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    12 January 2024, Volume 50 Issue 1
    • REVIEW
      Research progress of seed dormancy and germination regulation
      SONG Song-Quan, TANG Cui-Fang, LEI Hua-Ping, JIANG Xiao-Cheng, WANG Wei-Qing, CHENG Hong-Yan
      Acta Agronomica Sinica. 2024, 50(1):  1-15.  doi:10.3724/SP.J.1006.2024.34012
      Abstract ( 945 )   HTML ( 127 )   PDF (3074KB) ( 944 )   Save
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      Dormancy enables plant seeds to time germination until environmental conditions become favorable for seedling survival and growth. The dormancy characteristics of seeds are of important ecological adaptive significance and notable agricultural value. Phytohormone abscisic acid (ABA) and gibberellin (GA) are the key factors for seed dormancy and germination. Mature seeds in dormancy state contain high levels of ABA and low levels of GA. ABA induces and maintains seed dormancy, while GA antagonizes ABA and promotes seed germination. DELAY OF GERMINATION-1 (DOG1) is a major regulator of seed dormancy and had a synergistic effect with ABA to delay germination. DOG1 enhances ABA signal transduction by combining with PP2C ABA hypersensitive germination (AHG1/AHG3), and inhibits the action of AHG1 to increase ABA sensitivity and impose seed dormancy. Imprinted genes are regulated by epigenetic mechanisms before and after fertilization, and are closely related to the establishment and release of seed dormancy. In recent years, remarkable progress has been made in the regulation of seed dormancy. In the present paper, we reviewed the effects of phytohormones ABA and GA on seed dormancy and germination, the action mechanism regulating seed dormancy by DOG1, and the epigenetic regulation of seed dormancy and germination. In addition, we also propose some scientific issues that need to be further investigated in this field to provide some information for understanding the molecular mechanism of seed dormancy and germination, breeding in anti-preharvest sprouting in crop plants, and promoting the germination of dormant seeds.

      Genetic analysis of elite stripe rust resistance genes of founder parent Zhou 8425B in its derived varieties
      LI Yu-Jia, XU Hao, YU Shi-Nan, TANG Jian-Wei, LI Qiao-Yun, GAO Yan, ZHENG Ji-Zhou, DONG Chun-Hao, YUAN Yu-Hao, ZHENG Tian-Cun, YIN Gui-Hong
      Acta Agronomica Sinica. 2024, 50(1):  16-31.  doi:10.3724/SP.J.1006.2024.31013
      Abstract ( 365 )   HTML ( 45 )   PDF (4495KB) ( 226 )   Save
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      Zhou 8425B is a widely used dwarf large panicle and disease-resistant and stress-resistant Wheat Founder Parent in the wheat production areas of the Yellow-Huaihe-Haihe Rivers regions. The analysis of stripe rust resistance of derived varieties and the genetic transmission information of stripe rust resistance gene carried by Zhou 8425B is of great value for wheat new variety breeding. In this study, we used a highly toxic race of stripe rust, Tiaozhong 34 (CYR34), to identify the resistance of 222 collected Zhou 8425B derived varieties to stripe rust at seedling stage. The hybrid strain mainly CYR34 was used to identify the resistance to stripe rust at adult stage of the derived varieties. Subsequently, molecular markers closely linked to the stripe rust resistance genes (YrZH84, YrZH84.2, Yr30, YrZH22, and Yr9 carried by Zhou 8425B) were used for genotype detection of the derived varieties. The results showed that Zhou 8425B was highly resistant to stripe rust stripe rust at seedling stage and adult stage to the current virulent dominant race CYR34. Among the 222 derivative varieties of Zhou 8425B, 14 of them, including Changmai 9, Jiyanmai 10, Bainong 4199, Saidemai 7, and Zhengmai 103, etc., showed stable disease resistance in two years, accounting for 6.3%; 52 derivative varieties, include Zhoumai 11, Zhoumai 22, Zhoumai 26, Zhoumai 36, Lantian 36, Cunmai 16 and Zhengpinmai 8, etc., had stable disease resistance during the whole growth period, accounting for 23.4%. Many derivative varieties of Zhou 8425B were mainly derived from six offspring, including Zhoumai 11, Zhoumai 12, Zhoumai 13, Zhoumai 15, Zhoumai 16, and Zhoumai 17. In the first generation, Zhoumai 16 and Zhoumai 13 directly derived more varieties because of their good agronomic characters, while Zhoumai 15 and Zhoumai 17 derived fewer varieties. Zhoumai 12 and Zhoumai 13 bred the second generation, Zhoumai 22, and then derived 45 sub-generations, and Zhoumai 11 bred Aikang 58, and then derived 54 sub-generations. The excellent stripe rust resistance gene of Zhou 8425B was continuously separated and polymerized in the process of genetic breeding. The frequencies of YrZH84, YrZH84.2, YrZH22, Yr30, and Yr9 in the derived offspring were 34.7%, 14.9%, 41.9%, 66.2%, and 67.1%, respectively. Among the derivative varieties carrying only one disease resistance gene, the average severity of YrZH84 was the lowest (14.4%). Among the derived varieties that aggregate two disease resistance genes, the average severity of carrying YrZH84+YrZH22 was the lowest (20.0%); among the derived varieties that aggregate three disease resistance genes, the average severity of carrying YrZH84+YrZH22+Yr9 was the lowest (17.2%). Among the derived varieties that aggregate four disease resistance genes, the average severity of carrying YrZH84+YrZH22+Yr30+Yr9 was 16.9%, and the average severity of carrying YrZH84.2+YrZH22+Yr30+Yr9 was 38.4%. At seedling stage, derived varieties that carried the YrZH84 resistance gene or a combination of genes containing YrZH84 during the entire growth period had better disease resistance. These results provide the information of stripe rust gene for the continuous improvement and utilization of the Founder Parent Zhou 8425B in China, identify the new derived germplasm with high resistance and high yield to the highly virulent physiological race CYR34, which providing a reference for the genetic breeding of wheat resistance to stripe rust in China.

      Analysis of genetic model of sucrose content in peanut
      ZHI Chen-Yang, XUE Xiao-Meng, WU Jie, LI Xiong-Cai, WANG Jin, YAN Li-Ying, WANG Xin, CHEN Yu-Ning, KANG Yan-Ping, WANG Zhi-Hui, HUAI Dong-Xin, HONG Yan-Bin, JIANG Hui-Fang, LEI Yong, LIAO Bo-Shou
      Acta Agronomica Sinica. 2024, 50(1):  32-41.  doi:10.3724/SP.J.1006.2024.34050
      Abstract ( 176 )   HTML ( 18 )   PDF (425KB) ( 98 )   Save
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      Peanut (Arachis hypogaea L.) is one of the important cash crops in China, and about 40% of peanuts are used for food production. The sucrose content was positively correlated with the flavor and taste of peanut and its products, thus increasing sucrose content in kernel was significant for the promotion of peanut quality. In this study, two F2:3 populations (NYBP×SYT5-1 and 19-1934×JHT1) were constructed to investigate the genetic model of sucrose content, and analyze the correlations between sucrose content with oil content, protein content, and other seed traits. The results showed that the sucrose contents of peanut kernel in two populations were continuously distributed and abundantly varied, and the transgressive segregation phenomenon were observed. In the two populations, sucrose content was significantly negatively correlated with oil content but significantly positively correlated with protein content. However, the correlations between sucrose content and kernel length, kernel width, and 100-kernel weight were inconsistent. The genetic analysis in two populations showed that the sucrose content in peanut kernel was mainly regulated by two pairs of main genes with the additive effect and dominant effect. There was the interaction between the two main genes, as well as the additive effect. This study initially reveals the genetic regulation of sucrose content in peanut kernels, which is beneficial for the breeding of edible peanut varieties and the cultivation of special varieties.

      QTL mapping of alkaloids in tobacco
      LIU Ying-Chao, FANG Dun-Huang, XU Hai-Ming, TONG Zhi-Jun, XIAO Bing-Guang
      Acta Agronomica Sinica. 2024, 50(1):  42-54.  doi:10.3724/SP.J.1006.2024.34047
      Abstract ( 185 )   HTML ( 29 )   PDF (4340KB) ( 184 )   Save
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      Alkaloids are important chemical components in tobacco. In order to understand the genetic architecture of alkaloids in tobacco and identify major effect loci controlling alkaloids related traits, QTL mapping on tobacco alkaloids was performed. A set of 271 recombinant inbred lines (RIL) were constructed with Y3 and K326 as the parents. The RIL population was planted in Yanhe, Yuxi, Yunnan province and Shilin, Kunming, Yunnan province in 2018, 2019, and 2020, respectively. Five alkaloid phenotypes including total plant alkali (TPA), nicotine (NIC), nornicotine (NOR), anabasine (ANAB), and anatabine (ANAT) were measured. A linkage map of 46,129 markers was constructed by genome sequencing of the population. QTL mapping was performed by the software QTLNetwork 2.0 which was developed based on the mixed linear model. A total of 15 QTLs with significant additive effects were identified. The contribution rate of additive effect to the corresponding phenotypes varied from 0.58% to 11.57%. Four major QTLs, qTPA14 for total plant alkali, qNIC14 for nicotine, qANAB14 for anabasine, and qANAT14 for anatabine, accounted for more than 10% of phenotypic variation of the corresponding traits, which were located in linkage group 14. Six QTLs with significant additive-by-environment interaction effects were detected, their additive-by-environment interaction effects explained the phenotypic variation of 0.80%-1.81%. Five pairs of QTLs with significant additive-by-additive epistasis effects were detected, accounting for phenotypic variation from 0.15% to 2.31%, while two pairs of QTLs were detected with significant epistasis-by-environment interaction effects, which explaining the proportion of phenotypic variation from 0.81% to 1.16%. The results pave a foundation for further isolation of candidate genes, the dissection of genetic mechanism, and the molecular improvement of tobacco alkaloid traits.

      Mapping and cloning of plant height gene PHR1 in maize
      YANG Chen-Xi, ZHOU Wen-Qi, ZHOU Xiang-Yan, LIU Zhong-Xiang, ZHOU Yu-Qian, LIU Jie-Shan, YANG Yan-Zhong, HE Hai-Jun, WANG Xiao-Juan, LIAN Xiao-Rong, LI Yong-Sheng
      Acta Agronomica Sinica. 2024, 50(1):  55-66.  doi:10.3724/SP.J.1006.2024.33011
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      Plant height is an important index for the ideal maize breeding, which not only affects the mechanical harvest of maize, but also closely relates to the lodging resistance and the biological yield of maize. In this study, a plant height reducing mutant-1 (phr-1) was obtained by using low-dose fast neutron (4.19 Gy) irradiation to mutate the maize inbred line KWS39 at the low ear position. Phenotypic traits were investigated and analyzed in the field, and candidate genes were identified by mining and functional annotation of genes in the target region based on the B73 reference genome using the extreme trait pool sequencing analysis (BSA-seq) and the method of target region recombination exchange identification with F2 segregation population of phr-1×B73. These results indicated that there might be a variation site in Bin1.06 interval on chromosome 1, and the target region was precisely located between two markers, Umc1122 and Umc1583a, with a 600 kb interval by using the large segregation population and polymorphic markers. Brachytic2 (BR2), encoding a sugar protein regulating the polar transport of auxin in maize stems, is a known gene controlling plant height in this region. The sequencing of candidate genes revealed that phr-1 was a new allelic mutation of br2-1, with a 165 bp insertion in the fourth exon of the BR2 gene resulting in the amino acid at position 547 changing to a stop codon and premature protein translation termination. The mutation site and variation mode of phr-1 were completely different from those of br2-1 single base mutation site. The allelic hybridization experiment confirmed that the PHR1 candidate gene is a new allelic mutation of the BR2 gene. This study provides new germplasm resources for maize BR2 gene in plant height genetic improvement.

      Effects of small peptide Ospep5 on cadmium tolerance in rice
      LI Ming-Yue, ZHANG Wen-Ting, LI Yang, ZHANG Bao-Long, YANG Li-Ming, WANG Jin-Yan
      Acta Agronomica Sinica. 2024, 50(1):  67-75.  doi:10.3724/SP.J.1006.2024.32012
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      Cadmium (Cd) stress is one of the major heavy metal stresses which causes serious impacts on plant growth and development. Although studies have shown that plant small peptides have a mitigating effect on stress, research on their tolerance to cadmium toxicity in rice is limited. In previous studies, a few peptides were identified through translatome, transcriptome, and proteome analysis, among which Ospep5 was found to significantly improve salt tolerance in rice. In this study, the effects of Ospep5 on the growth of rice seedlings under cadmium stress were investigated using the japonica cultivar Nipponbare, Ospep5 overexpressing line Ospep5-OX, and the CRISPR/Cas9 mutant ospep5-3 as the experimental materials. The results showed that 500 μmol L-1 CdCl2 significantly inhibited the morphological growth and chlorophyll content of rice seedlings, while the activities of superoxide dismutase (SOD), contents of proline (Pro), malondialdehyde (MDA), and cadmium ion content were significantly increased. Compared to cadmium stress alone, exogenous application of synthetic Ospep5 effectively alleviated the inhibition of cadmium stress on the morphological growth of rice seedlings. Furthermore, SOD activity was significantly increased, and MDA content, Pro content, and cadmium ion content were significantly reduced. And the relative expression level of cadmium tolerance genes (OsHMA2, OsHMA3, OsCAL1) were upregulated. In conclusion, Ospep5 improves the tolerance of rice seedlings to cadmium stress by regulating various physiological and biochemical reactions and the expression of cadmium tolerance genes.

      Functional identification of maize transcription factor ZmMYB12 to enhance drought resistance and low phosphorus tolerance in plants
      WANG Li-Ping, WANG Xiao-Yu, FU Jing-Ye, WANG Qiang
      Acta Agronomica Sinica. 2024, 50(1):  76-88.  doi:10.3724/SP.J.1006.2024.33007
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      Maize usually suffers from various abiotic stresses such as drought, high temperature, high salt, and deficiency of nutrient elements during growth and development period, which will eventually lead to the decline of yield and quality, resulting in serious agricultural yield reduction. MYB transcription factors are widely distributed in plants and involved in the whole process of plant growth and development and environmental response. Therefore, the screening and identification of MYB transcription factors conferring stress resistance can provide the theoretical basis and technical support for genetic improvement in maize. In this study, a R2R3-MYB family transcription factor gene, ZmMYB12, was cloned from maize materials under drought treatment. This gene showed inducible expression in response to drought, ABA, and PEG treatments. Maize plants with virus induced gene silencing (VIGS) of ZmMYB12 had higher sensitivity to drought and accumulated more reactive oxygen species (ROS), as well as smaller roots. The survival rate of ZmMYB12 silencing plants was lower after re-watering than the wild type plants (WT), indicating that ZmMYB12 was a positive regulator of drought response. Overexpression of ZmMYB12 in Arabidopsis thaliana resulted in less ROS accumulation and more lateral roots than WT, thus elevating drought resistance. After low phosphorus stress, ZmMYB12 overexpression Arabidopsis had more lateral roots and stronger root system, as well as higher contents of chlorophyll and anthocyanin. The content of inorganic phosphorus in ZmMYB12 overexpression Arabidopsis was also higher than WT. In conclusion, this study indicates that ZmMYB12 positively regulates drought response and increases the uptake and utilization of phosphorus in plants, providing an elite gene for maize breeding to increase abiotic resistance.

      Acquisition and resistance analysis of transgenic Maize Inbred Line LG11 with insect and herbicide resistance
      YUE Run-Qing, LI Wen-Lan, MENG Zhao-Dong
      Acta Agronomica Sinica. 2024, 50(1):  89-99.  doi:10.3724/SP.J.1006.2024.33016
      Abstract ( 156 )   HTML ( 12 )   PDF (9127KB) ( 116 )   Save
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      Insect pests are an important limiting factor affecting the yield and quality of maize. Bacillus thuringiensis transgenic maize has good insect resistance and can reduce the use of chemical insecticides effectively, which is very popular among growers. However, a large area of continuous planting of transgenic corn will lead to the resistance of target pests. “Multi-gene” strategy is one of the management strategies to prevent or delay the occurrence of resistant populations of target pests. The anti-insect vector constructed in this study is a fusion protein M2cryAb-vip3Aa formed by combining the main structural domains of Cry1Ab and Vip3Aa proteins using synthetic methods. These two insecticidal proteins had no evolutionary homology and different insecticidal mechanisms, which could effectively reduce the resistance probability of target pests. In this study, to obtain a new transgenic maize material with insect resistance and herbicide resistance and excellent agronomic traits, and analyze its insect resistance and herbicide resistance, m2cryAb-vip3Aa and bar genes were introduced into the recipient materials in series by transgenic method, and Chang 7-2, a backbone inbred line, was used as the backcross parent for backcross transfer. The results of this study enriches the existing germplasm resources of insect-resistant and herbicide-tolerant maize, and provides new solutions for pest control and weed control in maize fields.

      Knockout of GmBADH1 gene using CRISPR/Cas9 technique to reduce salt tolerance in soybean
      SHI Yu-Xin, LIU Xin-Yue, SUN Jian-Qiang, LI Xiao-Fei, GUO Xiao-Yang, ZHOU Ya, QIU Li-Juan
      Acta Agronomica Sinica. 2024, 50(1):  100-109.  doi:10.3724/SP.J.1006.2024.34056
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      Functional identification of salt-tolerant genes is crucial for the improvement of soybean varieties and the development and utilization of saline-alkali land. Under salt stress, crops reduce the effect of salt damage on crop yield by synthesizing and accumulating betaine as osmotic protective agent. BADH gene is been proved to regulate plant response to stress in many plants, but the regulation mechanism in soybean is not clear. In this study, GmBADH1 gene was cloned, and qRT-PCR showed that the gene was expressed stems and leaves, especially in roots. The expression vector constructed by CRISPR/Cas9 system was transferred into soybean variety JACK by Agrobacterium-mediated soybean genetic transformation technology, and three targeted mutations that could regulate the salt tolerance of soybean were produced. They all occurred in the coding region, which had three difference of deletion 19 bp, insertion 1 bp, and insertion 9 bp to replace 2 bp. The first two, premature termination codons, resulting in truncated BADH1 protein, the latter occurred frameshift mutation. The mutant homozygous plants were treated with salt at the emergence stage and seedling stage, respectively. The results showed that the salt tolerance of the mutant homozygous plants was significantly lower than that of the wild type at seedling stage, and there was no significant difference compared with the wild type at seeding stage, which indicating that GmBADH1 gene may mainly regulate the salt tolerance of seedling stage. This study provides a basis for further excavation of soybean salt-tolerant genes and cultivation of soybean salt-tolerant varieties.

      Genome-wide identification of NAP transcription factors subfamily in Saccharum spontaneum and functional analysis of SsNAP2a involvement in leaf senescence
      WANG Heng-Bo, FENG Chun-Yan, ZHANG Yi-Xing, XIE Wan-Jie, DU Cui-Cui, WU Ming-Xing, ZHANG Ji-Sen
      Acta Agronomica Sinica. 2024, 50(1):  110-125.  doi:10.3724/SP.J.1006.2024.34037
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      NAP (NAC-like, Activated by APETALA3/PISTILLATA) is a subfamily of the transcription factor NAC gene family, which is widely involved in regulating plant growth and development, leaf senescence, and response to hormones and abiotic stress. Firstly, the NAP subfamily members were identified from the genomic database of Saccharum spontaneum, and phylogenetic analysis, conserved domains, and cis-regulatory elements were predicted using comparative genomics and various bioinformatics methods. Secondly, the allele SsNAP2a of the SsNAP2 member was isolated from the cDNA library of a wide accession SES208. The relative expression characteristics of the SsNAP2a were detected by qRT-PCR under hormone and abiotic stresses at different growth and development stages. Finally, transient overexpression and subcellular localization performed the function of SsNAP2a gene. The results showed that five NAP subfamily members were identified in the genome of S. spontaneum. The subcellular localization predicted that the encoded proteins of all members were localized in the nucleus. The Ka/Ks ratio of five gene pairs was less than 1, indicating that purifying selection was crucial in the evolution. Phylogenetic analysis revealed that 46 NAP members, including five representative angiosperms (Arabidopsis thaliana, Ananas comosus, Oryza sativa, Zea mays, and Sorghum bicolor), 12 reported species, and the S. spontaneum, were classified into four Clades. The evolution order was Clade I > Clade II > Clade III > Clade IV. In addition, the promoter regions of SsNAP members predicted many cis-acting elements in response to abscisic acid, jasmonic acid, low temperature, and other stresses. We speculated that they were involved in various hormone and abiotic stress-related response pathways. Furthermore, the full-length cDNA sequence of the SsNAP2a gene (GenBank accession number: OQ335094) was isolated from the wild accession SES208, with an open reading frame of 1173 bp and encoding 390 amino acid residues. The amino acid sequence similarity between SsNAP2 and SsNAP2a proteins was 97.70%. There was a difference of 10 amino acid residues, indicating that the autopolyploid allelic sequences of Saccharum species were significant difference. The qRT-PCR demonstrated that the SsNAP2a gene was constitutively expressed in various tissues of S. spontaneum, especially in the senescent bark and root, and its expression level was significantly induced under the treatment of ethylene, ET, abscisic acid (ABA), low temperature at 4℃, and high temperature at 40℃. However, the relative expression level of the SsNAP2a gene was significantly down-regulated under 8% polyethylene glycol (PEG) stress. Subcellular localization revealed that the SsNAP2a-GFP fusion protein was located in the cell nucleus of Nicotiana benthamiana leaves. After transient overexpression of the SsNAP2a gene for seven days, the leaves of N. benthamiana displayed obvious curling and shriveling phenotype. The relative expression level of ET synthesis-related genes (NtEFE26, NtAccdeaminase) was significantly up-regulated, while salicylic acid, jasmonic acid, and ABA synthesis-related genes (NtPR-1a/c, NtPR3, and NtAREB1) were significantly down-regulated, indicating that the SsNAP2a gene was involved in multiple hormone signaling pathways to induce leaf senescence. These results lay a foundation for exploring the biological functions of NAP subfamily gene members involved in sugarcane leaf senescence and provide candidate gene resources for breeding anti-senescence new cultivars.

      Function analysis of an AP2/ERF transcription factor GhTINY2 in cotton negatively regulating salt tolerance
      XIAO Sheng-Hua, LU Yan, LI An-Zi, QIN Yao-Bin, LIAO Ming-Jing, BI Zhao-Fu, ZHUO Gan-Feng, ZHU Yong-Hong, ZHU Long-Fu
      Acta Agronomica Sinica. 2024, 50(1):  126-137.  doi:10.3724/SP.J.1006.2024.34045
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      Cotton is a relatively salt-tolerant crop, but high salt stress leads to a significant decline in cotton yield and fiber quality. Mining the genes involved in salt-tolerance and illuminating the molecular mechanisms that underlie this resistance is of great importance in cotton breeding programs. Here, we identified an AP2/ERF transcription factor GhTINY2 in the transcriptome database from cotton treated with salt, and the relative expression level of GhTINY2 was reduced by salt. Subsequently, the salt-resistant phenotype and physiological indicators of the GhTINY2-overexpression Arabidopsis were analyzed. The results revealed that the GhTINY2-overexpression Arabidopsis had a significant decrease in seed germination rate, the content of proline, soluble sugar, and chlorophyll under salt stress, leading to more severe leaf wilting compared with WT. RNA-seq data from GhTINY2-transgenic Arabidopsis revealed that differentially expressed genes (DEGs) were enriched in a series of biological processes, including chlorophyll metabolism and response to stimulus, and the relative expression level of these DEGs significantly was down-regulated. Moreover, the silence of GhTINY2 in cotton through Virus-induced gene silencing (VIGS) assay showed that TRV:GhTINY2 had a significant increase in chlorophyll and proline content, leading to improved salt tolerance compare with TRV:00. In conclusion, these findings suggest that GhTINY2 was an important gene in cotton that negatively regulated salt stress resistance, and it was expected to create salt-tolerant cotton materials using GhTINY2 gene by modern genetic engineering technology in the future.

      Analysis of mutants developed by CRISPR/Cas9-based GhbHLH71 gene editing in cotton
      SHANG-GUAN Xiao-Xia, YANG Qin-Li, LI Huan-Li
      Acta Agronomica Sinica. 2024, 50(1):  138-148.  doi:10.3724/SP.J.1006.2024.34041
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      The basic/helix-loop-helix (bHLH) transcription factors play important regulatory roles in plant growth and development, secondary metabolism, signal transduction, and stress responses. The cDNA of cotton GhbHLH71 gene is 996 bp in length, encoding 331 amino acid residues. Protein sequence of GhbHLH71 contains a conserved bHLH structural domain, which is a member of the bHLH transcription factor family. The qRT-PCR showed that GhbHLH71 gene was relatively highly expressed at 3-12 DPA (days post anthesis, DPA) at rapid elongation stage in cotton fiber, implying that it mainly played a role in cotton fiber development. A CRISPR/Cas9 gene editing vector of GhbHLH71 gene was constructed, and then was transferred into upland cotton (Gossypium hirsutum L.). Six T0 mutant plants were obtained after PCR detection of Cas9 gene and mutation detection of target loci. The phenotypic traits of different T1 GhbHLH71 gene mutant plants were not significantly different from the control during cotton growth and development, but the fiber length of T1 mutants was significantly shorter compared with the control, and T2 generation mutant lines stably inherited the shortened fiber phenotype of T1 generation lines. The shortened fiber lengths of the 4# and 8# mutant lines in two consecutive generations were more than 20% shorter compared with the control, indicating that the mutation of GhbHLH71 gene mainly affected the elongation of cotton fiber cells. This study provides some insight into the biological functions of bHLH transcription factors in cotton and the molecular mechanism of cotton fiber development.

      Analysis of genotype × environment interaction and stability of yield components in faba bean lines
      SHAO Yang, GUO Yan-Ping, ZHOU Bing-Yue, ZHANG Feng, ZHANG Xin-Ming, WANG Yu-Ping
      Acta Agronomica Sinica. 2024, 50(1):  149-160.  doi:10.3724/SP.J.1006.2024.24265
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      The objective of this study is to evaluate the productivity components, stability, and adaptability of faba bean yield traits of advance lines in different pilots and select excellent lines adapted to different mage-environments. The representativeness and discriminating ability of each test-environment pilots were also evaluated, providing a basis for the selection of test-pilots. To measure plant height, grains of per plant, pods of per plant, the number of branches, 100-grain weight, and yield of per plot, a total of five advance lines [0215-1-4 (L1), 0208-3-1 (L2), 0208-3-2 (L3), 0323-2-1 (L4), and 0161-1 (L5)] and one control variety Henzheng ga candou (L6) were planted in 2017 and 2018 in Gansu province insix pilots including Countries of Hezheng, Kangle, Jishishan, Weiyuan, Linxia, and Zhang. The genotype and genotype with environment interactions of yield were analyzed by the combined analysis of variance and GGE biplot. The plot yield and plant height had no significant difference in genotype and environment interactions effect, all the other yield components had significant differences (P<0.01) in genotype effect, environmental effect, and genotype and environment interaction effect. There were very significant differences between genotype and year interaction on the number of branches, pods of per plant, grains per plant, and 100-weight grain (P<0.01) while there was no significant difference between plant height and grains of per plant. The correlation analysis showed that yield of per plot was significantly at P < 0.05 and positively correlated with pods of per plant, but negatively correlated with 100-grain weight. The GGE analysis demonstrated that the adaptability, yield, and stability of varieties (lines), as well as the ability of discrimination and the representativeness of pilots all had high GGE variation values, ranging from 78.54% to 97.38%. According to the adaptability analysis of faba bean varieties (lines), L3 had the highest yield adaptability in Kangle county, Jishishan county, Weiyuan county, Linxia county, and Hezheng county in 2018. According to the stability analysis, the varieties (lines) with the high yield in order were L3 > L2 > L6 > L4, and the varieties (lines) with the high stability in order were L4 > L1 > L5 > L3. The discriminating ability of the pilot was Kangle county in 2017, followed by Jishishan county in 2017 and 2018, and the representativeness of the pilot was Weiyuan county in 2017, followed by Kangle county in 2018 and Jishishan county in 2018. The lines with stable and high yield were L3 and L4. Combined with the ability of distinguishing and representativeness of the pilot, the most ideal mage-environment was Jishishan county. Based on GGE biplot data, faba bean with superior yield-trait components were identified in Gansu province and provided a reference base for comprehensive evaluation of faba bean varieties.

      Effects of bandwidth and plant spacing on biomass accumulation and allocation and yield formation in strip intercropping soybean
      YUAN Xiao-Ting, WANG Tian, LUO Kai, LIU Shan-Shan, PENG Xin-Yue, YANG Li-Da, PU Tian, WANG Xiao-Chun, YANG Wen-Yu, YONG Tai-Wen
      Acta Agronomica Sinica. 2024, 50(1):  161-171.  doi:10.3724/SP.J.1006.2024.34070
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      Reasonable field configuration can improve crop growth environment and increase system yield in intercropping systems. To further improve the field configuration technology for high yield and efficiency in the soybean-maize strip intercropping system in Southwest China, the soybean-maize strip intercropping system was used as the research object, and two factor split zone design was adopted. The effects of two bandwidths of 2.0 m (B1) and 2.4 m (B2) and four plant spacings of 9 cm (P1), 11 cm (P2), 14 cm (P3), and 18 cm (P4) on soybean biomass accumulation, allocation, grain filling, and yield were comprehensively analyzed. The results showed that the net photosynthetic rate of each plant spacing treatment under B2 bandwidth was higher than that of B1, and the two-year average increased by 14.26% compared with B1 under B2. At the same bandwidth, the net photosynthetic rate reached the maximum in B1P4 and B2P4, which increased by 13.57% and 25.21% compared with B1P1 and B2P1 at flowering stage, respectively. The biomass accumulation of soybean population increased and then decreased with the increase of plant spacing under the two bandwidths, and reached the maximum at B1P3 and B2P2, respectively, and B2 increased by 9.82%-22.08% compared with B1 at maturity stage. And the increase of bandwidth and plant spacing promoted the accumulation and transfer of soybean post-flowering matter to grain. Compared with B1, post-flowering dry matter accumulation and dry matter transfer increased by 13.82-28.01% and 13.38%-37.76% under B2 treatment, respectively, and the proportion of grain matter accumulation increased to 41.80%-44.26%. The increase of biomass accumulation improved the grain filling process, and the active grain-filling period (D) of soybean under B2 was extended by 2-3 days compared with B1. The mean grain-filling rate reached its maximum at P4 and increased by 5.80% and 6.58% compared with P1 under the two bandwidths. The yield results showed that, with the increase of bandwidth and plant spacing, the effective plants decreased, and the seeds per plan and 100-seed weight increased in the soybean-maize strip intercropping model. The soybean yield under B2 bandwidth increased by 22.32%-36.87% compared with B1, it reached the maximum in B1P3 and B2P2 under the two bandwidths, respectively, and increased by 17.83%-26.44% and 10.71%-10.76% compared with B1P1 and B2P1 for two years. In summary, the soybean plant spacing of 11 cm at the bandwidth of 2.4 m can effectively improve the post-flowering dry matter accumulation and allocation, promote grain filling, increase the number of seeds per plant and 100-seed weight, improve soybean population yield, and achieve the high yield and high efficiency in the soybean-maize intercropping system.

      Identification of heat tolerance of waxy maizes at flowering stage and screening of evaluation indexes in the middle and lower reaches of Yangtze River region
      SONG Xu-Dong, ZHU Guang-Long, ZHANG Shu-Yu, ZHANG Hui-Min, ZHOU Guang-Fei, ZHANG Zhen-Liang, MAO Yu-Xiang, LU Hu-Hua, CHEN Guo-Qing, SHI Ming-Liang, XUE Lin, ZHOU Gui-Sheng, HAO De-Rong
      Acta Agronomica Sinica. 2024, 50(1):  172-186.  doi:10.3724/SP.J.1006.2024.33019
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      Heat stress has acknowledged as one of the major threats to waxy maize production as a result of global warming gradually seriously. Heat resistance identification and indices screening could provide the theoretical basis in breeding heat-resistant cultivars. In the present study, 13 heat-related parameters of 10 waxy maize cultivars from the middle and lower reaches of Yangtze River area were measured under normal temperature treatment (CK) and two heat stress treatments (artificial warming treatment and delayed sowing treatment). Principal component analysis, membership function method, cluster analysis, and stepwise regression analysis were used to comprehensively evaluate the heat resistance of waxy maize at flowering stage. The results showed that most indices under heat stress decreased significantly compared with CK, except for chlorophyll a and PS II primary light energy conversion efficiency. Three independent comprehensive components were obtained from 13 single traits using a principal component analysis, and their contribution rates respectively were 64.46%, 15.06%, and 7.76%, which represented 87.28% information of the original data. Ten testing cultivars were divided into three heat-tolerance types that were heat tolerance category, medium heat tolerance category, and high temperature sensitive category based on comprehensive heat tolerance values (D) calculating by membership function methods. SYN2, ZN2, and SYN901 were identified as the heat-resistant cultivars, and planting these cultivars could partly alleviate the negative effects of heat stress on waxy maize production. Finally, stepwise regression method was used to establish a predictive equation to evaluate heat tolerance, which indicating that predictive values (PV) calculated by the predictive equation were basically consistent with D value. The results showed that yield, Fv/Fm, leaf area index, and pollen viability could be used to identify heat-resistant cultivars of waxy maize. The heat tolerant cultivars and identification index selected in the study could serve as a basis for subsequent breeding heat tolerant cultivars of waxy maize.

      Mechanism of cyclanilide enhanced the defoliation efficiency of thidiazuron in cotton by regulating endogenous hormones under low temperature stress
      SUN Shang-Wen, SHU Hong-Mei, YANG Chang-Qin, ZHANG Guo-Wei, WANG Xiao-Jing, MENG Ya-Li, WANG You-Hua, LIU Rui-Xian
      Acta Agronomica Sinica. 2024, 50(1):  187-198.  doi:10.3724/SP.J.1006.2024.34046
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      The technology of cotton leaf removal is an important prerequisite for realizing the mechanical harvesting of cotton, but low temperature affect the efficiency of TDZ-induced leaf abscission, which cannot meet the requirements of mechanized harvesting. In our laboratory, we found that cyclanilide (CYC) could improve the defoliation efficiency of TDZ at low temperature, but the mechanism of CYC promoting chemical defoliation (TDZ) under low temperature is unknown. Therefore, the content of plant hormones and the relative expression level of related genes in the abscission zone of cotton leaves were analyzed using Zhongmian 425 as the experimental material, setting two temperature levels (25℃ and 15℃) and three treatments [water (CK), single TDZ (T), and the compound of TDZ+CYC (TC)]. At 240 h after treatment under low temperature (15℃), the abscission rate of cotton leaves treated by T was only 53.0%, while the start time of cotton leaves abscission treated by TC was 24 h ahead of that treated by T and the abscission rate of cotton leaves treated by TC increased to 79.6%. At low temperature, compared with T, the relative expression level of auxin (IAA) transport genes (LAX2, PIN1), IAA response genes (IAA9, ARF3) and the content of IAA in the abscission zone of cotton leaves treated with TC were reduced significantly; the relative expression level of ethylene (ET) synthesis genes (ACS, ACO), and the content of ET synthesis precursor ACC were increased, the relative expression level of ET downstream signal gene(ERF1B) was up-regulated significantly in the abscission zone treated with TC. The jasmonate acid (JA) synthesis-related gene (AOC4) was up-regulated, and the content of JA was increased in abscission zone treated by TC. The inhibition of the IAA transport and signal transduction and the promotion of ET and JA synthesis and ET signal transduction caused by TC treatment in the abscission zones of cotton leaves could be the key reason that CYC could promote cotton leaves abscission effect of TDZ at low temperature.

      Identification of cotton low potassium tolerance based on AHP-membership function method at cotyledonary stage
      TAN Zhi-Xin, XIE Liu-Wei, LI Hong-Ge, LI Fang-Jun, TIAN Xiao-Li, LI Zhao-Hu
      Acta Agronomica Sinica. 2024, 50(1):  199-208.  doi:10.3724/SP.J.1006.2024.34057
      Abstract ( 146 )   HTML ( 12 )   PDF (626KB) ( 107 )   Save
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      In order to establish a rapid and reliable evaluation system for low-potassium (K) tolerance in cotton, the combination of decision evaluation model AHP and membership function method was used to evaluate the 384 cotton germplasm resources at the cotyledonary stage. The results showed that, under K deficiency (0.03 mmol L-1), the coefficient of variation of the relative area of K deficiency spots on cotyledons was the largest (96.24%), the coefficient of variation of plant dry weight was the smallest (14.73%). This trait of angle between cotyledons and hypocotyls, root length, root surface area, root volume, K+ concentration, and K+ accumulation ranged from 15.72% to 31.52%. The angle between cotyledons and hypocotyl was significantly negatively correlated with the relative area of K deficiency spots. The characteristics of root morphology were positively correlated with plant dry weight and K+ accumulation. According to the evaluation of AHP-membership function method, the comprehensive evaluation C-value of 384 cotton germplasms varied from 0.25 to 0.79. Among which Wujiyizhihua (C-value = 0.7868) had the strongest resistance to low K, and Brazil 014 (C-value = 0.2451) had the lowest resistance to low K. The results of systematic clustering showed that the proportions of highly (C-value > 0.74) and moderately (0.33 < C-value < 0.69) low-K tolerant germplasms were 1.30% and 95.31%, respectively; and the proportion of low-potassium-sensitive (C-value < 0.33) germplasms was 3.39%. The correlation between seed K+ concentration and low K tolerance of cotton at cotyledonary stage was not significant difference. In this study, the screening system of low-potassium tolerant germplasm resources of cotton was improved, the cotyledonary stage potassium tolerance of 384 test germplasms was identified, and 5 germplasms with strong potassium tolerance (Wujiyizhihua, Brown 1-61, Black Mountain Cotton 1, Qinli 514, Soviet Cotton 78) were screened, which provided the theoretical and application value.

      Ethephon ripening affects boll weight and fiber quality of machine-harvested cotton
      LIU Tao-Fen, LUO Dan, ZHANG Qi-Peng, SUN Yuan-Yuan, LI Pei-Song, TIAN Jing-Shan, ZHANG Wang-Feng, XIANG Dao, ZHANG Ya-Li, YANG Ming-Feng, GOU Ling
      Acta Agronomica Sinica. 2024, 50(1):  209-218.  doi:10.3724/SP.J.1006.2024.34039
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      The use of ethephon can achieve early and concentrated the machine-harvested cotton boll opening. The purpose of this study is to analyze the effect of the boll-period shortening on the boll weight, fiber quality, and relationship, when the ethephon was sprayed at different stages of the boll development with a selection of 18 materials. The results showed that when the period of cotton was 47-69 days, spraying ethephon at the age of boll was 40.5-49.7 days, which could advance boll opening by 3.5-5.7 days. It took 64.1-69.7 days for fiber to fully mature under ethephon, while it took 54.8-60.5 days for cotton seed. When boll was opened 1.5-6.2 days earlier, the probability of boll weight and fiber quality decreasing was up to 58.0%-76.5%. To achieve an early opening of the boll, ethephon can be applied at the boll age of 34.8-44.1 days, when the boll weight (< 0.26 g) and fiber quality damage (length reduction < 0.28 mm and fiber strength < 0.30 cN tex-1) at the top of cotton plant are reduced by 5% and 1%, respectively. In addition, the boll period was shortened in some materials, but the boll weight, fiber length, and strength were increased, with the 9.5%-23.5% probability. Therefore, the reasonable use of ethephon in some cotton varieties can shorten the boll period and improve the boll weight and quality.

      Effects and variability analysis of different salt and alkali stresses on the proteome of cotton leaves
      GUO Jia-Xin, YE Yang, GUO Hui-Juan, MIN Wei
      Acta Agronomica Sinica. 2024, 50(1):  219-236.  doi:10.3724/SP.J.1006.2024.34035
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      Salt stress and alkali stress are two different abiotic stresses that seriously affect agricultural production. Exploring the differences between salt stress and alkali stress in cotton provides a theoretical basis for cotton cultivation on different types of saline alkali land. In this experiment, three treatments of CK, NaCl (CS), and NaHCO3+Na2CO3 (AS) were conducted. Proteomics analysis and physiological index verification revealed that the tolerance mechanism of cotton to salt stress and alkali stress. Compared with CK, CS, and AS cotton plants decreased by 51.1% and 50.9% at P < 0.05 in biomass, respectively. The chlorophyll content and Pn under salt stress decreased significantly by 53.9% and 57.2%, respectively. Under salt stress, the activities of hexokinase, fructose phosphate kinase, pyruvate kinase, citrate synthase, glutamate dehydrogenase, and glutamic oxaloacetic transaminase in cotton leaves increased significantly by 13.8%, 14.4%, 4.7%, 4.5%, 36.6%, and 12.9%, respectively. Under alkali stress, the activities of hexokinase, phosphofructose kinase, pyruvate kinase, malate dehydrogenase, citrate synthase, glutamate dehydrogenase, and glutamic oxaloacetic transaminase in cotton leaves significantly increased by 4.8%, 38.8%, 15.1%, 4.3%, 3.4%, 15.2%, and 21.1%, respectively. Based on TMT proteomics, 458 differentially expressed proteins and 140 differentially expressed proteins in salt stress and alkali stress species were detected, respectively. These proteins were involved in photosynthesis, sugar metabolism, 2-oxocarbonic acid metabolism, amino acid synthesis and metabolism, and other life processes. These results showed that both salt stress and alkali stress inhibited cotton growth. The difference was that under salt stress, proteins expression related to photosynthesis decreased, photosynthesis was significantly inhibited, and carbon water and energy metabolism were enhanced, and more photosynthates were used for energy metabolism. Alkali stress had no significant effect on cotton photosynthesis, and more photosynthates might be transported to the roots to secrete organic acids.

      Relative expression profiles of genes response to salt stress and constructions of gene co-expression networks in Brassica napus L.
      YANG Chuang, WANG Ling, QUAN Cheng-Tao, YU Liang-Qian, DAI Cheng, GUO Liang, FU Ting-Dong, MA Chao-Zhi
      Acta Agronomica Sinica. 2024, 50(1):  237-250.  doi:10.3724/SP.J.1006.2024.34076
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      Brassica napus L. is an important oil crop. Salt stress is one of the major environmental conditions affecting the growth and development of B. napus, which may lead to yield reduction, quality deterioration, and even the death of B. napus. In this study, the B. napus semi-winter cultivar ZS11 was used as the experimental material to perform transcriptome sequencing on the leaf and root tissues with salt stress treatment (0, 0.25, 0.5, 1, 3, 6, 12, and 24 h). The measured 90 RNA-seq data provided the high-resolution time dynamic transcriptional expression spectrum of rapeseed tissues responding to salt stress was obtained. Correlation analysis showed that the samples exhibited significant clustering differences in early response and late response before and after 1 h salt stress treatment. Using DESeq2 for differential gene analysis, we identified 20,462 and 29,334 differential genes for the response of root and leaf tissues, respectively, indicating that the overall response of leaf tissue in rapeseed was more severe than the root. Furthermore, WGCNA was used to construct gene co-expression networks for the salt stress response in root and leaf tissues, respectively, and tan and yellow modules were significantly related to the early response to salt stress, and green and red modules were significantly related to the late response to salt stress. GO enrichment analysis was then performed, and 41 and 26 core transcription factors responding to salt stress at the early and late stages, respectively, were selected from these networks. Functional annotation showed that the known Arabidopsis homologous genes involved in salt stress responses at different stages existed in all four modules, and core genes, such as BnWRKY46 and BnWRKY57, had abundant SNPs variation and haplotypes in 505 salt stress-treated rape population variation data, suggesting that these core transcription factors might be key candidate genes for rape salt stress response. This study provides a reliable data reference and candidate gene resources for improving salt tolerance in B. napus.

      Crop growth characteristics and its effects on yield formation through nitrogen application and interspecific distance in soybean/maize strip relay intercropping
      YANG Li-Da, REN Jun-Bo, PENG Xin-Yue, YANG Xue-Li, LUO Kai, CHEN Ping, YUAN Xiao-Ting, PU Tian, YONG Tai-Wen, YANG Wen-Yu
      Acta Agronomica Sinica. 2024, 50(1):  251-264.  doi:10.3724/SP.J.1006.2024.33018
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      The objective of this study is to clarify relay intercropping crop growth characteristics and its effects on yield formation under nitrogen application and interspecific distance. In this study, the soybean/maize strip relay intercropping system was used as the main body, to explore the crop growth rate, dry mater accumulation, and distribution, and yield differences under different nitrogen levels (nitrogen application and no nitrogen application) and interspecific distance (the spacing of soybean 30, 45, 60, 75, and 100 cm mono-cropping), to fit the process of crop dry mater accumulation, and comprehensively analyze the crop growth law and yield benefit. The results showed that the maximum growth rate of maize was reached from male stage to milking stage, and the maximum growth rate was 30 cm (MS30) without nitrogen application, which was 34.99% higher than that of monoculture maize (MM100). The growth rate of relay intercropping was significantly lower than that of monoculture soybean (SS100) before the first flowering stage, but higher than that of SS100 after the first flowering stage, with the highest spacing of 60 cm (MS60), and 78.91% higher than that of SS100 from full flowering stage to full pod stage without nitrogen application. The logistic equation could fit well the matter accumulation process of maize and soybean, and the R2 were all above 0.95. Compared with no nitrogen application, nitrogen application delayed the accumulation peak and increased the biomass of maize. The matter accumulation of intercropping soybean was slower than monoculture at the early growth stage, but MS45 and MS60 were gradually equal to or even exceeded monoculture soybean at later growth stage. Nitrogen application increased the dry matter allocation rate to the grain, and maize yield increased by 10.05% and 40.90% in two years, respectively. With the increase of interspecific distance, soybean yield increased first and then decreased, with the highest value in MS60 and the lowest value in MS30. In the two years, soybean yield in MS60 was 23.88% and 31.77% higher than MS30 under no nitrogen application and nitrogen application, respectively. The land equivalent ratio under relay intercropping was all above 1.35, among which MS60 under nitrogen application was the largest (1.89). The suitable interspecific distance (interspecific distance 60 cm) under relay intercropping can realize the cooperative growth of maize and soybean, increase crop growth rate, promote the accumulation and distribution of matter, and improve the system yield and land equivalent ratio.

the Crop Science Society of China
the Institute of Crop Science, CAAS
China Science Publishing & Media Ltd.
Published: Science Press
Editor-in-chief: Wan Jian-min
Associate Editors-in-Chief:
Chen Xiao-ya Yang Jian-chang Zhang Xian-long Wang Jian-kang Xu Ming-liang Liu Chun-ming Wang Dao-wen Sun Chuan-qing Ding Yan-feng Jin Wei-wei Chu Cheng-cai Cheng Wei-hong
Director of the editorial department:
Yan Chun-ling
CN 11-1809/S
ISSN 0496-3490
Post subscription code: 82-336

  • Started in 2013
  • Covered by SCIE
  • Open access in ScienceDirect

Editor in chief: Wan Jian-min
CN 10-1112/S
ISSN 2095-5421, 2214-5141(online)
Online published:
Submission: https://www.editorialmanager.com/cj/
E-mail: cropjournal@caas.cn
Tel: 8610-82108548