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    01 November 2024, Volume 57 Issue 21
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Study on the Involvement of OsFWL3 in the Regulation of Metal Ion Transport and Accumulation in Rice
    LIU YiHan, MU QingShan, HE Xiang, CHEN Min, HU Jin, GUAN YaJing
    Scientia Agricultura Sinica. 2024, 57(21):  4161-4174.  doi:10.3864/j.issn.0578-1752.2024.21.001
    Abstract ( 286 )   HTML ( 54 )   PDF (5657KB) ( 228 )   Save
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    【Objective】FWL (Fruit Weight2.2-Like) gene is a negative regulator of cell proliferation, which not only regulates plant organogenesis and organ size, but also participates in the regulation of metal ion transport accumulation and signal transduction. Analyzing of the function of OsFWL3 gene is helpful to reveal the transport mechanism of trace metal elements in crops. It provides theoretical support for reducing heavy metal accumulation and improving crop quality. 【Method】The gene information, genome structure and phylogenetic tree of OsFWLs family were analyzed by bioinformatics method, and the expression profile of OsFWL3 gene was predicted. Two OsFWL3 knockout lines were obtained using CRISPR/Cas9 gene editing technology. Then wild type and Osfwl3 mutants were treated with ZnSO4 at seedling stage and filling stage, respectively. The phenotypes of plants and grains after treatment were analyzed, and the content variation of metal elements such as Zn was determined to explore the effects of OsFWL3 on the transport and accumulation of metal ions and seed quality. 【Result】The gene function of OsFWLs family is similar to some extent. OsFWL3 gene is highly expressed in anther and panicle, indicating that it is closely related to reproductive development of rice. The number of primary branches, grian length, grain thickness and 100-grain weight of Osfwl3 mutants are significantly larger than WT. OsFWL3 affects the content and distribution of Zn and other metal ions in rice seedlings and grains. The deletion of OsFWL3 gene affects the competitive transport of Zn, Cd and Mn from underground to above-ground, lower grain to central grain and husk to brown rice. 【Conclusion】OsFWL3 gene affects the distribution of Zn and other metal ions in rice grains and plants, and it plays an important role in regulating the growth and development of rice plants and grain size.

    QTL Mapping and Candidate Gene Screening for Nitrogen Use Efficiency in Maize
    HAN XuDong, YANG ChuanQi, ZHANG Qing, LI YaWei, YANG XiaXia, HE JiaTian, XUE JiQuan, ZHANG XingHua, XU ShuTu, LIU JianChao
    Scientia Agricultura Sinica. 2024, 57(21):  4175-4191.  doi:10.3864/j.issn.0578-1752.2024.21.002
    Abstract ( 387 )   HTML ( 32 )   PDF (7416KB) ( 264 )   Save
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    【Objective】Genetic improvement for efficient utilization of maize nutrients represents a crucial method to ensure national food security. Exploring quantitative trait locus (QTL) and related candidate genes of nitrogen use efficiency can provide a theoretical basis for improving the efficiency of nitrogen fertilizer in maize and cultivating high-yield and high-efficiency maize varieties. 【Method】In this study, QTL mapping analysis in one recombinant inbred line (RIL) population constructed by KA105 and KB024 was performed for grain yield under two different nitrogen treatments, including the derived traits partial factor productivity from applied nitrogen (PFPN), low nitrogen tolerance coefficient (LNTC) and nitrogen agronomic efficiency (NAE). Concurrently, integrating the seedling transcriptome data of the parent KA105 under nitrogen treatment, differentially expressed genes were identified, and candidate genes associated with maize nitrogen use efficiency were mined through co-expression analysis. Subsequently, the selected candidate genes were validated using qRT-PCR. 【Result】Through mapping analysis, a total of 36 QTLs distributed across different chromosomes were detected, explaining 1.63% to 17.26% of the phenotypic variation. Among these, eight major QTLs with a phenotypic variation explanation rate exceeding 10% were identified, along with seven genetically stable QTLs commonly identified across different traits or environments. Notably, qNNGYP1 located on chromosome 1 has been repeatedly detected in previous studies, with a phenotypic explanation rate of up to 11.73%. Additionally, other QTLs (qNNGYP1, qPFPN1) co-located in this interval across different environments, suggesting it as a focal region for further investigation. Combining transcriptome data of seedlings under low nitrogen stress, 39 differentially expressed genes within these QTL intervals were identified, and 6 key genes were identified through co-expression network prediction. The result of qRT-PCR indicated that the expression trends of the candidate genes under both nitrogen treatments were consistent with the transcriptome data. Specifically, GRMZM2G366873 was involved in the regulation of auxin homeostasis and may participate in maize responses to low nitrogen stress, drought stress, and boron stress through auxin signal transduction, also regulating ear length. GRMZM2G414192 was involved in the response of the photosynthetic system to low nitrogen stress and was regulated by brassinosteroids. GRMZM2G414043 was associated with maize grain length and biomass, while GRMZM2G040642 may be involved in the long-distance signal transduction of nitrogen. 【Conclusion】In summary, a total of 36 QTLs were identified, distributed across chromosomes 1, 4, 5, 7, 8, and 9, including eight major QTLs (PVE>10%). The candidate genes GRMZM2G366873, GRMZM2G414192, GRMZM2G414043, and GRMZM2G040642 were identified as potential genes for maize nitrogen efficiency.

    Comparison of the Genome Sequence Polymorphisms Between the Main Naked Barley Varieties Kunlun 14 and Kunlun 15 in Qinghai Province
    XU JinQing, BIAN HaiYan, CHEN TongRui, WANG Lei, WANG HanDong, YOU En, DENG Chao, TANG YouLin, SHEN YuHu
    Scientia Agricultura Sinica. 2024, 57(21):  4192-4204.  doi:10.3864/j.issn.0578-1752.2024.21.003
    Abstract ( 146 )   HTML ( 17 )   PDF (2478KB) ( 258 )   Save
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    【Objective】At present, Kunlun 14 and Kunlun 15 are the main varieties of the naked barley in Qinghai Province, and also the important backbone parents in the breeding of the naked barley. The genomic sequences of Kunlun 14 and Kunlun 15 were compared to provide references for the trace of important character regions/loci, pedigree analysis and their utilization in molecular design breeding of the naked barley. 【Method】In this study, the agronomic and grain traits of Kunlun 14 and Kunlun 15 were investigated in the field, and the whole genome resequencing were performed (sequencing depth ≥15×). The sequence differences of copy number variation (CNV), single nucleotide polymorphism (SNP) and insertion/deletion (InDel) were compared between the two varieties. According to the SNP distribution patterns, the polymorphism hotspot and genetic similar regions between them were identified. The mutation types of polymorphism hotspot and genetic similar regions were compared and analyzed. The gene enrichment analysis was carried out in the specific CNV regions and the polymorphism hotspots regions of the two varieties. 【Result】In addition to the plant height and peduncle internode length, Kunlun 14 and Kunlun 15 had high similarity in important agronomic and grain traits. Compared with the barley reference genome of Morex, the two varieties had a common CNV variation interval of 83 Mb, and the length of cultivar-specific CNV variation interval in Kunlun 14 and Kunlun 15 was 37 Mb and 38 Mb, respectively. There were 564 genes in the CNV region specific to Kunlun 14, which were significantly enriched in 15 GO terms, while 519 genes were in the CNV region specific to Kunlun 15 and were significantly enriched in 7 GO terms. Based on the SNP distribution patterns, 1 706 Mb polymorphism hotspots and 2 411 Mb sequence similarity intervals of Kunlun 14 and Kunlun 15 were identified at the whole genome level, and the polymorphism hotspots were mainly distributed on 3H, 6H and 7H. The polymorphism hotspots regions contained 16 768 genes, whose functions were mainly related to plant growth and development. There was no significant difference between polymorphism hotspot regions and genetic similar regions in SNP variation type, InDel length distribution and the proportion of mutations affecting coding function. The SNPs and InDel mutations in polymorphism hotspot and genetic similar regions were mainly missense mutations and followed by synonymy mutations. 【Conclusion】The field phenotypes of Kunlun 14 and Kunlun 15 were similar. At the whole genome level, a total of 75 Mb CNV variation regions between them and the polymorphism hotspot regions were mainly distributed at 3H, 6H and 7H.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    High Temperature and Drought Combined Stress Inhibited Photosystem Ⅱ Performance and Decreased Grain Yield of Summer Maize
    GUO Ya, REN Hao, WANG HongZhang, ZHANG JiWang, ZHAO Bin, REN BaiZhao, LIU Peng
    Scientia Agricultura Sinica. 2024, 57(21):  4205-4220.  doi:10.3864/j.issn.0578-1752.2024.21.004
    Abstract ( 191 )   HTML ( 19 )   PDF (745KB) ( 163 )   Save
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    【Objective】This study aimed to explore the underlying reasons for the reduction of maize photosynthesis under the high temperature and drought combined stress, so as to provide theoretical basis for alleviating the combined stress of high temperature and drought. 【Method】Maize cultivar “Denghai 605” was selected as the experimental material for this experiment. Two temperature levels were set, namely normal temperature control (30 ℃/22 ℃ for day (8:00-18:00)/ night (18:00- 8:00 the next day)) and high temperature treatment (38 ℃/28 ℃ for day/night). The two water conditions were normal water supply control (soil water content was 70%-80% of field capacity) and drought treatment (soil water content was set to 50%-60% of field capacity). There were four treatments in the experiment, including control (CK), high temperature stress (H), drought stress (D), high temperature and drought combined stress (HD), and the treatment began at VT stage (VT). The changes in leaf gas exchange parameters, photosystem Ⅱ (PSII) performance, key photosynthetic enzyme activity, plant biomass, and grain yield under different stress treatments were analyzed. 【Result】High temperature, drought and combined stress all led to the increase of chlorophyll fluorescence parameters, the ratio of a variable fluorescence FK to F0-FJ amplitude (WK) and variable fluorescence FJ to F0-FJ amplitude (VJ), and damaged the donor side and acceptor side of PSII. Compared with the control, PSII maximum quantum yield for primary photochemistry (φP0), the probability of captured excitons transferring electrons to other electron acceptors in the electron transfer chain beyond QA (Ψ0), quantum yield for electron transport (φE0), quantum yield of energy dissipation (φD0), quantum yield for reduction of the end electron acceptors at the PSI acceptor side (φR0), and performance index based on absorption of light energy (PIABS) were significantly decreased, and the absorption and transfer of light energy were inhibited; absorbed photon flux per active PSII (ABS/RC), trapped energy flux per active PSII (TR0/RC) and dissipated energy flux per active PSII (DI0/RC) increased significantly, but the electron flux from QA to the PQ pool per active PSII (ET0/RC) decreased significantly, which affected the energy distribution of reaction centers, reduced the number of PSII active reaction centers, and inhibited the performance of PSII. Combined stress could aggravate the inhibition of PSII performance by damaging the donor side, the acceptor side and the active reaction center. At the same time, the activities of ribose 1, 5-diphosphate carboxylase (Rubisco) and phosphoenolpyruvate carboxylase (PEPCase) decreased, which inhibited photosynthetic carbon assimilation. High temperature, drought, and combined stress reduced the net photosynthetic rate by reducing the performance of PSII and the activity of key photosynthetic enzymes. Compared with the control, the net photosynthetic rate of VT+5 d was reduced by 14.6%, 31.4%, and 39.9%, respectively. The decrease in photosynthetic rate inhibited the accumulation of biomass and its transport to grains. Under high temperature, drought, and combined stress, the grain yield decreased by 80.3%, 27.1%, and 84.0% than that under control, respectively. 【Conclusion】In summary, the combined stress of high temperature and drought mainly reduced net photosynthetic rate, hindered biomass, and reduced grain yield by inhibiting leaf PSII performance. The impact of combined stress on PSII performance and grain yield was greater than that of single stress under high temperature and drought.

    Interactive Effects of Planting Density and Nitrogen Application Rate on Plant Grain Yield and Water Use Efficiency of Two Maize Cultivars
    TIAN LongBing, SHEN ZhaoYin, ZHAO XiaoTian, ZHANG Fang, HOU WenFeng, GAO Qiang, WANG Yin
    Scientia Agricultura Sinica. 2024, 57(21):  4221-4237.  doi:10.3864/j.issn.0578-1752.2024.21.005
    Abstract ( 325 )   HTML ( 36 )   PDF (696KB) ( 288 )   Save
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    【Objective】Reasonably increasing planting density combined with appropriate nitrogen (N) application rate is an important technical approach for increasing maize yield and resource use efficiency. Understanding the interactive effects of planting density and N rate on maize growth, evapotranspiration (ET) and water use efficiency (WUE) during the growing season, could provide a basis for improving its use efficiency when increasing planting density and controlling N input in maize production. 【Method】Field experiments were conducted during 2022 to 2023 in Jilin Province. Two maize cultivars, Liangyu 99 (LY99) and Demeiya 3 (DMY3), were used in this study. Three planting densities of 50 000, 70 000 and 90 000 plants/hm2 and four N application rates of 0, 100, 200 and 300 kg N·hm-2 were designed to investigate the effects of planting density and N application rate on grain yield and water productivity of different maize cultivars, as well as the dry matter (DM), soil water content, ET and WUE at various growth stages. 【Result】Planting density significantly affected DM and grain yield of maize, but the response trends varied between cultivars. Grain yields of LY99 with 70 000 plants/hm2 was 11.1% and 18.3% higher than that with 50 000 and 90 000 plants/hm2, respectively. The average yield of DMY3 planted with 70 000 plants/hm2 and 90 000 plants/hm2 was 10.5% and 9.3% higher than that of 50 000 plants/hm2, respectively. Nitrogen fertilization significantly increased DM and grain yield of maize, and also showed significant interactive effects with cultivar or planting density. Compared with N0, grain yields of LY99 were increased by 38.0% to 60.7% under N1, and the yield increases for DMY3 were 24.4% to 38.2%. Notably, the yield responses to N rates were more pronounced for LY99 compared with DMY3. For both cultivars, the yield differences between low N rate and high N rate enlarged with increasing planting density, with LY99 showing a more distinct performance. The water consumption and utilization of maize plants were also significantly affected by planting density, N rate and their interaction. During the growing season, the total ET of DMY3 continually increased with increasing density, while that of LY99 showed the highest values with 70 000 plants/hm2 among different densities. In each density condition, the ET of both cultivars increased with increasing N application rates. The WUE of maize plants showed complex responses to planting density and N rate at different growth stages, due to the varied annual precipitation and distribution patterns. The average increase of water productivity of LY99 under planting 50 000 and 70 000 plants/hm2 was 8.6% and 10.4% compared with 90 000 plants/hm2 respectively. DMY3 had the highest water productivity when planting 70 000 plants/hm2, which increased by 5.8% and 5.3% compared with 50 000 and 90 000 plants/hm2, respectively. The water productivity showed different responses to N rate among the three densities. In general, the difference of nitrogen application under low density was small, but it increased significantly under medium and high density. Compared wtih DMY3, LY99 showed higher increases for water productivity when N fertilizer was applied under medium and high density conditions. The correlation analysis showed that interactive effects of planting density and N rate significantly affected maize yield and water productivity by influencing the water utilization at various growth stages. 【Conclusion】Planting density and N rate had significant interactive effects on maize yield and water utilization in the rain-fed region of Northeast China. The two maize cultivars used in this study could obtain high grain yield and water productivity under a moderately higher density of 70 000 plants/hm2 combined with 200 kg N·hm-2 rate.

    PLANT PROTECTION
    Functional Analysis of SlSnRK1.2 in Regulating Tomato Resistance to Grey Mould
    LI Jie, LIANG ZhiLin, SUN Yan, TAN GenJia, HUAI BaoYu
    Scientia Agricultura Sinica. 2024, 57(21):  4238-4247.  doi:10.3864/j.issn.0578-1752.2024.21.006
    Abstract ( 171 )   HTML ( 13 )   PDF (3358KB) ( 146 )   Save
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    【Background】Grey mould, caused by Botrytis cinerea, poses serious threats to tomato production. Sucrose non-fermenting-1-related protein kinase 1 (SnRK1) from plant is involved in the regulation of responses to biotic and abiotic stresses. However, whether tomato SnRK1 is involved in tomato resistance to grey mould remains unclear. 【Objective】In this study, SlSnRK1.2, which was up-regulated in the process of B. cinerea infecting tomato, was used as the research object to clone and analyze its function of regulating grey mould resistance, so as to provide theoretical basis and gene resources for the prevention and control of tomato grey mould. 【Method】The expression patterns of SlSnRK1.2 during the infection stage of B. cinerea and in different tomato tissues were examined through real-time fluorescence quantitative PCR (qRT-PCR); Subcellular localization of SlSnRK1.2 was analyzed using Agrobacterium-mediated transient transformation system; SlSnRK1.2 silencing plants were constructed by tobacco rattle virus (TRV)-mediated gene silencing (VIGS) technology, and the role of SlSnRK1.2 in the interaction between tomato and B. cinerea was preliminarily analyzed. The overexpression plants of SlSnRK1.2 were created by Agrobacterium-mediated tomato genetic transformation system, and the role of SlSnRK1.2 in regulating tomato resistance to grey mould was further clarified. NbSnRK1.2, a homologous gene of SlSnRK1.2, was silenced in N. benthamiana using TRV-mediated gene silencing technology to determine the function of NbSnRK1.2 during the interaction between N. benthamiana and B. cinerea. 【Result】Micro-Tom was used as the wild-type (WT) background, qRT-PCR technology was used to clarify that the transcriptional expression of SlSnRK1.2 was significantly induced by B. cinerea infection. Subcellular localization analysis revealed that SlSnRK1.2 was localized in the cytoplasm and nucleus. qRT-PCR analysis showed that SlSnRK1.2 was expressed in roots, stems, young leaves, mature leaves, flower buds, and flowers of tomato, with the highest relative expression level in the stems. Transient silencing of SlSnRK1.2 attenuated tomato resistance to grey mould, while overexpression of SlSnRK1.2 enhanced tomato resistance to grey mould. On this basis, transient silencing of NbSnRK1.2, a SlSnRK1.2 homologous gene, attenuated tobacco resistance to grey mould. 【Conclusion】SlSnRK1.2 positively regulates tomato resistance to grey mould and can be used as a genetic resource for molecular breeding of tomato resistance to grey mould.

    Isolation and Identification of Soybean Rhizosphere Growth-Promoting Bacteria and Their Salt Tolerance and Growth-Promoting Effects
    SHAO JiaZhu, LÜ Wen, LIAO XinLin, YUAN XinYu, SONG Zhen, JIANG DongHua
    Scientia Agricultura Sinica. 2024, 57(21):  4248-4263.  doi:10.3864/j.issn.0578-1752.2024.21.007
    Abstract ( 314 )   HTML ( 43 )   PDF (6181KB) ( 1066 )   Save
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    【Objective】Plant growth-promoting rhizobacteria (PGPR) can effectively promote plant growth through long-term mutualistic symbiosis with plants. The objective of this study is to screen and identify rhizosphere growth-promoting actinomycetes from soybean rhizosphere soil and clarify their growth-promoting mechanisms, and to lay a foundation for the development and utilization of microbial agents. 【Method】Different actinomycetes were collected using soil dilution coating method and plate streak inoculation method. The growth-promoting characteristics of isolates were analyzed using phosphorus solubilization, potassium solubilization, and siderophore assay media. Through microscopic observation, physiological and biochemical experiments, and multi-gene sequence analysis, the target growth-promoting actinomycetes were identified. Different salt concentrations (0-10%) were set to determine the salt tolerance of the target isolate. The growth-promoting ability of the isolate on soybean seedlings was detected by pot experiments and whether spraying the culture medium of the isolate under salt stress conditions could improve the stress resistance of soybean plants was evaluated. 【Result】Thirteen PGPR isolates were screened and obtained. Among then, the Sg-7 isolate not only had strong abilities to dissolve phosphorus, potassium, siderophore, and IAA production, but also effectively antagonized Pseudomonas syringae pv. glycinea (Psg). Based on morphological characteristics, biochemical, and multi-gene analyses, the soybean rhizosphere growth-promoting bacterium Sg-7 was identified as Streptomyces griseocarneus. The Sg-7 strain had certain salt tolerance characteristics, with a maximum tolerance to 7% NaCl salt concentration. Pot experiments showed that the diluted culture fluid after irrigation could effectively promote the germination of soybean seeds and improve growth indicators such as root length, stem diameter, fresh weight, and leaf area. Under 150 mmol·L-1 salt stress, irrigation with a 50× dilution of the Sg-7 culture fluid could effectively enhance SOD enzyme activity of soybean leaf, which was 1.84 times that of the salt stress treatment group. The CAT enzyme activity after irrigation with 100× dilution of culture fluid was 4.33 times higher than that of the salt stress treatment group. The POD enzyme activity after irrigation with 10× dilution of culture fluid was 1.10 times that of the salt stress treatment group. The culture fluid of Sg-7 strain could also enhance the root activity of soybean plants and promote the increase of carotenoid content in leaves to resist stress, which was 3.05 and 1.12 times that of salt stress treatment group, respectively. 【Conclusion】S. griseocarneus Sg-7 strain is a potential plant rhizosphere growth-promoting bacterium with broad development potential and practical application value.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    A Retrieval System for Great Soil Groups from China’s Provisional Soil Classification System for the 3rd National Soil Census
    LONG HuaiYu, LU ChangAi, JI HongJie, ZHANG RenLian
    Scientia Agricultura Sinica. 2024, 57(21):  4264-4275.  doi:10.3864/j.issn.0578-1752.2024.21.008
    Abstract ( 459 )   HTML ( 44 )   PDF (482KB) ( 725 )   Save
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    【Objective】The State Council’s Office of the Leading Group for the 3rd National Soil Census released China’s Provisional Soil Classification System for the 3rd National Soil Census (Trial), abbreviated as 3PSC-T, in 2023, without retrieval system, which is unable to address long-standing questions such as “one soil having multiple names” and “different soils sharing the same name”. Therefore, the retrieval system for great soil groups need be built. 【Method】The retrieval indicators for each great soil group were extracted from the soil classification basis in 3PSC-T by using multiple methods, such as referring to existing diagnostic horizons and characteristics, refining existing diagnostic horizons and characteristics, simplifying descriptive statements, and inducting characteristic soil layers. Four principles for retrieval order were set up, which included prioritizing the importance of agriculture, prioritizing ease over difficulty in identification, narrowing down meaning, clustering similar types, and reflecting the spatial evolution and development patterns of soil. 【Result】Consequently, the retrieval system for 62 great soil groups in 3PSC-T was established, and the retrieval order was: 1 Artificial soils, 2 Paddy soils, 3 Vegetable cultivated soils, 4 Irrigation silting soils, 5 Irrigated desert soils, 6 Solonetzs, 7 Frigid plateau solonchaks, 8 Desert solonchaks, 9 Acid sulphate soils, 10 Coastal solonchaks, 11 Meadow solonchaks, 12 Volcanic soils, 13 Frigid frozen soils, 14 Purplish soils, 15 Phospho-calcic soils, 16 Lithosols, 17 Skeletal soils, 18 Takyr, 19 Neo-alluvial soils, 20 Aeolian soils, 21 Loessial soils, 22 Red primitive soils, 23 Limestone soils, 24 Bleached baijiang soils, 25 Podzolic soils, 26 Brown coniferous forest soils, 27 Dark-brown earths, 28 Brown earths, 29 Yellow-brown earths, 30 Yellow-cinnamon soils, 31 Gray forest soils, 32 Black soils, 33 Gray-cinnamon soils, 34 Cinnamon soils, 35 Torrid red soils, 36 Chernozems, 37 Castanozems, 38 Castano-cinnamon soils, 39 Black loessial soils, 40 Brown calcic soils, 41 Sierozems, 42 Gray desert soils, 43 Gray-brown desert soils, 44 Brown desert soils, 45 Mountain meadow soils, 46 Shrubby meadow soils, 47 Shajiang black soils, 48 Meadow soils, 49 Fluvo-aquic soils, 50 Bog soils, 51 Peat soils, 52 Latosols, 53 Latosolic red earths, 54 Red earths, 55 Yellow earths, 56 Dark felty soils, 57 Felty soils, 58 Frigid calcic soils, 59 Cold calcic soils, 60 Cold brown calcic soils, 61 Frigid desert soils, 62 Cold desert soils, and 63 Cambosol. 【Conclusion】It was found that some soil groups in 3PSC-T had overlapping or inclusive associations with one another. This necessitates the creation of a retrieval system to effectively address the issue of “one soil having multiple names” and “different soils sharing the same name”. In general, the majority of the soil groups in 3PSC-T had the ability to provide retrieval indicators through the examination of profile morphology as well as physical and chemical properties of the soil. Nevertheless, the soil groups within the alpine soil order deviated from the classification principles of the overall classification system. This deviation posed challenges in differentiating them based on profile morphology and soil physicochemical properties. Instead, distinguishing these soil groups required consideration of non-soil properties, such as distribution area and altitude. Further efforts were required to enhance the categorization of soil types in 3PSC-T. It is suggested to incorporate specific groups and to establish a new group called Cambosol.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Study on Production Risk Assessment of Three Major Grain Crops in China Based on Multi-Source Data
    ZHAO SiJian, NIE Qian, ZHANG Qiao, CHEN AiLian, LI Yue
    Scientia Agricultura Sinica. 2024, 57(21):  4276-4289.  doi:10.3864/j.issn.0578-1752.2024.21.009
    Abstract ( 138 )   HTML ( 19 )   PDF (3837KB) ( 309 )   Save
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    【Objective】The extensive pricing model of “one province, one crop, one premium rate” has brought about problems, such as adverse selection, moral risk and disorderly operation, which seriously restricts the healthy and sustainable development of agricultural insurance in China. Accurate rate pricing cannot be achieved without agricultural risk assessment. Insurance rate pricing cannot be separated from risk assessment. Launching agricultural production risk assessment is an important task to achieve accurate rate pricing for grain insurance and to accelerate the high-quality development of agricultural insurance. 【Method】Aiming at the three major grain crops (rice, wheat, and maize) in China, three kinds of risk data sources (yield data, disaster loss data, and insurance data) were collected and organized for a long time series. With yield data as the core, combined with disaster and insurance data, the risk assessment modeling was carried out, throughout the adjustment for underestimation of county-level risks and rank correlation of provincial-level risks, to calculate the pure risk loss rate of the three crops at the county level, and then to use the quantile method in risk zoning for the three crops and produce risk maps. 【Result】The rank correlation adjustment of provincial-level risks was mainly based on disaster risk results, followed by insurance risk results. After adjustment, the rank correlation coefficient for rice was increased from 0.610 to 0.766, wheat was increased from 0.547 to 0.748, and maize was increased from 0.576 to 0.760. After adjustment, the average underestimation coefficient for the three major grain crops nationwide was between 20% and 40%, indicating that the average degree of risk underestimation using county-level yield nationwide is between 20% and 40%, with maize having a higher underestimation coefficient than rice and wheat. At the provincial level, the production risks of the three crops in Heilongjiang were all at an extremely high level. The production risks of rice and wheat in Inner Mongolia, rice and maize in Jilin and Liaoning, and wheat in Shanxi were at an extremely high level. At the county level, the extremely high risk of rice production (pure risk loss rate>4.4%) was mainly concentrated in the majority of planting counties in the three northeastern provinces, as well as in the planting counties bordered with the northeastern provinces in Inner Mongolia. The extremely high risk of wheat production (pure risk loss rate>6.3%) was mainly concentrated in the majority of wheat planting counties in Inner Mongolia. The extremely high risk of maize (pure risk loss rate>6.9%) production was mainly concentrated in the maize planting counties bordered with Inner Mongolia and the three northeastern provinces, Shanxi and Shaanxi, as well as most of the maize planting counties in Liaoning, Anhui, and Jiangxi. From the 833 major grain producing counties in China, the proportion of extremely high and high-risk counties of maize was the highest (accounting for 28.1%), followed by rice (accounting for 25.1%), and wheat was the lowest (accounting for 17.2%), indicating that the overall risk of maize was relatively high, while wheat was the lowest. 【Conclusion】The study revealed the magnitude and regional differences of production risks of the three major grain cops. In terms of national average levels, maize had the highest risk (average pure risk loss rate = 5.0%), followed by wheat (average pure risk loss rate = 3.1%), while rice had the lowest risk (average pure risk loss rate = 2.6%). In terms of spatial differences, rice had the highest risks in the northeast and central-south regions, wheat had the highest risks in North China and East China, and maize had the highest risks in North China, Northeast China, and East China. The spatial differences in risks for other levels of rice, wheat, and maize were also inconsistent.

    Structural Characteristics, Development Trends, and International Comparison of Greenhouse Gas Emissions in China’s Agri-Food System Under the Dual-Carbon Objectives
    NIU KunYu, GE RuoHao, CHEN MeiAn, JIN ShuQin, LIU Jing
    Scientia Agricultura Sinica. 2024, 57(21):  4290-4307.  doi:10.3864/j.issn.0578-1752.2024.21.010
    Abstract ( 216 )   HTML ( 11 )   PDF (1804KB) ( 121 )   Save
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    【Objective】This study adopted a comprehensive industry chain perspective to analyze the characteristics, structure, and trends of greenhouse gas emissions in the agricultural food system, contributing to the effective advancement of China’s low-carbon strategy. 【Method】Based on the latest statistics from the United Nations Food and Agriculture Organization, the statistical analysis, comparative analysis, and scenario analysis methods were used to analyze the trend and structural characteristics of greenhouse gas emissions in the agricultural food systems of China, the United States, Western Europe, India, Brazil, and globally from 1990 to 2020. Scenarios (baseline, 2 ℃ temperature control, and 1.5 ℃ temperature control) and specific stages were employed to predict China’s agricultural food system greenhouse gas emissions in 2050. This analysis formed the basis for identifying key areas for emission reduction and outlining achievable pathways. 【Result】Among the major economies mentioned, China was the only country with a significantly increasing per capita greenhouse gas emission in the agricultural food system, although it remained lower than that of developed economies. It was expected that, with the completion of industrialization, the advancement of low-carbonization in the energy industry, and increased difficulty in emission reduction in agricultural production, China’s share of greenhouse gas emissions from the agricultural food system in the total societal emissions would initially decrease and then rise. In this context, the food consumption stage was the fastest-growing segment in China’s agricultural food system greenhouse gas emissions, while the emission proportion from agricultural production was decreasing but still represented the largest share in the system. Unlike other major economies where enteric fermentation emissions were significantly higher, China’s agricultural production greenhouse gas emissions showed a diverse pattern, including enteric fermentation, rice cultivation, fertilizer application, and energy utilization. 【Conclusion】Future emission reduction efforts should focus on formulating comprehensive carbon-neutral strategies for the agricultural food system, low-carbonization of energy structures, promoting shifts in food consumption patterns, and advancing market-oriented emission reduction initiatives.

    HORTICULTURE
    Analysis of Glucosinolate Content and Component in Brassica rapa L.
    MA YuHe, PU YuanYuan, WANG JinXiong, WU JunYan, YANG Gang, ZHAO CaiXia, MA Li, LIU LiJun, WANG WangTian, MIAO ChunQing, GUAN ZhouBo, FAN TingTing, WANG XingRong, MA Rui, LIAN YinTao, SUN WanCang
    Scientia Agricultura Sinica. 2024, 57(21):  4308-4327.  doi:10.3864/j.issn.0578-1752.2024.21.011
    Abstract ( 137 )   HTML ( 11 )   PDF (5543KB) ( 129 )   Save
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    【Objective】 Brassica rapa L. originated from China, whose qualify analysis and evaluation was highly important for its development and utilization. 【Method】 In this study, rapeseed was used as the research object. After grinding extraction and ion exchange column treatment, the glucosinolate extract was obtained. The high performance liquid chromatography (HPLC) was utilized to determine and analyze the content and components of glucosinolates in 323 B. rapa L. accessions. 【Result】 The total content of glucosinolates in 323 B. rapa L. samples ranged from 26.19 to 238.21 µmol∙g-1, with an average of 137.22 µmol∙g-1. A total of 9 glucosinolate components were identified, consisting of five aliphatic glucosinolates (progoitrin, glucoraphanin, gluconapoleiferin, gluconapin and glucobrassicanapin), three indole glucosinolates (4-hydroxyglucobrassicin, glucobrassicin, and 4-methoxyglucobrassicin), and one aromatic glucosinolate (gluconasturtiin). The predominant glucosinolate in B. rapa L. was gluconapin, which belonged to the aliphatic glucosinolate group. It had an average concentration of 122.68 µmol∙g-1, making up 93.71% of the aliphatic glucosinolate content and 89.40% of the overall glucosinolate content. The comparison of the total glucosinolate content in accessions originated from various ecological zones indicated that the Yangtze River basin had the highest glucosinolate content, followed by the Huanghuai region, Xizang, and Gansu accessions. Among accessions, the content of 4-hydroxyglucobrassicin and glucobrassicin in Gansu accessions was more than that in accessions originated from the other three ecological zones. The content of gluconapin in Gansu accessions was the lowest compared to other accessions. The content of indole glucosinolates exhibited significant variations based on their sensitivity to temperature, with the highest levels observed in harsh winter types (11.50 µmol∙g-1), followed by winter types (7.60 µmol∙g-1), semi-winter types (6.77 µmol∙g-1), and spring types (3.87 µmol∙g-1). The findings demonstrated that, the content of progoitrin and indole glucosinolates differed significantly among the germplasm collection used in this study. Six accessions with high-indolyl glucoside content (ranging from 5.86 to 13.81 µmol∙g-1) and three accessions with progoitrin content (ranging from 50.14 to 68.38 µmol∙g-1) were selected by screening. 【Conclusion】 Aliphatic glucosinolates were the primary components of glucosinolates in B. rapa L., with gluconapin being the predominant component. The glucosinolate components exhibited significant variations depending on the genotype of the accessions. Different genotypes were selected based on their concentration of ultra-high glucosinolate, high indole glucosinolate, and high progoitrin.

    Evaluation of Fruit Aroma in Chinese Plum Germplasm Based on Electronic Nose Technology
    ZHAO HaiJuan, ZHANG YuPing, ZHANG YuJun, LIU Ning, XU Ming, LIU JiaCheng, WANG BiJun, LIU WeiSheng, LIU Shuo
    Scientia Agricultura Sinica. 2024, 57(21):  4328-4341.  doi:10.3864/j.issn.0578-1752.2024.21.012
    Abstract ( 128 )   HTML ( 12 )   PDF (5114KB) ( 162 )   Save
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    【Background】 The plum is one of the most widely distributed and cultivated fruit trees globally. The Chinese plum (Prunus salicina L.), originating from China, is known for its beauty, fragrance, juiciness, and rich nutritional value. It exhibits significant genetic heterogeneity and diversity in fruit traits. In recent years, there has been an increasing demand for the sensory quality of plum fruit aroma, prompting breeders to place greater emphasis on identifying and selecting germplasm with unique aromas. Those onventional methods for detecting fruit aroma involve complex preprocessing, high testing costs, and require high technical skills for operation. Additionally, these methods lack the advantages of rapid response and high detection speed, making non-destructive testing difficult and unable to accurately simulate consumer olfactory experiences. 【Objective】 The aim of this study aimed to use electronic nose technology to evaluate the diversity of aroma composition in Chinese plum germplasm, so as to provid essential materials and data support for plum breeding research and production practices in China. 【Method】 Using electronic nose technology, ten odor sensors were employed to identify the different fruit fragrances of 94 Chinese plum germplasm resources. The samples were then grouped and evaluated based on the results of K-means clustering. 【Result】Overall, the average values of different odors showed a normal distribution, but significant differences existed between different odor channels. The W1W channel exhibited the highest odor response value, while the W1C channel had the lowest value. Additionally, variance analysis of different odor channels revealed that W1W had the highest degree of dispersion in odor response values, followed by W5S, while W1C, W3C, W5C, and W3S showed lower dispersion and relatively concentrated distribution. The aroma data of 94 Chinese plum samples were divided into six groups using K-means clustering analysis, each representing plum fruits with specific aroma characteristics. Group one included germplasm sensitive to aromatic hydrocarbons, Group two to ethanol and nitrogen oxides, Group three to hydrogen, Group four to hydrogen and aromatic hydrocarbons, Group five showed low sensitivity to aromatic hydrocarbons, and Group six to hydrogen sulfide. These groupings provided important insights for further research on the aroma components and sensory quality of Chinese plum fruits. Significant differences in maximum, minimum, and median values of different odors were observed between groups. Further correlation analysis revealed significant positive or negative relationships between some odors. Principal component analysis (PCA), linear discriminant analysis (LDA), and uniform manifold approximation and projection (UMAP) were used for dimensionality reduction and visualization of aroma data, and the results indicated that these methods could distinguish the tested plum germplasm to varying degrees and correspond to their cluster groupings, each with different advantages and disadvantages in feature extraction and data visualization. Particularly, the independent use of LDA analysis had certain limitations and shortcomings. This study screened a batch of plum resources with prominent aroma characteristics, including ‘Wuxiangli’ ‘Zaoshuli’ ‘Lishuihong’ ‘Longnanli’ and ‘Xiangjiaoli (Fuxian)’. 【Conclusion】This study utilized electronic nose technology to analyze the aroma composition and distribution of Chinese plum germplasm, revealing that the W1W, W1S, and W5S channels had higher response values, primarily sensitive to volatile compounds such as hydrogen sulfide, methane, and nitrogen oxides. Six Chinese plum groups had different distinct aroma characteristics. In several response values, the mean difference between cluster 6 and the other five groups was significant, indicating that cluster 6 had unique characteristics in these response values.

    FOOD SCIENCE AND ENGINEERING
    Multidimensional Characterization of Astringency Quality in Dry Red Wine and Its Effects
    TAN FangDai, HE YingXia, LIU JiaYue, LI AiHua, TAO YongSheng
    Scientia Agricultura Sinica. 2024, 57(21):  4342-4355.  doi:10.3864/j.issn.0578-1752.2024.21.013
    Abstract ( 122 )   HTML ( 8 )   PDF (2356KB) ( 491 )   Save
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    【Objective】The astringency analysis method for dry red wine was designed by considering the time dependence of astringency, the sub-quality attributes of oral sensation and the instant facial expressions of panelists, so as to provide the methodological support for the multidimensional characterization of astringency quality. 【Method】 Astringency time dependence of dry red wine was characterized by the time intensity method, and the related parameters were measured, such as maximum astringency intensity (Imax), rate of intensity increase before Imax (Vi), rate of intensity decrease after Imax (Vd), area under the curve (AUC), and perception duration (Ttot). Astringency sub-qualities, such as drying, rough, and pucker, were evaluated using CATA and TDS methods. Likability of the wine was analyzed through immediate facial expressions. Twenty-seven Cabernet Sauvignon dry red wine samples from Gansu, Ningxia and Xinjiang were used to characterize the astringency intensity, sub-quality characteristics and perceived differences in liking. 【Result】 Significant variations in astringency were observed among the three regions, particularly in Imax, Vd, and AUC. Wine samples from Ningxia exhibited the highest values for Imax, AUC, and Vd compared to those from Gansu and Xinjiang. The sub-qualities of astringency, such as drying, numbing, rough, pucker, green, and grainy, were frequently identified, with a frequency exceeding 50%. These sub-qualities constituted the main attributes of astringency in the analyzed wines. Correlation analysis of multiple astringency indexes revealed that the astringency sub-qualities in dry red wines from the three regions primarily consisted of rough, pucker, and drying. Excessive roughness and numbing diminished positive emotions among panelists, while the graininess often elicited happy and surprised expressions. Principal component analysis (PCA) of the multivariate data on astringency in the sampled wines demonstrated that the multidimensional characterization technology method designed in this study had a strong ability to distinguish astringency of sampled wines. Wine samples from Ningxia had higher Imax, AUC, Vi, and Vd, while the rough, pucker, drying and numbing were more obvious. Xinjiang wine samples had stronger grainy sense, and the Imax, AUC, green and numbing were weaker, but the drinking comfort was better. The wine samples from Gansu had the strongest green astringency and the weakest rough dominance rate. 【Conclusion】The multidimensional characterization method for dry red wine developed in this study effectively captured the diversity of astringency in a concrete and detailed manner, which provided a more scientific evaluation of astringency differences, making it valuable for broader application and promotion.

    ANIMAL SCIENCE·VETERINARY SCIENCE
    Dissecting the Genetic Architecture of Yolk Ratio with Single-Step Genome-Wide Association Study
    GUO Jun, QU Liang, SHAO Dan, MA Meng, DOU TaoCun, LU Jian, HU YuPing, WANG XingGuo, WANG Qiang, LI YongFeng, GUO Wei, TONG HaiBing
    Scientia Agricultura Sinica. 2024, 57(21):  4356-4366.  doi:10.3864/j.issn.0578-1752.2024.21.014
    Abstract ( 202 )   HTML ( 19 )   PDF (2174KB) ( 580 )   Save
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    【Objective】 Genome-wide association study (GWAS) was used to dissect the genetic architecture of yolk ratio in the 60-week-old hens, with the aim of discovery and utilization of the excellent genetic resources of local chickens and establishing the foundation for the breeding layer to meet market demand. 【Method】 The phenotypic data was collected from the resource population of Jiangsu Institute of Poultry Science. The F2 population was used in this study, Dongxiang green-shelled chickens and White Leghorn were used as parents respectively, and the F1 and F2 generations were obtained through reciprocal crosses. The data set included age at the first egg, shank length, body weight, egg size, Haugh unit, and yolk ratio. Blood samples in F2 generation were collected, and 1 534 hens were genotyped using 600K gene microarray. SNPs data was quality controlled. Missing data was imputed by Beagle software. After data cleaning steps such as removing outliers from the phenotypic data, one-way analysis of variance was used to test the factors that affected the phenotypic value, and these factors enrolled in the fixed effects. The genetic parameters and variance components were determined with multivariate animal model by using pedigree relationship matrix. On the other hand, the genetic relationship matrix was set up with the pedigree and genomic data. Heritabilities were calculated with univariate animal model, and genetic correlation coefficients were determined with bivariate animal model. The BLUPF90 software was used to obtain the SNP effect value, and postGSf90 was used to obtain the P-value and weight corresponding to the SNP. Therefore, the significant SNPs related to the yolk ratio was obtained with GWAS method. Chromosome heritability and haplotype block size were calculated for the yolk ratio. 【Result】The yolk ratio of Dongxiang blue shelled chicken was significantly higher than that of White Leghorn. Fixed effects included the combination of laying batch-generation-breed level. Using the hybrid genetic relationship matrix, the heritability on the yolk ratio and egg size were 0.348±0.041 and 0.500±0.038, respectively. The genetic correlation coefficients between yolk ratio and egg size and Haugh unit were -0.463±0.075 and -0.165±0.111, respectively. Using the pedigree genetic relationship matrix, heritability on yolk ratio and egg size were 0.387±0.052 and 0.465±0.052, respectively. The genetic correlation coefficients between yolk ratio and egg size and Haugh unit were -0.405±0.091 and -0.166±0.121, respectively. The inflation factor on the yolk ratio association analysis was 1.007, indicating that no stratified structure was detected in the resource population. There was a locus on chicken chromosome 22 that was significantly associated with the yolk ratio, which was supported by additional SNPs. This QTL explained 2.18% of the phenotypic variance, and the haplotype block size reached 71 kb. In addition, the potential SNP was also detected on chromosome 2. Chromosome heritability analysis showed that the yolk ratio was controlled by small-effect polygenes, and the heritability was directly proportional to the number of genes harbored by the chromosome. 【Conclusion】The weighted single-step GWAS method was used to analyze the genetic architecture of yolk ratio at 60 weeks of age, and the ADAM9 gene was identified to associate with the yolk ratio. Compared with the pedigree genetic relationship matrix, the hybrid genetic relationship matrix reduced Mendelian sampling errors and obtained more accurate genetic parameters.

    Isolation of High-Efficient Ammonia-Removing Strains and Its Cultivated Condition Optimization
    DONG Qing, SONG LianJie, ZHANG HongWei, SU DongYao, ZHANG Ao, ZHANG Lu, ZHANG HuiWen, LI BoSen, GAO YuHong, SUN XinSheng
    Scientia Agricultura Sinica. 2024, 57(21):  4367-4375.  doi:10.3864/j.issn.0578-1752.2024.21.015
    Abstract ( 104 )   HTML ( 9 )   PDF (827KB) ( 85 )   Save
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    【Objective】Odor pollution from animal industry has become a pain point and a difficulty in recent years. Ammonia (NH3) is an important harmful gas with great contribution rate to odor pollution, and its cutting emission becomes an urgent task. The objective of this study was to isolate the efficient NH3-removing strains and to optimize its cultivated conditions, which would provide excellent strains for developing deodorizers. 【Method】Manure deposited in the sewage outlet of the farm was collected as the source of strain isolation. The NH3-removing strains were enriched, separated, and purified using NH3 enrichment culture medium. Then, the degradation ability of purified bacteria above was determined using NH3-degradating culture medium, and the efficient NH3-removing strains were screened. The isolated strains were identified by morphology (colony morphology and Gram staining) and 16S rRNA sequencing method. Further, the effects of various cultivated conditions on biomass of the strains, including pH (5.0, 6.0, 7.0, 8.0, and 9.0), carbon sources (xylose, sucrose, fructose, maltose, and glucose), salt contents (0%, 0.5%, 1.0%, 1.5%, and 2.0%), and inoculum amounts (1.0%, 5.0%, 10%, 15%, and 20%), were performed, and the optimal conditions were determined. 【Result】 (1) A total of 12 suspected NH3-degrading strains were isolated by enrichment culture (named as C1-C12), among of which, the strain named C5 had efficient NH3-removing ability. The logarithmic growth period of the strain C5 was 2 to 20 h. During the period, the degradation rate of C5 increased with the growing bacteria biomass, reaching 60.70% at 24 h and 75.49% at 48 h of culture, respectively, which was greater than that of the other 11 strains in the same time (P<0.05). From morphology and 16S rRNA identification, the C5 was characterized by stubby- and rod-shaded, pointless round at both ends, and no capsule, as well as Gram-negative bacteria. Combined with gene sequencing results and phylogenetic tree, the C5 was determined to be Klebsiella pneumoniae. (2) Various pH values, carbon sources, salt content, and inoculation amount showed different effects on the concentration of C5 after 48 h of culture. The C5 concentration at pH 7.0 was greater than that at pH 5.0, pH 6.0 and pH 9.0 (P<0.05). Compared with other carbon sources, the C5 concentration for glucose as carbon source was the greatest during the culture period of 8 to 48 h. Under different salt content and inoculation amount, there were differences (P<0.01) in bacterial concentration between groups, exhibiting the greatest concentration of bacterial solution at 1.5% salt and a peak value at 40 h of culture. Besides, the fast growth of bacteria was observed for 1.0% of inoculation amount and bacterial concentration increased, compared with other inoculation amounts. Therefore, the optimal growing conditions of the C5 were as follows: pH=7.0, carbon source of glucose, 1.5% salt content, and 1.0% of inoculum amount, where the growth rate and biomass of the C5 were optimal. 【Conclusion】Klebsiella pneumoniae (strain C5) isolated from manure possessed the ability to remove NH3 efficiently, which would have potential utilization value in livestock manure treatment.

    RESEARCH NOTES
    Screening of Interaction Proteins with AhSAP1 in Peanut Using the Yeast Two-Hybrid System
    ZHU YanTing, DANG Hao, NIU SiJie, LIN JingYi, YANG Hua, YANG Qiang, ZHANG Chong, CAI TieCheng, ZHUANG WeiJian, CHEN Hua
    Scientia Agricultura Sinica. 2024, 57(21):  4376-4390.  doi:10.3864/j.issn.0578-1752.2024.21.016
    Abstract ( 136 )   HTML ( 11 )   PDF (3959KB) ( 478 )   Save
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    【Objective】Seed size directly affects the yield of peanut. The key transcription factor AhSAP1, which regulates peanut seed size, was obtained by QTL mapping in the early stage, but the molecular mechanism of AHSAP1 regulating peanut seed size remains unclear. In this paper, a peanut embryo yeast two-hybrid cDNA library was constructed and AhSAP1 was used as bait to screen interacting proteins, and the spatial and temporal expression characteristics of candidate interacting protein genes were analyzed. It laid the foundation for further study on the molecular mechanism of AhSAP1 regulating peanut kernel development. 【Method】The peanut embryo Escherichia coli cDNA library was constructed and identified by SMART (switching mechanism at 5′ end of the RNA transcript) method. The decoy vector pGBKT7-AhSAP1 was constructed and its toxicity and self-activation to yeast cells were evaluated. The peanut embryo cDNA library plasmid and the bait plasmid pGBKT7-AhSAP1 were co-transformed into Y2H Gold yeast strains, and the positive colonies with good growth and blue color were screened and sequenced to obtain the candidate interacting protein gene sequences and predict the biological functions. The expression characteristics of candidate interacting protein genes in different tissues and organs of peanut, induced by exogenous plant hormones and induced by low calcium stress were determined by RNA-seq. According to the functional annotation results, the candidate factors that may be involved in plant seed development were selected, their CDS full-length sequences were amplified, and the target vector pGADT7 was constructed, and point-to-point yeast two-hybrid interaction was verified with pGBKT7-AhSAP1 co-transformed yeast cells. 【Result】The titer of peanut embryo Escherichia coli secondary cDNA library was 1.05×108 cfu/mL, the recombination rate was 98%, the average insert fragment size was more than 1 000 bp, and the library quality was high. The yeast two-hybrid decoy vector pGBKT7-AhSAP1 was successfully constructed, which had no self-activation in yeast cells and no toxicity to yeast. Sixty-eight yeast-positive clones were screened, and 60 candidate interacting proteins were obtained by sequence similarity comparison and removal of duplicating, which were mainly involved in energy production and metabolism, translation, ribosome structure and biological development, transcription, signal transduction mechanism, post-translational modification, inorganic ion transport and metabolism, chromatin structure, etc. Twelve candidate interacting proteins were selected for one-to-one yeast two-hybrid verification with AhSAP1, and 8 candidate interacting proteins were found to interact with AhSAP1. 【Conclusion】The mixed cDNA library of peanut embryo development at different stages was successfully constructed, and 60 candidate interacting proteins with AhSAP1 were screened. The candidate interacting proteins were mainly involved in energy production and metabolism, translation, ribosome structure and biogenesis, transcription, signal transduction mechanism, post-translational modification, inorganic ion transport and metabolism, chromatin structure, etc. These candidates interacting protein genes were expressed in root, stem, leaf, inflorescence, fruit needle, pericarp, seed coat and embryo, and the interaction between 8 candidate interacting proteins and AhSAP1 was confirmed.