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    26 July 2025, Volume 41 Issue 7
    Functional Mechanism of Plant CBL in Regulating the Responses to Abiotic and Biotic Stresses
    WANG Cong-huan, WU Guo-qiang, WEI Ming
    2025, 41(7):  1-16.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1265
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    Calcineurin B like protein (CBL) is a kind of specific Ca2+ sensor in plants, which plays a crucial role in plant growth and development, and the response to environmental stresses. CBL is composed of four typical elongation factor hands (EF-hands) domains for Ca2+ binding. Each EF-hand is α-helix-loop-α-helix structures composed of 12 relatively conserved amino acids. The cis-acting regulatory elements (such as W-box, MBS and G-BOX) in the promoter regionof CBL can bind with upstream transcription factors (TFs) to regulate transcription by activating or inhibiting the expression of downstream genes. CBL is involved in the regulation of signaling pathways such as abscisic acid (ABA), hormones, respiratory burst oxidases homolog (RBOH), and reactive oxygen species (ROS), to reduce water evaporation, thereby adapting various abiotic stresses. There are evidences that CBL can rapidly perceive changes in the transient intracellular Ca2+ signal when plants were subjected to abiotic and biotic stresses such as salt, drought, extreme temperature, nutrient stress and pathogen. CBLs not only interact with CBL-interacting protein kinase CIPK to phosphorylate downstream target proteins for calcium signal transduction, but also interact with other proteins (such as high-affinity K+ transporter 5, protein S-acyl transferase10 and type 2C protein phosphatases), thereby positively or negatively regulating stress tolerance in plant. Additionally, CBL mediates the growth and development of plant organs and tissues and promotes fruit ripening by regulating sugar signals. CBL also interacts with gigantea (GI) to affect the flowering time of plants. In this review, recent research findings on the discovery, structure, classification, regulatory mechanisms and roles of plant CBL in regulating the response to adversity stresses are summarized, and its future research directions are prospected, aiming to provide the gene resources and the theoretical basis for improving tolerance of crops and biological breeding to stress.

    Research Progress in the Involvement of Intracellular Transport Regulated by Endogenous Elicitors in Plant Growth and Development and Response to Adverse Stress
    LI Si-bo, QIAN Hong-ping, XU Chang-wen, WANG Xiao, LIN Jin-xing, CUI Ya-ning
    2025, 41(7):  17-27.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0124
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    Endogenous elicitors are endogenous signal molecules released by plants in response to pathogen infection or mechanical damage. When plants perceive the threat of infection by pathogens or suffer mechanical damage, they release endogenous elicitors. Once produced, these elicitors can be recognized by pattern recognition receptors (PRRs) located on the plasma membrane of the cell, triggering and amplifying an immune response (such as PTI). Endogenous elicitors can regulate the endocytosis of PRRs protein by means of protein modifications such as phosphorylation and ubiquitination, and regulate the intracellular cycle or degradation of PRRs, thus affecting the transmission of immune signals. These processes are not only involved in plant defense against pathogens, but also related to plant growth and development and regulation of resistance response.In this paper, the latest research progress of endogenous elicitors and their receptors is reviewed comprehensively and systematically. The regulation of endogenous elicitors on PRRs intracellular transport is discussed in detail, and the key role of endogenous elicitors in plant growth and development and resistance is also analyzed in depth, aiming to provide a new perspective for understanding the molecular and cytological mechanism of plant resistance response.

    Research Progress of Plant Phosphate Transporters in the Response to Stress
    ZHANG Xue-qiong, PAN Su-jun, LI Wei, DAI Liang-ying
    2025, 41(7):  28-36.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1153
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    Phosphorus (P) is an essential element for plant growth, development and many physiological functions. Phosphate transporters are responsible for the uptake and transport of phosphorus from the soil to plant organelles and play a significant role in plant growth and development as well as in responses to abiotic and biotic stresses. This article reviews the classification, structural characteristics, subcellular localization of plant phosphate transporters and their important roles in plant responses to abiotic and biotic stresses. Plant phosphate transporters can be divided into three major classes, with the first class further divided into five subfamilies. These proteins perform functions in different parts of the plant. Plant phosphate transporters not only participate in the absorption and transport of phosphate but also play a crucial role in stress responses. Under abiotic stress, plant phosphate transporters are induced to express, which can enhance the plant's tolerance to abiotic stress by increasing its antioxidant capacity. In addition, plant phosphate transporters interact with other proteins to regulate plant responses to abiotic stress. Under biotic stress, plant phosphate transporters regulate the transcriptional levels of defense-related genes in plants, thereby positively or negatively regulating plant resistance to pathogens. Existing studies have demonstrated the important role of plant phosphate transporters in stress responses, but their specific molecular mechanisms and regulatory networks still need further exploration. Future research directions will focus on screening and identifying upstream regulatory factors and downstream interacting target genes of plant phosphate transporters, and deeply elucidating the molecular mechanisms by which plant phosphate transporters regulate biotic and abiotic stress responses, with the aim of providing references for the efficient utilization of phosphorus in crops and high-yield and high-resistance molecular breeding.

    Research Progress in Melatonin in Plant Low-temperature Stress
    LIN Jia-yi, CHEN Qiang, ZHANG Lei, LIU Hong-xin, ZHENG Xiao-ming, PANG Hong-bo
    2025, 41(7):  37-48.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1061
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    Melatonin (MT), an indoleamine bioactive molecule prevalent in plants, has recently emerged as a focal point of research concerning its molecular mechanisms in plant responses to abiotic stress. This paper provides a comprehensive review of the latest advancements in understanding melatonin’s role in plant responses to low-temperature stress, with a particular emphasis on its molecular mechanisms for enhancing cold tolerance through multidimensional regulation. Physiologically, melatonin mitigates low-temperature-induced membrane lipid peroxidation and photoinhibition by stabilizing cell membrane lipid bilayers, safeguarding photosystem Ⅱ reaction centers, and scavenging excess reactive oxygen species (ROS). At the molecular level, melatonin enhances plant low-temperature tolerance through a complex signaling network: it activates the ICE1-CBF-COR transcriptional cascade to upregulate cold-responsive genes; transmits stress signals via receptor-mediated pathways; regulates the dynamics of secondary messengers such as Ca2+, NO, and H2O2; interacts with plant hormones (e.g., abscisic acid (ABA), jasmonic acid (JA), indole-3-acetic acid (IAA)) to form signaling networks; and triggers mitogen-activated protein kinase (MAPK) and calcium-dependent protein kinase (CDPK) cascades to amplify low-temperature stress signals. Melatonin holds significant potential for application in enhancing plant tolerance to low temperatures. The exogenous application of melatonin has been shown to enhance crop tolerance to low temperatures, while augmenting endogenous melatonin synthesis through gene editing represents a crucial strategy for improving crop resistance to low-temperature stress. Future research should adopt multidisciplinary approaches to investigate the role and potential applications of melatonin in plant responses to low-temperature stress, thereby facilitating the development of crop varieties with enhanced low-temperature tolerance.

    Regulation of Plant Tannin Synthesis and Mechanisms of Its Responses to Environment
    GAO Jing, CHENG Yi-cun, GAO Ming, ZHAO Yun-xiao, WANG Yang-dong
    2025, 41(7):  49-59.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0022
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    Tannins are an important class of secondary metabolites in plants, widely distributed in various tissues and organs of plants, and play a crucial role in plant growth and development. As compounds with strong biological activity, tannins not only help plants effectively cope with various environmental challenges but also play a core role in the plant immune defense system. The biosynthesis of tannins is complex and is typically divided into two main stages: Precursor synthesis and tannin polymerization. Precursor synthesis begins with aromatic amino acids, which are converted into the basic units of tannins through a series of enzymatic reactions. Tannin polymerization is involved in the cross-linking and polymerization of these units to form complex macromolecules with biological activity. Although extensive research has been conducted on the synthesis pathways, regulatory mechanisms, and environmental responses of tannins, there is still a lack of systematic summaries and comprehensive overviews. This paper provides a detailed introduction to the structural characteristics of plant tannins, analyzes their multiple functions in plant growth and development, and discusses the biosynthetic pathways of tannins. Additionally, this paper summarizes the regulatory effects of environmental factors on tannin synthesis, further revealing the potential of tannins in helping plants cope with stress, particularly in enhancing plant resistance to stress and ecological adaptability. Finally, this paper analyzes and provides an outlook on unresolved issues in current tannin research and future research directions, aiming to provide theoretical support for the rational development and application of plant tannins, and promoting their application in improving plant stress resistance, enhancing ecological adaptability, and optimizing plant ecological functions.

    Research Progress in Adaptor Protein Complex TPC in Plants
    WANG Yu-tong, ZHANG Ying-hui, XU Mei, YAN Xu, ZHAO Fei-yi, TIAN Dan
    2025, 41(7):  60-68.  doi:10.13560/j.cnki.biotech.bull.1985.2024-0947
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    Clathrin-mediated endocytosis (CME) is a predominant endocytic pathway for plasma membrane (PM) protein internalization in eukaryotic cells. In plants, CME is essential for growth and developmental processes, nutrient uptake, transduction between extra- and intra-cellular signals, polar auxin transport, and stress responses. Although clathrin is the core protein of the CME process, it lacks a transmembrane domain and relies on adaptor proteins to be recruited to the PM. It is known that the PM recruitment of clathrin mainly depends on the adaptor protein complex AP-2 in animal and plant cells. However, an increasing number of evidences on plant adaptor protein complexes show that there is also an evolutionary ancient adaptor protein complex TPC (TPLATE complex) in plants, which plays a critical role in plant growth and development as well as CME processes. In this review, we briefly introduce the composition, structure, and evolutionary history of TPC in plants. Then we analyze the domains of TPC subunits. Furthermore, we summarize the biological functions of TPC. Finally, we prospect the research on TPC in plant endocytosis and growth and development, aiming to provide new ideas and insights for understanding and clarifying the molecular mechanism of plant TPC in endocytic transport.

    Research Progress in Response of Rice miRNAs to Biotic Stress
    HOU Ying-xiang, FEI Si-tian, LI Ni, LI Lan, SONG Song-quan, WANG Wei-ping, ZHANG Chao
    2025, 41(7):  69-80.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0061
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    During the rice production process, rice (Oryza sativa) often faces various biotic stresses such as fungi, bacteria, viruses, and pests, which seriously threaten rice growth and yield. Improving the inherent resistance of rice is the most economical and effective way to resist pests and pathogens. MiRNAs are a class of endogenous non-coding small RNAs approximately 20-24 nucleotides in length,which negatively regulate gene expression by degrading target mRNAs or inhibiting translation. In recent years, significant progress has been achieved in the functional research of rice miRNAs in response to biotic stresses. Rice miRNAs participate in the pattern-triggered immunity (PTI) and effector-triggered immunity (ETI) signaling pathways through a variety of transcription factors, transporters, endogenous signaling molecules, signal receptors, oxidases, hydrolases, and kinases, etc., directly or indirectly regulating the tolerance of rice to biotic stresses. Several miRNAs, such as miR156, miR168, miR396, miR162a, miR1873, miR1871, and miR1432, play critical roles in balancing rice yield and resistance, offering important insights for breeding high-yielding and disease-resistant varieties. However, most current studies on rice miRNAs in responses to biotic stress focus on identifying and characterizing downstream target genes. Future efforts should intensify research on upstream regulatory elements and comprehensively dissect the signal transduction pathways mediated by miRNAs, specifically clarifying the functions of individual components and their interactive networks. This will lay a foundation for improving the efficiency of miRNA expression manipulation in rice varietal improvement. This paper mainly reviews the functions of rice miRNAs in response to biotic stresses such as rice blast, bacterial blight, stripe disease, viral diseases, and brown planthoppers. It also proposes scientific questions that need attention and further research in the future, aiming to provide strategies for rice molecular breeding.

    Association Analysis and Fingerprint Map Construction of Fruit Color Traits in Pepper via SSR Markers
    DUAN Min-jie, LI Yi-fei, WANG Chun-ping, HUANG Ren-zhong, HUANG Qi-zhong, ZHANG Shi-cai
    2025, 41(7):  81-94.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0031
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    Objective Genetic diversity of pepper germplasm materials was analyzed and molecular markers related to fruit color traits were further explored. Method The genetic relationship and diversity of 57 pepper germplasm materials was evaluated by 7 fruit color traits and 20 SSR markers. The generalized linear model (GLM) and mixed linear model (MLM) of Tassel 5.0 software were carried out to analyze the correlation between traits and SSR markers, which then were used for constructing the germplasm fingerprint maps. Result The range of genetic diversity index of the 7 fruit color traits was from 1.58 (peel-L) to 2.08 (pulp-a) and the range of coefficient of variation was 9.07%-51.84%. There was a certain correlation between E value and Lab values of peel and pulp. A total of 107 polymorphic loci were detected by 20 SSR markers, with an average of 5.1 loci per marker. Primer polymorphism information content (PIC) ranged from 0.221 to 0.864, with an average of 0.624. The average values of Shannon’s information index (I) and Nei’s genetic diversity index (H) were 1.3 and 0.646, respectively. The range of genetic similarity coefficient of the 57 pepper germplasms was from 0.069 to 0.915 1, with an average of 0.294. The 57 germplasms were divided into 2 subclusters by systematic clustering, with 28 and 29 germplasms contained in subcluster I and subcluster II, respectively. The population structure analysis showed that the germplasm was divided into 2 subgroups, with 33 and 24 germplasms in subgroup I and subgroup II, respectively, which had high overlap with the results of cluster analysis. A total of 16 markers related to pepper fruit color traits were detected using both GLM and MLM models, and the phenotypic explanation rate was from 6.6% to 13.8%. Marker SSR4 was extremely significantly associated with E value (P < 0.01). SSR6, SSR9 and SSR17 were detected to be significantly associated with the same trait in both models. The fingerprint maps of 57 pepper germplasms were constructed using 9 pairs of core primers. Conclusion This study comprehensively analyzes the genetic diversity of fruit E value and Lab value of 57 pepper germplasms and the genetic diversity based on SSR markers, and constructes the germplasm fingerprint maps, which may provide a certain theoretical basis for the identification of pepper germplasm resources, the screening of excellent germplasm and molecular-assisted breeding.

    Identification and Induced Expression Analysis of Transcription Factors NAC in Soybean Resistance to Soybean Mosaic Virus Based on WGCNA
    NIU Jing-ping, ZHAO Jing, GUO Qian, WANG Shu-hong, ZHAO Jin-zhong, DU Wei-jun, YIN Cong-cong, YUE Ai-qin
    2025, 41(7):  95-105.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0109
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    Objective To identify the hub genes of transcription factors associated with soybean resistance to soybean mosaic virus (SMV), which may provide a theoretical foundation for elucidating molecular mechanism and developing resistant germplasm. Method Using the transcriptome data of soybean disease-resistant material X149 and disease-susceptible material X97 induced by SMV SC15, PlantTFDB v5.0 database was used to predict the transcription factor genes of the whole genome. Weighted gene co-expression network analysis (WGCNA) was applied to identify the resistance-associated modules and hub transcription factors based on differentially expressed transcription factor genes. String 12.0 was for predicitng transcription factor interacting proteins. RT-qPCR was to analyze the expression patterns of transcription factor genes induced by hormones (ETH, SA, MeJA and ABA). Result The whole genome contained 3 170 genes encoding transcription factors, of which 1 727 were differentially expressed. WGCNA analysis revealed that 1 727 genes were divided into six co-expression modules. Notably, the brown and turquoise modules demonstrated significant correlations with X149 resistance. Within the brown and turquoise modules, GmNAC030 in turquoise module, a NAC transcription factor, showed the highest connectivity and was identified as a core transcription factor. Other NAC transcription factor genes in the turquoise module expression were similar to GmNAC030 included GmNAC043, GmNAC085, GmNAC092, and GmNAC101. Prediction of interacting protein indicated that the interacting proteins of GmNAC030, GmNAC043, and GmNAC092 were primarily transcription factors. Specifically, only the interacting proteins Glyma.02g131700 (bZIP1), Glyma.16g164800 (AP2-EREBP), and Glyma.08G118200 (WRKY48) were located within the module. RT-qPCR analysis demonstrated that GmNAC030 expression was predominantly induced by MeJA, while the other four NAC transcription factors were mainly induced by both MeJA and ETH. Conclusion The expression of five NAC transcription factors, GmNAC030, GmNAC043, GmNAC085, GmNAC092 and GmNAC101, is associated with soybean resistance to SMV and can be induced by MeJA and ETH.

    Cloning and Preliminary Functional Analysis of HvERECTA Gene in Hordeum vulgare
    HAN Yi, HOU Chang-lin, TANG Lu, SUN Lu, XIE Xiao-dong, LIANG Chen, CHEN Xiao-qiang
    2025, 41(7):  106-116.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1077
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    Objective Transmembrane protein ERECTA, a protein kinase (LRR-RLK) rich in leucine receptor -like serine/threonine, controls cell proliferation, regulates stomatal structure and responds to various stresses. The aim of this study is to elucidate the structure and functional expression of the HvERECTA gene in barley and lay a foundation for its biological function identification and crop molecular breeding. Method Using barley Morex as material, the HvERECTA gene of barley was cloned and its biological information and gene function were analyzed. RT-qPCR was used to analyze the functional characteristics of barley HvERECTA gene in different organs and under different stresses, and to study the expression patterns of HvERECTA gene in barley and typical bHLH-transcription factors HvSPCH, HvMUTE and HvFAMA related to barley stomatal development. Result The length of open reading frame of barley HvERECTA was 2 934 bp, encoding 977 amino acids, having typical LRR-RLK structure, localization predicted in the plasma membrane and containing signal peptide. The phylogenetic tree showed that it belonged to the ERECTA subfamily of Gramineae, and protein interaction predicted that it was a key gene regulating stomatal development. The results of RT-qPCR showed that the expression of HvERCTA was the highest in early leaves and lowest in the roots, and significantly different in abiotic stress of ABA, high temperature, low temperature and simulated drought (20% PEG6000). Under simulated drought stress, the gene expression of HvERECTA and HvSPCH showed similar trend, but opposite trend with the expression of HvMUTE gene. Conclusion HvERECTA gene is cloned from barley. It is a key gene to regulate the stomatal development. It is expressed in early leaves of barley, participates in ABA signal transduction pathway, and responds the abiotic stresses such as high- or low- temperature and drought.

    Identification of 4CL Gene Family in Arachis hypogaea L. and Expression Analysis in Response to Drought and Salt Stress
    ZHANG Ze, YANG Xiu-li, NING Dong-xian
    2025, 41(7):  117-127.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1266
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    Objective To analyze the basic characteristics of peanut 4CL (4-coumarate:CoA ligase) gene family members and their responses to drought and salt stress in order to provide important target genes for breeding peanut varieties tolerant to drought and salt. Method The members of peanut 4CL gene family were identified by HMM file, NCBI CDD, and Pfam databases at the whole genome level. ExPASy-ProtParam was used to analyze the physicochemical properties of proteins. MEGA7 and itol tools were used for phylogenetic analysis. The conserved motifs and conserved domains of proteins were analyzed by MEME and CD-search tools in NCBI, respectively. The cis-acting elements were analyzed and visualized by PlantCARE and TBtools, respectively. The transcriptional changes of peanut 4CLs were analyzed by RNA-seq data and RT-qPCR. Result Based on the reference data of peanut Tifrunner genome, 56 peanut 4CL genes were identified, with amino acid length ranging from 239 to 1 208, pI ranging from 5.5 to 9.22, and aliphatic index ranging from 80.2 to 103.13. The instability index (II) was 25.51-48.79, and GRAVY was -0.367-0.139. The Ah4CLs were unevenly distributed on 20 chromosomes of peanut A and B genome, and the Ah4CLs gene density was the highest on chromosomes 5 and 15. In the same clustering branch, Ah4CLs had similar conserved motif composition and intron-exon distribution structure, with exon number ranging from 1 to 18. Ah4CLs promoter regions were rich in light, abiotic stress, hormone and growth response elements. The expression of Ah4CLs was tissue-specific and higher in the root, flower and seed. Under ABA, salt and drought stress, the transcription levels of some Ah4CLs significantly increased, especially Ah4CL28 was significantly up-regulated under ABA, drought, and salt stress. These genes may play an important role in peanut response to abiotic stress. Conclusion The 56 identified peanut 4CL gene family members have different structures and characteristics, and some motifs and domains are conserved. Ah4CLs not only affects plant growth and development but also participates in abiotic stress response.

    Identification of LEA Gene Family and Analysis on Its Response to Aluminum Stress in Ricinus communis L.
    LI Kai-yue, DENG Xiao-xia, YIN Yuan, DU Ya-tong, XU Yuan-jing, WANG Jing-hong, YU Song, LIN Ji-xiang
    2025, 41(7):  128-138.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1255
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    Objective To identify members of the RcLEA gene family in the Ricinus communis genome and analyze their gene characteristics, potential functions, which lays a foundation for exploring the role of LEA in regulating tolerance to aluminum stress. Method Bioinformatics was used to identify the LEA gene family of R. communis, then their physicochemical properties, phylogenetic relationships, gene structure, conserved motifs, promoter cis-acting elements of its proteins were analyzed, and finally the expressions of some RcLEA genes under aluminum stress was analyzed. Result A total of 52 LEA family gene members were identified from the R. communis genome. By phylogenetic analysis, they were classified into eight subgroups: RcLEA_1, RcLEA_2, RcLEA_3, RcLEA_4, RcLEA_5, RcLEA_6, Rc_DHN, and Rc_SMP, distributed across ten chromosomes. The number of amino acids of family members ranged from 91 to 406, with isoelectric points between 4.54 and 10.29, and molecular weights ranging from 9 951.42 to 44 515.78 Da. Most LEA proteins were hydrophilic. Motif analysis revealed significant variations among different subfamilies, while maintaining similarity within the same subclass. Gene structure analysis indicated that most genes contained one or more introns. Promoter regions of all 52 genes contained one or more cis-acting elements related to plant hormones and stress responses. Collinearity analysis identified eight pairs of duplicated genes. Additionally, real-time quantitative PCR results demonstrated that the expressions of the RcLEA genes changed under aluminum stress treatment compared to the control, suggesting their involvement in the responses to aluminum stress. Conclusion A total of 52 RcLEA genes are identified from the R. communis genome, which are classified into eight subgroups. Expression analysis of selected RcLEA genes from these eight subgroups under aluminum stress reveals significant alterations in the expressions of most genes, indicating their potential crucial roles in R. communis responding to aluminum stress.

    Transcriptome Analysis of the Effect of Ca 2+ Treatment on the Seed Germination of Flax
    GUO Xiu-juan, FENG Yu, WU Rui-xiang, WANG Li-qin, YANG Jian-chun
    2025, 41(7):  139-149.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1269
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    Objective The effect of Ca2 + on the germination characteristics of flax (Linum usitatissimum L.) seeds was identified, and the regulation mechanism of flax seed germination was deeply understood, which may provide a theoretical basis for flax seed germination and new insights for further understanding the influence of Ca2 + on flax seed germination. Method The seeds of the flax variety Jinya 10 were treated with a 0.5% calcium chloride solution and then dried in air for germination experiment. Bioinformatics was used to analyze the expressions of genes before and after Ca2 + treatment at 5, 10 and 15 h. Result The time of dew-white and budding of flax seeds treated with Ca2+ was significantly advanced. Transcriptome sequencing was utilized to analyze gene expression before and after Ca²⁺ treatment at 5, 10, and 15 h time points. Following a 5-hour Ca2 + retreatment of the flax seeds, a total of 1 357 differentially expressed genes were identified compared to the control group, with 558 genes upregulated and 799 genes downregulated. As the duration of Ca2 + treatment extended, the number of differentially expressed genes significantly decreased. After a 10-hour Ca2+ treatment, a total of 641 differentially expressed genes were discovered, with 385 upregulated and 256 downregulated. When the flax seeds were treated with Ca2 + for 15-hour, only 168 genes showed differential expression compared to the control, of which 151 were upregulated and 17 were downregulated. Gene co-expression network analysis was employed to select the most relevant modules and hub genes in each group. Conclusion In the H5 group (seeds treated with Ca2+ for 5 h), the Meblue module was significantly associated, a total of 147 genes were identified and enriched in the pathways of carbon metabolism, plant hormone signaling transduction and carbon fixation in photosynthetic organisms. The Mebrown module in the H15 group (seeds treated with Ca2+ for 15 h) contained a total of 141 genes, which were enriched in carbon metabolism pathway, phenylpropanoid biosynthesis pathway and linolenic acid metabolism pathway. Furthermore, 35 genes related to plant hormone signal transduction pathways in flax seeds are specifically selected for differential expression due to Ca2 + treatment.

    Identification of the PMEI Gene Family of Pectin Methylesterase Inhibitor in Foxtail Millet and Analysis of Its Response to Abiotic Stress
    LI Xin-ni, LI Jun-yi, MA Xue-hua, HE Wei, LI Jia-li, YU Jia, CAO Xiao-ning, QIAO Zhi-jun, LIU Si-chen
    2025, 41(7):  150-163.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0018
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    Objective PMEI (pectin methylesterase inhibitor) is a key component regulating the structure and properties of the cell wall, playing a significant role in plant stress response mechanisms. Therefore, studying the response mechanism of PMEI gene in foxtail millet under abiotic stress may provide a theoretical basis for the mechanism of resistance to stress in millet. Method Bioinformatics was used to identify PMEI gene family in foxtail millet, and quantitative real-time PCR was employed to analyze the expression patterns of family members under low temperature, drought, MeJA and ABA stress. Result A total of 68 SiPMEI gene family members were identified in the foxtail millet (Setaria italica) genome, unevenly distributed on nine chromosomes, and most of them were localized to the cell wall or chloroplasts. There were two main domains of SiPMEI family members, PMEI domain and pectinesteras + PMEI domain, as well as similar physicochemical properties, subcellular localization and gene structure of the family members containing the same domain. Promoter cis-acting element analysis suggested that the SiPMEI gene family members contained multiple abiotic stress and hormone responsive elements. In this study, eight SiPMEI family members containing more than two stress response elements were selected for quantitative real-time expression analysis. The results showed that SiPMEI was differentially expressed in the root, stem, leaf and ear of the foxtail millet. Under abiotic stress (low temperature, drought), hormone stress (MeJA, ABA), the expression of SiPMEI gene tended to increase within 0-24 h, and the highest expressions of member SiPMEI30, SiPMEI32, SiPMEI36 and SiPMEI63 containing PMEI domain and localized on the cell wall responded to stress at 8‒24 h. The highest expression distribution of member SiPMEI22, SiPMEI31, SiPMEI38, and SiPMEI47 with subcellular localization on chloroplasts was more dispersed and had more upregulation sites. Conclusion SiPMEI has a positive response under low temperature, ABA and MeJA stresses, and has a similar expression trend under drought and MeJA stresses. These differential expressions suggest that SiPMEI may respond to abiotic stress through different molecular mechanisms.

    Biological Basis Study for Grain Shattering in Proso Millet and Identification of Genes Regulating Grain Shattering
    WANG Yue-chen, HAN Xin-qi, WEI Wen-min, CUI Zhao-lan, LUO Yang-mei, CHEN Peng-ru, WANG Hai-gang, LIU long-long, ZHANG Li, WANG Lun
    2025, 41(7):  164-171.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0150
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    Objective Proso millet (Panicum miliaceum L.) plays an important role in dryland agriculture, saline alkali land development and utilization, and disaster relief and replanting in China. Grain shattering is a crucial factor affecting yield of proso millet. Investigating the mechanism of grain shattering in proso millet is very important for breeding new varieties with low grain shattering and improving crop production. Method Having the proso millet gerplasm 'Yemizi' with strong grain shattering and 'Hongnianmi' with low grain shattering by paraffin sections, we studied the structural differences of abscission layer cells among millet varieties with different grain shattering characteristics. In addition, we identified proso millet grain shattering genes using reverse genetics strategies. Result Our results showed that 'Yemizi' had a distinct abscission layer, while no complete abscission layer was observed in 'Hongnianmi'. In addition, the homologous genes of rice (Oryza sativa) grain shattering genes were identified in proso millet and their expression in 'Yemeizi' at 1, 20, and 35 d after flowering was analyzed. The results showed that most genes had the highest gene expression at 1 d after flowering. Finally, PmSh1-1, was further analyzed, and the results showed that the expression of the PmSh1-1 in 'Yemizi' and 'Hongnianmi' was not significantly different, but the cDNA sequence of the PmSh1-1 in 'Hongnianmi' was shorter compared with it in 'Yemizi'. Conclusion We found that incomplete abscission layer may lead to a reduction of grain shattering and PmSh1-1, and a homologous gene of rice OsSh1 could be a potential grain shattering gene in proso millet.

    Cloning and Interaction Analysis of StPTST2a Gene in Potato
    HUANG Dan-dan, WU Yun-yi, ZOU Jian-hua, YU Ting, ZHU Yan-hui, YANG Mei-hong, DONG Wen-li, GAO Dong-li
    2025, 41(7):  172-180.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0043
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    Objective PROTEIN TARGETING TO STARCH (PTST) plays an important role in starch biosynthesis. This study aims to investigate the functional role of StPTST2a in potato (Solanum tuberosum L.). Method RT-PCR was used to clone StPTST2a gene and its amino acid sequence was analyzed by the bioinformatic method. RT-qPCR was performed to examine the expressions of StPTST2a in various organs. Prokaryotic expression system was used to express MBP-tagged StPTST2a in vitro and subsequently the recombinant protein was tested for its binding to starch. Seven starch biosynthesis-related genes were cloned, and the protein-protein interaction analysis was performed with yeast two-hybrid (Y2H) assays and luciferase complementation imaging (LCI) assays. Result StPTST2a encoded a protein of 532 amino acids, and it harboured a CBM48 domain at the C-terminus. StPTST2a was expressed at various tissues, and the transcript level was higher in the leaves than that in other organs. In vitro starch binding assays showed that StPTST2a bound to the starch. Y2H and LCI assays demonstrated that StPTST2a interacted with StSS4, StSS6 and StISA1.1 in yeast cells and in plants. Conclusion StPTST2a form a complex with starch biosynthesis-related genes, which may collaboratively control starch biosynthesis and formation of starch granules.

    Cloning and Functional Study of Transcription Factor StMYB96 in Potato
    LI Xia, ZHANG Ze-wei, LIU Ze-jun, WANG Nan, GUO Jiang-bo, XIN Cui-hua, ZHANG Tong, JIAN Lei
    2025, 41(7):  181-192.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0105
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    Objective MYB transcription factors (TFs) play critical roles in plant responses to abiotic stress by regulating the expressions of stress response genes. This study is aimed to investigate the role of StMYB96 in responses to abiotic stress in potato. Method The StMYB96 gene was cloned and subjected to bioinformatics analysis, subcellular localization, and expression pattern analysis. By expressing StMYB96 in vitro and combining it with DNA affinity purification sequencing (DAP-seq), the target genes of StMYB96 and the abiotic stress-responding pathways in which they participate were identified genome-wide. Result The length of open reading frame (ORE) of StMYB96 was 1 002 bp, it encoded a peptide of 333 amino acids. Multiple sequence alignment revealed that StMYB96 was closely related to SlMYB306 (Solanum lycopersicum), CaMYB306 (Capsicum annuum), and LbMYB306 (Lycium barbarum). Subcellular localization analysis indicated that StMYB96 was localized in the nucleus. The quantitative reverse transcription-PCR (RT-qPCR) analysis showed that StMYB96 was expressed in the leaves, stems, and roots, with the highest expression observed in the stems. The expression of StMYB96 was significantly up-regulated under drought stress but down-regulated under low-temperature treatment. DAP-seq analysis revealed that StMYB96 mainly bound to three specific binding sites in the promoters of 8 837 target genes, which were involved in various metabolic pathways. Notably, genes associated with the flavonoid biosynthesis pathway and the fatty acid elongation pathway were directly regulated by StMYB96. Conclusion StMYB96 may play a role in responding to drought and low-temperature stresses by regulating different metabolic pathways.

    Genome-wide Identification of the IbNRT2 Gene Family and Its Expression in Sweet Potato
    WANG Fang, QIAO Shuai, SONG Wei, CUI Peng-juan, LIAO An-zhong, TAN Wen-fang, YANG Song-tao
    2025, 41(7):  193-204.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1277
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    Objective To identify the high-affinity nitrate transporter NRT2 family members of sweet potato (Ipomoea batatas (L.) Lam.), and to analyze the physicochemical properties, structure and expression analysis under different stresses, thus providing theoretical support for the functional identification of NRT2 family members in sweet potato. Method The IbNRT2 gene members were systematically identified by bioinformatics, transcriptome analysis and low-nitrogen phenotype screening using NCBI and sweet potato genome databases, and the gene structure, conserved motifs and expression characteristics were analyzed. Result Seven IbNRT2 genes were identified in the whole genome of sweet potato. The results of physicochemical property analysis showed that the residual number of amino acids encoded by the members of the IbNRT2 family was 462-536, the theoretical isoelectric points were greater than 7, and all the encoded proteins were hydrophobic proteins. Seven IbNRT2 and nine closely related wild diploid (Ipomoea trifida) ItfNRT2 genes were distributed on 5 chromosomes. Phylogenetic tree analysis showed that the seven IbNRT2 family members were divided into three subgroups, and sweet potato was the most closely related to the wild diploid. The cis-actingelement analysis showed that there were numerous environmental and hormone response elements in the promoter region of IbNRT2 gene, among which the light response element was the most. The intraspecific collinearity relationship of sweet potato showed that there were two pairs of collinearity between the seven IbNRT2 genes. The results of interspecific collinearity showed that 7 pairs of collinearity were formed between sweet potato and wild diploid ItfNRT2 genes, and 4 pairs of collinearity were formed with ArabidopsisAtNRT2s. The results of tissue and salt stress expression analysis showed that IbNRT2.7 was widely expressed, with the highest expression in the seeds and leaves, and the expression was up-regulated by salt stress. IbNRT2.1 and IbNRT2.2 were mainly expressed in the roots and down-regulated by salt stress. RT-qPCR analysis of different nitrogen-treated sweet potato varieties showed that IbNRT2.1 and IbNRT2.2 were most significantly induced by low nitrate, and the induced expression folds of IbNRT2.1 and IbNRT2.2 were more pronounced in nitrogen-efficient sweet potato varieties (Chuanshu 221, Chuanshu 228, Xichengshu 007) than in nitrogen-inefficient sweet potato varieties (Pushu 32, Mianzi 9). Conclusion Seven IbNRT2 genes are identified in sweet potato, and the expression patterns of IbNRT2 vary in different parts, salt stress and low nitrogen stress, which lays a foundation for further study of the function of IbNRT2 in different parts.

    Cloning of SOS1 Gene Promoters from Poplar and Analysis of Its Response to Salt Stress
    FU Bo-han, MAO Hua, ZHAO Xin-cheng, LU Hong, OU Yong-bin, YAO Yin-an
    2025, 41(7):  205-213.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0069
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    Objective SOS1 (salt overly sensitive 1), a Na+/H+ antiporter located on the plasma membrane, plays a crucial role in plant responses to salt stress. Comparative analysis of SOS1 gene promoters from different Populus species would provide an important foundation for understanding and applying SOS1 gene and its promoters for stress resistance improvement. Method The study used different tree species of the genus Populus as experimental materials and the expression patterns of the SOS1 gene was analyzed by real-time quantitative PCR. The promoter fragments of the SOS1 gene were cloned from P. alba, P. euphratica, and P. russkii, and were ligated with the GUS (β-glucuronidase) reporter gene and transformed into Populus tomentosa to obtain transgenic plants. The tissue specificity and response to salt stress of the promoter were studied by GUS histochemical staining and enzyme activity quantification using transgenic P. tomentosa. Result The expression pattern of SOS1 genes varies significantly among different tree species, e.g. under salt stress, the expression of SOS1 gene in the stem of P. alba is upregulated, while there is no significant change in P. euphratica, and is downregulated in P. russkii. All three cloned promoter fragments can drive the expression of the GUS gene in the leaves, stems, and roots of transgenic P. tomentosa, presenting promoter activity, with higher activity in the stems and roots. The SOS1 promoter from P. alba is active in the epidermis and cortex of the stem, but has very low activity in the phloem, cambium, and xylem; the SOS1 promoter from P. russkii shows higher activity in the xylem, but very low activity in the cambium and bark; the SOS1 promoter from P. euphratica is active in all parts, with the highest activity in the cambium. Under salt stress conditions, the activity of all three promoters increases. Conclusion Different poplar SOS1 gene promoters may respond to salt stress but have different tissue specificity.

    Identification and Expression Profile Analysis of the TRB Gene Family in Tea Plant
    GONG Yu-han, CHEN Lan, SHANGFANG Hui-zi, HAO Ling-ying, LIU Shuo-qian
    2025, 41(7):  214-225.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0048
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    Objective Telomere repeat-binding (TRB) proteins, a class of double-stranded telomeric DNA-binding proteins, play crucial roles in plant growth and development. This study is aimed to identify the TRB gene family in tea plant [Camellia sinensis (L.) O. Kuntze], clone a key gene CsTRB1 and characterize its molecular properties, and investigate the expression patterns of CsTRBs under abiotic stresses. These findings may establish a molecular foundation for elucidating the functional mechanisms of TRB genes in C. sinensis. Method Genome-wide identification of CsTRB gene family was performed, with bioinformatics analyses of conserved motifs, gene structures, chromosomal localization, gene collinearity, and cis-acting elements, as well as predicting. Proteins' physicochemical properties and structures were predicted. Using 'Bixiangzao' as experimental material, the CDS sequence of CsTRB1 was cloned. Transcriptomic data and RT-qPCR were combined to analyze expression patterns of CsTRB family members across tissues, under low-temperature stress, and following hormone treatments. Result Seven CsTRB genes were identified. CsTRB1 was successfully cloned, containing an 885 bp coding sequence encoding 295 amino acids with a typical TRB domain. Family members were distributed across six chromosomes. CsTRBs were classified into two subfamilies via phylogenetic analysis. Collinearity analysis revealed gene duplication event of CsTRB gene within the tea genome and identified seven orthologous gene pairs between tea TRB family and Arabidopsis thalianaTRB family. RT-qPCR demonstrated that under low-temperature stress, the expressions of CsTRB1, CsTRB2, CsTRB6, and CsTRB7 significantly decreased, while the expressions of CsTRB3, CsTRB4, and CsTRB5 markedly increased. All CsTRB members except CsTRB5 showed high expression under ABA treatment. The expressions of CsTRB1, CsTRB4, and CsTRB5 was significantly upregulated by IAA treatment. Conclusion The expression of CsTRB gene may be regulated by cis-acting elements responsding to light, low temperature, and hormones, suggesting their critical roles in tea plant development and abiotic stress responses. The successful cloning and molecular characterization of CsTRB1 provide essential materials for subsequent protein interaction studies and functional validation.

    Cloning of Plastidial PfLPAT1B Gene from Perilla frutescens and Its Functional Analysis in Oil Biosynthesis
    HUANG Xu-sheng, ZHOU Ya-li, CHAI Xu-dong, WEN Jing, WANG Ji-ping, JIA Xiao-yun, LI Run-zhi
    2025, 41(7):  226-236.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1056
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    Objective Lysophosphatidic acid acyltransferase (LPAT) plays a crucial role in plant growth, development, and lipid metabolism. This study aims to investigate the biological function of the PfLPAT1B gene of Perilla frutescens, providing a scientific foundation for the genetic improvement and breeding of new cultivars of Perilla and other oilseed crops. Method The PfLPAT1B gene sequence was identified from the Perilla genome database, and its sequence characteristics and phylogenetic relationships were analyzed using omics tools. The expression patterns of PfLPAT1B in various tissues and different developmental seeds were evaluated by RT-qPCR. The enzyme activity of PfLPAT1B protein was assessed using the Escherichia coli LPAT-deficient strain SM2-1. The function of PfLPAT1B in oil biosynthesis was performed via genetic transformation of Saccharomyces cerevisiae and Nicotiana tabacum. Result PfLPAT1B gene encodes a total of 369 amino acid residues and it is a basic unstable hydrophilic protein, containing a typical conserved domain of lysophosphatidic acid acyltransferase. PfLPAT1B gene was expressed in different tissues and seeds at different developmental stages of Perilla, with the highest expression in flowers and an increasing trend during seed development. Subcellular localization showed that PfLPAT1B is localized in chloroplast. Complementation assays in the SM2-1 strain demonstrated that PfLPAT1B possesses LPAT enzymatic activity. Overexpression of PfLPAT1B gene in S. cerevisiae and N. tabacum significantly enhanced the total oil content, accompanied by increased levels of C16:0 and C16:1. Moreover, transgenic tobacco showed a notable increase in starch content and a decrease in soluble sugar content. Conclusion The PfLPAT1B gene from Perilla encodes a functional LPAT enzyme. Heterologous overexpression of PfLPAT1B can significantly enhance oil biosynthesis and accumulation and alter the content of major fatty acids in host tissues.

    Metabolic Response Analysis of Brachypodium distachyon to Photoperiods
    JIANG Tian-wei, MA Pei-jie, LI Ya-jiao, CHEN Cai-jun, LIU Xiao-xia, WANG Xiao-li
    2025, 41(7):  237-247.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1099
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    Objective To analyze the effects of different photoperiods on the metabolome of Brachypodium distachyon (Bd21), and to investigate how photoperiod variation influences Bd21 growth and development. This study aims to provide insights into the photoperiod adaptation mechanisms of temperate grasses and assist in breeding widely adaptable new varieties. Method Leaves of B. distachyon (Bd21) were collected at three time points (ZT0, ZT12, and ZT24) under long-day (16 h light∶8 h dark, LD) and short-day (8 h light∶16 h dark, SD) conditions. Metabolic differences between LD and SD conditions in B. distachyon were compared using ultrahigh-performance liquid chromatography-mass spectrometry (UPLC-MS). Result A total of 739 metabolites were detected, including 135 organic acids and their derivatives, 93 organic oxygen-containing compounds, and 92 lipids and lipid-like molecules. Metabolomic separation between LD and SD conditions was observed, with an overall increase in metabolic levels and upregulation of amino acid-related metabolites under SD, including aspartate, isoleucine, and histidine. The S24 vs L24 comparison enriched 19 metabolic pathways, including glutamate, aspartate, and glutamine metabolism, lysine degradation, and the biosynthesis and degradation of valine, leucine, and isoleucine. Several amino acid pathways were closely linked to the TCA cycle. Conclusion The metabolome of B. distachyon is sensitive to short-day (SD) conditions. B. distachyon may adapt to the changes of shorter photoperiod and insufficient photosynthesis by upregulating the downstream metabolic network of aspartic acid and the synthesis and degradation of branched-chain amino acids, aiming to maintain metabolic balance. This indicates that amino acid metabolism regulation plays an important role in the adaptation of B. distachyon to the shortened photoperiod.

    Cloning and Expression Analysis of the CrMYB4 Gene in Carex rigescens
    WEI Yu-jia, LI Yan, KANG Yu-han, GONG Xiao-nan, DU Min, TU Lan, SHI Peng, YU Zi-han, SUN Yan, ZHANG Kun
    2025, 41(7):  248-260.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0080
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    Objective To explore the role of CrMYB4 in the growth, development and abiotic stress of Carex rigescens may provide theoretical guidance for further research on its function in the stress resistance mechanism of C. rigescens. Method Based on the previous results of salt-responsive omics of C. rigescens, a CrMYB4 protein was screened, and its bioinformatics characteristics, subcellular localization, promoter cis-acting element analysis, and expression patterns under different tissues, abiotic stresses, and exogenous hormone treatments were evaluated. Result The coding region of the CrMYB4 sequence was 747 bp, encoding 248 amino acids. Phylogenetic analysis revealed that it belonged to the typical R2R3-MYB group and was closely related to plants in the Cyperaceae family. Subcellular localization analysis demonstrated that the CrMYB4 protein was localized in the nucleus. CrMYB4 was highly expressed in new and old leaves, while its expressions were relatively low in tissues such as roots, young buds, and leaf sheaths. Analysis of promoter cis-acting elements showed that the promoter region of CrMYB4 contained multiple functional elements associated with hormone response, plant physiology, and transcriptional recognition. Expression analysis under different abiotic stresses showed that the CrMYB4 gene responded rapidly to salt, drought, and low-temperature stresses, with its expression increasing rapidly after 1 h of treatment. It is speculated that CrMYB4 may be involved in the response of C. rigescens toabiotic stress. Under treatments with different plant hormones (ABA, SA, IAA, and GA3), the expression of CrMYB4 was significantly upregulated and reached its maximum value at 6 d. It is speculated that CrMYB4 may be involved in the hormone response of C. rigescens. Conclusion CrMYB4 has potential roles in the growth and development, stress responses to adversity, and hormone responses of C. rigescens. It plays a crucial role in salt stress and the IAA signaling pathway.

    Biocontrol Characteristics of Strain JB7 against Fusarium graminearum
    ZHANG Yue, BI Yu, MU Xue-nan, ZHENG Zi-wei, WANG Zhi-gang, XU Wei-hui
    2025, 41(7):  261-271.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1187
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    Objective Bacillus methylotrophicus JB7 may inhibit Fusarium graminearum (Fg) growth and control Fusarium head blight in wheat; however, the mechanisms underlying its antifungal activity and disease management remain unclear. In this study, the antifungal and disease control mechanisms of B. methylotrophicus were explored, which may provide a theoretical basis for its further application. Method The effects of cell-free supernatants (CFS) from strain JB7 on the number of Fg spores, mycelium growth and antioxidant enzyme activity of Fg were studied through assessment of spore germination, antifungal activity and antioxidant enzyme activity. Transcriptomic sequencing was employed to analyze differentially expressed genes (DEGs), while amplicon sequencing was used to evaluate the effects of JB7 spraying on wheat ears mycotoxin content and microbial community structure. Result The CFS from strain JB7 showed a strong inhibitory effect on mycelial growth and germination of Fg. Additionally, the CFS led to an increase in the activity of superoxide dismutase (SOD) and catalase (CAT) in Fg mycelium, while causing a reduction in peroxidase (POD) and glutathione reductase (GR) activities. Transcriptome analysis indicates that the CFS from strain JB7 affected the expression of genes related to oxidative stress and toxin production. The application of JB7 resulted in an alteration of microbial community structure in wheat ears, leading to a decrease in Fg density and mycotoxin content. Conclusion The CFS from strain JB7 shows antifungal effects on Fg by triggering oxidative stress and reducing mycotoxin levels, and strain JB7 may change the microbial community structure in wheat ears and reduce the density and mycotoxin content of Fg.

    Effect of Exogenous Salicylic Acid on Wheat Infested with Blumeria graminis f. sp. tritici and Its Transcriptome Analysis
    LI Cheng-hua, DOU Fei-fei, REN Yu-zhao, LIU Cai-xia, LIU Feng-lou, WANG Zhang-jun, LI Qing-feng
    2025, 41(7):  272-280.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1201
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    Objective Powdery mildew, one of the most devastating diseases affecting wheat, poses a significant threat to wheat yield and quality. This study aims to investigate the regulatory effects of exogenous salicylic acid (SA) on Blumeria graminis f. sp. tritici (abbreviated as Bgt below) infection and to reveal the molecular mechanisms underlying Bgt infection and SA-mediated resistance to disease through transcriptomic analysis. Method In this experiment, we analyzed the transcriptome of wheat powdery mildew fungus under Bgt-infested conditions for 1-4 d using powdery mildew-susceptible common wheat Zhongzuo 9504 as the material. Concurrently, salicylic acid was applied externally to observe the effect of phytohormones on powdery mildew-infected wheat. Result Transcriptomic analysis revealed that, compared to 1 d, 399 genes were upregulated and 1 110 genes were downregulated at 4 d. The upregulated genes were enriched in pathways such as secondary metabolite biosynthesis, metabolic pathways, and proteasome, while the downregulated genes were primarily involved in the degradation of valine, leucine, and isoleucine. Protein-protein interaction network analysis identified three core hub genes of Bgt: BGTH12_LOCUS642, BGTH12_LOCUS3045, and BGTH12_LOCUS5497, indicating their crucial roles during wheat infection. Furthermore, exogenous SA significantly upregulated the expressions of six hub genes in wheat (ERF109, PP2C30, TIFY6B, HSP70, At4g15970, and HERK1), thereby reducing the damage caused by Bgt. Conclusion During the infection process, Bgt demonsttrates extensive differential gene expression, with BGTH12_LOCUS642, BGTH12_LOCUS3045, and BGTH12_LOCUS5497 identified as core hub genes. The external application of salicylic acid may induce the expressions of resistance genes and inhibit the growth and development of conidia under the conditions of powdery mildew infection, thereby slowing down the infection of powdery mildew to a certain extent and increasing the resistance of wheat to powdery mildew.

    Modulation of the Growth, Quality, and Cadmium Content of Lily Bulbs by Bacillus velezensis XY40-1
    ZHANG Jin-hao, DENG Hui, ZHANG Qing-zhuang, TAO Yu, ZHOU Chi, LI Xin
    2025, 41(7):  281-291.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1245
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    Objective This study aims to investigate the effects of Bacillus velezensis XY40-1 microbial agent on the yield and quality of lily bulbs. Additionally, it seeks to elucidate its impact on soil physicochemical properties, microbial community structure, and heavy metal transport functions, with the ultimate goal of providing a scientific foundation for efficient lily cultivation and soil health management. Method A drip irrigation system was used to apply the microbial agent during the lily growth process. The fresh weight, dry weight, as well as the protein, polysaccharide, total saponin content, and cadmium accumulation in the lily bulbs were measured. Additionally, high-throughput sequencing technology was employed to analyze the changes in the rhizosphere soil microbial community structure, while metagenomic analysis was used to examine the expression patterns of functional genes related to nitrogen metabolism and cadmium transport. Result After the application of XY40-1 microbial agent, the soil pH increased to 5.41, and the content of available potassium rose by 31.15%. The fresh weight and dry weight of individual lily bulbs increased by 18.89% and 19.49%, respectively, while the yield per mu was enhanced by 16.47%. Moreover, the protein content in the lily bulbs increased by 15.1%, polysaccharide content by 11.5%, and total saponin content by 21.4%, while cadmium accumulation decreased by 11.45%. Microbial community analysis revealed that the relative abundance of Firmicutes and Bacteroidota significantly increased in the treatment group, while the abundance of Proteobacteria and Actinobacteria decreased. Metagenomic data indicated a significant upregulation of nitrogen fixation genes (nifD, nifH), nitrate reduction genes (narG, napA), and cadmium resistance genes (czcA, czcD). Conclusion B. velezensis XY40-1 significantly enhances lily yield and quality, while reducing cadmium accumulation in the bulbs, by improving soil physicochemical properties, optimizing microbial community structure, and activating key metabolic function genes.

    Effects of Co-treatment of Nano-silica and Bacillus cereus SS1 on the Growth of Tobacco
    DONG Xu-kun, CHE Yong-mei, WANG Ming-shuo, LUO Zheng-gang, GUAN En-sen, ZHAO Fang-gui, YE Qing, LIU Xin
    2025, 41(7):  292-298.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0089
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    Objective A growth-promoting Bacillus cereus strain SS1 was identified previously in our laboratory. The objective of this study is to examine the combined effects of silicon nanopowder and B. cereus SS1 on the growth of tobacco, as well as to explore the related functional mechanism. Method Zhongyan 101 was used as experimental material. Tobacco seedlings were treated with silicon nanopowder and SS1 alone or in combination, the growth and photosynthetic indexes, as well as the activities and gene expressions of nitrogen metabolism-related enzymes of each treatment were detected. Result The nanometer silica and SS1 at certain concentration, and co-treatment of nanopowder and SS1 all promoted the growth of tobacco, and the promotion effect of co-treatment was the most significant. The above-ground growth indexes of co-treated tobacco, such as plant height, stem diameter, leaf number, and leaf area, were higher than those of the seedlings treated with silicon nanopowder or SS1 alone and the non-treated control plants; moreover, the root length, root weight and root-shoot ratio also significantly increased by co-application. Meanwhile, the results showed that the co-treatment of SS1 and silicon nanopowder significantly increased the chlorophyll content, net photosynthetic rate, stomatal conductance and other photosynthetic indexes of tobacco plants. The activities of nitrate reductase (NR) and glutamine synthetase (GR) as well as the expressions of related genes NtNIA1 and NtGS1 also more significantly increased by SS1 and silicon nanopowder co-treatment compared to treatment with SS1 or silicon nanopowder alone. Conclusion The combination of silicon nanopowder and SS1 may promote plant growth by ameliorating the photosynthetic performance and promoting nitrogen metabolism of plant.

    Isolation and Identification of Endophytic Bacteria from Saposhnikovia divaricata and Analysis of Its Growth-promoting Characteristics
    SUN Meng-xue, ZHANG Yi-ying, Xu Peng, SUN Zhuo, WANG Yun-he, HAN Zhong-ming
    2025, 41(7):  299-311.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0070
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    Objective The growth-promoting strains were isolated and screened from the healthy roots of Saposhnikovia divaricata, and their growth-promoting effects on S. divaricata were investigated. The potential of endophytic bacteria for development and utilization was mined. Method Endophytic bacteria were isolated from healthy S. divaricata roots by dilution coating method and growth-promoting bacteria with nitrogen fixation, phosphorus solubilization, IAA production and siderophore production were screened by selective medium. The growth-promoting strains were classified by morphological, physiological and biochemical identification and 16S rRNA analysis. The colonization ability of endophytic bacteria in soil and its effect on the growth of S. divaricata were investigated based on antibiotic labeling method and pot experiment. Result A total of 202 endophytic bacteria were isolated from the roots of S. divaricata, 26 strains had the ability of nitrogen fixation, 21 strains had the ability to dissolve phosphorus, 24 strains had the ability to produce IAA, and 27 strains had the ability to produce siderophores. Two strains of endophytic bacteria MX-56 and MX-31 had three or more different growth-promoting functions, were identified as Klebsiella michiganensis and Pseudomonas koreensis. Two rifampicin-resistant strains stably colonized in the rhizosphere soil of S. divaricata. In the pot experiment, compared with CK, MX-56 had the most significant effect on plant height, root length, root diameter, aboveground fresh weight, root fresh weight and root dry weight of S. divaricata, increased 13.47%, 43.04%, 42.75%, 31.43%, 63.21% and 77.12% (P<0.05). Compared to CK, the total contents of two chromones in S. divaricata plants treated with strain MX-31 and MX-56 respectively increased by 33.72% and 30.23%. Conclusion In this study, two strains of P. koreaensis MX-31 and K. michiganensis MX-56 with good growth-promoting effect are isolated and screened from healthy S. divaricata, it may significantly increase the biomass and the content of effective components of S. divaricata.

    Effects of the Mycovirus BbOCuV1 on the Growth and Development of Host Beauveria bassiana and Its Pathogenicity to Ostrinia furnacalis Larvae
    JIA Xue, SUI Li, ZOU Xiao-wei, LU Yang, ZHANG Zheng-kun, LI Qi-yun
    2025, 41(7):  312-325.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0053
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    Objective By clarifying the effects of the mycovirus Beauveria bassiana orthocurvula virus 1(BbOCuV1) on the growth and development of the host Beauveria bassiana and on the pathogenicity of insect pests, we proved whether the mycovirus is a key factor that cause the decline of the pathogenicity of B. bassiana. Method Detoxification and horizontal transmission were used to study the effects of BbOCuV1 on the growth rate, spore production and biomass of B. bassiana colonies. The pathogenicity of B. bassiana on the second instar larvae of the Ostrinia furnacalis was determined. Transcriptome analysis was used to define the molecular mechanism of fungal viruses affecting the host strains. Result The growth rate, spore production and biomass of host strain significantly increased after virus BbOCuV1 infection, while the pathogenicity towards the insect pest significantly decreased. Transcriptome analysis showed that growth and development-related pathways and genes expression significantly upretulated, after virus infection, while the expressions of genes related to insect epidermis penetration and toxin metabolism significantly downregulated. Conclusion The growth and development as well as biomass of B. bassiana infected with virus BbOCuV1 increase, but pathogenicity against pests reduces, indicating that mycovirus infection is an important factor in the pathogenicity decline of B. bassiana to pests.

    Screening of Cellulose-degrading Bacteria in Pleurotus ostreatus Cultivation Substrate and Evaluation of Degradation Effect of Microbial Consortium
    LIU Jiao-jiao, MU De-mei, XIA Li-ming, FANG Yong, LIU Zuo-jun
    2025, 41(7):  326-335.  doi:10.13560/j.cnki.biotech.bull.1985.2024-1167
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    Objective To screen cellulose-degrading bacteria from the cultivation substrate of Pleurotus ostreatus and construct an efficient microbial consortium for the degradation of agricultural waste. Method Bacterial strains were isolated and purified using sodium carboxymethyl cellulose (CMC-Na) as the sole carbon source from the cultivation substrate of P. ostreatus, and those with cellulose-degrading capabilities were preliminarily identified through Congo red staining. Molecular biology techniques were employed for their identification and to construct a phylogenetic tree. Eleven microbial consortiums were established from strains without mutual antagonism and the cellulase activities of these communities were measured at different fermentation times. The optimal cellulose-degrading microbial consortium was then used in degradation experiments with rice straw and cottonseed hulls to determine its cellulose-degrading capability. Result Four symbiotic bacteria with efficient cellulose-degrading capabilities were identified: Streptomyces sp., Paenibacillus lautus, Microbacterium arborescens and Microbacterium aoyamense. The results indicated that composite microbial system H, composed of Streptomyces sp. Q5 and P. lautus Q6, showed as the most effective cellulose-degrading strain. On the fourth day of culture, its filter paper cellulase (FPase) enzyme activity increased by 229.97% and 134.29%, respectively, compared to the single strains. Additionally, the carboxymethyl cellulase (CMCase) enzyme activity increased by 92.81% and 21.94%, respectively, when compared to the individual strains. After 15 d of treatment with composite microbial system H, the degradation rate was 38.72% for rice straw and 35.76% for cottonseed hulls. Specifically, the degradation rates of lignin, cellulose, and hemicellulose in the rice straw were 33.94%, 31.17%, and 22.43%, respectively, while the corresponding rates for cottonseed hulls were 27.95%, 25.56%, and 53.86%. Further analysis of the surface morphological changes of both rice straw and cottonseed hulls before and after degradation confirmed that composite microbial system H possessed a superior cellulose-degrading capability. Conclusion The microbial consortium constructed in this study can efficiently degrade straw and cottonseed hulls, providing a theoretical basis for improving the effective utilization of agricultural waste resources.

    Enhancing the Thermostability of Lysozyme RPL187 Based on Protein Intelligence Models
    WANG Hui, FAN Ling-xi, SUN Ji-lu, WANG Yuan, WU Ning-feng, TIAN Jian, GUAN Fei-fei
    2025, 41(7):  336-346.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0008
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    Objective Lysozyme may be widely used as bacteriostatic agent in food, biomedicine and other fields. However, the stability of lysozyme as a biologically active substance is greatly affected by temperature, which makes it difficult to meet the needs of different industries. Therefore, a modification strategy incorporating the use of artificial intelligence models to design and screen protein mutants was used to improve the thermostability of lysozyme and expand the practical applications of lysozyme. Method The research material is lysozyme RPL187 from rumen protozoa genome. The variation of residual enzyme activities of lysozyme RPL187 at different temperatures (37, 45, 50, and 55 ℃) for different treating times (0, 1, 2, 4, and 8 h) were detected by the national standard method through the heterologous expression of the protein in Escherichia coli. The multipoint mutant of RPL187 was generated and screened based on the AI model, and the variations of residual enzyme activities of lysozyme mutant were detected at different temperatures and different times by the national standard method. The mechanism of the improved thermostability was investigated by determining the changes in Tm value, free energy, number of hydrogen bonds, and content of secondary structure between the wild type and the mutant. Result The specific activity of the RPL187 was (142 000±2 000) U/mg at 37 ℃ and pH 6.5, which was 5 times that of egg white lysozyme. RPL187 was more stable at 37 ℃ and 45 ℃, but with the increase of temperature and the prolongation of the heat treatment time, there was a significant decrease in the specific activity of the enzyme. After incubation for 1 h at 55 ℃, the specific activity of the enzyme decreased by about 88%. In order to improve the thermostability of RPL187, a total of 11 RPL187 multipoint mutants were screened based on the artificial intelligence model; seven mutants were successfully expressed solubilistically in E. coli, among which, RPL187-592 and RPL187-209 had the inhibitory activity against Micrococcus garciniae. Further results of the thermostability assay showed that RPL187-592 and RPL187-209 were stable at 50 ℃. The residual enzyme specific activities after heat treatment at 50 ℃ for 8 h were 4.43 times and 2.29 times that of the wild type, the Tm values were 2.06 ℃ and 2.41 ℃ higher than that of the wild type, and the free energies were 1.57 kcal/mol and 0.43 kcal/mol lower than that of the wild type, which demonstrated a more stable conformation and a higher thermostability than those of the wild type. Compared with the wild type, mutant RPL187-592 showed an increase of three intramolecular hydrogen bonds and the shift of amino acids from hydrophilic to hydrophobic (K2V and K137V) both contributed to the increase in thermostability of the protein; whereas, in RPL187-209, the increase in thermostability may be due to the shift of amino acids from flexible to rigid (K78P and K108P) and from hydrophilic to hydrophobic (K137A). Conclusion This modification strategy may effectively improve the thermostability of proteins, which is of great significance for the practical needs of expanding the application range of lysozyme, and also provides a referable basis for related research.

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    2025, 41(7):  347. 
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    2025, 41(7):  348. 
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    2025, 41(7):  349. 
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