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    16 March 2022, Volume 55 Issue 6
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Research Progress on the Physiology and Its Molecular Mechanism of Seed Desiccation Tolerance
    SONG SongQuan,LIU Jun,TANG CuiFang,CHENG HongYan,WANG WeiQing,ZHANG Qi,ZHANG WenHu,GAO JiaDong
    Scientia Agricultura Sinica. 2022, 55(6):  1047-1063.  doi:10.3864/j.issn.0578-1752.2022.06.001
    Abstract ( 1003 )   HTML ( 118 )   PDF (971KB) ( 459 )   Save
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    Dehydration tolerance (DT) is defined as the ability of an organism or tissue to survive the removal of all, or almost all the cellular water without irreversible damage. DT of seeds is an adaptive mechanism to ensure the survival and reproduction of plant species in the long-term evolution process, and plays a key role in the conservation of plant seeds and germplasm resources. However, the DT of seeds is a complex trait, and its molecular mechanism is not now largely understood. Therefore, in the present paper, the research progresses on the physiological and molecular mechanisms of seed DT were reviewed. It was found that the DT of orthodox seeds was gradually formed during development, and reached the peak at physiological maturity. Recalcitrant seeds do not undergo the development stage of maturity dehydration, and are very sensitive to dehydration throughout development. Mature orthodox seeds maintained their resistance to re-dehydration at the initial stage of imbibition. With the time course of germination, the DT decreased gradually, and finally lost completely. The DT of seeds and embryos can be re-established during the early stage of germination, and of different tissues is different. The DT of seeds and embryos was inversely correlated with the decrease in mitochondrial respiratory activity. Respiratory activity of recalcitrant axis mitochondria was higher than that of orthodox embryo ones. During dehydration, the H2O2 content, the production rate of superoxide anion radical (·O2-) and the content of thiobarbituric acid reactive substance in desiccation-tolerant embryos (axes) were significantly lower than those of desiccation-sensitive embryos (axes), while the reactive oxygen species scavenging system in desiccation-tolerant embryos (axes), including enzymatic and non-enzymatic activities, was significantly higher than that in desiccation-sensitive embryos (axes). During the maturation of seeds, the accumulation of late embryogenesis abundant (LEA) proteins, small heat shock proteins and non-reducing oligosaccharides is closely related to the formation of DT. The AFL subfamily of B3 transcription factors (including ABI3 (ABA INSENSITIVE 3), FUS3 (FUSCA3) and LEC2 (LEAFY COTYLEDON 2)) increase the DT of seeds and embryos by positively regulating the accumulation of storage materials and protective proteins. The level of DNA methylation increased significantly throughout seed development and then decreased gradually during seed germination. Compared with embryos during the early stage of development and seedlings, mature embryos had a higher level of genomic methylation. In seeds, the parallel ABA and DOG1 (DELAY OF GERMINATION 1) signaling pathways activate synthesis of raffinose family oligosaccharides, and expression of LEA and HSP (heat shock protein) genes, thus regulating the onset of DT and transit to dormancy. Finally, the scientific issues that require to be further studied in this field are proposed, including the re-establishment of their model research system by using seeds and their tissues with different DT. Germinability, DT and dormancy characteristics of seeds are initiated and completed during development, and the relationship among them is still now unclear. There are both core ABA signaling pathway and DOG1 signaling pathway in seeds, and they converge at the ABI3 or downstream of ABI3. Which pathway will response preferentially and how these two pathways coordinate during dehydration of seeds? This paper will provide a reference for comprehensively understanding of the physiology and molecular mechanism of seed DT, increasing the stress resistance and yield of plant crops, improving the storage conditions of the resource bank and long-term preserving plant seed (germplasm) resources.

    Genome-Wide Association Analysis of Lead Tolerance in Wheat at Seedling Stage
    ZHI Lei,ZHE Li,SUN NanNan,YANG Yang,Dauren Serikbay,JIA HanZhong,HU YinGang,CHEN Liang
    Scientia Agricultura Sinica. 2022, 55(6):  1064-1081.  doi:10.3864/j.issn.0578-1752.2022.06.002
    Abstract ( 585 )   HTML ( 82 )   PDF (2881KB) ( 363 )   Save
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    【Objective】With the advancement of industrialization, the pollution of arable land by heavy metals, especially lead, has become a worldwide problem. Wheat is an important food crop, and its safe cultivation is critical to maintaining food security. Screening wheat varieties with strong tolerance to lead, low lead accumulation and mining relevant regulatory genes or QTL regions would lay foundation for further elucidating the genetic mechanism of lead tolerance in wheat. 【Method】The tolerance to lead of 102 wheat varieties (advanced lines) were evaluated with a 140 mg·kg-1 lead nitrate solution at the seedling stage, by the weighted membership function value (D Value) of the lead tolerance coefficients of maximum root length, root biomass and growth rate under three replicates, combining with the 335 438 high-quality SNPs (single nucleotide polymorphism)markers by wheat 660K SNP chip, a genome-wide association study (GWAS) for lead tolerance in wheat was conducted, to mine the candidate genes for lead tolerance in wheat. 【Result】The lead among between wheat varieties (advanced lines) under different replicates showed rich variation, with the coefficient of variation of 44.8%-46.2%, and the correlation coefficient was between 0.87-0.97 (P<0.001). It was found that varieties with strong lead tolerance showed low lead accumulation characteristics. The genotyping results showed that the SNP polymorphic information content (PIC) was 0.28-0.32, the population structure analysis showed that these wheat materials could be divided into 7 subgroups; a total of 20 SNPs and 8 key chromosomal segments that were significantly associated with lead tolerance in wheat (P≤0.001) were detected by two GWAS methods respectively distributed on chromosomes 1B, 2A, 2D, 3A, 3B, 5A, and 7A. A single locus explains 15.33%-19.75% of the phenotypic variation, and 10 and 8 key chromosomal segments were detected in two and more environments. Analysis of candidate genes for significant and stable association sites and intervals revealed that the functions of the candidate genes were mainly related to transmembrane transport, protein modification and oxidative stress response, including 7 genes related to transporters, including TraesCS1B02G433800, TraesCS7A02G118800, TraesCS7A02G117900 etc. Two genes involved in ubiquitination and deubiquitination (TraesCS2A02G550900 and TraesCS7A02G477300), three genes encoding transmembrane proteins (TraesCS2D02G570500, TraesCS3B02G039900 and TraesCS3B02G466000), one gene related to peroxidase (TraesCS7A02G474200). 【Conclusion】Seven wheat materials with strong lead tolerance were screened, 20 SNPs in 8 candidate regions significantly related to lead tolerance in wheat were detected, and 13 candidate genes related to lead tolerance in wheat were finally screened.

    Comprehensive Evaluation of Phenotypic Characters of Nature Population in Upland Cotton
    WANG XiuXiu,XING AiShuang,YANG Ru,HE ShouPu,JIA YinHua,PAN ZhaoE,WANG LiRu,DU XiongMing,SONG XianLiang
    Scientia Agricultura Sinica. 2022, 55(6):  1082-1094.  doi:10.3864/j.issn.0578-1752.2022.06.003
    Abstract ( 614 )   HTML ( 75 )   PDF (731KB) ( 269 )   Save
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    【Objective】To analyze the fiber quality, yield, and early maturity of Upland cotton germplasm, screen the evaluation indicators comprehensively, establish a reliable evaluation model, and provide theoretical support for developing new Upland cotton varieties.【Method】 A total of 630 Upland cotton accessions selected from various sources were used to investigate 17 traits in 8 environments in three major cotton-growing regions in China. The best linear unbiased prediction (BLUP) was estimated for phenotypic traits by utilizing R package “lme4” and used for further analysis, including correlation analysis, principal component analysis, affiliation function method, cluster analysis and stepwise regression to evaluate population characteristics.【Result】This upland cotton population had high genetic diversity with a diversity index ranging from 1.961 to 2.084, and significant regional specificity existed. The boll number, fiber elongation, spinning consistent index, and short fiber index of this population had considerable variation, while that of fiber length, fiber strength, and growth period traits was lower. Correlation analysis showed significant or highly significant correlations among most traits, and there was a strong correlation among some fiber quality traits. The 17 traits were converted into 6 independent composite indices through principal component analysis with a contribution range of 5.860%-31.044% and a cumulative contribution of 82.642%. Principal component analysis can classify the fiber quality traits, yield traits and agronomic traits in this upland cotton population. The comprehensive evaluation value (F value) of phenotypes was calculated using the affiliation function method, the phenotypes of 17 traits were all significantly correlated with F values. The result demonstrated that the accessions with high F values (mean value of 0.668) had significantly higher yield traits (boll number, boll weight, lint percentage, and seed index), fiber quality traits (fiber length, fiber length uniformity, fiber strength, spinning consistent index, and short fiber index), plant height and fertility traits than those with low F values (mean value of 0.396). Regression equations for eight traits (boll number, boll open date, boll weight, flowering date, lint percentage, plant height, sympodial brand node, and spinning consistent index) as independent variables were established using stepwise regression. Based on the F value, the 630 germplasm were clustered into four categories, the first category was the high-quality fiber type, containing 118 accessions; the second category was the high-yield type, comprising 250 accessions; the third category was the early-maturing type, comprising 51 accessions; the characteristics of the fourth category was between the second and third categories. Finally, 23 better fiber quality and 135 high-yield germplasms were selected for breeding and production.【Conclusion】The phenotypic traits of Upland cotton are geographically specific; it is feasible to use multivariate statistical analysis to comprehensively evaluate Upland cotton germplasm; the whole population can be classified into four categories (high-quality, high-yield, early maturity and other types).

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Increasing Effects of Wheat Yield Under Mulching Cultivation in Northern of China: A Meta-Analysis
    QIN YuQing,CHENG HongBo,CHAI YuWei,MA JianTao,LI Rui,LI YaWei,CHANG Lei,CHAI ShouXi
    Scientia Agricultura Sinica. 2022, 55(6):  1095-1109.  doi:10.3864/j.issn.0578-1752.2022.06.004
    Abstract ( 453 )   HTML ( 4 )   PDF (698KB) ( 244 )   Save
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    【Objective】The aim of this study was to identify the impact of straw mulching and plastic film mulching on wheat production in Northern China, so as to explore the suitable promotion areas of two mulching systems.【Method】In this study, a total of 165 literature was retrieved and screened in recent 40 years, which were put into the framework of Meta-analysis, and were carried out through different forms of overall effect analysis (such as change rate and response ratio) for theoretical research. Generally, by adopting random effect model, the changes of wheat agronomic indexes and farmland moisture conditions under different mulching patterns were analyzed. Then, the response rules of mulching yield increasing effect to various environmental conditions (altitude, precipitation, temperature, and sunshine) and field management measures (mulching period, planting density, tillage, and fertilization) were revealed by subgroup analysis, while the function fitting, weight analysis and statistical test were carried out. The correlation between the variables involved in this study was quantitatively analyzed by Pearson correlation coefficient method.【Result】Compared with the open field cultivation, the straw and plastic film mulching significantly increased wheat yield by 19.53% (95%CI =0.55%-38.52%) and 24.91% (95%CI =3.18%-46.64%), which also inhibited field evapotranspiration. Furthermore, it was found that there were some differences in contribution rate of yield components to yield under different mulching patterns, which were: effective spike number > grain number per spike > 1000 grain weight (straw mulching); effective spike number > 1 000 grain weight > grain number per spike (plastic film mulching). The increase of grain number per spike under straw mulching was higher, that about 5.7% (95%CI = -4.10%-15.50%); while increase of effective spike number and 1000 grain weight under film mulching was more significant, which were 25.2% (95%CI = 14.11%-36.29%) and 6.4% (95%CI = 1.50%-11.30%), respectively. In addition to the advantages of promoting production, the biomass and water use efficiency of film mulching were also 18.17% and 14.39% higher than that of straw mulching, respectively. Specifically, the yield increase rate of plastic film mulching was 0.89%-23.34% higher than that of straw mulching in most meteorological subregions. Meanwhile, with the declined of terrain height, the yield increasing effect of plastic film showed the growth trend, compared with non-mulching treatments, and the yield increase rate could reach 34.26% in low altitude area (< 800m). However, the yield increasing advantage of plastic film mulching over straw mulching was declined gradually with the increase of mulching years. In the more than 8 years of mulching experiments, the overall yield increasing rate of straw mulching was higher. The yield of straw mulching was also affected by fertilization and tillage measures, especially in the three treatments of no tillage, no fertilization and applying phosphate fertilizer, the yield increase rates were 32.68%, 25.94% and 21.71%, respectively. According to statistical test, among the three subgroups of altitude, average annual sunshine hours and planting density, their inter group heterogeneity Q test statistics were larger, indicated that the variation degree of each effect quantity in these groups was higher. Finally, it was found that under the conditions of straw and plastic film mulching, the factors with the highest correlation to yield were effective spike number (r = 0.808) and water use efficiency (r = 0.718), while the most primary factors affecting the soil water content in two mulching systems were evapotranspiration (r = -0.859) and water use efficiency (r = 0.856), respectively.【Conclusion】In conclusion, these two mulching patterns possessed obvious effect on yield increase, while plastic film mulching had more advantages in low altitude, relative drought and cold regions; The straw mulching was more suitable for long-term conservation tillage system, so as to achieve the coordinated development of production and ecology. Therefore, the key to the success of wheat mulching technology in northern China is to choose scientific mulching methods according to local and time conditions.

    Early Detection on Wheat Canopy Powdery Mildew with Hyperspectral Imaging
    CAI WeiDi,ZHANG Yu,LIU HaiYan,ZHENG HengBiao,CHENG Tao,TIAN YongChao,ZHU Yan,CAO WeiXing,YAO Xia
    Scientia Agricultura Sinica. 2022, 55(6):  1110-1126.  doi:10.3864/j.issn.0578-1752.2022.06.005
    Abstract ( 431 )   HTML ( 11 )   PDF (3572KB) ( 180 )   Save
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    【Objective】In this study, the near-ground imaging spectrometer was used to obtain time-series images of wheat canopy after inoculation with powdery mildew, which aimed to explore the ability and performance of the combination of spectral feature and texture feature in the early detection of wheat powdery mildew at canopy scale. 【Method】 Based on the field trials of wheat varieties with different disease resistance in different years, the wavelet features sensitive to wheat powdery mildew were extracted by continuous wavelet transform (CWT) method, and the corresponding texture features were extracted based on spectral features to construct normalized difference texture index (NDTI). Meanwhile, the representative traditional vegetation indices (Vis) were selected. Then, based on these features and combinations, the partial least squares discriminant analysis (PLS-LDA) model was used to establish wheat canopy healthy and disease recognition model. The partial least squares regression (PLSR) was used to estimate the severity of wheat canopy disease. The technique was used to distinguish the healthy and disease wheat at different days after inoculation based on the optimal features and combinations. 【Result】 Based on CWT, the selected four wavelet features were 595 nm (yellow region) at 6 scales, 614 nm (red region) at 5 scales, 708 nm (near infrared region) at 3 scales, and 754 nm (near infrared region) at 4 scales respectively. The following texture features were selected for the best texture index combination: ENT754, MEA754, ENT708, ENT595, ENT614, HOM708, HOM595, HOM614, DIS595, HOM754 and DIS614. Besides, it was found that the texture feature MEA754 had the superior performance among all the texture, with the highest correlation between the severity of disease and texture (R2=0.67). The PLS-LDA model based on the combination of wavelet feature and texture feature had the highest accuracy, with the overall classification accuracy of 81.17% and the Kappa coefficient of 0.63. In addition, the PLSR model based on spectral index and texture index was the best, and the R 2 of modeling and testing was 0.76 and 0.71, respectively. The severity of wheat canopy powdery mildew was about 26% (about 24 days after inoculation), which was identified in this study at the earliest time. 【Conclusion】 The wheat healthy and disease recognition model based on the combination of wavelet feature and texture feature could significantly improve the accuracy of disease classification, and the combination of spectral index and texture index could significantly improve the accuracy and stability of disease severity estimation. The method and results of this study could provide the reference for disease monitoring of other crops and technical support for accurate application of modern intelligent agriculture.

    Prediction of Maize Yield in Relay Strip Intercropping Under Different Water and Nitrogen Conditions Based on PLS
    TAN XianMing,ZHANG JiaWei,WANG ZhongLin,CHEN JunXu,YANG Feng,YANG WenYu
    Scientia Agricultura Sinica. 2022, 55(6):  1127-1138.  doi:10.3864/j.issn.0578-1752.2022.06.006
    Abstract ( 444 )   HTML ( 1 )   PDF (2429KB) ( 176 )   Save
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    【Objective】 This study was designed mainly to provide technical means for non-destructive prediction of intercropped maize yield. The prime objective of our study was to construct a “hyperspectral parameter-photosynthetic pigment-yield” model from the hyperspectral data. 【Method】 Based on field trials of different years, locations, varieties, and treatments (nitrogen fertilizer, moisture), the relationships among photosynthetic pigment parameters, canopy hyperspectral parameters, and maize yield at each growth period and the entire growth period of intercropping maize were comprehensively analyzed. In addition, the optimal growth period and photosynthetic pigment parameters for maize yield prediction were also clarified. Then, the prediction model of yield was constructed based on linear function, quadratic function and partial least squares regression (PLS). 【Result】 Among the photosynthetic pigment-yield prediction models, the PLS prediction model of yield based on canopy carotenoid density had the best effect (R 2=0.882, RMSE=0.669 t·hm -2). In the spectral parameter-photosynthetic pigment analysis, the chlorophyll content during the tasseling stage had the best correlation with the band free combination index rRVI (534, 546) (r=0.927). The correlation between the other photosynthetic pigment parameters and the corresponding spectral index was above 0.797. In the hyperspectral parameter- photosynthetic pigment-yield prediction model, the chlorophyll content, carotenoid content, canopy chlorophyll density, and canopy carotenoid density were used as connection points, and using the spectral indices of rNDVI (534, 546), rRVI (531, 555), rNDVI (532, 546), and rNDVI (531, 555) as independent variables, the PLS output prediction model had better effect (R2=0.509, RMSE=1.352 t·hm -2). 【Conclusion】 In intercropping maize, the pigment parameters were used as a bridge between spectral data and yield. A prediction model was established through PLS regression, which could achieve a better estimation of maize yield and provide the theoretical and technical reference for field management and growth monitoring of maize in intercropping.

    PLANT PROTECTION
    The Role of Chalcone Synthase Gene in Grape Resistance to Gray Mold and Downy Mildew
    GUO ZeXi,SUN DaYun,QU JunJie,PAN FengYing,LIU LuLu,YIN Ling
    Scientia Agricultura Sinica. 2022, 55(6):  1139-1148.  doi:10.3864/j.issn.0578-1752.2022.06.007
    Abstract ( 400 )   HTML ( 29 )   PDF (1810KB) ( 134 )   Save
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    【Objective】The objective of this study is to clone the chalcone synthase gene CHS1 in different resistant grape varieties, clarify the expression pattern under the infection of Botrytis cinerea and Plasmopara viticola, verify the subcellular localization and function of CHS1, and to provide a basis for exploring the broad-spectrum resistance mechanism of CHS1.【Method】CHS1 genes were cloned from the leaf cDNAs of Vitis quinquangularis ‘Yeniang-2’, Vitis vinifera ‘Thompson Seedless’ and Vitis amurensis ‘Shuanghong’. The differences in the nucleic acid sequence and physicochemical properties of CHS1 protein were analyzed by bioinformatics. Expression patterns of CHS1 in different varieties during B. cinerea and P. viticola infection were also detected by qRT-PCR. The expression vector was constructed, and the transient expression technology mediated by Agrobacterium was used to verify the subcellular localization and disease resistance function in the leaves of tobacco and ‘Thompson Seedless’ tissue culture seedlings.【Result】The CHS1 is highly conserved among different grape varieties. The full length of ORF is 1 182 bp, which encodes 393 amino acids. The predicted molecular weight of the encoded protein is 42.92 kD, which is a stable hydrophilic protein. After being infected by B. cinerea, the expression patterns of VqCHS1 and VvCHS1 were similar, but the expression level of VqCHS1 in the resistant variety ‘Yejiang-2’ was significantly higher. After being infected by P. viticola, VaCHS1 and VvCHS1 showed different expression pattern changes. The expression level of VaCHS1 in the resistant variety ‘Shuanghong’ continued to increase, while the expression level of VvCHS1 gradually decreased. The results of subcellular localization showed that CHS1 protein is distributed in the cytoplasm and nucleus, but it is mainly localized in the nucleus. Compared with the negative control, the ‘Thompson Seedless’ leaves overexpressing VaCHS1 showed stronger resistance to gray mold and downy mildew after artificial inoculation. 【Conclusion】The expression level of CHS1 in resistant varieties is higher than that in susceptible varieties, and overexpression of CHS1 can increase the resistance of susceptible grape varieties to gray mold and downy mildew.

    Widely Targeted Metabolomics Analysis of the Effects of Myzus persicae Feeding on Prunus persica Secondary Metabolites
    YAN LeLe,BU LuLu,NIU Liang,ZENG WenFang,LU ZhenHua,CUI GuoChao,MIAO YuLe,PAN Lei,WANG ZhiQiang
    Scientia Agricultura Sinica. 2022, 55(6):  1149-1158.  doi:10.3864/j.issn.0578-1752.2022.06.008
    Abstract ( 473 )   HTML ( 37 )   PDF (2043KB) ( 236 )   Save
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    【Objective】The objective of this study is to clarify the underlying biochemical mechanisms related to the resistance and susceptibility of peach towards Myzus persicae, and to identify the key secondary metabolites of peach responding to the M. persicae infection.【Method】New shoots of resistant (‘96-5-1’ (9651), ZaoYouTao (ZYT)) and susceptible (Zhong You 13 (CN13), Zhong Nong Jin Hui (ZNJH)) peach trees were inoculated with M. persicae for 3 days and used for secondary metabolite extraction and UPLC-MS/MS analysis. Differentially altered metabolites (DAMs) were screened with |log2 fold change|≥1, P-value≤0.01 as threshold. The VIP value of the OPLS-DA model was used to perform differences between resistant and susceptible peach.【Result】To illustrate the biochemical mechanisms of M. persicae resistance in peach, aphid-resistant/aphid-susceptible peach varieties infested with M. persicae for 3 days. A total of 528 metabolites were identified in the treated samples through widely targeted metabolomics analysis. Using principal component analysis (PCA), hierarchical cluster analysis (HCA) and Venn diagram analysis, it was found that 7 DAMs were identified from the susceptible variety CN13, and 2 of them were significantly decreased, 5 of them were significantly increased. 7 DAMs were identified from the susceptible variety ZNJH, and 3 of them were significantly decreased, 4 of them were significantly increased. 33 DAMs were identified from the resistant variety ‘96-5-1’, and 1 of them was significantly decreased, 32 of them were significantly increased. 55 DAMs were identified from the resistant variety ZYT, and 12 of them were significantly decreased, 43 of them were significantly increased. The majority of the DAMs were identified from the two resistant varieties, and the overall magnitude of change was greater in the resistant varieties than that in the susceptible varieties. Finally, 15 secondary metabolites (6 amino acids and their derivatives, 5 phenolic acids, 3 nucleotides and their derivatives, and 1 organic acid) were considered to be involved in M. persicae resistance of ‘Shou xing tao’.【Conclusion】The significantly up-regulated secondary metabolites obtained in M. persicae-resistant peach varieties were mainly involved in the response to M. persicae feeding. The regulation of these secondary metabolites (amino acids and their derivatives, phenolic acids, nucleotides and their derivatives, organic acid) is the important mechanism of defense reaction to M. persicae.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    The Spatial Characteristics of Soil Organic Carbon Sequestration and N2O Emission with Long-Term Manure Fertilization Scenarios from Dry Land in North China Plain
    WANG ShuHui,TAO Wen,LIANG Shuo,ZHANG XuBo,SUN Nan,XU MingGang
    Scientia Agricultura Sinica. 2022, 55(6):  1159-1171.  doi:10.3864/j.issn.0578-1752.2022.06.009
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    【Objective】To provide theoretical guidance for mitigating global warming, the comprehensive effects of manure amendment on soil organic carbon sequestration and greenhouse gas emissions were studied.【Method】Based on the long-term experiment, the validated process-based model-SPACSYS, combined with regional database and ArcGIS, was used to clarify the spatial characteristics of average annual soil organic carbon sequestration rate (SOCSR), average annual soil N2O emission and average annual net global warming potential (NGWP) under three long-term fertilization scenarios (equal nitrogen fertilization), namely, chemical fertilizers only (NPK), 50% of chemical fertilizers combined with 50% of manure (NPKM(5:5)) and 30% of chemical fertilizers combined with 70% of manure (NPKM(3:7)), from dry land in the North China Plain from 2010 to 2050.【Result】The SOCSR from dry land in the North China Plain was higher in the east and lower in the west, and the higher regions mainly included Jiangsu Province and Shandong Province. Correlation analysis showed that there was a significant negative correlation between SOCSR and initial soil organic carbon content. Stepwise linear regression analysis further indicated that initial soil organic carbon content, mean annual temperature and soil pH were three important factors affecting SOCSR, which accounted for 24% of the variation of SOCSR. The average annual soil N2O emission was higher in the central part of the North China Plain, lower in the north and south, and the higher regions included parts of Shandong Province and Jiangsu Province. Correlation analysis showed that there was a significant positive correlation between average annual soil N2O emission and initial soil organic carbon content. In general, compared with NPK, NPKM(5:5) and NPKM(3:7) both increased SOCSR and decreased average annual soil N2O emission from dry land in the North China Plain, of which SOCSR (233 and 236 kg C·hm-2·a-1) increased by 79% and 82%, respectively, the average annual soil N2O emission(15.8 and 14.4 kg N·hm-2·a-1) decreased by 21% and 28%, respectively. As a result, NGWP (6.6 and 5.9 t CO2-eq·hm-2·a-1) decreased by 26% and 34%, respectively.【Conclusion】Compared with the application of chemical fertilizers, the application of chemical fertilizers combined with manure was beneficial to SOC sequestration, soil N2O emission reduction and mitigation of glowing warming from dry land in the North China Plain over the long-term.

    Characteristics of Efficient Nitrogen Uptake and Transport of Rapeseed at Seedling Stage
    CHAO ChengSheng,WANG YuQian,SHEN XinJie,DAI Jing,GU ChiMing,LI YinShui,XIE LiHua,HU XiaoJia,QIN Lu,LIAO Xing
    Scientia Agricultura Sinica. 2022, 55(6):  1172-1188.  doi:10.3864/j.issn.0578-1752.2022.06.010
    Abstract ( 476 )   HTML ( 36 )   PDF (2059KB) ( 155 )   Save
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    【Objective】Nitrogen (N) uptake efficiency is one of the important factors affecting crop N efficiency, investigating characteristics of efficient N uptake and transport was the purpose to provide the theoretical basis for improving N efficiency and yield of rapeseed (Brassica napus L.) varieties.【Method】To explore the mechanisms underlying high N uptake and transport in rapeseed, two rapeseed germplasms with contrasting N efficiency (N efficient germplasm ‘498’ and N inefficient germplasm ‘428’) were used in this study under normal N (9.5 mmol·L-1) and low N (0.475 mmol·L-1) conditions at three different growth stages (Phenological growth stages 12, 14 and 16) in hydroponic culture. At the same time, the 15N isotope tracer technique was applied to study the uptake and transport capacity of NO3- and NH4+. Additionally, the expression level of genes (BnNPFs, BnNRT2s and BnAMTs)related to N uptake and transport in rapeseed germplasms with contrasting N efficiency were further analyzed by real-time quantitative PCR (RT-qPCR).【Result】Rapeseed germplasm ‘498’ showed superior advantages in plant growth and root development under different N concentrations, and the root morphological indexes (main root length, total root length, root surface area, root volume and lateral root number), biomass, N accumulation and N uptake efficiency were all significantly greater than those of germplasm ‘428’.15N isotope tracer test also showed that ‘498’ showed greater advantage in the uptake and accumulation of NO3 - and NH4+, especially for NH4+, as indicated by the significant differences in the accumulation of 15NH4+ between two germplasms. The RT-qPCR analysis further found that under normal N conditions, the relative expressions of BnNPF6.3a, BnNRT2.1e, BnNPF7.2a, BnNPF7.2c, BnNPF6.2c, BnAMT1;2a, BnAMT1;3c, BnAMT1;4a, BnAMT2;1a and BnAMT2;1b (involved in the uptake and transport of NO3 - and NH4+) was significantly higher in ‘498’ than that in ‘428’. While under low N stress, the relative expressions of BnNRT2.4a, BnNRT2.5a and BnNRT2.5b (involved in NO3 -uptake and transport) was significantly lower in the root of ‘498’ than that of ‘428’, but the expression level of BnNPF7.3a and BnNPF6.2c (referred to NO3 - transport and redistribution) was significantly higher in ‘498’ than that in ‘428’, as well as the expression level of BnAMT1;1a, BnAMT1;2a, BnAMT1;3c, BnAMT1;4a, BnAMT2;1a and BnAMT2;1b (involved in NH4 + uptake and transport).【Conclusion】Compared with N-inefficient germplasm ‘428’, N-efficient germplasm ‘498’ were superior in root length, root surface area (volume) and lateral root number, additionally with greater ability in N (especially NH4 +) uptake and accumulation. Under normal N application conditions, the expression of genes involved in NO3- and NH4+ absorption and transport were relatively higher in ‘498’, while the relative expression of genes involved in the NO3 - transport and redistribution as well as NH4+ absorption and transport were significantly higher in ‘498’ than that in ‘428’ under low N stress, illustrating the relative higher N uptake efficiency of ‘498’ possibly linked to the higher expressions of several BnNPFs sand BnAMTs.

    HORTICULTURE
    Effects of Supplemental Far-Red Light on Growth and Abiotic Stress Tolerance of Pepper Seedlings
    DONG SangJie,JIANG XiaoChun,WANG LingYu,LIN Rui,QI ZhenYu,YU JingQuan,ZHOU YanHong
    Scientia Agricultura Sinica. 2022, 55(6):  1189-1198.  doi:10.3864/j.issn.0578-1752.2022.06.011
    Abstract ( 438 )   HTML ( 41 )   PDF (1286KB) ( 189 )   Save
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    【Objective】This study analyzed the effects of supplementary far-red light (FR) on the growth and abiotic stress tolerance of pepper seedlings, aiming to provide a theoretical basis on precise light environments for cultivating high quality vegetable seedlings.【Method】In this study, Bola Hongshuai pepper cultivar was used as the research material. The 7-day-old seedlings were cultivated under two LED light environments, including the control spectrum (NL; R/B = 3/1, 150 μmol∙m -2∙s-1 PPFD) and the NL with an extra 10 μmol∙m -2∙s-1far-red light (6% FR), 20 μmol∙m -2∙s-1far-red light (13% FR), and 30 μmol∙m -2∙s-1far-red light (20% FR). Chilling and drought were imposed when the seedlings were 21 days old. Biomass, resistance-related gene expression, antioxidant enzyme activity, hormone content, chlorophyll fluorescence parameters, and leaf relative electrolyte leakage (REL) were analyzed to explore the effects of supplemental FR on growth and abiotic stress tolerance of pepper seedlings. 【Result】 Compared with the control, the supplementation of 6% FR was beneficial to increase the height, stem thickness, dry weight, fresh weight and seedling indexes of pepper seedlings. Moreover, the supplementation of 6% FR significantly increased the expression of the cold response gene CBF1 and antioxidant enzyme-related genes, such as Cu/Zn-SOD, GR, APX, CAT and DHAR under chilling stress. The activity of SOD, APX, DHAR, CAT and GR as well as the ABA content of pepper seedlings under low temperature increased by 25.2%, 53.6%, 55.8%, 72.7%, 33.4% and 69.5%, respectively, following the treatment with supplemental FR. The PSII maximum photochemical efficiency (Fv/Fm) of pepper leaves after supplementation of 6% FR under low temperature stress significantly increased compared with the control, while REL decreased obviously, indicating that supplementation of 6% FR alleviated the low temperature-induced PSII photoinhibition and damage in leaves and enhanced the cold tolerance of pepper seedlings. Under the drought stress condition, compared with the control, the supplementation of 6% FR improved the activity of SOD, GR, APX, CAT and DHAR by 13.7%, 38.0%, 37.2%, 27.6% and 23.7%, respectively. The ABA content and PSII actual photochemical efficiency (ΦPSII) were both increased while REL was decreased significantly. The results showed that the supplementation of 6% FR reduced the degree of PSII photoinhibition and membrane lipid peroxidation caused by drought stress, and improved the drought tolerance of pepper seedlings.【Conclusion】The study found that supplementation of 6% FR could not only improve the quality of pepper seedlings, but also enhance the resistance of pepper seedlings to low temperature and drought stresses by improving antioxidant defense and ABA homeostasis.

    Identification of R2R3-MYB Subfamily in Chinese Jujube and Their Expression Pattern During the Fruit Development
    LI ShiJia,LÜ ZiJing,ZHAO Jin
    Scientia Agricultura Sinica. 2022, 55(6):  1199-1212.  doi:10.3864/j.issn.0578-1752.2022.06.012
    Abstract ( 699 )   HTML ( 56 )   PDF (6642KB) ( 305 )   Save
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    【Objective】As one of the largest gene families in plants, MYB family plays an important role in plant growth and fruit development. This study would identify the members of the R2R3-MYB subfamily of Chinese jujube and analyze their bioinformatics and expression patterns, aiming to provide the reference for further exploring their function in jujube growth, especially in fruit development.【Method】The R2R3-MYB genes of Arabidopsis thaliana were used as probes, and this subfamily members in the jujube genome were screened out by BLAST and HMMER. Based on the Plant Transcription Factors Database, Uniprot, Gcorn Plant and other databases, R2R3-MYB genes were analyzed by MEGA-X, ExPASy, TBtools, and PheatMap. Transcriptome data and real-time quantitative PCR (qRT-PCR) were used to evaluate the expression of these genes, and the interaction proteins of key candidate proteins were predicted and verified. 【Result】In this study, 118 R2R3-MYB subfamily members were identified in jujube genome, all of which had a typical MYB domain. They were divided into 18 subgroups, named ZjMYB1-ZjMYB118. The number of amino acids encoded by the genes ranged from 200 to 400. Among them, 95 genes were located on 12 chromosomes, and which on Chr. 4 was the most widely distributed. The phylogenetic analysis showed that orthologous and paralogous events occurred during the evolution of these genes. By transcriptomic and qRT-PCR analysis, some R2R3-MYB genes were involved in the fruit development. In particular, ZjMYB59 and ZjMYB104 might participate in fruit enlargement and color development. Protein interaction prediction and yeast two-hybrid test confirmed that ZjMYB104 and ZjMPK3 were interacted each other. 【Conclusion】118 members of the R2R3-MYB subfamily members were identified and distributed on 12 chromosomes. The structure of these subfamily members was highly conservative. The orthologous and paralogous events occurred in jujube genome evolution and the candidate genes closely related to fruit development were screened out. This study provided clues and references for further functional analysis of R2R3-MYB subfamily in jujube.

    FOOD SCIENCE AND ENGINEERING
    Risks and Their Prevention and Control of Modified Mycotoxins in Grain and Its Products
    YANG Hong,CAO WenMing,CHEN HeYan,WEI XueQing,SHU LiDan,LI Tong
    Scientia Agricultura Sinica. 2022, 55(6):  1213-1226.  doi:10.3864/j.issn.0578-1752.2022.06.013
    Abstract ( 400 )   HTML ( 31 )   PDF (562KB) ( 112 )   Save
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    Modified mycotoxins are a principal group of food safety hazards. In this study, based on reviews of the formation mechanism, classification, pollution status, toxicological research, metabolic rule, and analytical and detoxification methods of modified mycotoxins, the known and potential risks were identified, and risk prevention and control measures were analysed. The known risks were: in grain and its products, there was a high positive detection rate of modified mycotoxins, with a wide range of concentrations, which increased the risk of exposure to mycotoxins. Toxicological research on modified mycotoxins demonstrated that some modified mycotoxins transform back into their parent form during the process of metabolism and produced toxic effects equal to their parent form. Additionally, some other modified mycotoxins were toxic in themselves. The potential risks were: the coexistence of a diversity of metabolized toxins might lead to obscure toxicological mechanisms and effects. Some new modified mycotoxins with unknown structures, properties, or toxicities could be produced in the process of crop metabolism or food processing. Because it was restricted by a lack of analytical technology and quality control methods, it was difficult to obtain accurate and reliable qualitative and quantitative results for modified mycotoxins. Thus, the toxicological data for modified mycotoxins were extremely scarce, resulting in disputes over toxicological properties. The risk prevention and control measures were: starting from the source, the new varieties should be bred that were resistant to mycotoxin infection, normalize standards and testing systems should be built, and the purchase, storage, and production of agriculture products should be strictly controlled. In the food processing stage, more detailed research into detoxification technology after mycotoxin infection needs was required.

    ANIMAL SCIENCE·VETERINARY SCIENCE·RESOURCE INSECT
    Definition and Genetic Parameters Estimation for Health Traits by Using on-Farm Management Data in Dairy Cattle
    WANG Kai,ZHANG HaiLiang,DONG YiXin,CHEN ShaoKan,GUO Gang,LIU Lin,WANG YaChun
    Scientia Agricultura Sinica. 2022, 55(6):  1227-1240.  doi:10.3864/j.issn.0578-1752.2022.06.014
    Abstract ( 390 )   HTML ( 40 )   PDF (1798KB) ( 117 )   Save
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    【Objective】This study was conducted to define health traits and to estimate their genetic parameters by using on-farm management data in dairy cattle.【Method】In this study, the health event records were collected by extracting from farm management software from 46 large-scale dairy farms in Northern China. Totally, 1 326 kinds of health events were grouped into five categories by standardizing the acronyms of on-farm records, 18 kinds of most frequent health events were selected from five categories. According to whether a health event occurred at least once within a lactation, 18 binary individual health traits corresponding to the 18 selected health events were defined (observations being 0 or 1). Furthermore, in order to define cow’s general resistance to certain type of health problem, five binary general health traits were defined according to whether health problems within a health category occurs at least once in a lactation (observations being 0 or 1). The single trait and two traits linear animal models were used to estimate the genetic parameters for 23 newly defined health traits. 【Result】The estimated heritabilities for 23 health traits ranged from 0 (rumen acidosis) to 0.03 (milk fever). Udder health, reproductive disorders and metabolic disorders had the highest heritability (approximately 0.02) among five general health traits, whereas digestive disorders and hoof health had relatively low heritability (less than 0.01). Clustering health events into categories resulted in higher heritability for reproductive disorders and digestive disorders, while heritabilities of udder health, metabolic disorders and hoof health were lower than that of single health traits with the highest heritability in their respective category. The low genetic correlations between different health category traits were found; however, the moderate genetic correlations among some health traits from same category were observed. The health traits within the hoof health had the high genetic correlations with each other, ranged from 0.63 (Laminitis and Footrot) to 0.99 (Laminitis and Sole ulcer). For reproductive disorders, retained placenta had medium genetic correlations with metritis (0.47) and endometritis (0.46), respectively. For digestive disorders, relatively high genetic correlations were found between diarrhea and enteritis (0.94) as well as dyspepsia and antony of forestomachs (0.80). The ketosis and abomasum displacement had a genetic correlation of 0.42.【Conclusion】Based on the current data quality of health records in Chinese dairy farm and the results from the current study, it was suggested that five individual health traits (clinical mastitis, metritis, ketosis, abomasum displacement and milk fever) and two general health traits (digestive disorders and hoof health) could be considered as target traits, and they should be intensively considered in research and breeding practice for improving health traits genetically in Chinese Holstein population. This study proved the feasibility of defining health traits using on-farm management data in Chinese dairy cattle. The results from the current study provided a reference for research and genetic selection of disease resistance in dairy cattle, and could help to promote balanced breeding in Chinese dairy cattle population.

    Effects of FB1 on Apoptosis and Autophagy of Porcine Oocytes in vitro Maturation
    LI WenHui,HE YiJing,JIANG Yao,ZHAO HongYu,PENG Lei,LI Jia,RUI Rong,JU ShiQiang
    Scientia Agricultura Sinica. 2022, 55(6):  1241-1252.  doi:10.3864/j.issn.0578-1752.2022.06.015
    Abstract ( 371 )   HTML ( 33 )   PDF (4024KB) ( 121 )   Save
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    【Objective】The purpose of this study was to investigate the effects of fumonisin B1 (FB1) on porcine oocyte in vitro maturation and its potential mechanism, so as to provide the theoretical reference for effective prevention and treatment of reproductive toxicity injury caused by FB1 in the clinic.【Method】Porcine cumulus oocyte complexes (COCs) were collected and randomly divided into groups, and treated with different concentrations of FB1 (0, 10, 20 and 30 μg·mL-1) for 44 h during in vitro maturation, respectively. Then, the first polar body (PB1) extrusion and embryo development after activation of oocytes were analyzed, and the effects of FB1 on meiotic progression and cytoskeleton structure of oocytes were further detected by immunofluorescence staining and combined with confocal microscopy. In order to further explore the mechanism of FB1 on toxic injury of porcine oocytes, the JC-1, Annexin V-FITC and LC3A/B fluorescence staining were used to detect mitochondrial function, early apoptosis and autophagy levels in oocytes, respectively. On this basis, the expression of apoptosis/autophagy related to proteins were further analyzed by Western blotting.【Result】FB1 treatment had significant inhibitory effects on oocyte maturation, while PB1 extrusion rate decreased in a concentration-dependent manner. When the concentration of FB1 reached more than 20 μg·mL -1, the PB1 extrusion was significantly decreased (P<0.01), and the cleavage rate and blastocyst rate of the embryos after oocytes parthenogenetic activation were significantly decreased (P<0.01), damaging the development potential of oocytes to a certain extent. And the cell cycle analysis showed that FB1 treatment also disordered the meiotic cycle progression, resulting in a significant increase in the proportion of oocytes that arrested at germinal vesicle breakdown (GVBD) (P<0.01), and a significant decrease in the proportion of oocytes that successfully developed to the Metaphase II (MII) (P<0.01), with an increase in spindle abnormal rate (P<0.01) and a decrease in actin distribution at the plasma membrane (P<0.05). Further studies showed that, compared with control group, the mitochondrial membrane potential was significantly decreased (P<0.05), impairing mitochondrial function. At the same time, the rate of early apoptosis was obviously increased (P<0.01), and the level of autophagy was also significantly increased (P<0.01) in FB1-treated oocytes. Western blotting analysis showed that the expressions of pro-apoptotic protein BAX and autophagy protein LC3A/B II in FB1-treated oocytes were significantly up-regulated (P<0.05), and the expression of anti-apoptotic protein BCL2 was significantly down-regulated (P<0.05), indicating the occurrence of early apoptosis and autophagy.【Conclusion】FB1 had obvious toxic effects on porcine oocyte maturation in vitro and embryo development after activation, resulting in meiotic cycle arrest, spindle structure disorder, actin distribution reduction and mitochondrial injury, and its toxic mechanism might be related to the induction of apoptosis and autophagy in oocytes.

    Screening and Identification of HSP90 Interacting Proteins in Silkworm (Bombyx mori)
    LONG YanBi,WU YunFei,ZHANG Qian,CHEN Peng,PAN MinHui
    Scientia Agricultura Sinica. 2022, 55(6):  1253-1262.  doi:10.3864/j.issn.0578-1752.2022.06.016
    Abstract ( 400 )   HTML ( 36 )   PDF (2531KB) ( 131 )   Save
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    【Objective】HSP90 is a member of the heat shock protein family and plays an important role in insect resistance and metamorphosis. Studies have shown that HSP90 can promote the proliferation of Bombyx mori nucleopolyhedrovirus (BmNPV), but the mechanism of action is unclear. The objective of this study is to identify the interacting proteins of BmHSP90, and to provide a reference for the analysis of its mechanism of promoting BmNPV proliferation.【Method】The BmHSP90 HA eukaryotic overexpression vector linked to pIZ/V5-His was constructed. After transfection in BmN-SWU1 cells for 48 h, BmNPV was infected and cultured for 48 h to collect the proteins. After the total protein was retained, the proteins were divided into two tubes and co-immunoprecipitation was performed. The interacting protein was caught with anti-HA antibody and IgG antibody, respectively, after staining the protein gel with silver nitrate, the different bands were obtained and mass spectrometry analysis was performed. The mass spectrometry results were combined with the information analysis to screen candidate interacting proteins, and then the interacting proteins were cloned and identified. The co-localization of HSP90 and the interacting protein was verified by immunofluorescence, and the co-immunoprecipitation experiment was further used to determine whether they had an interaction relationship.【Result】The results of silver nitrate staining showed that the experimental group and the control group had different bands near 90, 70 and 60 kD, and verified that the different bands at 90 kD were bait proteins. The other two different bands were analyzed by mass spectrometry, and a total of 7 candidate interacting proteins were identified. Two of the candidate proteins were selected for follow-up study through analysis, namely Tubulin-specific chaperone E (Tbce) and Golgin subfamily A member 5 (Golga5). The maximum open reading frame length of the BmTbce is 1 728 bp, which encodes 576 amino acids, and the maximum open reading frame length of the BmGolga5 is 1 854 bp, which encodes 618 amino acids. The homology alignment and phylogenetic tree showed that the microtubule binding domain of BmTbce (cytoskeleton-associated protein-glycine-rich, CAP-Gly) was located at the N-terminus and was highly conserved among different species. The transmembrane domain (TMD) of BmGolga5 was located at the C-terminus and was also conservative. The fluorescence co-localization showed that BmHSP90 co-localized with BmTbce and BmGolga5 in the cytoplasm, and it was further proved by co-immunoprecipitation that BmHSP90 HA and BmTbceFlag, BmHSP90HA and BmGolga5Flag had an interaction relationship.【Conclusion】After screening and identification, in the process of BmNPV infection of B. mori cells, the proteins that interact with the B. mori heat shock protein HSP90 are BmTbce and BmGolga5.