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    12 September 2023, Volume 49 Issue 9
    • CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
      ALGWAS: two-stage Adaptive Lasso-based genome-wide association study
      YANG Wen-Yu, WU Cheng-Xiu, XIAO Ying-Jie, YAN Jian-Bing
      Acta Agronomica Sinica. 2023, 49(9):  2321-2330.  doi:10.3724/SP.J.1006.2023.23072
      Abstract ( 481 )   HTML ( 37 )   PDF (874KB) ( 823 )   Save
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      As mainstream methods for genome-wide association analysis, mixed linear model methods have been widely used. However, the existing methods still have the problem of low detection power. In this study, a two-stage Adaptive Lasso-based genome-wide association analysis (ALGWAS) method was proposed. In the first stage, single nucleotide polymorphism (SNP) associated with target traits were screened by Adaptive Lasso, a variable selection method. In the second stage, SNPs selected from the first stage were put into the linear model as the covariates for genome-wide scanning. Compared with fastGWA, GEMMA and EMMAX, the ALGWAS method had the highest detection power and lower false discovery rate (FDR) in the simulation experiments. The above four methods were applied to genome-wide association analysis of Complete-diallel plus Unbalanced Breeding-like Inter-Cross (CUBIC) population of 1341 individuals in maize. ALGWAS method can detect the genes (ZmMADS69, ZmMADS15/31, ZmZCN8, and ZmRAP2.7) related to days to tasseling, the genes (ZmBRD1 and ZmBR2) related to plant height, and the genes (ZmUB2, ZmKRN2, and ZmCLE7) related to yield, while the other three commonly used genome-wide association analysis methods had low detection efficiency. In this study, a non-mixed linear model class of genome-wide association analysis method was proposed, which had higher detection advantage for microeffect polygenes and provided a new way for genetic analysis of complex traits.

      Identification of ZmC2s gene family and functional analysis of ZmC2-15 under heat tolerance in maize
      HUANG Yu-Jie, ZHANG Xiao-Tian, CHEN Hui-Li, WANG Hong-Wei, DING Shuang-Cheng
      Acta Agronomica Sinica. 2023, 49(9):  2331-2343.  doi:10.3724/SP.J.1006.2023.23069
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      The objective of this study is to identify the members of maize ZmC2s gene family, to analyze the association between their genetic variations and heat tolerance, and to lay a foundation for clarifying its function and molecular mechanism in maize heat tolerance. Using the C2 protein domain PF00168, hmmsearch was applied to search for the members of the ZmC2s gene family from maize B73 genome. The protein isoelectric point, molecular weight, phylogenetic evolution, and gene family replication were analyzed. Using the method of candidate gene association analysis, the association between the natural variations of ZmC2s and the heat tolerance of maize seedlings was conducted, and the important heat-tolerant candidate genes of maize ZmC2s gene family were found. The relative gene expression level of the heat-tolerant candidate gene under stress was identified by Real time fluorescent quantitative PCR (RT-qPCR). The subcellular expression sites of heat-tolerant candidate gene were identified by transforming maize protoplasts. A total of 95 maize ZmC2s genes were identified from the reference genome B73 in maize. According to the order of their physical coordinates, 95 maize ZmC2s genes were named from ZmC2-1 to ZmC2-95, respectively. The length of the 95 proteins was 130-2141, the isoelectric point was 4.1-10.8, and the molecular weight was 14.1-230.1. The evolution tree of C2 gene in maize, rice, and sorghum genomes was constructed. We found that C2 genes can be divided into three major cluster branches, and each cluster branch can be subdivided into two small cluster branches. Analyzing the whole genome collinearity data of maize, rice, and sorghum, 59 ZmC2s genes were detected to have corresponding replication genes in rice and sorghum genomes. A candidate-gene based on the association analysis of ZmC2s showed that ZmC2-15/60/91 were important candidate genes for heat tolerance in maize (P ≤ 0.001, MLM), among which ZmC2-15 was the most significantly associated to heat tolerance at seedling stage (P ≤ 0.000,01, MLM), and the relative expression level of ZmC2-15 was up-regulated under various stress treatments. Subcellular localization indicated that ZmC2-15 was localized in the cytoplasm, nuclear membrane, and endoplasmic reticulum. The overexpression of ZmC2-15 improved plant heat tolerance. ZmC2-15 can be used as an important candidate gene for regulating heat tolerance in maize.

      Relative expression patterns of laccase gene family members in upland Gossypium hirsutum L.
      ZUO Chun-Yang, LI Ya-Wei, LI Yan-Long, JIN Shuang-Xia, ZHU Long-Fu, ZHANG Xian-Long, MIN Ling
      Acta Agronomica Sinica. 2023, 49(9):  2344-2361.  doi:10.3724/SP.J.1006.2023.24246
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      Laccase, a member of the blue copper oxidase protein family, plays an important role in plant lignin synthesis and improving plant resistance to stress. In this study, 104 members of the Laccase gene (GhLAC) family were identified from the upland cotton genome. Phylogenetic tree and tissue expression map were constructed. Twenty genes were randomly selected for qRT-PCR analysis to verify the results of expression heat map. To further explore the role of laccase in cotton, promoter-GUS fusion vectors were transformed into Arabidopsis thaliana. The detailed expression patterns of six members of the Laccase gene family (GhLAC12A, GhLAC14A, GhLAC20A, GhLAC25D, GhLAC59D, and GhLAC63D) were studied by GUS staining in different tissues during different developmental period of transgenic Arabidopsis thaliana. To explore the role of laccase in stress, the expression of the six laccase genes was analyzed by cutting and piercing, and the corresponding genes were analyzed by qRT-PCR using the anther of two cotton strains ‘84021’ (high temperature tolerant) and ‘H05’ (high temperature sensitive) at different stages under normal and high temperature conditions. The results showed that 20 randomly selected genes were differentially expressed in six tissues of root, stem, leaf, petal, anther, and stigma, and the relative expression levels of most genes were consistent with the transcriptome data. The promoter of six laccase genes could drive GUS gene expression in different levels at germination, two-leaf, and four-leaf stages. The trauma treatment indicated that the promoter of GhLAC12A and GhLAC14A significantly improved the ability to drive GUS protein expression in leaves after trauma induction, suggesting that the two genes might be involved in traumatic stress response. In addition, the relative expression levels of the six GhLACs genes were significantly down-regulated after high temperature stress at the tetrad stage and anther dehiscence stage of cotton strain ‘84021’, suggesting GhLACs gene might negatively regulate the high temperature tolerance of cotton anthers. The results of this study provide the reference for further exploring the function of laccase family genes.

      Mutation effects of OsCDF1 gene and its genomic variations in rice
      HU Yan-Juan, XUE Dan, GENG Di, ZHU Mo, WANG Tian-Qiong, WANG Xiao-Xue
      Acta Agronomica Sinica. 2023, 49(9):  2362-2372.  doi:10.3724/SP.J.1006.2023.22062
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      Flowering time (heading date) affects yield, quality, and regional adaptability of rice. The Cycling DOF Factor 1 (CDF1) protein is a transcriptional repressor of CONSTANS (CO) and negatively regulates flowering time in Arabidopsis. However, the biological functions of OsCDF1 in rice is not quite clear. To explore the biological functions of OsCDF1 and its effects on flowering time control in rice, we constructed two binary vectors carrying guide RNAs targeting OsCDF1 gene via CRISPR/Cas9 system. The resultant plasmids were transferred into SN9816 which was the variety widely cultivated in northern China by using an Agrobacterium-mediated transformation, and the mutations of OsCDF1 was firstly generated in SN9816. The flowering time and yield related traits of SN9816 and oscdf1 mutants were investigated in the paddy field. The main results were as follows: Two homozygous oscdf1 lines were identified, including a five bp deletion at 16th bp of the first exon and a single base pair A insertion at 338th bp of the second exon. Sequence alignment analysis revealed that the two types of mutations resulted in frame-shift and premature translation termination. Mutations of OsCDF1 delayed flowering time, but increased yield under natural long day conditions in rice. Analysis of OsCDF1 genetic variations and haplotype networks revealed that the rice accessions had evolved high genomic diversity in OsCDF1 locus. The knockout mutants of OsCDF1 created by CRISPR/Cas9 provided the theoretical basis to further study the role of OsCDF1 gene in rice and the potential gene and germplasm resources for genetic improvement in rice.

      Effects of salt stress on ion dynamics and the relative expression level of salt tolerance genes in peanut seedlings
      XU Yang, ZHANG Dai, KANG Tao, WEN Sai-Qun, ZHANG Guan-Chu, DING Hong, GUO Qing, QIN Fei-Fei, DAI Liang-Xiang, ZHANG Zhi-Meng
      Acta Agronomica Sinica. 2023, 49(9):  2373-2384.  doi:10.3724/SP.J.1006.2023.24242
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      Different peanut varieties have different salt tolerance. In this study, to determine the flow rates of ions Na+, K+, Ca2+, NH4+, NO3-, and Cl- in root tips of peanut seedlings under salt stress via Non-invasive Micro-test technique, a salt-tolerant peanut variety Huayu 25 (HY25) and a salt-sensitive variety Huayu 20 (HY20) were used as the experimental materials. The growth traits, the relative expression level of major salt tolerance genes, and the contents of osmotic regulatory substances (soluble sugar and proline) were also measured to establish the difference of ion absorption and stress resistance regulation in different varieties. The results showed as follows: (1) Under NaCl stress, Na+ influx was inhibited, and its efflux increased, but promoted the influx of K+. The efflux rate of Na+ and influx rate of K+ in HY25 were higher than HY20, which may improve salinity tolerance by preserving K+ and discharging Na+. (2) Salt stress promoted Ca2+ influx, and the Ca2+ influx rate of salt-tolerant varieties was higher than the salt-sensitive varieties, which might be related to salt tolerance. (3) NO3- exhibited efflux in both varieties under salt stress, but the efflux rate of the salt-tolerant variety HY25 was lower, indicating that HY25 could resist the harm of salt stress by slowing the loss of NO3-. (4) Salt stress induced Cl- efflux in HY25 but influx in salt-sensitive variety, indicating that HY25 could reduce the toxicity of Cl- by accelerating the efflux of Cl-. (5) Salt stress significantly up-regulated the relative expression level of salt-tolerant genes AhNHX1, AhHA1, AhSAMDC1, and AhLeaD in salt-tolerant variety HY25, which could help improve its salt tolerance. Clarifying the dynamic changes of root ion flow and resistance mechanism under salt stress can provide the theoretical support for improving the emergence, establishment, and development of peanut seedlings in saline-alkali land and the establishment of regulation technology.

      Cloning and functional analysis of sucrose transporter protein SsSWEET11 gene in sugarcane (Saccharum spontaneum L.)
      DU Cui-Cui, WU Ming-Xing, ZHANG Ya-Ting, XIE Wan-Jie, ZHANG Ji-Sen, WANG Heng-Bo
      Acta Agronomica Sinica. 2023, 49(9):  2385-2397.  doi:10.3724/SP.J.1006.2023.24232
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      SWEET (Sugars Will Eventually be Exported Transporter) proteins are widely involved in the physiological and biochemical processes of plant growth and development and response to pathogen stress by regulating the transportation, distribution, transformation, and storage of sugar in plants. This study revealed the biological function of SWEET genes in the growth and development of sugarcane and its interaction with red stripe pathogen Acidovorax avenae subsp. avenae (Aaa). Firstly, based on the PacBio full-length transcriptome cDNA library of S. spontaneum SES208 and comparative genomics, the specific primers were designed according to the re-annotated SsSWEET11 gene sequence. The full-length sequence was mined from the cDNA library by quantitative RT-PCR technology. The characteristics of the SWEET proteins were analyzed using various biological information tools, and the SWEET proteins from some plants were constructed a phylogenetic tree. Secondly, RT-qPCR detected the relative expressions of the SsSWEET11 gene with different tissues and two cultivars, ROC22 (resistant to red stripe) and MT11-610 (susceptible to red stripe). Finally, transient overexpression and subcellular localization performed the function of the SsSWEET11 gene. The results showed that the full-length cDNA sequence of the SsSWEET11 gene (GenBank accession number: OP554214) was cloned from S. spontaneum SES208, with an open reading frame of 927 bp and encoding 308 amino acid residues, which contained two MtN3_saliva domains and seven transmembrane domains. Phylogenetic analysis revealed that the SWEET protein family could be divided into four subfamilies, and SsSWEET11 belonged to subfamily III. The amino acid sequence similarity between SsSWEET11 and SbSWEET11 protein from sorghum is 97.99%. qRT-PCR demonstrated that the SsSWEET11 gene was constitutively expressed in different tissues of S. spontaneum and the relative expression level in leaves and roots was significantly higher than that in other tissues. Under the stress of Aaa, the SsSWEET11 gene presented a different expression pattern between sugarcane cultivars (ROC22) and (MT11-610) and was significantly reduced in the resistant sugarcane cultivar compared with the blank control. However, the expression of ShSWEET11 was significantly up-regulated in the susceptible sugarcane cultivar after 48 hours post-inoculation (hpi) and 72 hpi, which were 5.90 times and 5.43 times higher than the control, respectively. Subcellular localization indicated that the SsSWEET11-GFP fusion protein was located in the plasma membrane. After transiently overexpression of the SsSWEET11 gene for one day, the color of Nicotiana benthamiana leaves remained unchanged by DAB staining, and seven days after inoculation with Pseudomonas solanacearum, and Fusarium solani var. coeruleum, the incidence of transient overexpression of ShSWEET11 gene in N. benthamiana leaves were more susceptible than that of the control. Allergic reaction-related genes, jasmonic acid, and salicylic acid metabolism pathway-related genes were up-regulated, but ethylene pathway-related genes did not respond, suggesting that the SsSWEET11 gene is involved in jasmonic acid and salicylic acid signal transduction pathways, and the infection of N. benthamiana leaves by pathogens can induce an allergic reaction. These results not only provided an accumulation for the development of molecular markers associated with sugarcane resistance to Aaa but also laid a foundation for in-depth analysis of the molecular mechanism in sugarcane in response to Aaa infection.

      CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS
      Dynamic change profile of histone H3K18cr on rice whole genome under cold stress
      LIU Kai, CHEN Ji-Jin, LIU Shuai, CHEN Xu, ZHAO Xin-Ru, SUN Shang, XUE Chao, GONG Zhi-Yun
      Acta Agronomica Sinica. 2023, 49(9):  2398-2411.  doi:10.3724/SP.J.1006.2023.22059
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      Histone modifications play an important role in response to abiotic stresses in rice. Crotonylation is a novel histone modification, and the changes of crotonylation in rice subjected to low temperature stress are rarely reported. In this study, RNA-seq and ChIP-seq were performed on normal-grown and cold-treated rice seedlings of rice variety ‘Nipponbare’, followed by combined analysis of the regulatory characteristics of the crotonylation modification at the histone H3 lysine 18-specific site (H3K18cr) on gene expression under cold stress. The results showed that H3K18cr was mainly enriched in the first exon and intergenic region and had positive correlation with gene expression and gene length. The global distribution of H3K18cr in rice genome did not change under cold stress, but WB and ChIP-seq results indicated that the overall modification level decreased. The differential modification analysis revealed that there were significant increase and decrease of 899 genes and 409 genes in the modification after cold stress, respectively. Association analysis with RNA-seq showed that a total of 199 genes had increased levels of H3K18cr modification and up-regulated expression levels. GO enrichment revealed that these genes were mainly involved in the processes such as the regulation of transcriptional activity. Further analysis indicated that histone H3K18cr was involved in the response process of cold stress by regulating the expression of transcription factors such as OsDREB1A, OsEATB, OsAP2-39, and OsNAC9 in rice. The results provide a theoretical basis for further understanding the epigenetic mechanisms of histone crotonylation in response to cold stress in plants.

      Comparative transcriptome analysis of elite ‘ROC’ sugarcane parents for exploring genes involved in Sporisorium scitamineum infection by using Illumina- and SMRT-based RNA-seq
      HU Xin, LUO Zheng-Ying, LI Chun-Jia, WU Zhuan-Di, LI Xu-Juan, LIU Xin-Long
      Acta Agronomica Sinica. 2023, 49(9):  2412-2432.  doi:10.3724/SP.J.1006.2023.24228
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      Sugarcane smut, caused by the fungus Sporisorium scitamineum, is the most challenging disease of sugarcane, causing significant losses in cane yield. There is a dearth of information on smut resistance mechanism in elite parents for the development of smut-resistant varieties. In the present study, we adopted joint Illumina- and Single Molecule Real-Time (SMRT)-based RNA-seq analysis to identify transcript expression in an smut-resistant and -susceptible parents (ROC25 and ROC22) infected with S. scitamineum. A total of 79,885 high-quality transcripts was obtained, including 60,115 open reading frames, 3692 alternate splicing isoforms, 1799 long non-coding RNAs, 29,139 simple sequence repeats, and 7794 transcription factors. About 92.72% of the total transcripts were annotated, which should have increased the available data amount for transcriptome profile analysis. There were 2033 and 9716 differentially expressed transcripts (DETs) in ROC22 and ROC25, respectively. The analyses of GO and KEGG enrichment showed that more GO terms and KEGG pathways were observed in ROC25 than ROC22. It was found that MAPK signalling pathway-plant, phenylpropanoid biosynthesis, plant-pathogen interaction, linoleic acid metabolism, and starch and sucrose metabolism were enriched both in resistant and susceptible parents. In addition, MAPK superfamily genes were differentially regulated in different parents, more DETs of MEKK1 and MKK4 were detected in resistant parent, and the relative expression levels of MKK5, MPK10, and MPK12 genes were specifically altered in resistant parent. It suggested that MAPK superfamily genes might play the important roles in the regulation of sugarcane response to S. scitamineum infection. Moreover, lots of transcription factors (TFs) associated with plant disease resistance were found to respond to S. scitamineum infection in both ROC22 and ROC25 parents, including WRKY, MYB, NAC, and AP2/ERF-ERF. Majority of the TFs were up-regulated. Compared to the susceptible ROC22 parent, the number of activated transcription factors in the resistant ROC25 parent was higher, indicating that these extra TFs might have positive effects in the defense against S. scitamineum. This study provides a comprehensive set of reference transcripts for sugarcane and thus increases our understanding on the interactions between sugarcane and S. scitamineum, which should be helpful in guiding on exploitation and utilization of smut-resistance gene resources.

      Response of maize transcriptional factor ZmEREB211 to abiotic stress
      AI Rong, ZHANG Chun, YUE Man-Fang, ZOU Hua-Wen, WU Zhong-Yi
      Acta Agronomica Sinica. 2023, 49(9):  2433-2445.  doi:10.3724/SP.J.1006.2023.23071
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      AP2/ERF (APETALA2/ethylene-responsive factor) transcription factor is one of the largest transcription factor families in plants, which plays an important role in regulating plant growth and development and responding to various stresses. Exploring the function of AP2/ERF family genes in maize (Zea mays L.) will provide the important genetic resources for the creation of new maize germplasm. In this study, ZmEREB211 (Gene ID: 103647485) gene was cloned and its basic characteristics, tissue expression characteristics, and the relative expression patterns in response to stress were analyzed by bioinformatics and qRT-PCR. The transgenic Arabidopsis lines were subjected to corresponding stress treatment and phenotypic identification. The results showed that the gene contained only one exon and the full-length cDNA was 792 bp which encoding 263 amino acids. The ZmEREB211 protein had a molecular weight of 27.9 kD and a theoretical isoelectric point of 6.01. It had a conserved domain unique to the AP2/ERF family. The relative expression level of ZmEREB211 gene was the highest in maize root system and the relative expression level in young roots was higher than mature roots. At the same time, the gene had different degrees of induced expression under dehydration, high salt, drought, and low temperature treatment conditions. On 1/2 MS medium containing different concentrations of NaCl, mannitol, and jasmonic acid (JA), root length of ZmEREB211 transgenic Arabidopsis lines was significantly longer than wild type. Under drought and high salt treatments, transgenic Arabidopsis lines had stronger tolerance than wild type, and the number of green leaves at seedling stage was significantly higher than wild type and the peroxidase (POD) activity and chlorophyll content were significantly higher than wild type. ZmEREB211 may be involved in the regulation of root growth and development in maize, and can play a positive regulatory role in high salt, drought, osmotic stress, and JA hormone treatments. This study provides an important reference for further analysis of the biological function of ZmEREB211 in maize.

      Comparative transcriptome profiling of dormancy regulatory network in peanut
      WANG Fei-Fei, ZHANG Sheng-Zhong, HU Xiao-Hui, CHU Ye, CUI Feng-Gao, ZHONG Wen, ZHAO Li-Bo, ZHANG Tian-Yu, GUO Jin-Tao, YU Hao-Liang, MIAO Hua-Rong, CHEN Jing
      Acta Agronomica Sinica. 2023, 49(9):  2446-2461.  doi:10.3724/SP.J.1006.2023.24186
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      Seed dormancy is an important and complex agronomic trait affecting yield and quality of peanut (Arachis hypogaea L.). Seed dormancy and germination was reported to be regulated by the balance between abscisic acid (ABA) and gibberellic acid (GA). In this study, transcriptomic sequencing was performed with Huayu 52 (HY52), a peanut cultivar with strong dormancy, and two EMS mutant lines from HY52 with a weak level of dormancy. Seeds from these three lines were imbibed for 0, 12, and 24 h before tissue harvesting and RNA seq analysis. GA content of M23 and M67 was significantly higher than HY52 at 12 h after imbibition, however, the ABA content and ABA/GA ratio were lower than HY52. A total of 31,374 differentially expressed genes (DEGs) including biosynthesis and signal transduction related genes of plant hormones such as ABA and GA were discovered. We identified 50 genes related to ABA, 8 genes related to GA, 49 genes related to ethylene, and 13 genes related to auxin. Expression profiles of ABA and GA related genes was consistent with the higher GA and lower ABA content in the mutants compared with HY52 after 12 h and 24 h imbibition. In addition, many DEGs involved in carbohydrate and lipid metabolism, amino acid metabolism, and glutathione metabolism pathway were also identified. There were 5 carbohydrate metabolism related genes (GPT) and 4 lipid metabolism related genes. In addition, differentially regulated circadian rhythm pathways were found to involve in the process of peanut seed dormancy release. These results suggested that the regulation of dormancy maintenance and release was more complicated than phytohormone balance.

      Function analysis of potato StCYP85A3 in promoting germination and root elongation
      LIU Jie, CAI Cheng-Cheng, LIU Shi-Feng, DENG Meng-Sheng, WANG Xue-Feng, WEN He, LI Luo-Pin, YAN Feng-Jun, WANG Xi-Yao
      Acta Agronomica Sinica. 2023, 49(9):  2462-2471.  doi:10.3724/SP.J.1006.2023.24236
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      Brassinolide (BR), a kind of phytosterol hormone, plays the significant role in regulating plant growth and development and stress response. The function of StCYP85A3 as the encoding gene of BR synthease in potato remains to be further explored. In this study, the gene was cloned from potato ‘Chuanyu 10’. Bioinformatics analysis showed that this protein had a typical aerobic binding domain for catalyzing BR synthesis and belonged to the CYP85As family. Tissue expression analysis indicated that the relative expression level of this gene was the highest in the bud eye of the tuber and the root, and the expression amount in the bud eye increased with the extension of storage time, and the expression increased rapidly when the dormancy of the tuber bud eye was released. At the same time, the exogenous BR could induce the relative expression level of StCYP85A3 in potato bud eye and root, which significantly promoted the germination of seed potato and the elongation of seedling root. The overexpression of StCYP85A3 in Arabidopsis wild-type and mutant cyp85a2 demonstrated that the seed germination and root elongation of the overexpression strains were earlier than wild type, and the defects of mutant seed germination and plant root growth retardation were remedied by supplementing strains. In addition, exogenous BR did not significantly promote seed germination and root elongation of over expressed strains, but significantly promoted seed germination and root elongation of wild type, mutant, and complementary strains. The above potato and Arabidopsis thaliana experiments showed that StCYP85A3 had the function of promoting germination and root elongation.

      Interaction of sugarcane VAMP associated protein ScPVA12 with SCMV P3N-PIPO
      YU Quan-Xin, YANG Zong-Tao, ZHANG Hai, CHENG Guang-Yuan, ZHOU Ying-Shuan, JIAO Wen-Di, ZENG Kang, LUO Ting-Xu, HUANG Guo-Qiang, ZHANG Mu-Qing, XU Jing-Sheng
      Acta Agronomica Sinica. 2023, 49(9):  2472-2484.  doi:10.3724/SP.J.1006.2023.24244
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      PVA12 (plant vesicle-associated membrane protein (VAMP)-associated proteins homolog 12) is a member of the VAP27 family of proteins that mediate endoplasmic reticulum (ER) vesicle transport and membrane fusion in cells. Sugarcane (Saccharum spp. hybrid) PVA12 responding to Sugarcane mosaic virus (SCMV) infection has not been reported. In the present study, the coding gene of PVA12 was cloned from sugarcane cultivar ROC22 and designated as ScPVA12. The open reading frame (ORF) of ScPVA12 was 735 bp in length, which encoded a protein with 244 amino acids. Bioinformatics analysis showed that ScPVA12 was an unstable hydrophilic liposoluble protein with a transmembrane at the C-terminal domain. The ratio of the random coil ranked the highest in the secondary structure. Phylogenetic tree analysis revealed that the ScPVA12 was differentiated in monocotyledons and dicotyledons. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays showed that ScPVA12 interacted with SCMV-P3N-PIPO. Subcellular localization experiments indicated that ScPVA12 was localized in the ER. Co-localization experiments demonstrated that ScPVA12 and SCMV-P3N-PIPO co-localized in the ER. Real-time quantitative PCR analysis showed that ScPVA12 gene was expressed in all sugarcane tissues, with the lowest expression level in the eighth internode and the highest expression level in the 7th leaves. The relative expression level of ScPVA12 gene was significantly affected under the stress of SCMV. ScPVA12 was down-regulated upon SCMV infection and then recovered to the regular level compared with the control.

      RT-PCR cloning and functional analysis of ScbHLH13 in sugarcane
      MO Guang-Ling, YU Chen-Jing, LIANG Yan-Lan, ZHOU Ding-Gang, LUO Jun, WANG Mo, QUE You-Xiong, HUANG Ning, LING Hui
      Acta Agronomica Sinica. 2023, 49(9):  2485-2497.  doi:10.3724/SP.J.1006.2023.24233
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      The transcription factors in bHLH family are important regulators in controlling plant growth and development, secondary metabolism, and anthocyanin biosynthesis. In this study, we successfully cloned ScbHLH13 using sorghum bHLH13 (XM_002440799.2) as the probe sequence by RT-PCR in sugarcane. Meanwhile, the physicochemical properties of the gene and its encoded protein were predicted. Phylogenetic relationship, subcellular localization, and the relative expression pattern of ScbHLH13 were investigated. ScbHLH13 contained 586 amino acids, and mainly consisted of random coils and α-helice. It was hydrophilic, unstable, and weakly basic. Moreover, ScbHLH13 had the typical nuclear localization signals and no transmembrane. The sequence contained two typical conserved domains, bHLH-MYC, and HLH, which belonged to the bHLH superfamily. Analysis of sugarcane transcriptomes under low nitrogen, and Sorghum mosaic virus, and Sporisorium scitamineum invasion revealed that the transcriptional levels of ScbHLH13 were altered in response to various abiotic and biotic stresses. When transiently expressed in tobacco, ScbHLH13-YFP was observed in nuclei and on cell membrane, whereas it specifically localized in nuclei of sugarcane protoplasts. QRT-PCR assays showed that relative expression level of ScbHLH13 in sugarcane protoplasts could not induce transcription of the relative genes in anthocyanin synthesis and the metabolism pathway, suggesting that ScbHLH13 may not involve in anthocyanin biosynthesis and metabolism. In conclusion, this study explores the basic features and functions of ScbHLH13, which will be helpful for the future research on characterizing the biological roles of bHLH members in regulating sugarcane growth and defense.

      TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY
      Establishment of near-infrared reflectance spectroscopy model for predicting sucrose content of single seed in peanut
      HU Mei-Ling, ZHI Chen-Yang, XUE Xiao-Meng, WU Jie, WANG Jin, YAN Li-Ying, WANG Xin, CHEN Yu-Ning, KANG Yan-Ping, WANG Zhi-Hui, HUAI Dong-Xin, JIANG Hui-Fang, LEI Yong, LIAO Bo-Shou
      Acta Agronomica Sinica. 2023, 49(9):  2498-2504.  doi:10.3724/SP.J.1006.2023.24241
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      Sucrose content is an important factor to determine the taste and flavor of peanuts. Breeding sweet peanut varieties with high sucrose content has been a main objective for the edible peanut genetic improvement. Developing a near-infrared reflectance spectroscopy model for predicting the sucrose content in single peanut kernel will accelerate the process of sweet peanut breeding. In this study, 128 representative materials with abundant genetic diversity were selected, the near-infrared spectral data were collected, and the sucrose content of each seed was determined by high performance liquid chromatography (HPLC) with RID detector. Based on spectral data and chemical value of sucrose content, a calibration model for predicting the sucrose content in single seed with a coefficient of determination (R2) of 0.913 and a root mean square error of cross validation (RMSECV) of 0.750, was built up by partial least squares (PLS) method. 50 peanut seeds were analyzed by both NIR and HPLC for external validation and the correlation coefficient between the prediction value and the chemical value reached 0.92, indicating that this model could predict sucrose content with adequate accuracy and reliability. This calibration model for sucrose content in single seed could be applied to the selection of high sucrose lines in the early generations of hybrid progenies and in the purity monitoring of seed and raw material, which will be a supportive technique for the edible peanut variety breeding and industrial application.

      Effects of increased nitrogen on Bt protein expression and nitrogen metabolism in the leaf subtending to cotton boll
      LI Yi-Yang, LI Yuan, ZHAO Zi-Xu, ZHANG Ding-Shun, DU Jia-Ning, WU Shu-Juan, SUN Si-Qi, CHEN Yuan, ZHANG Xiang, CHEN De-Hua, LIU Zhen-Yu
      Acta Agronomica Sinica. 2023, 49(9):  2505-2516.  doi:10.3724/SP.J.1006.2023.24217
      Abstract ( 266 )   HTML ( 8 )   PDF (543KB) ( 453 )   Save
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      The objective of this study is to explore the effect of soil nitrogen increase on the expression of insecticidal proteins in the leaf subtending to cotton boll and the related physiological mechanism of nitrogen metabolism and to provide the theoretical and technical support for agronomic regulation of Bt cotton boll stage insect resistance. A split-plot experiment was conducted. The conventional cultivar Sikang 1 and hybrid cultivar Sikang 3 were used as the experimental materials. Enhanced nitrogen fertilizer rates of increased 25% to 100% nitrogen [300 (CK), 375, 450, 525, and 600 kg hm-2 as pure nitrogen] were designed to study the effect on Bt protein content and nitrogen metabolic physiological in the leaf subtending to cotton boll. The results showed that the content of Bt protein in the leaf subtending to cotton boll of two types of cultivars showed a constant increase with the increase of nitrogen application rate. Compared with the control (300 kg hm-2), Bt protein content in the leaf subtending to cotton boll increased by 6.1%-96.9% with the increase of 25%-100% nitrogen application. The physiological mechanism of nitrogen metabolism showed that the trend of soluble protein (SP), free amino acid (AA), and key enzymes of protein synthesis [Glutamic Oxalacetic Transaminase (GOT) and Glutamine Synthetase (GS)] in the leaf subtending to cotton boll were consistent with those of Bt protein. The activities of key enzymes of protein decomposition (protease and peptidase) decreased with the increase of nitrogen application. The leaf area index (LAI) increased with the increase of nitrogen application, while the yield increased first and then decreased with the increase of soil nitrogen. The optimum LAI and the maximum yield were both 1.25 times (375 kg hm-2) that of conventional nitrogen application. In conclusion, on the basis of conventional nitrogen application, an appropriate amount of nitrogen fertilizer was beneficial to the synthesis of Bt protein, the formation of the optimal LAI, and the increase of yield in the counter-position leaves of cotton boll, which was conductive to the synergistic expression of high yield and insect resistance of Bt cotton.

      Effects of coconut bran application rate on soil physicochemical properties and sweet-potato yield
      YANG Yi, HE Zhi-Qiang, LIN Jia-Hui, LI Yang, CHEN Fei, LYU Chang-Wen, TANG Dao-Bin, ZHOU Quan-Lu, WANG Ji-Chun
      Acta Agronomica Sinica. 2023, 49(9):  2517-2527.  doi:10.3724/SP.J.1006.2023.24212
      Abstract ( 393 )   HTML ( 22 )   PDF (370KB) ( 396 )   Save
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      The objective of this study is to study the effects of different application rate of coconut bran on soil physical and chemical properties and sweet-potato yield, and to explore the correlation between the application rate of coconut bran and soil physical and chemical properties, which can provide the theoretical basis and practical production guidance for the improvement and maintenance of farmland soil fertility and the high-yield and high-quality cultivation of fresh-eating sweet potato. In 2020 and 2021, a randomized block design was used to study the effects of coconut bran (dry weight) application rate of 0, 20,250, 40,500, and 60,750 kg hm-2 on soil and sweet potato. The results showed that, with the increase of coconut bran application, soil bulk density decreased gradually, but soil porosity, soil mass water content, soil organic matter content, soil available nitrogen, phosphorus and potassium contents increased gradually. Among them, soil available potassium content increased the most, followed by alkali-hydrolyzed nitrogen and the least available phosphorus. The number of bacteria, fungi, and actinomycetes in rhizosphere soil all increased with the increase of coconut bran application. Moreover, the application of coconut bran can increase the number of storage root per plant, increase the number of 200-400 g, 100-200 g, and 50-100 g of storage root, and improve the commodity rate of sweet-potato. The yield of sweet-potato storage root increased first and then decreased with the increase of coconut bran application, and the highest was 40,500 kg hm-2 of coconut bran application, while the starch content of sweet-potato storage roots decreased with the increase of coconut bran application. In conclusion, the application of coconut bran can effectively improve the soil structure and fertility, promote the formation of sweet-potato storage roots and large potatoes, increase the yield of storage root and the commodity rate of sweet potato.

      Effect of ethylene-chlormequat-potassium on root morphological structure and grain yield in sorghum
      FANG Meng-Ying, REN Liang, LU Lin, DONG Xue-Rui, WU Zhi-Hai, YAN Peng, DONG Zhi-Qiang
      Acta Agronomica Sinica. 2023, 49(9):  2528-2538.  doi:10.3724/SP.J.1006.2023.24243
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      Shaping good root structure is the key problem to play the high yield potential of sorghum and increase the yield of grain sorghum. To investigate the effect of ethylene-chlormequat-potassium (ECK) on sorghum root morphology and yield, field trials were conducted in 2020 and 2021 using the medium to tall sorghum variety Liaoza 19 (LZ19) and the dwarf variety Liaoza 37 (LZ37). A randomized zonal experimental design was used to set up the ECK treatment with a foliar spray of 0.75 L hm-2 at the five-leaf stage of sorghum and a control spray with an equal amount of clean water (CK). The results showed that compared with CK, sorghum root dry weight showed a gradual decrease from the flowering stage. ECK significantly increased root dry weight of LZ19 and LZ37 at filling and maturity stages by 11.4% and 19.7%, 10.6% and 9.9%, respectively. ECK increased root length and root surface area of fine and medium roots (root system diameter < 4 mm) of LZ19 and root length, root surface area, and root volume of LZ37 at different diameters at grain filling stage. At maturity stage, ECK increased root length, root surface area, root volume, and root mean diameter of LZ19 and that of LZ37, and the increase of system morphological indexes was the largest in the two varieties. ECK treatment increased the yield of LZ19 by 3.3% and 13.4%, and LZ37 by 11.4% and 9.8% in 2020 and 2021, respectively, compared with CK. In conclusion, ECK treatment promoted sorghum root development and significantly increased sorghum yields, which can be used as an important cultivation measure for sorghum root promotion and yield increase in summer sown areas.

      Effect of water management on yield and its components of winter wheat in different precipitation years
      ZHANG Li-Hua, ZHANG Jing-Ting, DONG Zhi-Qiang, HOU Wan-Bin, ZHAI Li-Chao, YAO Yan-Rong, LYU Li-Hua, ZHAO Yi-An, JIA Xiu-Ling
      Acta Agronomica Sinica. 2023, 49(9):  2539-2551.  doi:10.3724/SP.J.1006.2023.21062
      Abstract ( 372 )   HTML ( 19 )   PDF (329KB) ( 762 )   Save
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      To clarify the irrigation strategy for improving yield and its components, the relationship among precipitation, irrigation, yield, and its components was systemically studied under different rainfall conditions. In this experiment, Jimai 585 used as the experimental materials, and five irrigation treatments including W0 (rain fed), W1 (jointing), W2 (jointing, flowering), W3 (jointing, flowering, filling), and W4 (overwintering, jointing, flowering, filling) in the wheat season were set up from 2010 to 2017. The results were as follows: (1) wheat grain yield varied from 6400 to 6800 kg hm-2 under different irrigation treatment in most rainfall years, and spike numbers was positively correlated with grain yield (r = 0.860*), the grain yield, grain numbers per spike, and thousand grain weight increased with the increase of irrigation amount, however, the yield increase rate decreased with each additional water (from 13.8% to 1.7%). (2) There was no obvious correlation between the total precipitation and stage precipitation and grain yield, but the impact of total precipitation on the thousand grain weight was higher than the other factors. Under the conditions of W0 and W1, the effect of total precipitation on the numbers per spike was greater than spike numbers, and vice versa after increasing irrigation. At the same time, the grain numbers per spike was significantly positively correlated with the precipitation before jointing, and the correlation between the grain numbers per spike and the precipitation from February 1 to jointing under W0 was greater than that from sowing to jointing, but it decreased with the increase of irrigation amount. With the exception of the correlation between W4 irrigation and grain numbers per spike and thousand grain weight was higher than that of precipitation, the correlation between two factors and precipitation was higher under W1-W3 conditions, which indicating that irrigation alleviate the adverse effect of insufficient precipitation on grain number per spike. (3) The number of grains per spike and thousand grain weight were the highest in a rainfall year characterized with more precipitation at early growth stages and less precipitation at later growth stages (precipitation before and after jointing 88.2 mm + 29 mm), and the highest yield was observed under 3 irrigation times, and there was no significant difference between 3 irrigation times and 2 irrigation times. In years characterized balanced rainfall (precipitation before and after jointing < 60 mm, 30-80 mm), lower and relatively stable yield and its component appeared, and the grain numbers per spike and thousand grain weight increased slightly after irrigation, but the difference in grain yield between 2 and 3 irrigation times was significant (yield increase rate: 10.5% and 22.9%) (the rainfall after jointing < 36 mm). In year characterized with less precipitation at early growth stages and more precipitation at later growth stages in (precipitation before and after jointing < 25 mm, 40 to 90 mm), the maximum grain number per spike increased by 1.5 to 7.1 grains when irrigation increased, and the grain yield of irrigation 3 times was 13.4% higher than that of irrigation 2 times when inadequate effective rainfall happened in April. In conclusion, when the precipitation before jointing was less than 60 mm (especially less than 25 mm), irrigation at jointing had an obvious effect on the increase of grain number per spike, 3 times irrigation promoted grain yield when the precipitation after jointing is less than 36 mm in a relatively balanced rainfall year type and no sufficient effective rainfall in April in years characterized with less precipitation at early stages and more precipitation at late stages, and irrigation 2 times was suggested to achieve the maximum grain yield in other rainfall years.

      Relationship between the starch properties and its surface-bound proteins in grains with hardness in Avena nuda L.
      NAN Jin-Sheng, AN Jiang-Hong, CHAI Ming-Na, JIANG Yu-Lian, ZHU Zhi-Qiang, YANG Yan, HAN Bing
      Acta Agronomica Sinica. 2023, 49(9):  2552-2561.  doi:10.3724/SP.J.1006.2023.21064
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      Kernel hardness can reflect the texture of wheat kernels and it is closely related to its milling performance and edible quality. The objective of this study is to explore the relationship between naked oat (Avena nuda L.) grain starch properties and its surface-bound proteins with grain hardness. In this experiment, to determine the content of total starch, amylopectin and amylose, observe the characteristics and particle size of starch granules, and identify the surface-bound proteins of starch granules by mass spectrometry, five soft and five hard naked oat germplasms were used as the materials. The results showed that the shapes of starch granules in mature grains were round, oval and irregular, and the number, volume and surface area of starch granules showed a unimodal distribution. The size of starch granules could be divided into small (particle size < 6 μm), medium (6-40 μm), and large starch granules (particle size > 40 μm). Naked oat grains were mainly composed of small and medium starch granules. The grain size distribution of starch in naked oat grains with different hardness was different. The number and volume percentage of small starch granules in soft naked oat were higher than those in hard, and the number and volume percentage of medium and large starch grains were lower than those in hard. The percentage of surface area of soft naked oat small and medium starch granules was higher than that of hard, and the percentage of surface area of large starch granules was lower than that of hard. There was a significant positive correlation between the grain hardness of naked oat and the amylopectin content. The results showed that the protein content of the starch granule surface with a size of 14 kD was higher in soft oats than in hard ones. A total of 41 proteins were identified by HPLC/MS for starch granule surface-binding proteins at 14 kD, including Vromindoline protein and oat alpha amylase trypsin inhibitor. The above results showed that the starch content, particle size distribution and starch surface-bound protein in naked oat grains were all related to grain hardness. The higher the amylopectin content, the greater the number, surface area and volume distribution of small starch granules. The higher the protein content such as Vromindoline, and the softer the naked oat kernel. This study laid the foundation for revealing the formation mechanism of grain hardness in Avena nuda L.

      Effects of soil microbes on rice allelopathy and its mechanism of wild rice (Oryza longistaminata) and its descendants
      XU Gao-Feng, SHEN Shi-Cai, ZHANG Fu-Dou, YANG Shao-Song, JIN Gui-Mei, ZHENG Feng-Ping, WEN Li-Na, ZHANG Yun, WU Ran-Di
      Acta Agronomica Sinica. 2023, 49(9):  2562-2571.  doi:10.3724/SP.J.1006.2023.22047
      Abstract ( 298 )   HTML ( 11 )   PDF (1467KB) ( 462 )   Save
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      Soil microbes may affect weed inhibitory effects of allelopathic crops that it is great significant to understand their causes and mechanisms for green ecological management of weeds in paddy fields. Oryza longistaminata (OL), a wild rice with strong allelopathic potential, is excellent cultivars for breeding allelopathic rice. However, the effect of soil microbes on allelopathy of this wild rice and its descendants is still unclear. In this study, the allelopathic effects of two allelopathic rice genotypes (OL and its descendant-RL169) and non-allelopathic Asian cultivated rice cultivar (RD23) affected by soil microbes on barnyardgrass were studied, and characteristics of rhizosphere soil microbes, and absorption and utilization of nutrients of barnyardgrass were analyzed. The results showed that: 1) Soil microbes significantly increased weed suppression of wild rice (OL) and its descendants (RL169) (P < 0.05) and had no significant effect on RD23. Multivariate analysis of variance showed that plant height, root length and biomass of barnyardgrass were significantly increased with interaction of soil microbes, root exudates and different allelopathic rice genotypes (P < 0.05). 2) Wild rice (OL) and its descendants (RL169) changed soil microbe community structure, reduced diversity and richness of bacteria in barnyardgrass rhizosphere soils, which the number of bacteria was significantly lower than that of RD23 at the family, genus and species levels (P < 0.05). 3) Soil microbes significantly reduced absorption and utilization of N, P and K of barnyardgrass which co-cultured with wild rice and its descendants (RL169). In the presence of soil microbes, the absorption of N and P of barnyardgrass which co-cultured with wild rice(OL) and the absorption of N of barnyardgrass which co-cultured with rice genotypes (RL169) were significantly reduced. Multivariate analysis of variance revealed that soil microbes and allelopathic rice genotypes significantly affected N (P < 0.01) or K (P < 0.05) absorption of barnyardgrass and had no significant effect on P absorption; but nutrient utilization of barnyardgrass was only obviously affected by allelopathic rice genotypes (P < 0.05). In conclusion, the allelopathic suppression of wild rice (OL) and its descendants (RL169) was significantly improved through soil microbe community structure changing of barnyardgrass rhizosphere soils and nutrient absorption and utilization (N, P, and K) reducing of barnyardgrass. Our study could increase further understanding of effect of soil microbes on rice allelopathy and provide a theoretical basis for the development and utilization of wild rice germplasm resources.

      RESEARCH NOTES
      Effects of multiple cropping green manure on grain quality and yield of wheat with different irrigation levels
      ZHANG Diao-Liang, YANG Zhao, HU Fa-Long, YIN Wen, CHAI Qiang, FAN Zhi-Long
      Acta Agronomica Sinica. 2023, 49(9):  2572-2581.  doi:10.3724/SP.J.1006.2023.21067
      Abstract ( 346 )   HTML ( 10 )   PDF (562KB) ( 362 )   Save
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      It is proved that the effect of multiple cropping green manure after wheat harvest on wheat grain quality and yield with different irrigation levels has an important theoretical and practical supporting role in building a stable and high-yield and high-quality production mode of wheat based on green manure in arid irrigation areas. In this study, a split plot design was adopted. Two cropping patterns were set up in the main area: wheat with multiple cropping green manure (W-G) and wheat with autumn fallow (W). Three irrigation levels were set in the sub district during wheat growth period: low irrigation quota (I1: 190 mm), medium irrigation quota (I2: 240 mm), and high irrigation quota (I3: 290 mm). The quality indexes, such as protein content, starch content, unit weight, and other quality, and yield performance of wheat were measured and analyzed in 2020 and 2021. The results showed that with the same irrigation level, compared with W, the grain protein content of W-G wheat increased by 5.8%-26.5%, the wet gluten content increased by 9.3%-26.4%, and the unit weight increased by 0.4%-2.1%. Between different irrigation levels in the same cropping pattern, the protein content of wheat grains increased with the decrease of irrigation level, while the starch content and wet gluten content of wheat grains decreased with the decrease of irrigation level. The grain unit weight of I2 wheat was significantly greater than I1 and I3. Among the treatments of each combination, the protein content of wheat grain at the medium irrigation level of multiple cropping green manure (W-GI2) and the low irrigation level of multiple green manure (W-GI1) had no significant difference, which were higher than other treatments, increased by 15.1%-35.0% compared with the high irrigation level of wheat with autumn fallow (WI3), and the starch content and wet gluten content of wheat grain at W-GI2 had no significant difference compared with the high irrigation level of wheat with multiple cropping green manure (W-GI3), which were 3.2%-3.4% and 7.5%-12.9% higher than WI3, respectively. The unit weight of wheat grains was maximized. Compared with leisure after wheat harvest treatment, wheat grain yield of W-G increased by 7.0%-13.2%. The grain yield of wheat decreased with the decrease of irrigation level, but the grain yield of W-GI2 wheat was not significantly different from that of W-GI3, and increased by 6.0% on average compared with that of WI3. Compared with W, W-G increased the content of soil organic matter before wheat sowing by 5.6%-31.5%, but the content of soil organic matter of W-GI2 was not significantly different from that of W-GI3 in 20-40 cm soil layer, and increased by 7.8% on average compared with that of WI3. In conclusion, multiple cropping green manure after wheat harvest can significantly increase the content of soil organic matter in wheat field compared with leisure after wheat harvest treatment, enable wheat to obtain higher grain yield with medium irrigation conditions, and improve grain protein, starch, wet gluten content, and other qualities, which can be used as a recommended agronomic measure for sustainable and stable yield and high yield and quality optimization of wheat under limited irrigation in arid irrigation areas.

      Genetic diversity and population structure analysis of 378 introduced sweetpotato germplasm collections
      SU Yi-Jun, ZHAO Lu-Kuan, TANG Fen, DAI Xi-Bin, SUN Ya-Wei, ZHOU Zhi-Lin, LIU Ya-Ju, CAO Qing-He
      Acta Agronomica Sinica. 2023, 49(9):  2582-2593.  doi:10.3724/SP.J.1006.2023.24195
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      Sweetpotato is originated from South and Central America, and the introduction of foreign species can not only increase the total quantity amount of germplasm resources of China, but also dramatically change the breeding bottleneck of sweetpotato with narrow genetic base. In this study, 378 sweetpotato germplasm collections from the national sweetpotato in vitro Genebank in Xuzhou were genotyped using 30 pairs of simple sequence repeat (SSR) primers by high-throughput nucleic acid fragment analyzer. Phylogenetic affinities among the introduced sweetpotato germplasm were analyzed to provide a theoretical basis for sweetpotato germplasm innovation and utilization. The bands were read using PROSize 3.0 software to establish the 01 database. The 01 database were imported into Darwin software to calculate Jaccard genetic distance matrix, clustered analysis of genetic distance matrix using MEGA 10, and then annotated with iTOL. Population structure of sweetpotato germplasm was constructed using Structure 2.3.4. The results showed that a total of 120 polymorphic loci were amplified in 378 sweetpotato germplasm, with an average of four polymorphic loci per primer pair. The genetic distance matrix revealed that the average genetic distance was 0.4027, the maximum genetic distance was 0.6207 and the minimum genetic distance was 0.0448. Clustering analysis of 378 sweetpotato germplasm were divided into three groups in 0.2960 genetic distance. Group I, II, and III contained 29, 104, and 245 collections, respectively. Population structure analysis indicated two clades were identified base on the largest value at ΔK=2. Clade II (95.4%) was dominated by newly introduced germplasm after 2015, indicating that the newly introduced germplasm enriched the genetic diversity of sweetpotato collections and broadening the population structure in the original resource base. Therefore, the genetic diversity and population structure analysis of 378 introduced sweetpotato germplasm can provide a reference for sweetpotato germplasm innovation, genetic breeding, and elite alleles mining and utilization.

      Interaction identification between protein kinase MeSnRK2.12 and transcription factor MebHLH1 and its relative expression level in cassava
      YU Xue-Ting, LI Ke, LI Meng-Tao, BAO Ru-Xue, CHEN Xin, WANG Wen-Quan
      Acta Agronomica Sinica. 2023, 49(9):  2594-2600.  doi:10.3724/SP.J.1006.2023.24225
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      Protein kinase SnRK2s (Sucrose Non-fermenting Related Protein Kinase 2) is a key component of plant stress resistance mechanism. Cassava is an important food and industrial crop in the world, with the characteristics of high starch accumulation and stresses resistance. So far, the mechanism of MeSnRK2 family member involved in the regulation of starch synthesis under stress is unclear. MeSnRK2.12 was chosen in this study, which is a member of the weakly ABA-induced SnRK2s. Firstly, some stress-response cis-elements distributed in its promoter region, such as drought stress cis-element MBS and ABA response element ABRE. Secondly, its amino acid sequence was highly homologous to AtSnRK2.8 and OsSAPK1/2. MeSnRK2.12 could respond quickly to ABA and PEG treatments within two hours, and its transcription activity was inhibited in roots, but was induced in stem, and the highest values were 15 times and 8 times of the control, respectively. There was also an significant up-regulation trend in leaves, but the degree was lower than that in stems. Subcellular localization showed that MeSnRK2.12 located in the cytoplasm and cell nucleus. The interaction between MeSnRK2.12 and MebHLH1 was verified by yeast two-hybrid and bimolecular fluorescence complementation (BiFC) experiments. Previous studies revealed that MebHLH1 could up-regulate the transcription activity of MeSus1, and the activity of sucrose synthase was directly related to plant sink strength. Therefore, it is speculated that MeSnRK2.12 not only plays an important role in response to stress, but also may be involved in the regulation of starch synthesis mediated by ABA signal, which helps cassava achieve relatively high starch yield under stress conditions.

Co-sponsored:
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

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  • 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:
https://www.sciencedirect.com/journal/the-crop-journal
Submission: https://www.editorialmanager.com/cj/
E-mail: cropjournal@caas.cn
Tel: 8610-82108548

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