iron Spraying During Different Fruit Developmental Stages SONG ZhiZhong, WANG JianPing, SHI ShengPeng, CAO JingWen, LIU WanHao, XU WeiHua, XIAO HuiLin, TANG MeiLing Scientia Agricultura Sinica    2023, 56 (18): 3629-3641.   DOI: 10.3864/j.issn.0578-1752.2023.18.011 Abstract （104）   HTML （3）    PDF （1897KB）（142）       Knowledge map    Save 【Objective】Ferritin plays an important role in plant growth and development, and its biological function in fruit trees are essentially unknown. Cloning of grape Ferritin family genes and revealing their expression patterns at different fruit developmental stages and their response differences to foliar spraying of amino acid iron (Fe) compound fertilizers could provide a theoretical basis for studying the molecular mechanisms of Fe nutrition and metabolism in fruit trees. 【Method】The Ferritin family genes were screened and identified in grape genome by homologous cloning. The detailed characteristics of Ferritin genes and their encoded proteins were analyzed by using bioinformatical methods. The tissue-specific expression patterns of Ferritin family genes during distinct fruit developmental stages and differential response to foliar spraying of amino acid-iron compound fertilizer were determined by carrying out real-time fluorescent quantitative PCR. 【Result】In total, 4 Ferritin family genes were retrieved and cloned from grape, entitled with VvFer1-VvFer4, which were distributed on No. 6, 8 and 13 chromosomes, containing 7 introns with different lengths. VvFer proteins were mainly located in chloroplast and nucleus. The amino acid sequence identity of Ferritin homologs from 16 plant species was as high as 61.48%. Phylogenetic tree analysis indicated that Ferritin homologs belonging to the same genus, such as Arabidopsis and turnip of Cruciferae, tobacco and potato of Solanaceae, soybeans, peanuts and chickpeas of Leguminosae, rubber trees, cassava and castor of Euphorbiaceae, and apples, peaches and strawberries of Rosaceae, possessed a closer genetic distance during evolution. Grape VvFer3 was closely clustered with Solanaceae homologs. The expression levels of VvFer genes were different among distinct tissues or organs of 5-year-old Mathelan grape trees during different fruit developmental stages. In particular, the expression level of VvFer3 was the most abundant, and the maximum expression was observed in fruits from hard core stage to mature stage, followed by VvFer2 and VvFer4. The content of Fe Marselan fruits was slightly different among distinct grape developmental stages, which was gradually increased from young fruit stage, and reached the highest value at veraison stage, and then slightly decreased until mature stage, but still higher than that of young fruit stage and hard core stage. Foliar spraying treatment significantly enhanced Fe content of fruits at mature stage, accompanied by ACO (aconitase), NIR (nitrate reductase) and SDH (succinate dehydrogenase). Genes of VvFer2-4 were significantly up-regulated by foliar spraying of amino acid-iron compound fertilizer, which was closely related to distinct grape tissues/organs and different fruit developmental stages. In details, the expression of VvFer2 in fruits was sensitive to foliar spraying treatment during the whole period of grape development. The expressions of VvFer3 in all tested tissues were sensitive to foliar spraying treatment from young fruit stage to veraison stage. The expressions of VvFer4 in phloem and leaves were continuously induced by foliar spraying treatment during the whole period of grape development, whereas in fruits from young fruit stage to veraison stage. The expression of VvFer1 was relatively low, but very uniform, and there was no response to foliar spraying treatment at the transcription level. 【Conclusion】Four Ferritin family genes were cloned and identified in grape, whose expression were significantly different among distinct tissues during different fruit developmental stages and were prone to be up-regulated under foliar spraying treatment of amino acid-iron compound fertilizer. The overall expression level of VvFer3 gene was the highest in all tested tissues (especially in fruits) during the whole fruit development stage, and was up-regulated in fruits under foliar spraying treatment from young fruit stage to the verason stage. Table and Figures | Reference | Related Articles | Metrics

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Genome Wide Association Study for Resistance to Citrus Brown Spot Disease YANG ShengNan, CHENG Li, TAN YueXia, ZHU YanSong, JIANG Dong Scientia Agricultura Sinica    2023, 56 (18): 3642-3654.   DOI: 10.3864/j.issn.0578-1752.2023.18.012 Abstract （188）   HTML （13）    PDF （2918KB）（221）       Knowledge map    Save 【Objective】The study objective was to explore candidate genes associated with resistance to Alternaria brown spot disease in citrus mandarin, so as to provide the basis for developing suitable molecular markers for further citrus resistance breeding work. 【Method】In summer and autumn of 2022, the young leaves of 136 citrus mandarin accessions were picked and inoculated with the fungus mycelium of Alternaria alternata in the laboratory. The combined results of two experiments were used to obtain the reliable results of 121 citrus mandarins responding to Alternaria brown spot. The phenotypes of 121 citrus mandarins were selected to verify the effectiveness of the CAPS marker developed in a previous study. Then, in order to obtain the candidate SNPs related to Alternaria brown spot resistance, the phenotypes of 121 citrus mandarins and corresponding SNP genotype data from GBS sequencing were analyzed with PCA, GWAS, and Fst methods, respectively. Candidate genes had been selected from the flanking region of 25 kb sequences surrounding the candidate SNPs site, and then they were screened out according to phytozome annotations. The expressions of candidate genes were analyzed after inoculating with the pathogen fungus on leave of Chengtuo hongju and Clementina (Algeria) for 24, 48, and 72 h. 【Result】Among 121 citrus mandarins, 67 varieties such as Clementine and Satsuma were resistant to Alternaria brown spot disease, whereas 54 varieties such as Hong Ju and Ponkan were susceptible. Some varieties resistance to Alternaria brown spot disease could discriminated by CAPS, but its accuracy only accounted for 76.86% in the study. GWAS analysis identified six significant SNPs highly associated with disease resistance, among which SNP1 located in Ciclev10021676 at 24 838 146 bp of chromosome 3 could be used to predicate the resistance of varieties, and their genotype showed a strong correlation with phenotype. Eight significant SNPs highly associated with disease resistance selected by Fst analysis. Finally, five genes of Ciclev10018604, Ciclev10023485, Ciclev10023486, Ciclev10024586 and Ciclev10019874 were screened out. The expression levels of these five genes in Chengtuo Hongju were up-regulated extensively after inoculating leaves with the pathogen fungus, and their expression levels reached the highest 48 hours later. 【Conclusion】Through GWAS, the SNP at 24 838 146 bp on chromosome 3 was found to be the most significant one with high resistance correlation to Alternaria brown spot disease, and the genotype at this location could be effectively used to distinguish the resistant varieties. The candidate genes responding to Alternaria brown spot disease in mandarin were discussed, i.e., Ciclev10019874, Ciclev10018604, Ciclev10023485, Ciclev10024586, and Ciclev10023486. Table and Figures | Reference | Related Articles | Metrics

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Effects of Different Distances from Original Planting Row on Tree Growth and Fruit Yield of Young Trees of G935 Dwarf Rootstock Miyato Fuji Under Continuous Cropping LI MinJi, LI XingLiang, ZHANG Qiang, ZHOU Jia, YANG YuZhang, ZHOU BeiBei, ZHANG JunKe, WEI QinPing Scientia Agricultura Sinica    2023, 56 (17): 3412-3419.   DOI: 10.3864/j.issn.0578-1752.2023.17.014 Abstract （104）   HTML （5）    PDF （448KB）（297）       Knowledge map    Save 【Objective】 The effects of five different planting distances from the original planting line on the growth of young apple trees of G935 dwarf self-heeling rootstock Miyato Fuji were investigated and studied for four consecutive years, and the resistance to repeated cropping of G935 dwarf self-heeling rootstock grafted Miyato Fuji was evaluated, so as to provide a theoretical basis for the renewal of old and inefficient apple orchards and the upgrading of cultivation models in China. 【Method】 In the spring of 2018, the 12-year-old apple tree (Miyato Fuji/SH6/seedling stock) was planted, without soil sterilization, adding organic fertilizer, chemical fertilizer and biological microbial fertilizer, and the G935 dwarf self-rooting stock Miyato Fuji seedlings (2-year root and 1-year dry) were directly planted at different distances (0, 0.5, 1, 1.5 and 2 m) from the original planting line, and the fine spinning hammer tree shape was used for pruning. The differences in tree growth, leaf function, early flowering and early fruiting, and fruit yield and quality of young trees of G935 dwarf self-rooting rootstock Miyato Fuji were investigated under 5 treatments for 4 consecutive years after planting. 【Result】 Under the condition of replanting, there was no significant difference in the growth, leaf function, early flowering and early fruiting, and fruit yield and quality of G935 dwarf self-rooting rootstock Miyato Fuji at different distances from the original planting line. Within 4 years after replanting, with the growth of tree age, the height, trunk diameter and number of main branches of Fuji trees under five treatments increased year by year. In the fourth year after replanting, the average number of main branches in each treatment reached more than 30. From the second year of planting, the proportion of long branches under each treatment decreased year by year, and the proportion of short branches increased year by year. In the fourth year of continuous cropping, there was no significant difference in the growth of new shoots, chlorophyll content of leaves, net photosynthetic rate and one hundred leaves weight (fresh weight and dry weight) of Fuji trees under different treatments; the average yield per plant and fruit quality (average fruit weight, fruit shape index, titratable acid content, soluble solid content, and fruit solid-acid ratio) of Fuji fruit under all treatments were similar, without significant difference. 【Conclusion】 Under the condition of replanting in continuous cropping, there was no significant difference in the growth, leaf function, early flowering and early fruiting, fruit yield and quality of the young trees planted with G935 dwarf self-rooting rootstock Miyato Fuji at different distances from the original planting line four years before planting. The branch composition of the trees in each treatment was reasonable, the tree vigor was moderate and not weak, the flowering was early, and the fruit quality was good. G935 was suitable for replanting in continuous cropping, and the effect of resistance to repeated cropping was not affected by the distance from the original planting line. Table and Figures | Reference | Related Articles | Metrics

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Genetic Variation Analysis and Candidate Genes Mining of Regulating Flesh Color in Pomelo CHENG Li, YANG ShengNan, ZHU YanSong, WANG Xu, ZHAO WanTong, LI RenJing, LI Pei, YUAN ZhongJie, JIANG Dong Scientia Agricultura Sinica    2023, 56 (17): 3420-3434.   DOI: 10.3864/j.issn.0578-1752.2023.17.015 Abstract （134）   HTML （5）    PDF （2837KB）（147）       Knowledge map    Save 【Objective】 The flesh color is an important appearance and quality trait of pomelo varieties. The aim of this study was to excavate variation loci and genes significantly related to the flesh color of pomelo varieties, so as to lay a foundation for further understanding the flesh color development mechanism and marker-assisted breeding of pomelo varieties. 【Method】 100 pomelo accessions were used to identify their flesh color with a color difference meter, and classified based on their phenotype results. The simplified genome sequencing of 100 pomelo accessions was carried out using GBS (genotyping-by-sequencing) technology. The eigenvalues and eigenvectors of the genotype data obtained from sequencing were calculated by GCTA software to analyze the population structure, and the genetic differentiation index (Fst) of two different flesh color populations were calculated using Plink 2.0 software. The GLM model in GEMMA software was adopted to perform genome-wide association analysis, and the variation loci significantly associated with color phenotype were identified for further allelic variation analysis. According to the citrus linkage disequilibrium (LD) distance, the candidate genes located in the 25 kb adjacent region around the variation loci that might be related to the forming of flesh color were screened out, four white and four red pomelo accessions were randomly selected to perform qPCR to obtain the candidate genes expression at different fruit development stages. 【Result】 According to the phenotype of flesh color, 100 accessions were divided into two categories corresponding to white and red pomelo accessions, including 58 white flesh pomelos and 42 red flesh pomelos. Six SNP sites with Fst index greater than 0.4 and -log10 (P)>9 were screened out after Fst genetic differentiation analysis and GWAS genome-wide association analysis. The genotypes of six variation SNPs sites were extracted from 100 pomelos accessions and the genotypes of these SNPs could be used to predict the flesh color of pomelos. A total of 14 genes that function annotated as isoprene synthesis, plastid development, abscisic acid signal regulation, ethylene response related genes and transcription factors were screened and selected. Genes expression analysis showed that Cg7g013760 (mitogen-activated protein kinase kinase MAPKKK17), Cg7g013840 (chloroplast porphyrinogen deaminase), Cg7g014020 (transcription factor TCP7), Cg7g014120 (chloroplast superoxide dismutase SOD3), Cg7g014190 (FAD synthetase 2) were related to the forming of flesh color in pomelos. 【Conclusion】 A total of 6 SNP sites significantly associated with flesh color in pomelos accessions were identified by GWAS and Fst analysis. A total of 14 genes within the 25 kb regions around the mutation SNPs site were screened and annotated. The gene expression analysis showed that five genes might be involved in the forming of flesh color in pomelo accessions. Table and Figures | Reference | Related Articles | Metrics

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Mining Genes Related to Fruit Quality in Sweet Oranges Based on Specific Locus Amplified Fragment Sequencing LI RenJing, SHEN WanXia, ZHAO WanTong, CHENG Li, LI Pei, JIANG Dong Scientia Agricultura Sinica    2023, 56 (16): 3168-3182.   DOI: 10.3864/j.issn.0578-1752.2023.16.010 Abstract （183）   HTML （14）    PDF （2593KB）（111）       Knowledge map    Save 【Objective】By using specific locus amplified fragment sequencing (SLAF-seq) sequencing method and combined with fixation indices analysis on different sweet orange populations, the candidate genes related with some excellent traits in sweet orange were mined, so as to provide some candidate genes for the further molecular breeding in sweet orange. 【Method】SLAF-seq technology was used to genotype 240 sweet orange germplasm resources, which represented extensive genetic diversity and from different geographical origins accessions preserved in the National Citrus Germplasm Repository in Beibei of Chongqing. The genomic SNP genotype data were obtained using the sweet orange genome as the reference, and the Fst and XP-CLR values between different subpopulations were calculated. 【Result】A total of 497.82 Mb-reads were obtained by sequencing 240 sweet oranges accessions with SLAF-seq technology, after the sequencing reads were mapped on to the reference genome by BWA software, and a total of 1 467 968 SNPs were identified by GATK and samtools. The Fst and XP-CLR indices were used to screen out candidate genes related to three traits, including fruit navel occurrence, fruit weight, and titratable acid content. Those genes adjacent to the selected SNPs were called. The result showed that orange1.1G044639m and orange1.1g023641m which annotated auxin efflux vector were related to the formation of navel in sweet orange. SNP in orange1.1G023641m which encoded E3 ubiquitin ligase had the highest Fst score between two different single-fruit weight population, and orange1.1G046891m might affect fruit weight by regulating carbohydrate formation; orange1.1G011684m encoding dihydrolipoamide transacetylase of PDH, and the SNP caused a nonsynonymous mutation in its CDS region, which might determine the citric acid content in fruit; orange1.1G034502m encodes Serine/threonine protein kinase, annotated as phosphorylation dephosphorylation function also related with the citric acid content in fruit. 【Conclusion】In this study, the Fst indices were used to screen six candidate genes associated with three fruit traits in sweet orange by using SNPs from SLAF-seq data. Finally, the relevant genes were screened out for three horticultural traits, including navel occurrence, fruit weight and titratable acid content, and the identified candidate genes should be giving further investigation to explore their regulation mechanism in these traits. Table and Figures | Reference | Related Articles | Metrics

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Identification of Callose Synthetase Gene Family and Functional Analysis of PlCalS5 in Paeonia lactiflora HE Dan, YOU XiaoLong, HE SongLin, ZHANG MingXing, ZHANG JiaoRui, HUA Chao, WANG Zheng, LIU YiPing Scientia Agricultura Sinica    2023, 56 (16): 3183-3198.   DOI: 10.3864/j.issn.0578-1752.2023.16.011 Abstract （142）   HTML （17）    PDF （6172KB）（132）       Knowledge map    Save 【Objective】 The CalS family plays an important role in regulating callose synthesis in plants. In this study, the members of the CalS family were identified and their bioinformatics and expression patterns were analyzed, which provided an evidence for distant cross incompatibility of Paeonia lactiflora (P. lactiflora). 【Method】The pollen germination and tube growth in self- and cross-pollinated stigma were observed by fluorescence microscope. The callose content, endogenous Abscisic acid (ABA) content and β-1,3-glucanase activity in stigma were measured. Eight members of the CalS family were cloned, and Expasy, MEME, TBtools, MEGA 7.0 and so on were used to predict the basic physicochemical properties and conserved motifs of proteins of the PlCalS family members and to construct a phylogenetic evolutionary tree. The expression of eight PlCalS in stigma at 24 h of self-pollination, 24 h of cross-pollination and 36 h of cross-pollination were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). Multiple sequence alignment of CalS5 was performed, a phylogenetic tree was constructed, and the expression characteristics of PlCalS5 in response to ABA treatment at different concentrations were analyzed. 【Result】Fluorescence microscopic observation of pollen tube showed that callose plugging occurred in the stigma of hybrid, which restricted the pollen germination and tube growth. Callose content in cross-pollinated stigma was found to be higher in most periods than that in self-pollinated stigma. Then, the callose activity of β-1,3-glucose and ABA content in stigma with different pollination affinity showed regular differences. The members of the PlCalS family of P. lactiflora were identified and named by integrity analysis of the structural domains and sequence matching, and 15 conserved motifs with stable distribution in the PlCalS family were encoded by every eight genes. Multi-species phylogenetic relationships showed that the CalS family could be divided into three branches, with the PlCalS family distributed in only two branches, of which PlCalS5 was more closely related to CalS5 in Paeonia suffruticosa, Arabidopsis thaliana and Solanum lycopersicum. The bioinformatics analysis showed that the eight family members encoded 1 745-1 951 amino acids, with a total number of 28 583-31 870 atoms and isoelectric points of 7.99-9.13. The analysis of the FPKM values in transcriptome showed that the PlCalS family members were highly expressed in the same period of hybridization and at 36 h of hybridization under the same treatment. qRT-PCR showed that the relative expression levels of 8 genes at 24 h of self-pollination were lower than those at 24 h and 36 h of cross-pollination. In addition, the PlCalS5 gene was found to be more sensitive to high ABA treatment. 【Conclusion】 There were eight gene members in the CalS family of P. lactiflora, which played an important role in regulating the callose formation of P. lactiflora. The expression level of the PlCalS gene in cross-pollinated stigma was higher than that in self-pollinated stigma in most periods, which might be involved in callose abnormal deposition. Heterologous pollen stimulation to the stigma of P. lactiflora could enhance a certain ABA synthesis pathway. ABA induced callose deposition by positively regulating the expression of callose synthesis genes, thus inhibiting pollen germination and tube elongation, and finally affecting pollination compatibility. Table and Figures | Reference | Related Articles | Metrics

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Development of Ogura CMS Restorers of Broccoli via Genetic Transformation of Rfo XING MiaoMiao, XU YuanYuan, LU YuYu, YAN JiYong, ZENG AiSong Scientia Agricultura Sinica    2023, 56 (15): 2966-2976.   DOI: 10.3864/j.issn.0578-1752.2023.15.011 Abstract （113）   HTML （4）    PDF （3102KB）（240）       Knowledge map    Save 【Objective】 The broccoli restorers were developed by transforming the matching fertility restorer gene of Rfo from radish into the leading cultivar Ogura CMS line of Nai Han You Xiu (named as SFB45), so as to efficaciously use Ogura cytoplasmic male sterility (CMS) hybrids in breeding and to provide resources for genetic improvement of germplasm.【Method】 The sequence of CDS with its preceding promoter of Rfo were synthesized and the pRfo::Rfo plant expression vector was constructed to infect the cotyledon with petiole, and hypocotyl explants of the SFB45 through Agrobacterium-mediated transformation method. After regeneration and screening of herbicide resistant seedlings, transgenic plants were identified by detection of the Bar resistant marker. The fertility of transgenic plants were observed at flowering stage. The pollen viability of SFB45 and the transgenic plants were analyzed using the alexander stain. Total RNA of <3 mm and >3 mm flower buds from SFB45 and the transgenic plants were isolated and transcribed into cDNA. The Rfo specific primers were designed, and RT-PCR was performed to analyze the expression levels of Rfo and also the genes related to tapetum and pollen wall development in flower buds of SFB45 and its restorer lines.【Result】 A total of 10 transgenic plants were obtained by genetic transformation. Among which, the fertilities of 8 were restored to varying degrees with the average pollen viability ranging from 84.2% to 90.4%. RT-PCR analysis showed that Rfo was expressed in flower buds of fertility restored plants. The key regulators of tapetum development (DYT1 and TDF1) and the essential gene (ACOS5) for the synthesis of sporopollenin, a major component of the pollen wall were up-regulated in <3 mm flower buds of Ogura CMS restore lines. The tapetum degradation related gene AMS and tetrad callose wall and pollen outer wall development related genes CalS5 and CYP703 were up-regulated in >3 mm flower buds of the restore lines. The analysis of the self- and hybrid-crossing progenies of the positive transgenic lines R-1, R-3 and R-6 showed that the introduced Rfo could be stably inherited, which was consistent with the Mendelian inheritance.【Conclusion】 The fertility restorer lines of Ogura CMS SFB45 were obtained by genetic transformation of Rfo, and the integration of Rfo into the genome of SFB45 recovered the expression of genes associated with tapetum and pollen wall development. Table and Figures | Reference | Related Articles | Metrics

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Analysis of the Interaction Between VvGAI1 and VvJAZ9 Proteins in Grape and Its Expression Pattern Under Low Temperature LIU DeShuai, FENG Mei, SUN YuTong, WANG Ye, CHI JingNan, YAO WenKong Scientia Agricultura Sinica    2023, 56 (15): 2977-2994.   DOI: 10.3864/j.issn.0578-1752.2023.15.012 Abstract （135）   HTML （9）    PDF （9800KB）（79）       Knowledge map    Save 【Objective】 DELLA protein belongs to plant-specific GRAS protein family, which is a significant regulatory factor in GA signal transduction pathway and plays important roles in plant growth, development and resistance to different stresses. In this study, the European grapevine VvGAI1 gene was cloned and the analysis of subcellular localization, protein interaction and expression were performed, so as to lay the foundation for the further study of the function of DELLA protein in response to cold stress in grapevine.【Method】 The VvGAI1 gene sequence was obtained by homologous cloning from the leaves of Vitis vinifera cv. Chardonnay. The VvGAI1 sequence was analyzed by bioinformatics method, and the multiple sequence alignment and phylogenetic trees were performed by DNAMAN and MEGA7.0, respectively. The location of VvGAI1 protein in cells was determined by subcellular localization, and the transcriptional activation activity of the VvGAI1 protein was confirmed by a yeast assay. The interaction between VvGAI1 protein and VvJAZ9 protein was verified by yeast two-hybrid and BiFC assays. The VvGAI1 protein polyclonal antibodies from rabbit was prepared by VvGAI1 protein purified from prokaryotic expression. The VvGAI1 protein expression at low temperature was detected by Western blot method. The effect of exogenous MeJA and GA3 on the cold resistance in grape was analyzed by relative electrical conductivity.【Result】 The VvGAI1 gene was cloned from Chardonnay leaves, with carrying an ORF of 1 773 bp, encoding 590 amino acids, locating on chromosome 1, and containing only one exon. The VvGAI1 protein had a molecular weight of 64.87 kDa and pI of 5.31, which was an acidic unstable hydrophilic protein. The VvGAI1 belonged to GRAS family and had the conserved DELLA and GRAS domains. Protein clustering analysis showed that VvGAI1 was closely related to Arabidopsis GAI and tobacco GAI1. The results of subcellular location and transcriptional activation showed that VvGAI1 was a transcription factor localized in the nucleus and had transcriptional self-activation activity. The interaction between VvGAI1 and VvJAZ9 was confirmed by yeast two-hybrid and BiFC assays. The VvGAI1 gene sequence was cloned into the prokaryotic expression vector to form a pET28b-VvGAI1 recombinant vector, and the Escherichia coli BL21 carrying pET28b-VvGAI1 recombinant vector was incubated in culture medium with 1.0 mmol∙L-1 isopropyl-β-d-thiogalactoside (IPTG) at 16 ℃ to obtain VvGAI1-His fusion protein. The anti-VvGAI1 polyclonal antibody (rabbit-derived) was prepared by antigen immunization and serum purification and used to specifically detect VvGAI1 protein in Chardonnay grapes. Western blot results showed that the VvGAI1 protein in grape protoplasts under low temperature treatment showed an increasing first and then decreasing trend, which indicated the expression of VvGAI1 protein was induced at a low temperature. For 50 μmol·L-1 MeJA and 50 μmol·L-1 GA3 treatments, compared with the control group exogenous MeJA treatment could improve the cold resistance of grape, whereas GA3 treatment made grapes more sensitive to cold.a【Conclusion】 Grape VvGAI1 protein was a transcription factor and interacts with VvJAZ9. The VvGAI1 responded to low temperature stress, and exogenous MeJA was able to positively regulate the cold stress, while GA3 negatively regulated the cold response. Table and Figures | Reference | Related Articles | Metrics

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Background Selection and Comparison of Marker Superiority and Inferiority of Aphid-Resistant Seedlings in an Interspecific Cross Peach Population LIU SuNing, BIE HangLing, WANG JunXiu, CHEN XueJia, WANG XinWei, WANG LiRong, CAO Ke Scientia Agricultura Sinica    2023, 56 (15): 2995-3005.   DOI: 10.3864/j.issn.0578-1752.2023.15.013 Abstract （115）   HTML （9）    PDF （2020KB）（116）       Knowledge map    Save 【Objective】 To establish a background selection system in peach, the seedlings contained aphid-resistance locus and high female parent recovery rate were screened from an F2 population crossed by Xiang Pi You Tao peach (big fruit and susceptible to aphid) and Zhou Xing Shan Tao peach (small fruit and resistant to aphid). 【Method】 Firstly, three methods were used to select background markers, including the high polymorphic single nucleotide polymorphism (SNP) obtained from the previous study (Pre-work SNP), SNP randomly selected in the whole genome (Random SNP), and functional SNP affecting the start and stop codon (Functional SNP). The number of final SNP selected by the above methods were 775. Then, using these SNPs, the parents recovery rate for all 121 individuals of the F2 population were calculated, respectively. The repeatability of the selection methods was evaluated by comparing whether the top 10 seedlings with different selection markers were coincident or not. After completing the evaluation of aphid resistance, single fruit weight, and soluble solids content of F2 population, 10 seedlings with extreme phenotypes for the single fruit weight and soluble solids content were selected, respectively. And the superiority and inferiority of different selection methods were estimated by comparing the significance of the differences in Xiang Pi You Tao recovery rates between the two types of phenotypes. Finally, the SNPs in the aphid-resistant location area were used as the foreground markers to screen the elite seedlings with high maternal genetic background and aphid resistance. 【Result】 The background recovery rates of the F2 seedlings which calculated by the three methods were 36.34%-71.99%, 31.75%-74.92%, and 4.51%-66.53%, respectively. Among the top 10 seedlings with high Xiang Pi You Tao recovery rates screened by the three background markers, Pre-work SNP and Random SNP had two duplicate single plants, and so do Pre-work SNP and Functional SNP, and there were 6 repetitive single plants in Random SNP and Functional SNP. This result indicated that the repeatability between the Random SNP and Functional SNP was the highest among all comparisons. When single fruit weight was selected as the breeding target, among the extreme phenotypic monocots, the three background markers, such as Pre-work SNP, Random SNP, and Functional SNP, had a significant Xiang Pi You Tao background recovery rate of 0.069, 0.26, and 0.092, respectively, which meant high relativity was found between the background recovery rate calculated by Pre-work SNP and their fruit weight, followed by Functional SNP, and Random SNP difference was not significant. When soluble solids content was selected as the target, the Xiang Pi You Tao background recovery rates among extreme phenotypic monocots were significant at 0.77, 0.65 and 0.31, respectively, and the differences among the three background markers were not significant. Finally, two individuals with high recovery rate of Xiang Pi You Tao peach were screened, including N20 and N36. Among them, N20 comprised the aphid-resistant markers, and this individual showed aphid resistance with an average fruit weight of 34.42 g and soluble solids content of 16.1%, which was considered to be the superior single strain of this population. 【Conclusion】 In this study population, Pre-work SNP showed a stronger correlation between single fruit weight and Xiang Pi You Tao background recovery rates than Functional SNP and Random SNP, confirming the superiority of this background marker selection method, and the superior performance of N20 plants, which selected with this background marker in the target traits also supported this result. This study provided an idea of background selection and a method to judge the superiority and inferiority of different background markers in the study population, which could effectively improve the efficiency of resistance breeding in fruit crops. Table and Figures | Reference | Related Articles | Metrics

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Comprehensive Evaluation of Fruit Texture and Taste Quality of Pineapple Based on Multiple Methods FU Shan, LIANG Ye, XU JiuLiang, RUAN YunZe, LUO Jian, LI TingYu Scientia Agricultura Sinica    2023, 56 (15): 3006-3019.   DOI: 10.3864/j.issn.0578-1752.2023.15.014 Abstract （157）   HTML （11）    PDF （573KB）（152）       Knowledge map    Save 【Objective】 This study adopted different methodologies to identify the key texture characteristics and physicochemical indexes affecting the total sensory quality of pineapple (Ananas comosus (Linn.) Merr.), and to establish a new comprehensive evaluation method for the precise testing of fruit texture and taste quality.【Method】 In this study, seven different varieties of pineapple were selected for the measurement of sensory attributes, texture characteristics, and physicochemical compositions. Based on the variance and correlation analysis, the key texture properties and physicochemical indexes that affect sensory quality were identified. Further, the principal component regression analysis was performed, with key texture characteristics and physicochemical indexes as independent variables, and the total score of sensory evaluation as dependent variables, to obtain a statistically significant prediction model for the comprehensive evaluation of pineapple quality.【Result】 There were significant differences in some texture properties and physicochemical indexes among different varieties of pineapple, such as hardness, chewiness, maximum shear force, sugar-acid ratio, soluble protein, vitamin C and soluble pectin; the coefficient of variation among varieties was greater than 25%, while the difference in elasticity and cohesiveness was not significant among varieties. The overall satisfaction score of different pineapple varieties from the highest to lowest was Tainong 17>Tainong 16>Tainong 4>MD-2>Tainong 11>Smooth Cayenne>Comte de Paris. Tainong 17 showed the best quality of fruit texture and taste, its total soluble solid content was 16.23%, sugar to acid ratio was 31.82, soluble pectin content was 23.72 mg∙g-1, hardness was 1 826.55 N, Chewiness was 789.77 mJ, and the maximum shear force was 3 491.37 N. Correlation analysis showed that there were six key indexes affecting the overall satisfaction of sensory significantly, including hardness, chewiness, maximum shear force and physicochemical index of soluble solids, sugar-acid ratio and soluble pectin. The determination coefficient R2 of the sensory quality prediction model based on the principal component regression analysis was 0.916, and the standard deviation was 0.11. 【Conclusion】 The texture and taste quality of pineapple vary greatly among different varieties, and a single evaluation method could not accurately evaluate its comprehensive quality. A prediction model was established for pineapple sensory evaluation based on the key texture properties and physicochemical indexes, which could accurately predict the texture and taste quality of pineapple, and made up for the lack of objectivity in artificial sensory analysis. Table and Figures | Reference | Related Articles | Metrics

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Screening and Comprehensive Evaluation of Tomato Rootstocks with High Efficiency of Phosphorus Utilization GAO ZiYuan, HU JingAng, ZHANG BeiBei, GONG Biao Scientia Agricultura Sinica    2023, 56 (14): 2761-2775.   DOI: 10.3864/j.issn.0578-1752.2023.14.011 Abstract （126）   HTML （3）    PDF （2195KB）（299）       Knowledge map    Save 【Objective】Phosphorus is a non-renewable resource, and the uptake and utilization efficiency of soil phosphorus by plants is low. It is of great economic and ecological value to improve the efficiency of phosphorus absorption and utilization by grafting. The aim of this study was to screen the varieties of grafting tomato rootstocks with efficient use of phosphorus and to establish a light, simple and efficient evaluation technology system, which had theoretical and practical guiding effects on the breeding, demonstration and promotion of new rootstocks with efficient use of phosphorus.【Method】The scion tomato varieties of Qinglove No.1 were grafted on 25 rootstocks (G1-G25) or self-grafted (G0). The experiment was conducted in tomato seedling stage and whole growth stage. In the seedling stage test, the grafted seedlings were treated with normal phosphorus (Hoagland nutrient solution, NP) and low phosphorus (Hoagland nutrient solution with 10% phosphorus content, LP) by hydroponics. 15 days later, 16 indexes were measured, including growth and development, phosphorus absorption and utilization efficiency of grafted seedlings. In the whole growth period experiment, the grafted seedlings were still used as the test material. The phosphorus application level of the control group was set at 1 272 kg∙hm-2, and that of the LP treatment group was 50% of that of the control group. Nine indexes such as fresh weight of stem and leaf, yield and fruit quality were determined. Then, the correlation analysis, principal component analysis, membership function value analysis, cluster analysis, multiple regression analysis and other mathematical analysis models were used to screen the test materials with phosphorus efficient utilization type grafting tomato rootstock and to evaluate them comprehensively.【Result】The average coefficient of variation of indexes at seedling stage and whole growth stage under NP treatment was 9.74% and 2.85%, respectively, and the average coefficient of variation of indexes at seedling stage and whole growth stage under LP treatment was 16.10% and 5.84%, respectively. The variation coefficient of each index under LP treatment was generally higher than that under NP treatment, indicating that the difference of influence of rootstock genotype on grafted tomato was amplified under LP condition. Correlation analysis showed that yield (I-17) under NP condition was positively correlated with stem and leaf dry weight (I-1), stem diameter (I-5), stem and leaf P mass fraction (I-7), stem and leaf P absorption efficiency (I-9), whole plant P absorption efficiency (I-11), and stem and leaf P transport efficiency (I-12); under the LP condition, the tomato yield (I-17) was positively correlated with stem and leaf dry weight (I-1), root dry weight (I-2), stem diameter (I-5), strong seedling index (I-6), stem and leaf P mass fraction (I-7), stem and leaf P absorption efficiency (I-9), root and leaf P absorption efficiency (I-10), whole plant P absorption efficiency (I-11), and stem and leaf P transport efficiency (I-12). The ranking of principal component and membership function values showed that the ranking rules of the two analysis methods were basically consistent, and both of them were consistent with the performance pattern of cluster analysis. However, there were slight differences in the ranking of individual rootstock varieties, so this paper used the comprehensive average performance of the two rankings as the final ranking, and calculated the top five grafting combinations (G24, G1, G8, G3, and G25). Through multiple regression analysis, the seedling stage evaluation indexes suitable for low phosphorus tolerance of tomato rootstock were obtained, and the regression equations of key seedling stage indexes affecting yield and quality were established: YI-17=1354.630-5.552XI-4, YI-20=2.956XI-5-7.949XI-14+2.927, and YI-23=48.807+0.005XI-11.【Conclusion】In this study, a set of simple, objective and relatively objective technology system for screening and comprehensive evaluation of tomato rootstock with efficient utilization of phosphorus was established. Three rootstocks, including Korean rootstock No. 1, Jinpeng Rootstock No. 1 and Western Tomato rootstock, were identified to have comprehensive advantages of efficient utilization of phosphorus fertilizer. Table and Figures | Reference | Related Articles | Metrics

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Mechanism Underlying the Improved Quality of Bagged Fruits in Ponkan SHI Ying, CHEN SiYi, ZENG YiKe, TANG Jun, LI DiPing, LI GuoJing, HUANG XianBiao, LI ChunLong, XIE ZongZhou, LIU JiHong Scientia Agricultura Sinica    2023, 56 (14): 2776-2786.   DOI: 10.3864/j.issn.0578-1752.2023.14.012 Abstract （138）   HTML （3）    PDF （631KB）（208）       Knowledge map    Save 【Background】 Ponkan (Citrus reticulata) is an important loose-skin tangerine in China. Egan No. 1 Ponkan is widely grown in Dangyang City, Hubei Province. When this cultivar is used for producing fruit, the major problem is that advanced harvesting leads to undesirable fruit quality, and thus, to a negative impact on consumer appeal. Therefore, the industry has a critical need for the technologies that allow growers to delay harvest until Egan No. 1 Ponkan fruit is fully mature. 【Objective】 In this study, whether bagging improves the quality of fruit produced by Egan No. 1 Ponkan was tested, and the underlying mechanism by examining the expression levels of genes involved in sucrose and citric acid metabolism was analyzed. 【Method】The experiment was conducted in orchards that produce Egan No. 1 located in Banyue Town, Dangyang City, Hubei Province, to compare the qualities of fruit that were produced after bagging to control fruit that was not bagged. The indexes were compared, including peel brightness (L*), color parameters a*, b*, citrus color index (CCI), total soluble solids, titratable acids, soluble sugars and organic acids. In addition, qRT-PCR was used to analyze the expression levels of the genes associated with sucrose and citric acid metabolism. 【Result】The bagging induced changes in peel color parameters, including changes in a*, b*, CCI, increased L*, and significantly reduced peel firmness. The bagging significantly increased the total soluble solid content when the fruit was harvested, and the sucrose, fructose and glucose levels were significantly elevated. A gas chromatography analysis showed that the levels of citric acid, the major organic acid in citrus fruits, was slightly higher in the bagged fruits relative to the control fruit. The qRT-PCR analysis showed that the expression levels of three genes associated with sucrose biosynthesis (CsSPS3, CsSPS4 and CsSS2) were significantly higher in the bagged fruit relative to the control fruit. In contrast, the expression levels of genes implicated in citric acid catabolism, such as CsACO1, were remarkably downregulated in the bagged fruit. 【Conclusion】Taken together, these data indicated that the bagging improved the quality of Ponkan fruit possibly by changing the ambient temperature and humidity in the micro-milieu of the bagged fruits, which might have elevated the expression levels of genes that influence soluble sugar content and led to better sugar accumulation and improved fruit quality of Ponkan. Table and Figures | Reference | Related Articles | Metrics

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Response Characteristics of Rhizosphere and Root Endosphere Bacteria and Rhizosphere Enzyme Activities to Soil Compaction Stress in Young Apple Tree LI JiaQi, XUN Mi, SHI JunYuan, SONG JianFei, SHI YuJia, ZHANG WeiWei, YANG HongQiang Scientia Agricultura Sinica    2023, 56 (13): 2563-2573.   DOI: 10.3864/j.issn.0578-1752.2023.13.010 Abstract （176）   HTML （20）    PDF （756KB）（226）       Knowledge map    Save 【Objective】The aim of this study was to identify the main factors causing bacterial abundance changed in the apple roots microenvironment under compaction stress, so as to provide a reference for further revealing the biological characteristics of apple rhizosphere and orchard soil management under soil stress.【Method】The experimental materials were potted young apple (Malus domestica Borkh.cv. Red Fuji) tree with the rootstocks of Malus hupehensis Rehd. and Malus robusta Rehd., respectively. After pressing the potted soil to form compaction stress, the rhizosphere mineral nutrient content, soil enzyme activity and the bacterial abundance of rhizosphere and root endosphere were measured.【Result】In both M. huphensis and M. robusta, the soil compaction stress significantly increased rhizosphere available phosphorus content, available potassium content and catalase activity, however, significantly decreased rhizosphere alkaline-hydrolyzed nitrogen content, sucrase activity, urease activity, acid phosphatase activity and root endosphere bacterial abundance. Furthermore, the soil compaction stress also changed the composition and structure of rhizosphere microorganisms. Under soil compaction stress, the amount and abundance of rhizosphere bacterial and the activity of fluorescein diacetate (FDA) hydrolase changed with different rootstocks; however, which were significantly decreased in Red Fuji/M. hupehensis, with the reduction rates were 46.88%, 50.50% and 29.13%, respectively, and significantly increased in Red Fuji/M. robusta, with the increases were 51.41%, 20.22% and 13.76%, respectively. In compacted soil, rhizosphere alkali- hydrolyzed nitrogen content, hydrolase (sucrase, urease, acid phosphatase) activities and the bacterial abundance of root endosphere in Red Fuji/M. hupehensis decreased more than that in Red Fuji/M. robusta. Compared with Red Fuji/M. hupehensis, Red Fuji/M. robusta had stronger FDA hydrolase activity and recruited more rhizosphere bacteria. Redundancy analysis showed that rhizosphere available potassium, alkali-hydrolyzed nitrogen, and FDA hydrolase activities had the highest explanatory rate for variation of bacterial abundance in rhizosphere and root endosphere of apple under soil compaction stress.【Conclusion】The soil compaction stress significantly affected rhizosphere microbial composition, then changed soil enzyme activity and mineral nutrient content in apple rhizosphere. Rhizosphere bacteria and FDA hydrolase activity were different with rootstock, and highly inhibited by soil compaction stress in Red Fuji/M. hupehensis. Under soil compaction stress, the content of rhizosphere available potassium and alkali-hydrolyzed nitrogen, and the activity of rhizosphere FDA hydrolase were more closely related to the abundance of rhizosphere and root endosphere bacteria. Table and Figures | Reference | Related Articles | Metrics

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The Function of PmABCG9 Transporter Related to the Volatilization of Benzyl Alcohol in Prunus mume HAO RuiJie, QIU Chen, GENG XiaoYun, JIA HaoTian, ZHANG YaJing, CHANG Jun, FENG XinXin Scientia Agricultura Sinica    2023, 56 (13): 2574-2585.   DOI: 10.3864/j.issn.0578-1752.2023.13.011 Abstract （120）   HTML （9）    PDF （2638KB）（345）       Knowledge map    Save 【Objective】The aim of this study was to improve the volatilization mechanism of floral scent by exploring the gene related to the transmembrane transport of important aromatic components in P. mume. 【Method】The P. mume Caizhiwufen was applied as material plant. Based on the analysis of volatile and endogenous contents of benzyl alcohol at different blooming stages, the ABC transporters (ATP-binding cassette transporter) involved in the transmembrane transport of benzyl alcohol were identified by weighted gene co-expression network analysis (WGCNA), phylogenetic analysis, and gene expression profiles. Finally, the functions of PmABCG9 were validated by subcellular localization, transgenic technology and incubation assay with substrates. 【Result】The flowering process of P. mume was divided into seven blooming stages, and the total relative content of benzaldehyde, benzyl alcohol and benzyl acetate reached more than 80% at the initial blooming stage, indicating that they were the key aromatic components of Caizhiwufen. The quantitative analysis showed that the endogenous content of benzaldehyde was the highest among the above three, while its volatile content was only 13.17 ng·g-1·h-1 at the highest level. The endogenous and volatile content of benzyl alcohol reached the highest at wither stage, up to 82.28 ng·g-1·h-1 of the volatile content. The highest volatile content of benzyl acetate was at full blooming stage and up to 280.21 ng·g-1·h-1. WGCNA between the transcriptome data of different blooming stages and the volatile content of aromatic components showed that the turquoise module highly correlated with the volatile content of benzyl alcohol, importantly. 11 ABCG subfamily genes were identified. Moreover, based on the expression patterns of PmABCG9 in different flower organs and blooming stages of P. mume, it was inferred that PmABCG9 was related to the transmembrane transport of benzyl alcohol in P. mume. Therefore, PmABCG9 gene was cloned, and the typical NBD-TMD domain of PmABCG9 gene was confirmed. Subcellular localization showed that PmABCG9 was localized on the cell membrane. The leaves of transgenic PmABCG9 Nicotiana benthamiana and wild-type plants were incubated with benzyl alcohol solution, and the volatile content of benzyl alcohol from each line were analyzed by GC-MS. It was noted that the volatile content of benzyl alcohol from transgenic tobaccos was significantly higher than that of wild-type ones, demonstrating that PmABCG9 had the function of transporting benzyl alcohol. 【Conclusion】 Benzaldehyde, benzyl alcohol and benzyl acetate were the key aromatic components at florescence of P. mume Caizhiwufen. PmABCG9 belonged to the semi-molecular transporters type from ABCG subfamily, PmABCG9 was closely related to volatile content benzyl alcohol, furthermore, it could effectively transport benzyl alcohol into the air through cell membrane. Table and Figures | Reference | Related Articles | Metrics

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