Table of Content

16 February 2024, Volume 57 Issue 4

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CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS

Genome-Wide Association Studies for Canopy Activity Related Traits and Its Genetic Effects on Yield-Related Traits

LI FaJi, CHENG DunGong, YU XiaoCong, WEN WeiE, LIU JinDong, ZHAI ShengNan, LIU AiFeng, GUO Jun, CAO XinYou, LIU Cheng, SONG JianMin, LIU JianJun, LI HaoSheng

Scientia Agricultura Sinica. 2024, 57(4):  627-637.  doi:10.3864/j.issn.0578-1752.2024.04.001

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【Objective】Canopy activity is an important indicator of wheat growth and development. Identification the loci for canopy activity related traits and their relationships with grain yield (GY) related traits can provide theoretical support for the dissection of genetic structure of yield trait and assisted wheat breeding.【Method】A total of 166 wheat varieties originating from both domestic and international sources were planted in Anyang of Henan province and Suixi of Anhui province in cropping seasons. With the integrated physical map containing 326 570 SNP markers from the wheat 90K and 660K chips, genome-wide association studies for normalized difference vegetation index at seedling stage (NDVI-S) and 10 days after flowering (NDVI-10), and chlorophyll content in flag leaf at 10 days after flowering (Chl-10) were carried out. The results were compared with the previous study for GY related traits using the same material. 【Result】Analysis of variance (ANOVA) showed highly significant effects (P<0.01) of genotypes, environments and genotype×environment interactions on NDVI-S, NDVI-10 and Chl-10, with broad-sense heritabilities (h2 b) of 0.81, 0.81 and 0.91, respectively. Thirteen, 12 and 15 loci were detected to be significantly correlated with NDVI-S, NDVI-10 and Chl-10, respectively, among which 12, 11 and 12 were new, and five loci were associated with two or more traits. The number of favorable alleles for NDVI-S, NDVI-10 and Chl-10 ranged from 4 to 11, 3 to 11 and 4 to 12, respectively, in the 166 wheat varieties, and the phenotypic values increased with the accumulation of favorable alleles. NDVI-S showed significant (P<0.01) and positive correlations with thousand-kernel weight, kernel length and kernel width. Chl-10 was significant positively correlated with GY and flag leaf width (P<0.01), whereas significant negatively correlated with spike number per unit area, plant height and uppermost internode length (P<0.01). Seven pleiotropic loci were detected co-related with both GY and canopy activity related traits.【Conclusion】NDVI-S can be directly used for selection of yield traits. The stable and pleiotropic loci detected in this study can be used for marker-assisted selection.

Red and Blue Light Promotes Cotton Callus Induction and Proliferation

LI KaiLi, WEI YunXiao, CHONG ZhiLi, MENG ZhiGang, WANG Yuan, LIANG ChengZhen, CHEN QuanJia, ZHANG Rui

Scientia Agricultura Sinica. 2024, 57(4):  638-649.  doi:10.3864/j.issn.0578-1752.2024.04.002

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【Objective】Upland cotton (Gossypium hirsutum) genetic transformation faces series of challenges such as a prolonged cycle and low efficiency, with the relatively slow proliferation rate of callus tissue being a critical factor contributing to the extended transformation period. This study aims to investigate the optimal light conditions for upland cotton callus growing. The establishment of this research endeavor is poised to accelerate callus proliferation and ultimately provide a technical foundation for shortening the period of cotton genetic transformation. 【Method】The hypocotyls of the upland cotton line WC were used as explants to induce callus tissues under four different lights: red, blue, red-blue (1:1), and white light (CK). The study aimed to investigate the varied effects of the different lights on callus induction and proliferation, determine the optimal light condition by comparing with the callus proliferation rate, morphological characteristics et al. under different light treatments. 【Result】Different light treatments had a significant impact on the callus induction and growth. The red-blue light treatment exhibited a most positive effect on both callus inducing and proliferation, which observed on the 7th and 15th day. The fresh weight of callus at 7 d under the red-blue light (0.39 g) was the heaviest, followed by the blue light (0.34 g), then the white light (0.24 g) and the red light (0.23 g). The same fresh weight order of the callus was observed at 15 d, with the treatment of the red-blue light (1.15 g) > blue light (0.98 g) > white light (0.69 g) > red light (0.51 g). The callus weight under red-blue light was 1.65 times heavier than the control, increased by 16.5% and 125.5% compared to the treatment of only blue or red light, respectively. In line with this, the callus proliferation rate in the second week under red-blue light was as high as 14.67%, which is twice to that of the red-light treatment (7.17%). The expression level of those genes promoted to cell proliferation and somatic embryo regeneration was consistent with the phenotype, the highest level was under the red-blue light treatment. Furthermore, the activity of the catalase (CAT) was significantly increased under the red-blue light treatment, while the content of reactive oxygen species (ROS) was lower than control. 【Conclusion】Employing different light conditions could result in the varied proliferation rate of cotton callus. The optimal light treatment is by red-blue light in a 1﹕1 ratio, followed by blue light and then white light. However, employing red light only does not favor the callus growth. The treatment of red-blue light (1﹕1) induces the expression of phytochromes and cryptochromes in the callus, increases the expression level of those genes promoted to callus growth. It also enhances the activity of catalase, reduces the content of ROS, and finally promotes the callus proliferation.

Genetic Analysis and Gene Mapping of Glossy Leaf in Brassica napus

GUAN ZhiLin, JIN FengWei, LIU TingTing, WANG Yi, TAN YingYing, YANG ChunHui, LI RuiTong, WANG Bo, LIU KeDe, DONG Yun

Scientia Agricultura Sinica. 2024, 57(4):  650-662.  doi:10.3864/j.issn.0578-1752.2024.04.003

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【Objective】Epicuticular waxes is a hydrophobic layer covering plant leaves and stems, playing an important role in stress resistance and affecting photosynthesis, growth and development. Rapeseed (Brassica napus L.) is one of the main oil crops in the world, but the mechanism of epicuticular waxes formation on its leaves is still unclear. This study aims to reveal the genetic mechanism of epicuticular wax on leaves through a mutant, which will help to achieve high and stable yield of rapeseed.【Method】In this study, leaf of glossy leaf mutant M8 and common waxy leaf inbred lines Zhongshuang 11 (ZS11) and the C20 were characterized, and its photosynthetic rate were measured by portable plant photosynthesis measurement system. Two F₁ hybrids and F₂ segregating populations were constructed by crossing the M8 with ZS11 and C20, respectively, and were used to analyze the heredity of leaf characters in rapeseed. The F₂ population hybridized by M8 and ZS11 was constructed to analyze the genetic regulation of the glossy leaf trait in rapeseed by bulked segregation analysis (BSA) combined with Target-sequencing (Target-seq). Combined with comparative genome and transcriptome database, candidate genes were predicted and then verified by RT-PCR.【Result】The leaf stomatal conductance and photosynthetic efficiency were higher in the glossy leaf mutant M8, compared with ZS11 in rapeseed. The genetic analysis results showed that the glossy leaf was controlled by one pair of recessive genes and glossy leaf was recessive compared with waxy leaf. It was finally fine mapped in the physical region of 0.134-0.699 Mb on chromosome A08 by map based cloning method. Further analysis revealed that there was a large segmental deletion in the 0.22-0.58 Mb region of chromosome A08 in the M8 glossy leaf mutant compared to ZS11, and BnaA08G0006900ZS (BnaA08.SAGL1) in this region of the genome was identified as the candidate gene for the glossy leaf trait. This gene encodes a Kelch-F-box protein, and its deletion may lead to the observed glossy leaf trait in the M8 mutant, as it was highly expressed in ZS11 but none in M8. 【Conclusion】Compared with wild-type waxy leaves, the photosynthetic rate of the new glossy leaf mutant M8 was higher, and the glossy leaf phenotype was controlled by a recessive gene. In the picture, the glossy leaf phenotype regulatory gene was identified as BnaA08.SAGL1, and deletion of the gene resulted in glossy leaf in rapeseed.

TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY

Remote Sensing Monitoring of Cropping Patterns Based on Phenology Information Atlas

TAO JianBin, WANG Yun, ZHANG XinYue, JIANG QiYue, WU WenBin

Scientia Agricultura Sinica. 2024, 57(4):  663-678.  doi:10.3864/j.issn.0578-1752.2024.04.004

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【Objective】 Cropping patterns are a summary of crop sequence which reflects the use patterns and efficiency of cropland resources. Through analyzing the phenological differences of different cropping patterns, the cropland phenology information atlas and cropping pattern spectrum for croplands on the Jianghan Plain were constructed, and the major cropping patterns in this area were mapped. 【Method】 The cropland phenology information atlas including different cropland use patterns was formed through expressing graphically the spatial difference between vegetation index states and cropland use patterns, according to the prior knowledge of crop planting and the phenological differences of different cropping patterns under the framework of geo-information atlas. Taking the major cropping patterns on the Jianghan Plain as the study cases, the vegetation index states in the key phenological periods were arranged and combined to establish the information remapping rule from the vegetation index states to the cropping patterns, their phenological characteristics were explored, and the cropping pattern spectrum was constructed. Then the data during the key phenological periods and phenological knowledge were integrated to map cropping patterns on the Jianghan Plain by using the Naive Bayes Networks. The vegetation index states of the key phenological periods were quantitatively expressed by using the knowledge probability coding method.【Result】The cropping pattern spectrum on the Jianghan Plain was constructed, and it was found that the cropping pattern spectrum on the Jianghan Plain was composed of eight cropping patterns: Spring single-cropping, Summer single-cropping, Spring-and-Summer double-cropping, Summer-and-Autumn double-cropping, double-cropping paddy-rice, cash crops, aquaculture ponds, trees or abandoned croplands. The results showed that the proposed cropping pattern spectrum and the method of mapping cropping patterns based on the key phenological periods and the Naive Bayesian Networks could extract all cropping patterns accurately, at the same time, which had good performance and suitability. There was a significant trend of the expansion of Summer-and-Autumn double-cropping and the shrink of Spring-and-Summer double-cropping and the Summer single-cropping on the Jianghan Plain during the study period.【Conclusion】The cropping pattern spectrum gave a picture of the overall situation of intensive utilization of croplands on the Jianghan Plain, for improving the accuracy of monitoring the use of croplands and enriching the connotation of the use of cropland resources.

Stacking Ensemble Learning Modeling and Forecasting of Maize Yield Based on Meteorological Factors

LI QianChuan, XU ShiWei, ZHANG YongEn, ZHUANG JiaYu, LI DengHua, LIU BaoHua, ZHU ZhiXun, LIU Hao

Scientia Agricultura Sinica. 2024, 57(4):  679-697.  doi:10.3864/j.issn.0578-1752.2024.04.005

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【Objective】In the context of intensified global climate change and frequent meteorological disasters, exploring the significance of meteorological factors on maize yield and accurately predicting maize yield is crucial for enhancing agricultural production and field management. This paper aims to quantitatively analyze the importance of meteorological factors during various growth stages of maize on yield and to establish a highly accurate and reliable maize meteorological yield stacking ensemble learning estimation model for yield prediction.【Method】Using the HP filter method and moving average method, trend yield models for various counties were determined, and county-level meteorological yields were isolated. Three ensemble learning methods (light gradient boosting machine (LightGBM), Bagging, and Stacking) were employed. By analyzing daily meteorological data and maize yield data over 34 years from 596 county-level administrative regions and meteorological observation stations across 12 provinces in China, three maize meteorological yield prediction models based on different ensemble learning frameworks (LightGBM, Bagging, and Stacking) were established.【Result】The HP filter method as the trend yield model was mainly applicable in the regions of Shaanxi, Henan, Jiangsu, and Anhui. Compared to the HP filter method, more counties were suitable for the moving average method, with most counties having the R² distribution above 0.8. Based on a 5-year sliding forecast and model accuracy evaluation indicators, the mean absolute percentage error (MAPE) for the three models on maize yield was below 6%. The Stacking model achieved a MAPE of 4.60%, indicating high prediction accuracy and strong generalizability. The results demonstrate that the maize meteorological yield stack-integrated learning prediction model has higher accuracy and stronger robustness. It effectively utilizes the characteristics and advantages of each base learner to improve prediction accuracy, making it the optimal model for predicting maize yield based on meteorological factors. Furthermore, a quantitative analysis of the impact of 27 meteorological factors during the maize growth stages in 12 provinces, using the random forest feature importance score, is of reference value for crop monitoring and field management.【Conclusion】The three ensemble learning methods, especially the stack-integrated learning model (Stacking), can accurately reflect the spatiotemporal distribution changes in maize yield. The stack-integrated learning model for maize yield based on meteorological factors provides a new method for field management and accurate prediction of maize yield.

PLANT PROTECTION

Identification of Tea Plant Viruses in Fujian Province and Establishment of Multiplex PCR Detection Assay

CHEN XiHong, CAI Wei, YU Yun, LI Min, WANG NianWu, DU ZhenGuo, SHEN JianGuo, GAO FangLuan

Scientia Agricultura Sinica. 2024, 57(4):  698-710.  doi:10.3864/j.issn.0578-1752.2024.04.006

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【Objective】This study endeavors to explore the species diversity and prevalence of viruses within tea plantations in Fujian Province. Furthermore, it aims to devise a multiplex PCR assay capable of swiftly detecting multiple viruses simultaneously.【Method】From 2019 to 2023, a total of 1 869 samples were gathered from tea plants displaying virus-like symptoms, including chlorosis, blotch, and necrosis, across nine regions, encompassing Fuzhou, Nanping, Ningde, Quanzhou, Zhangzhou, Xiamen, Sanming, Putian, and Longyan cities. High-throughput sequencing technology, combined with PCR and RT-PCR detection methods, was used to identify the pathogens causing viral diseases in tea plants. Specific target fragments amplified by PCR were cloned, sequenced, and subjected to phylogenetic analysis. In addition, specific primers were designed based on reported virus sequences in GenBank, and a multiplex PCR assay was established for detecting major tea plant viruses in Fujian Province by optimizing reaction conditions, including annealing temperature, primer concentration, and cycle number. The assay’s specificity, sensitivity, and practical application were subsequently determined.【Result】Three viruses were detected in tea plant samples from Fujian tea plantations, with detection rates ranked as follows: oil tea associated geminivirus (OTaGV) at 48.90%, tea plant necrotic ring blotch virus (TPNRBV) at 26.75%, and camellia cryptic virus 1 (CCV1) at 17.98%. Among the 1 258 samples that tested positive for viruses, 807 samples were infected with only one of the three viruses (OTaGV, TPNRBV, or CCV1), corresponding to detection rates of 37.20%, 21.38%, and 5.56%, respectively. The remaining 451 samples were co-infected with two or three viruses, resulting in a co-infection rate of 35.85%. The most common co-infection types were OTaGV+CCV1 (17.49%), TPNRBV+CCV1 (0.40%), TPNRBV+OTaGV (14.71%), and OTaGV+CCV1+TPNRBV (3.26%). Geographically, OTaGV was distributed in all nine regions, with the highest detection rate in Zhangzhou at 96.77%. CCV1 was present in eight regions (excluding Xiamen), with the highest detection rate in Sanming at 66.00%. TPNRBV was found in five regions (Fuzhou, Nanping, Ningde, Quanzhou and Zhangzhou), with the highest detection rate in Quanzhou at 79.13%. Among the nine regions, only OTaGV was detected in Xiamen, OTaGV and CCV1 were detected in Sanming, Putian, and Longyan, and all three viruses were detected in the other five regions. The viral co-infection rate was highest in Zhangzhou at 85.00% and lowest in Ningde at 23.03%. Phylogenetic tree construction based on CCV1 and TPNRBV gene sequences indicated that the CCV1 isolate FW obtained in this study had the closest resemblance to the reported Fujian isolate FJ_SH104 (GenBank accession number: ON807095), and the TPNRBV isolate FU had the closest resemblance to the Fujian isolate QZHA92 (GenBank accession number: OQ948454). The established multiplex PCR detection assay exhibited strong specificity, as it only amplified specific target fragments of OTaGV, TPNRBV, and CCV1, while no amplification product was observed in other viruses or healthy tea plant samples. The lowest sensitivity of the multiplex PCR was detecting OTaGV and TPNRBV at dilutions of 10^-4 and CCV1 at a dilution of 10^-3. The multiplex PCR assay successfully detected all three viruses in 60 disease samples collected from tea plantations, and the results were consistent with those obtained by single PCR detection.【Conclusion】OTaGV, CCV1, and TPNRBV are the major virus species in tea plants in the Fujian, with OTaGV being reported for the first time in this province. This study also revealed that OTaGV can infect tea plants. Among the detected viruses, OTaGV has the widest distribution, followed by CCV1 and TPNRBV. Presently, tea plants in Fujian are still mainly infected by single virus infections. However, viral co-infections are prevalent, showcasing combinations such as TPNRBV+CCV1, TPNRBV+OTaGV, CCV1+OTaGV, and OTaGV+CCV1+TPNRBV. The established multiplex PCR assay exhibits strong specificity and high sensitivity, making it suitable for the rapid detection of the three viruses (OTaGV, CCV1 and TPNRBV) in tea plantations. Taken together, the findings obtained in this study will provide a theoretical basis and technical support for prevention and control of viral diseases in tea plants in Fujian Province.

Effects of Apolipophorin on Ovarian Development and Lipid Deposition in Locusta migratoria

ZHAO YiYan, GUO HongFang, LIU WeiMin, ZHAO XiaoMing, ZHANG JianZhen

Scientia Agricultura Sinica. 2024, 57(4):  711-720.  doi:10.3864/j.issn.0578-1752.2024.04.007

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【Background】Lipids are one of the important nutrients required by organisms, which play important physiological functions, including diapause, flight, embryonic development and energy regulation in insects. Apolipophorins (apoLps) are the main components of insect lipophorin particles, which are involved in the transport of lipids among different tissues.【Objective】The objective of this study is to investigate the function of apolipophorins (LmapoLp-II/I and LmapoLp-III) by RNA interference (RNAi) in ovarian lipid transport in Locusta migratoria, and to provide a new molecular target for pest control.【Method】RNAi was performed to silence two apolipophorin genes (LmapoLp-II/I and LmapoLp-III) at 1 day post adult eclosion (1 PAE), respectively. dsGFP was used as control, 15 μg dsRNA was injected into each insect. The ovaries at 4, 6 and 8 PAE were dissected and observed after dsRNA injection. The silencing efficiency of LmapoLps in the ovaries at 8 PAE was analyzed by reverse-transcription quantitative PCR (RT-qPCR), and EF1α was used as internal reference gene. Lipidomics technique was used to determine and analyze the differences of lipid metabolites in the ovaries between dsGFP- and dsLmapoLp-II/I-treated groups. The approach combining the fold change, the P value and the VIP value of the OPLS-DA model was adopted to screen for differential abundance of lipids. The contents of neutral lipids and total triglycerides were determined by Bodipy staining and triglyceride assay kit.【Result】After injection of dsLmapoLp-II/I and dsLmapoLp-III, the expression level of target genes could be significantly silenced by 80.84% and 92.89%, respectively. RNAi-mediated silencing of LmapoLp-II/I led to a retarded ovarian development, with significantly decreased neutral lipid content and triglyceride content, while the ovaries of L. migratoria injected with dsLmapoLp-III were the same as those in the control group, which could develop normally and gradually grow larger, with the color gradually changing from white to yellow. After LmapoLp-II/I silencing, a total of 1 166 up-regulated metabolites and 1 384 down-regulated metabolites were detected by lipidomics analysis, of which 20 triglycerides were significantly down-regulated.【Conclusion】LmapoLp-II/I is a major apolipophorin gene that affects ovarian development and is involved in the accumulation and transport of ovarian lipids. These research results not only enrich the research content of lipid metabolism in insects, but also help to find new targets for pest control.

SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT

Investigation on Potential of Replacing Chemical Fertilizer for Crop Straw and Livestock Manure Organic Fertilizer in Shandong Province

MA RongHui, YANG WuJie, YU Lei, YANG ZeLong, WANG Jian, GUO YueSheng

Scientia Agricultura Sinica. 2024, 57(4):  721-739.  doi:10.3864/j.issn.0578-1752.2024.04.008

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【Objective】The quantity and total nutrient of crop straw, livestock and poultry manure resources were calculated, and their potential to replace chemical fertilizers was estimated, so as to provide the data support for the overall and rational utilization of organic fertilizer resources in Shandong. 【Method】Based on the crops (wheat, corn, rice/rice, soybean, potato, peanut and cotton) and livestock/poultry (cattle, pig, sheep, poultry and rabbit), the grass-grain ratio and excretion coefficient methods were used to calculate the total amounts of organic fertilizer and total nutrients in Shandong in 2020, and the potential of organic fertilizer replacing chemical fertilizer was estimated. 【Result】 (1) In 2020, the total amount of straw resources was 7 616.8×10⁴ t, and the total amount of nutrients was 163.2×10⁴ t, in Shandong. The theoretical potential of straw nutrients to replace chemical fertilizers was 42.9%, while the actual potential was 20.9%. The theoretical potential of N, P₂O₅ and K₂O in straw to replace chemical fertilizers was 35.3%, 9.0% and 92.2%, respectively, while the actual potential was 12.5%, 4.2% and 53.7%, respectively. The total amount of straw resources increased by 16.8% from 2016 to 2020. (2) In 2020, the total amount of livestock and poultry manure resources was 9 131.2×10⁴ t, the total amount of nutrients was 82.7×10⁴ t, in Shandong. The theoretical potential of livestock and poultry manure nutrient resources to replace chemical fertilizers was 21.7%, and the actual potential was 19.5%; the theoretical substitution potential of N, P₂O₅ and K₂O resources in livestock manure was 19.6%, 11.4% and 36.4%, respectively, while the actual substitution potential was 17.7%, 10.3% and 32.9%, respectively. From 2016 to 2020, the total amount of livestock and poultry manure resources decreased by 25.4%. (3) The theoretical potential of crop straw and livestock manure nutrients in different regions of Shandong to replace chemical fertilizers was as followed: Northwest Shandong>South Shandong>South Central Shandong>Central Shandong>East Shandong. Northwest Shandong and South Shandong were with the highest potential of 77.0% and 75.6%, respectively; East Shandong was with the lowest of 46.5%. Among them, Dezhou had the greatest potential of 95.3%, followed by 92.6% of Dongying and 91.2% of Binzhou. Yantai and Weihai were with the smallest of 35.5% and 40.6%, respectively. (4) The theoretical potential of different types of resources of crop straw and livestock/poultry manure nutrients to replace chemical fertilizers were as followed: corn (16.7%)>wheat (16.2%)>potato (4.9%)>peanut (3.6%)>rice (0.7%)>cotton (0.4%), soybean (0.4%)>cow manure (8.7%)>pig manure (6.5%)>sheep manure (3.2%)>poultry manure (3.2%)>rabbit manure (0.1%).【Conclusion】In 2020, the total amount of straw and livestock manure resources was 16 748.0×10⁴ t, and the total amount of nutrients was 245.9×10⁴ t, in Shandong. The theoretical potential of straw and livestock manure nutrient resources to replace chemical fertilizers was 64.6%, and the actual potential was 40.4%. The theoretical substitution potential of N, P₂O₅ and K₂O in straw and livestock manure was 54.8%, 20.4% and 128.6%, respectively, and the actual substitution potential was 30.2%, 14.5% and 86.6%, respectively. The organic fertilizer resources of crop straw and livestock manure in western, northern and southern Shandong were very rich, while those in central and eastern Shandong were relatively few. There were more organic fertilizer resources of crop straws, such as corn and wheat, and more organic fertilizer resources of livestock, and poultry manure, such as cattle and pigs. Efficient and rational use of organic fertilizer resources, such as crop straws and livestock and poultry manure, was an important guarantee for reducing fertilizer application and increasing efficiency.

Water Utilization and Economic Benefit of Wheat Multiple Cropping with Green Manure Under Nitrogen Reduction in Hexi Irrigation Area of Northwest China

MA BiJiao, CHEN GuiPing, GOU ZhiWen, YIN Wen, FAN ZhiLong, HU FaLong, FAN Hong, HE Wei

Scientia Agricultura Sinica. 2024, 57(4):  740-754.  doi:10.3864/j.issn.0578-1752.2024.04.009

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【Objective】 In view of the problems of excessive application of nitrogen fertilizer and lower water resource utilization efficiency and economic benefits in crop production in the irrigation area of Hexi Oasis, the aim of this study was to explore the effects of multiple green manures after wheat and moderate reduction of chemical nitrogen fertilizer on the water consumption characteristics and economic benefit of wheat field. 【Method】 From 2019 to 2020, a split plot experiment was conducted in the oasis irrigation area of Hexi, Gansu province. Two planting patterns were set up in the main area, namely, multiple cropping of green manure after wheat harvest (W-G) and single cropping of wheat (W). In the sub-plot, there were five nitrogen application levels, namely no nitrogen application (N₀), conventional nitrogen application level 180 kg·hm^-2 (N₄), reduced 45% nitrogen application (N₁), reduced 30% nitrogen application (N₂), and reduced 15% nitrogen application (N₃).【Result】Grain yield of wheat and systematic biothermal energy were significantly increased by multiple green manure after wheat (W-G) and the moderate reduction of chemical nitrogen fertilizer, in 2019 and 2020, which increased by 10.8% and 12.4%, respectively, and the yield of systematic biothermal energy increased by 37.8% and 40.3%, respectively. Compared with nitrogen reduction of 15% (W-G-N₃) in sole wheat and traditional nitrogen application (W-N₄) in sole wheat, the grain yield increased by 6.9%-16.7% and 7.9%-13.6%, respectively, and the biothermal energy yield increased by 52.0%-62.2% and 27.1%-58.9%, respectively. The water consumption of W-G decreased by 6.3%-16.0% compared with that of W wheat growing stage, and W-G-N₃ decreased the seasonal water consumption of W-N₃ and W-N₄ by 13.4%-20.5% and 20.8%-29.0%, respectively. Repeated planting of green manure could improve the water use efficiency of wheat, and W-G increased by 7.9% and 19.2%, respectively. In 2019, compared with W-N₃ and W-N₄, the WUE of W-G-N₃ increased by 23.5% and 5.1%, respectively. Compared with W-N₃ and W-N₄, W-G-N₃ could improve the energy yield of per unit water efficiency of the system, which was increased by 2.7%-14.5% and 9.3%-17.5%, respectively. Compared with the W and W-G increased the cost input, and the gross output also increased. In 2019, the net return of W-G-N₃ increased by 9.8% and 9.5% compared with W-N₃ and W-N₄, respectively; in 2020, the net return of W-G-N₃ decreased by 15.6% and 15.7% compared with W-N₃ and W-N₄, respectively. In 2019 and 2020, the output/input of multiple cropping green manure after wheat harvest reduced by 20.7% and 23.1% compared with sole wheat, and the output/input of W-G-N₃ was 14.8%-23.1% compared with W-N₃ and W-N₄, and W-G reduced the benefit per cubic meter water of the system due to more resources input. 【Conclusion】In the Hexi oasis irrigation areas, multiple green manure after wheat combined with moderate reduction of chemical nitrogen fertilizer could improve crop yield and economic benefit, as well as water resource utilization efficiency, among which the comprehensive effect of multiple green manure after wheat combined with 15% nitrogen treatment is outstanding, which could be used as an ideal planting pattern and nitrogen application level to improve water resource utilization and farmers' income.

HORTICULTURE

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

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【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.

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

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【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.

FOOD SCIENCE AND ENGINEERING

Effect of Pleurotus eryngii Powder on Quality Characteristics of Extruded Rice

JIANG Wen, LIANG WenXin, PEI Fei, SU AnXiang, MA GaoXing, FANG Donglu, HU QiuHui, MA Ning

Scientia Agricultura Sinica. 2024, 57(4):  779-796.  doi:10.3864/j.issn.0578-1752.2024.04.012

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【Objective】The twin-screw extrusion process was used to produce extruded rice products with P. eryngii powder and rice flour as raw materials, and the predicted glycemic index of the products was analyzed to provide technical support for the development of edible mushroom extruded rice products with comprehensive nutrition and low glycemic index. 【Method】 The content of protein, crude fiber, amino acid and fat of extruded rice with 20%, 40% and 60% P. eryngii powder were determined. RVA rapid viscosity analyzer and rotary rheometer were used to analyze the gelatinization properties and rheological properties of P. eryngii powder with different additive amounts. The internal structure, color, texture properties, starch hydrolysis rate, predicted glycemic index (pGI) and sensory score of extrusion-rice with different dosage of P. eryngii were analyzed by scanning electron microscopy, colorimeter, texture analyzer, and in vitro digestion and sensory evaluation. 【Result】Compared with blank extruded rice, the content of protein, crude fiber and amino acid in extruded rice was significantly increased by adding P. eryngii powder, and the content of protein was increased by 71.84%, 70.19% and 96.70%, and the content of crude fiber was increased by 850%, 2350% and 3750%, respectively. The total amino acid content increased by 40.98%, 58.96% and 66.03%, respectively. The gelatinization and rheological properties of the mixed powder system showed a decreasing trend with the increase of the added amount of P. eryngii powder, and the peak viscosity, valley viscosity, final viscosity, disintegration value and recovery value gradually decreased, and G' and G" gradually decreased. The typical weak gel accounted for a large proportion of elasticity, and the added amount of 20% P. eryngii powder was the closest to the powder parameters of rice flour. Compared with the blank extruded rice, it was found by scanning electron microscopy that the cross section pores of extruded rice increased with the increase of the powder content, and the structure tightness decreased. Among them, 20% of extruded rice had tight structure and less cracks. The values of L^* and b^* in extruded rice of P. eryngii by colorimeter decreased significantly, and a^* increased first and then decreased. The water absorption rate and cooking loss rate after cooking increased with the addition of P. eryngii powder, but the expansion rate had no significant effect, and the cooking characteristics of 20% P. eryngii extruded rice were the best. The hardness, elasticity, adhesiveness and chewability of the extruded rice increased with the addition of P. eryngii powder, while the cohesiveness and resilience first decreased and then increased. Starch digestibility, rapidly digestible starch (RDS), slowly digestible starch (SDS) and predicted glycemic index (pGI) also increased with the increase of P. eryngii powder addition, but they were all lower than that of normal rice and blank extruded rice, while resistant starch (RS) content was greater than that of the two groups and increased with the increase. The predicted glycemic index (pGI) value of 20% P. eryngii extruded rice was the lowest 60.18, which was 20.60 lower than that of normal rice, and the content of resistant starch (RS) was the highest. Sensory evaluation showed that the appearance structure, palatability, taste, cold rice texture and comprehensive score of P. eryngii extruded rice decreased with the increase of the amount of P. eryngii powder, while the odor score decreased first and then increased. From the point of view of the score, 20% of P. eryngii extruded rice was 66.75 scores, which was the most acceptable to consumers. 【Conclusion】 20% of extruded rice with P. eryngii was nutrient-rich and had suitable texture indexes. The predicted glycemic index (pGI) was 60.18, which was 20.60 lower than that of normal rice. The nutritional value of extruded rice was significantly improved by adding P. eryngii powder, and the food quality was better.

ANIMAL SCIENCE·VETERINARY SCIENCE

lncRNA-MSTRG.7889.1 Competitively Binds to bta-miR-146a Targeting Smad4 to Regulate Apoptosis of Yak Granulosa Cells

MENG ZhaoYi, WANG YunLu, YAO YiLong, XI GuangYin, NIU JiaQiang, SOLANGSIZHU, GUO Min, XU YeFen

Scientia Agricultura Sinica. 2024, 57(4):  797-809.  doi:10.3864/j.issn.0578-1752.2024.04.013

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【Background】 Apoptosis of granulosa cells (GCs) is an important cause of follicular atresia, and competing endogenous RNA mechanism has been proven to be involved in regulating the apoptosis process of GCs. The previous transcriptome sequencing study of yak ovary showed that lncRNA-MSTRG.7889.1 had binding sites with bta-miR-146a, while bta-miR-146a had a targeting relationship with Smad4. 【Objective】The aim of this experiment was to explore the molecular regulatory mechanisms of ceRNA that affected the apoptosis of follicular GCs in yak, so as to lay a foundation for revealing the mystery of the reproductive regulation of the yak.【Method】 Healthy and atretic follicles of adult female yaks were collected and isolated, and the expressions of lncRNA-MSTRG.7889.1, bta-miR-146a and Smad4 in follicles were detected by RT-qPCR. Healthy and atretic follicles were sliced, and GCs apoptosis in healthy and atretic follicles was detected by TUNEL. The localizations of lncRNA-MSTRG.7889.1, bta-miR-146a and Smad4 in follicles were analyzed by fluorescence in situ hybridization. The yak GCs were isolated and cultured in vitro, and transfected with Smad4 overexpression vector and bta-miR-146a mimics, respectively. The apoptosis rate of GCs cells was detected by flow cytometry, and the expressions of pro-apoptotic proteins CASPASE3 and BAX and anti-apoptotic proteins BCL-2 were detected by Western blot. After co-transfecting Smad4 overexpression vector and bta-miR-146a mimics with GCs, it was investigated whether bta-miR-146a targeting Smad4 affects GCs apoptosis in yaks. lncRNA-MSTRG.7889.1 overexpression lentiviral vector was used to infect GCs, and the effect of lncRNA-MSTRG.7889.1 on GCs apoptosis was detected by flow cytometry and Western blot. lncRNA-MSTRG.7889.1 vector and bta-miR-146a mimics were co-transfected with GCs to further analyze whether lncRNA-MSTRG.7889.1 competitive binding of bta-miR-146a targeting Smad4 affected the apoptosis of GCs in yaks. 【Result】 The expression of lncRNA-MSTRG.7889.1 and Smad4 gene mRNA in healthy yak follicles was significantly higher than that in atresia follicles (P<0.01), while the expression of bta-miR-146a was opposite (P<0.01). TUNEL test results showed that the fluorescence intensity of GCs in atretic follicles was significantly higher than that in healthy follicles (P<0.01), indicating that the apoptosis of GCs in atretic follicles was significantly higher than that in healthy follicles. The results of fluorescence in situ hybridization showed that Smad4, bta-miR-146a and lncRNA-MSTRG.7889.1 were co-expressed in the follicles of yaks, and their expressions in healthy and atretic follicles were basically consistent with the results of RT-qPCR, indicating that ceRNA mechanism might be involved in the development of healthy follicles and follicle atresia in yaks. Overexpression of Smad4 in yaks GCs significantly decreased the apoptosis rate of GCs (P<0.01), and the expressions of CASPASE3 and BAX protein were significantly decreased (P<0.01), while the expression of BCL-2 was significantly increased (P<0.01). Overexpression of bta-miR-146a significantly increased the apoptosis rate of GCs (P<0.01), increased the expressions of CASPASE3 and BAX proteins (P<0.01), and decreased the expression of BCL-2 (P<0.01). bta-miR-146a targeted inhibition of Smad4 expression (P<0.01); Smad4 and bta-miR-146a were co-transfected into GCs, and the results showed that bta-miR-146a targeted inhibition of Smad4 reduced the former's promoting effect on GCs apoptosis (P<0.01). Overexpression of lncRNA-MSTRG.7889.1 significantly decreased the apoptosis rate of GCs in yaks (P<0.01), the expressions of CASPASE3 and BAX protein were significantly decreased (P<0.01), and the expression of BCL-2 was significantly increased (P<0.01). lncRNA-MSTRG.7889.1 significantly inhibited the expression of bta-miR-146a (P<0.01). lncRNA-MSTRG.7889.1 and bta-miR-146a were co-transfected into GCs, and lncRNA-MSTRG.7889.1 targeting bta-miR-146a decreased the promotion effect of the latter on GCs apoptosis (P<0.01), and the results of RT-qPCR and Western blot showed that lncRNA-MSTRG7889.1 competitively bound bta-miR-146a to promote the expression of Smad4 gene at mRNA and protein levels (P<0.01). 【Conclusion】 lncRNA-MSTRG.7889.1 competitively binds to bta-miR-146a, promotes the expression of Smad4 and inhibits the apoptosis of yak GCs.

Development of a Full-Automated Magnetic Particle Chemiluminescence Immunoassay Assay for Quantitative Detection of Antibodies Against Foot and Mouth Disease Virus Serotype O

BAO YanFang, JIANG Tao, HE Li, LÜ Lü, LI TaoShan, LI Xin, SUN YanYan, YANG Guang, WEI Ting, PAN XiaoLe, LIN Mi

Scientia Agricultura Sinica. 2024, 57(4):  810-819.  doi:10.3864/j.issn.0578-1752.2024.04.014

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【Background】 Foot-and-mouth disease (FMD) is an acute, febrile and virulent infectious disease caused by foot-and-mouth disease virus (FMDV), and vaccination is an effective measure to prevent the spread of FMD. The level of immune antibody monitoring is an important basis of evaluating the effectiveness of vaccine immunization, and developing immunization procedures. It is an indispensable step in the prevention and control of FMD. Therefore, to establish an efficient, rapid and fully automated antibody detection method is of great significance. 【Objective】The aim of this study was to establish a novel, fully-automatic and quantitative antibody detection method for FMD virus type O based on the chemiluminescence immunoassay (CLIA) technology of magnetic particles (MPs), in order to provide the technical support for the immunization monitoring of FMD and epidemic prevention and control.【Method】In this study, the magnetic nanoparticles were used for solid-phase carriers and separation carriers to encapsulate the captured antibody, the detection antibodies was labelled by using alkaline phosphatase (ALP), and AMPPD was used for the luminescent substrate. Finally, a new magnetic particle Chemiluminescence CLIA method (MP-CLIA) was established after optimization of conditions. In this method, firstly, the magnetic particle-polyclonal antibodies (MPs-pAbs), the samples to be tested, and the antigen of FMDV type O were added to a reaction system and incubated at 37 ℃, then the appropriate amount of enzyme-labeled antibodies (ALP-pAbs) was added and incubated at 37 ℃, finally, the chemiluminescence substrate AMPPD was added to detect the relative light unit (RLU). In this study, a standard curve was fitted by testing the standards, and thereceiver operating characteristic (ROC) curve was applied to determine the determination criteria of the assay. The quality control samples were utilized for methodological evaluation, and the field samples were also tested and compared with the liquid phase blocking ELISA (LPB-ELISA) to validate the assay results. 【Result】 The optimized reaction conditions were 0.25 mg·mL^-1 of magnetic particles, 1:1 000 dilution of FMD O antigen, 1:2 000 dilution of enzyme-labeled antibody, and 20 µL of spiked sample volume. The entire detection process was completed in a fully automated chemiluminescence immunoassay analysis apparatus, with a reaction time of 20 min, The quantitative detection could be conducted within the range of antibody content of 0-1 280 U (potency of 0-1:2 048), and the standard curve R²>0.99. The sensitivity of assay was 94.66%, the specificity was 97.10%, and there was no cross reactivity with the antibodies of six different pathogens which include SVV, PRRSV, BEFV, PCV2, QRFV, and PPRV. The repeatability of the assay showed the coefficient of variation less than 10%. The detection of field samples demonstrated the accordance rate was 94.69% between CLIA and LPB-ELISA, and the result of quantitative detection showed a correlation (R²=0.8473, P<0.0001) during two method. 【Conclusion】 The established MP-CLIA method was time-consuming, easy to operate, and could be fully automated by matching with the domestic automatic chemiluminescence instrument, which was a new and highly efficient method for the quantitative detection of antibody to FMD virus type O, and had a high value for clinical application.

RESEARCH NOTES

Characteristics of Good Taste and High Yield Type Japonica Rice in the Lower Reaches of the Yangtze River

ZHU TianCi, MA TianFeng, KE Jian, ZHU TieZhong, HE HaiBing, YOU CuiCui, WU ChenYang, WANG GuanJun, WU LiQuan

Scientia Agricultura Sinica. 2024, 57(4):  820-830.  doi:10.3864/j.issn.0578-1752.2024.04.015

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【Objective】In order to explore the common agronomic and physiological characteristics of good taste and high yield type japonica rice in the lower reaches of the Yangtze River, the study could provide theoretical basis and technical support for realizing synergistic improvement of rice yield and quality in this region. 【Method】The cultivar screening tests were conducted in 2018 and 2022 using 14 and 13 conventional japonica rice cultivars, respectively, to systematically compare the yield and composition, taste quality and textural characteristics, as well as agronomic and physiological indicators such as biomass, stem non-structural carbohydrates (NSC), and leaf area duration (LAD) among the different cultivars.【Result】The taste value and yield varied across the different cultivars, so through clustering by taste value and yield, they can be further divided into three types: medium taste and low yield (ML), medium taste and high yield (MH), good taste and high yield (GH). The average taste value and yield of GH in two years were 68.5 and 10.2 t·hm^-2, respectively, which were 6.8% higher in taste value and 14.6% higher in yield than MH and ML. In terms of performance for yield and quality traits, GH showed the highest grain filling percentage and taste scores compared to MH and ML. In terms of the dry matter accumulation, GH increased the proportion of dry matter accumulation at the early stage of grain filling stage, and showed the highest dry matter accumulation from heading stage to 15 days after heading stage. Further analysis of the causes of dry matter accumulation at this stage revealed that GH significantly increased the NSC remobilization rate while steadily increasing LAD. Correlation analyses showed highly significant positive correlations between grain filling percentage and taste, dry matter accumulation and LAD from heading stage to 15 days after heading stage, and NSC remobilization rate.【Conclusion】 While maintaining a high LAD from heading stage to 15 days after heading stage (100.4 m²·m^-2·d), further increasing NSC remobilization rate during grain filling stage (79.9%) to promote the initiation of grain filling, increasing the dry matter accumulation from heading stage to 15 days after heading stage (3.6 t·hm^-2), then improving the grain filling percentage (95.4%), and taste (9.6), which are the common characteristics of good taste and high yield type japonica rice in this region. In addition, the development of water and fertilizer management techniques targeting the enhancement of NSC remobilization at the grain filling stage and NSC accumulation at heading stage is expected to further exploit the yield and taste quality potential of the above good taste and high yield type japonica rice.