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Research Progress in the Control of Multidrug Resistant Bacteria Based on in CRISPR-Cas System ZHOU Qian, TANG Meng-jun, ZHANG Xiao-yan, LU Jun-xian, TANG Xiu-jun, YANG Xing-xing, GAO Yu-shi Biotechnology Bulletin    2025, 41 (5): 42-51.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0872 Abstract （3690）   HTML （17）    PDF（pc） （1394KB）（135）       Knowledge map    Save Antimicrobial resistance of animal-derived bacteria affects the safety of animal breeding and causes an important threat to human public health and food safety. The abuse of antibiotics intensifies the spread of drug-resistant bacteria, and the research and development of new antibacterial drugs is becoming increasingly difficult. Drug-resistant bacteria contamination of animal-derived bacteria has become a global public crisis. Once drug-resistant bacteria spread from animals to humans, it will greatly threaten human health, and we urgently need new methods and strategies to deal with bacterial resistance. CRISPR-Cas (clustered regularly interspaced short palindromic repeats CRISPR-associated) is the third generation of "genome point editing technology". This technology can target the cutting of exogenous DNA and protect the genetic stability of microbial genetic material. Compared with traditional multi-drug-resistant bacteria control strategies, CRISPR-Cas system has a unique DNA sequence targeting and sensitivity. The sensitivity and detection timeliness and other performance indicators can be improved through accurate, simple and efficient gene editing technology, combined with nucleic acid amplification technology, colorimetric technology. This paper introduces the origin, systematic classification, and mechanism of gene editing of CRISPR-Cas system. Based on this brief introduction, it focuses on the research progress and application of the system in the field of multi-drug-resistant bacteria prevention and control, and have case studies and current challenges in the elimination of drug-resistant pathogens, the elimination of drug-resistant genes, and the diagnosis and detection of pathogens. In general, sequence-specific antimicrobials based on CRISPR-Cas system may reduce bacterial multidrug resistance, and the combination of nucleic acid amplification technology and real-time monitoring equipment may improve the accuracy and efficiency of detection, providing a new idea for the prevention and control of drug resistance and monitoring of animal-derived bacteria. Table and Figures | Reference | Related Articles | Metrics

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Integrated Transcriptome and Metabolome Analysis to Explore the Carotenoid Synthesis and Metabolism Mechanism in Anoectochilus roxburghii under Different Shading Conditions HU Ruo-qun, ZENG Jing-jing, LIANG Wan-feng, CAO Jia-yu, HUANG Xiao-wei, LIANG Xiao-ying, QIU Ming-yue, CHEN Ying Biotechnology Bulletin    2025, 41 (5): 231-243.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-1007 Abstract （3490）   HTML （9）    PDF（pc） （1955KB）（114）       Knowledge map    Save Objective To explore the effects of different shading conditions on the biosynthesis and metabolism of carotenoids in Anoectochilus roxburghii and to decipher the molecular mechanisms underlying carotenoid accumulation. Method Anoectochilus roxburghii was used as the experimental material, and the leaves from different shading treatments (T1: 25% shading, T2: 50% shading, T3: 75% shading) were selected. High-throughput RNA sequencing technology (RNA-Seq) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were employed to obtain transcriptomic and metabolomic data of A. roxburghii. Bioinformatics analysis was conducted to identify and quantify differentially expressed genes and metabolites, and to explore their relationships. Result Under different shading conditions, a total of 20 differentially expressed genes and 21 differential metabolites were identified. The significant expressions of key enzyme genes such as ZEP, LUT5, LUT1, LCYE, and NCED1 regulated the changes in metabolites like zeaxanthin, neoxanthin, and violaxanthin, with the highest content under the 75% shading treatment. Additionally, 22 transcription factors were predicted to regulate 14 differentially expressed genes, including ZEP, ZDS, and CYP707A, with 15 transcription factors responding to abscisic acid, gibberellins, and other plant hormone elements involved in the regulation of carotenoid biosynthesis. Eight differentially expressed genes involved in carotenoid biosynthesis were selected for RT-qPCR validation, and the results indicated that the expression trends of these 8 genes were consistent with the sequencing results. Conclusion Under different shading conditions, a total of 15 transcription factors in the leaves of A. roxburghii responded to five types of plant hormone elements, thereby regulating the expression of differentially expressed genes such as ZEP, NCED1, and CCD7, leading to significant accumulation of carotenoid-related metabolites such as zeaxanthin and lutein under 75% shading treatment. Table and Figures | Reference | Related Articles | Metrics

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Isolation and Application of Soybean Rhizobia and Symbiosis-promoting Rhizobacteria from Heilongjiang Province LIU Ke-han, YANG Sheng-hui, HUANG Qiao-yun, CUI Wen-jing Biotechnology Bulletin    2025, 41 (1): 252-262.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0564 Abstract （3224）   HTML （22）    PDF（pc） （3374KB）（396）       Knowledge map    Save 【Objective】Heilongjiang province is the main area of producing soybeans in China, with rich resources of soybean-nodulating rhizobia. Selecting high-quality stress-resistant soybean-nodulating rhizobia and rhizobacteria for soybean quality and yield improvement in this region was of great importance.【Method】In this experiment, rhizobia and symbiosis-promoting rhizobacteria were isolated and purified from root nodules and rhizosphere soil in Heilongjiang province, respectively. Species identification was conducted by rpoB and 16S rRNA gene sequencing. The tolerances of rhizobial strains to salt stress（1.2% NaCl）and drought stress（15% PEG6000）were evaluated in tube culture conditions. Highly efficient symbiotic rhizobial strains were selected by inoculating with stress tolerant rhizobial strains to soybean HH43. Representative rhizobial strains and symbiosis-promoting rhizobacterial strains were co-inoculated to HH43 to obtain highly-efficient symbiotic rhizobial strains and rhizobacterial strains with significant symbiosis-promoting effects.【Result】1）A total of 136 rhizobial strains were isolated, of which 129 were slow growing rhizobia（belonging to Bradyrhizobium elkanii, B. japonicum, B. diazoefficiens, B. yuanmingense and B. liaoningense）and 7 were fast-growing rhizobia（belonging to Sinorhizobium fredii and Sinorhizobium sp.）. 2）Six rhizobacterial strains that inhibited the pathogenic fungus of soybean root rot（Fusarium solani）were isolated（B1-B6, belonging to Bacillus velezensis, B. subtilis, B. licheniformis, B. cereus and B. megaterium）and four of them（B1, B2, B4 and B5）had the ability of producing IAA. 3）A total of 28 representative rhizobial strains with the tolerance to salt or drought were obtained, among which two strains, B. japonicum GN1 and B. diazoefficiens GN10, demonstrated dual resistance to salt and drought stress. Additionally, GN10 was determined to be the most efficient symbiotic strain among all representative strains through screening of their relative symbiotic phenotypes. 4）The results of co-inoculation phenotype of soybean indicated that B. velezensis B4 significantly enhanced the symbiotic performance of B. diazoefficiens GN10 compared to other tested bacillus strains.【Conclusion】We successfully obtaine one slow-growing soybean-nodulating rhizobial strain with dual-tolerance to salt and drought and one B. velezensis strain B4 that significantly enhance the symbiotic performance, providing strong support for the development and application of high-efficiency composite rhizobia inoculants. Table and Figures | Reference | Related Articles | Metrics

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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 Biotechnology Bulletin    2025, 41 (7): 128-138.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-1255 Abstract （3211）   HTML （7）    PDF（pc） （3539KB）（65）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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Mining the Core Genes Being Tolerant to Cadmium in Wild Soybean by WGCNA ZHU Li-juan, ZHANG Kai, WEN Xiao-lei, CHU Jia-hao, SHI Feng-yu, WANG Yan-li Biotechnology Bulletin    2025, 41 (8): 124-136.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-1113 Abstract （3003）   HTML （10）    PDF（pc） （11908KB）（80）       Knowledge map    Save Objective Wild soybean has characteristics of tolerance to stress, gradually becoming a germplasm resource for improved cultivated soybeans. It is significant to clarify the molecular regulation mechanism of tolerance to cadmium in wild soybeans and provide a basis for cultivating tolerant soybean varieties. Method In this study, 200 wild soybeans in eastern Hebei were used as experimental materials. Wild soybean seedlings were treated with Hoagland nutrient solution containing 75 mol/L CdCl2, and the dry weight of the seedlings was measured. Transcriptome sequencing was performed on Cd-tolerant wild soybean R and sensitive material S at 24 h and 48 h under Cd treatment, respectively. KEGG and GO enrichment analyses were performed on differentially expressed genes (DEGs), and weighted gene co-expression network analysis (WGCNA) was used to detect Cd-tolerant core genes. Result The 200 wild soybean seedlings have shown remarkable differences under stress. Compared with the control, the shoot dry weight and root dry weight of seedlings under cadmium treatment significantly reduced. Transcriptome analysis showed that 6 443 and 4 496 DEGs were identified in R and S materials, respectively. GO and KEGG analyses found that these DEGs were enriched in photosynthesis and stress response pathways. Combined with WGCNA, the key turquoise and blue modules were significantly correlated with cadmium tolerance in wild soybeans. According to the connectivity and functional annotation of the genes in the module, it was predicted that 8 genes, such as LOC114376469, LOC114412091, LOC114388638, and LOC114399512 may play a role in the process of cadmium stress in wild soybean. Conclusion Two specific modules related to the tolerance of wild soybean to cadmium are identified, and LOC114376469, LOC114412091, LOC114388638, and LOC114399512 are screened to be the core genes related to the tolerance of wild soybean to cadmium. Table and Figures | Reference | Related Articles | Metrics

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Development of Functional Molecular Markers for Tomato Fruit Weight Gene and Population Genotyping Analysis HOU Ya-tao, LI Ying-hui, DENG Lei, LI Chang-bao, LI Chuan-you, SUN Chuan-long Biotechnology Bulletin    2025, 41 (4): 98-105.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0101 Abstract （2884）   HTML （14）    PDF（pc） （2376KB）（158）       Knowledge map    Save Objective This study is aimed to develop and apply molecular markers to identify and analyze the patterns of genetic variation in Fruit weight 3.2 (Fw3.2) and Fruit weight 11.3 (Fw11.3), two pivotal genes involved in the regulation of tomato (Solanum lycopersicum) fruit weight, at different domestication stages. It is expected to provide effective tools for the precise identification and utilization of tomato genetic resources, as well as the rapid domestication of wild germplasm. Method Fragment length polymorphism molecular markers were designed specifically based on the copy number variation (CNV) of Fw3.2 and the 1.4 kb large fragment insertion/deletion (Indel) variation at the 3′ end of Fw11.3. These markers were used to conduct genotype analysis on a collection of 259 tomato germplasms (including 79 Solanum pimpinellifolium (PIM) accessions, 95 Solanum lycopersicum var. cerasiforme (CER) accessions and 85 big-fruited S. lycopersicum accessions (BIG)), determining variation of Fw3.2 and Fw11.3 and their distribution within different tomato populations. Result By the molecular markers developed in this study, the CNVs (fw3.2WT and fw3.2dup ) of the Fw3.2 gene and the large fragment Indel variants of the Fw11.3 gene (fw11.3-WT and fw11.3-D) were successfully identified. Within PIM population, the frequencies of the large-fruit alleles fw3.2dup and fw11.3-D were 0 and 2.53%, respectively. In the CER population, these allele frequencies increased to 10.53% for fw3.2dup and 8.42% for fw11.3-D. Notably, in the BIG population, the prevalence of fw3.2dup and fw11.3-D alleles rose significantly to 69.41% and 92.94%, respectively. Conclusion Both fw3.2dup and fw11.3-D alleles significantly increase tomato fruit weight. Furthermore, these alleles lead to a synergistic effect that further enhances fruit weight in tomatoes. During the domestication and genetic improvement of tomatoes, the frequency of these large-fruit alleles has progressively increased, mirroring the evolutionary trajectory toward larger fruit size. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in the Biological Metabolic Pathway and Functions of Plant Carotenoids LIU Yuan, ZHAO Ran, LU Zhen-fang, LI Rui-li Biotechnology Bulletin    2025, 41 (5): 23-31.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-1057 Abstract （2468）   HTML （47）    PDF（pc） （1104KB）（313）       Knowledge map    Save Carotenoids are a group of lipophilic isoprenoid metabolites, which are widely distributed in animals, plants, and microorganisms. It is demonstrated that α-carotene, β-carotene and lutein are the main components of carotenoids. In addition to influencing the ornamental quality of plants as natural pigments, carotenoids play a crucial function in photosynthesis. Moreover, they are also critically important for humans as precursors of vitamin A synthesis. The carotenoid metabolic pathway, including the precursor synthesis pathway, the biosynthesis and degradation pathway, is well established and has been widely explored in plants. With the intensive studies of carotenoid metabolism, the key genes and their functions have been identified, facilitating a deeper understanding of the regulatory mechanisms underlying plant carotenoid metabolism. This review focuses on the carotenoid metabolic pathway, with emphasis on recent progresses in biological functions of carotenoids, particularly in photoprotection, the agricultural and horticultural application, the resistance to abiotic stresses, and the regulation of plant hormone signaling. This review will offer a valuable information for the elucidation of carotenoids metabolism and facilitating selective molecular breeding. Table and Figures | Reference | Related Articles | Metrics

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Gene Cloning and Functional Analysis of the Anthocyanin-related VcGSTF19 Gene in Blueberry （Vaccinium corymbosum L.） ZHANG Yong-yan, GUO Si-jian, LI Jing, HAO Si-yi, LI Rui-de, LIU Jia-peng, CHENG Chun-zhen Biotechnology Bulletin    2025, 41 (9): 139-146.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0190 Abstract （2432）   HTML （13）    PDF（pc） （2662KB）（81）       Knowledge map    Save Objective Glutathione S-transferase Phi (F) subfamily members (GSTFs) play key roles in anthocyanin accumulation and transportation in many plants. To study the function of blueberry (Vaccinium corymbosum L.) GSTF, the blueberry anthocyanin-related GSTF19 gene was cloned and its function was studied. Method Reverse transcription PCR was used to clone the coding sequence (CDS) of VcGSTF19 related toanthocyanin from blueberry and construct overexpressing vector. Based on the blueberry fruit transient overexpression and Arabidopsis tt19 complementary transgenic transformation assays, the functions of VcGSTF19 were further studied. Result Transient overexpression of VcGSTF19 significantly promoted the accumulation of anthocyanins in blueberry peel. The transient overexpression of VcGSTF19 improved significantly the anthocyanin accumulation by 6.61-fold in blueberry fruit peel, compared to the empty vector control. Moreover, its transient overexpression significantly upregulated the expressions of anthocyanin biosynthesis related structural genes, including VcCHS, VcCHI, VcF3H, VcDFR, VcANS and VcUFGT. Heterologous transformation of Arabidopsis tt19 mutant with VcGSTF19 restored the anthocyanin accumulation in the mutant, with anthocyanin contents in rosette leaves accounting for approximately 6.21-fold of the tt19 mutant. Conclusion Blueberry VcGSTF19 gene plays an important role in anthocyanin accumulation and transport. Table and Figures | Reference | Related Articles | Metrics

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Mechanisms and Application Research Progress of Bacterial Genomic Homologous Recombination Mediated by Single-stranded DNA Annealing Protein YIN Hao, YOU Liu-chao, HAN Rui, GAO Peng-cheng, FU Lei, CHU Yue-feng Biotechnology Bulletin    2025, 41 (1): 39-48.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0457 Abstract （2091）   HTML （20）    PDF（pc） （2451KB）（256）       Knowledge map    Save Genome editing technology is an important tool for studying the gene function, drug resistance mechanism, and pathogenic mechanism of bacteria and other microorganisms. Homologous recombination is one of the key methods for bacterial genome editing. The conventional endogenous pathways in bacteria suffer from low efficiency. However, a single-stranded DNA annealing protein（SSAP）from bacteriophages, which has shown gene editing efficiencies far surpassing those of endogenous pathways. This protein has the characteristics of single-stranded DNA binding activity and mediates genome directed recombination, making it an extremely promising tool for genome editing. This article provides an overview of the basic principles of homologous recombination, the fundamental components of the SSAP-mediated homologous recombination pathway from phages, recombination mechanism models, and application strategies. The aim is to assist in further elucidating the process of homologous recombination mediated by SSAP to provide technical support for studying the functions and pathogenic mechanisms of more bacterial genes, and to develop engineering strains. It also provides technical support for bacteria lacking gene editing methods. Table and Figures | Reference | Related Articles | Metrics

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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 Biotechnology Bulletin    2025, 41 (7): 226-236.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-1056 Abstract （2087）   HTML （4）    PDF（pc） （6181KB）（67）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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Improving the Efficiency of Gene Editing by Optimizing sgRNA in Aspergillus tubingensis LIANG Li-cun, WANG Ke-fen, SONG Zu-huan, LIU Meng-ting, LI Jia-yu, LUO Hui-ying, YAO Bin, YANG Hao-meng Biotechnology Bulletin    2025, 41 (3): 62-70.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0784 Abstract （2066）   HTML （12）    PDF（pc） （2071KB）（83）       Knowledge map    Save Objective In the CRISPR/Cas9 gene editing system, sgRNA is one of the important gene editing elements. It binds to the Cas9 protein and complements the genomic DNA through the spacer sequence to guide the Cas9 protein to accurately cut the genome. In order to improve the gene editing efficiency of Aspergillus tubingensis, sgRNA optimization is a feasible strategy. Method The promoter and hairpin structure of sgRNA were optimized, and the gene editing efficiency was verified in A. tubingensis. Result The gene editing efficiency of sgRNA with “lock” structure was 9.37% higher than that of the control group when gene editing was performed by RNP method. When sgRNA was expressed in vivo, the gene editing efficiency of tRNAGly15 and tRNAGly17 promoters was 14%-16% higher than that of 5S rRNA promoter, respectively. Using the tRNAGly15 promoter to express sgRNA with a “lock” structure increased the white spore rate and increased the gene editing efficiency of A. tubingensis to 96%. Conclusion The “lock” structure and two promotors of tRNAGly15 and tRNAGly17 may improve the gene editing efficiency of A. tubingensis. Table and Figures | Reference | Related Articles | Metrics

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Genome-wide Identification and Expression Analysis of bZIP Transcription Factor Family in Dendrobium officinale ZENG Dan, HUANG Yuan, WANG Jian, ZHANG Yan, LIU Qing-xia, GU Rong-hui, SUN Qing-wen, CHEN Hong-yu Biotechnology Bulletin    2025, 41 (8): 197-210.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0094 Abstract （2048）   HTML （25）    PDF（pc） （6925KB）（94）       Knowledge map    Save Objective To identify the bZIP gene family of Dendrobium officinale,and clarify the biological function of DobZIP transcription factors through bioinformatics and gene expression analysis, thus aiming to provide theoretical guidance for further exploration of the molecular mechanism of DobZIP transcription factors. Method Based on the genome data of Dendrobium officinale in the IMP database, we identified the members of DobZIP gene family, and performed multiple of bioinformatics analysis. Furthermore, we used transcriptome data and real-time fluorescent quantitative PCR technology to analyze the tissue expression pattern and abiotic stress expression of DobZIP transcription factors. Result A total of 54 DobZIP genes were identified. None of the DobZIP proteins had signal peptides, and only DobZIP54 had a transmembrane domain. The subcellular localization of the DobZIP proteins were mainly in the nucleus. Members of the DobZIP family were classified into 10 subfamilies, and members of the same subfamily contained similar structures and motifs. DobZIP genes were unevenly distributed on 18 chromosomes, with two pairs of genes showing tandem relationships. There were multiple collinear relationships between species, and DobZIP family had a high homology with the OsbZIP family in rice. Cis-acting elements analysis indicated that the DobZIP family members all contained multiple cis-acting elements involved in growth and development and response to abiotic stress. Gene expression pattern analysis showed that the DobZIP family members had tissue-specific expression, and DobZIP4、DobZIP10、DobZIP28、DobZIP32 and DobZIP37 were induced by cold, drought and NaCl stress to varying degrees. In addition to DobZIP32 and DobZIP37, DobZIP4, DobZIP10 and DobZIP28 were also induced by high temperature stress. Conclusion A total of 54 DobZIP genes are identified in the genome of Dendrobium officinale. DobZIP genes was in tissue-specific expression, and DobZIP4, DobZIP10, DobZIP28, DobZIP32 and DobZIP37 are induced to varying degrees under cold, drought, and NaCl stress. Table and Figures | Reference | Related Articles | Metrics

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Functional Analysis of Soybean GmPM31 Gene Promoter Involvement in Response to High Temperature and Humidity Stress LIN Zi-yi, WU Yi-zhou, YE Fang-xian, ZHU Shu-ying, LIU Yan-min, LIU Su-shuang Biotechnology Bulletin    2025, 41 (3): 90-97.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0435 Abstract （2004）   HTML （13）    PDF（pc） （3977KB）（160）       Knowledge map    Save Objective To study the function of GmPM31 promoter on seed vigor formation in response to high temperature and high humidity stress, and to lay a foundation for comprehensively unravelling the function of soybean small heat shock proteins involved in the response to high temperature and high humidity stress. Method Homozygous T3 transgenic Arabidopsis thaliana with GmPM31 promoter was treated with high temperature and high humidity stress (40℃/100% relative humidity), and wild-type A. thaliana was used as control to detect its tolerance and stomatal opening. Analysis of GUS gene expression driven by the soybean GmPM31 promoter by GUS histochemical staining and quantitative real-time PCR. The seed vigor was detected by 2, 3, 5-triphenyltetrazolium chloride (TTC) staining. Result Compared with the control group, the tolerance of GmPM31-transgenicpromotor A. thaliana to high temperature and high humidity increased. The activity of GUS increased, and they were expressed in the leaves, roots and flowers, and germination rate and vigor increased. Conclusion GmPM31 promoter improve the resistance of seeds to high temperature and high humidity stress and the ability to resist deterioration. Table and Figures | Reference | Related Articles | Metrics

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Advances in Spatial Metabolomics in Medicinal Plants LIU Yu-shi, LI Zhen, ZOU Yu-chen, TANG Wei-wei, LI Bin Biotechnology Bulletin    2025, 41 (9): 22-31.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0276 Abstract （1992）   HTML （24）    PDF（pc） （838KB）（167）       Knowledge map    Save Mass spectrometry imaging (MSI) is an innovative molecular imaging technique offering significant advantages, including label-free analysis, high molecular coverage, and exceptional sensitivity. MSI has been widely used in studies on the spatial distribution of metabolites in tissues. The continuous advancement of this technique has brought spatial metabolomics, a methodology that integrates MSI with metabolomics. This approach allows for simultaneous localization and imaging analysis of metabolites in tissues across diverse biological samples, encompassing animals, plants, and microorganisms. Recently, spatial metabolomics has demonstrated great potential in medicinal plant research as a crucial technique for precise spatial localization of metabolites in tissues. This review first presents the basic principles and experimental procedures of spatial metabolomics, followed by comparisons of several major MSI techniques, including matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI), desorption electrospray ionization mass spectrometry imaging (DESI-MSI), and secondary ion mass spectrometry imaging (SIMS-MSI), where distinctive advantages and limitations of each approach are delineated, as well as key aspects in sample preparation and data processing are addressed. The review subsequently focuses on the applications of spatial metabolomics in medicinal plants, including visualizing the spatial distribution and accumulation pattern of metabolites, clarifying the biosynthesis and transport sites of metabolites, and discovering functional genes associated with biosynthetic pathways in medicinal plants. Finally, this review discusses the current challenges and prospects of spatial metabolomics, aiming to provide a new perspective for investigations in medicinal plant research. Table and Figures | Reference | Related Articles | Metrics

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Gravity-driven Microfluidic Chip Based on RPA-CRISPR/Cas12a for the Rapid Detection of Mycobacterium tuberculosis GAO Chang, ZHUANG Tian-chi, LI Ning, LIU Yun, GU Peng-fei, ZHAO Xin-yi, JI Ming-hui Biotechnology Bulletin    2025, 41 (5): 62-69.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0955 Abstract （1974）   HTML （14）    PDF（pc） （1982KB）（624）       Knowledge map    Save Objective To develop a rapid and efficient method for detecting Mycobacterium tuberculosis (MTB), providing a technical solution for rapid screening of tuberculosis (TB) in grassroots units and remote areas, thereby enhancing TB prevention and control efforts. Method RPA (recombinase polymerase amplification) primers and crRNA were designed, and a gravity-driven microfluidic chip was introduced to establish an RPA-CRISPR/Cas12a on-chip detection method. The sensitivity and specificity of this method were further analyzed using samples from suspected TB patients for on-chip detection and sputum culture, comparing the established method with the sputum culture method for consistency. Result The established RPA-CRISPR/Cas12a on-chip detection method achieves MTB detection within 30 min, with a limit of detection (LOD) of 1 copy/μL. Using sputum culture identification as a reference standard, the on-chip detection method demonstrated a sensitivity of 91.11%, specificity of 94.34%, positive predictive value of 93.19%, negative predictive value of 92.59%, and accuracy of 92.86%, with a Kappa value of 0.856. Conclusion The developed RPA-CRISPR/Cas12a on-chip detection method presents high sensitivity and specificity, characterized by rapid, simple, and convenient. Table and Figures | Reference | Related Articles | Metrics

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Effects of Exogenous 2，4-Epibrassinolide on Physiological Characteristics of Daucus carota L. Seedlings under Cadmium Stress ZHOU Zhi-guo, FAN Shuang-hu, DENG Chen, FENG Xue Biotechnology Bulletin    2025, 41 (5): 165-174.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-1066 Abstract （1907）   HTML （8）    PDF（pc） （2236KB）（49）       Knowledge map    Save Objective This study is aimed to investigate the impact of varying concentrations of 2,4-epibrassinolide on growth indices, physiological parameters, and the expression of cadmium (Cd) uptake and transporter-related genes in carrot (Daucus carota L.) seedlings subjected to Cd stress. The findings provide theoretical insights into Cd resistance research in carrots. Method The "Hongxin 4" carrot cultivar (Daucus carota L.) was selected for the experimental materials. Using a hydroponic system, we examined the effects of different concentrations (0.1, 0.5, 1, and 1.5 μmol/L) of 2,4-epibrassinolide on the growth traits, physiological indicators, and photosynthesis of carrot seedlings under 0.1 mmol/L CdCl2 stress. Result The growth of carrot seedlings was significantly inhibited under 0.1 mmol/L Cd stress. However, the exogenous application of 1 μmol/L 2,4-epibrassinolide inhibited cadmium ion absorption and transport, promoted the root length and plant height, elevated the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) in the leaves, increased the contents of photosynthetic pigments, enhanced the photosynthetic efficiency, and suppressed the expressions of Cd uptake and transport-related genes (DcHMA3, DcHMA4, DcCAX2 and DcCAX4),thereby maintaining the balance of redox metabolic pathways in the plants. Conclusion 2,4-epibrassinolide promoted carrot root growth, enhanced antioxidant enzyme activity, improved photosynthetic efficiency, regulated the glutathione-phytochelatin (GSH-PC) metabolic pathway, and modulated the expressions of related genes to alleviate Cd stress-induced damage in carrot seedlings. The optimal concentration of 2,4-epibrassinolid for carrots was found to be 1 μmol/L. Table and Figures | Reference | Related Articles | Metrics

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Efficient CRISPR Gene Editing in Rice Assisted by RUBY DENG Mei-bi, YAN Lang, ZHAN Zhi-tian, ZHU Min, HE Yu-bing Biotechnology Bulletin    2025, 41 (8): 65-73.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0339 Abstract （1850）   HTML （19）    PDF（pc） （2682KB）（209）       Knowledge map    Save Objective To develop a RUBY-assisted CRISPR/Cas9 gene-editing vector (GCR) enabling visual screening of transgenic plants and gene-editing events. Method The GCR vector was constructed by coupling RUBY with the Cas9/gRNA expression cassette. Using OsAGO2, OsAGO3, and OsAGO7 in rice (Oryza sativa) as target genes, two multi-gene editing vectors (GCR-237-1 and GCR-237-2) were designed. These vectors were introduced into rice (Zhonghua 11, ZH11) via Agrobacterium-mediated genetic transformation to generate gene-edited plants. Result After the transformation of GCR-237-1 and GCR-237-2 into rice callus, RUBY markers effectively indicated transgenic positive events, and the target genes of stable transformed red plants were efficiently edited. Conclusion The GCR vector enables effective gene editing in rice, and transgenic or gene-edited plants can be rapidly identified through direct visual observation. Table and Figures | Reference | Related Articles | Metrics

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Analysis of Soybean Pod Dehiscence Habit Based on Whole Genome Association Analysis and Genetic Diversity SONG Ying-pei, WANG Can, ZHOU Hui-wen, KONG Ke-ke, XU Meng-ge, WANG Rui-kai Biotechnology Bulletin    2025, 41 (2): 97-106.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0358 Abstract （1801）   HTML （7）    PDF（pc） （2722KB）（342）       Knowledge map    Save Objective Investigating molecular mechanisms and germplasm resources of pod dehiscence resistance would provide evidence for uncovering the genetic and domestication mechanism of pod dehiscence resistance in soybean as well as accelerating the breeding of new soybean varieties with pod dehiscence resistance on southern China. Method With genome-wide association studies (GWAS) and genetic diversity analysis between wild and cultivated soybean populations, the genes related to soybean pod dehiscence habit were explored. The phenotype determination of 302 soybean varieties were conducted, and GWAS was performed using 95 ‍744 single nucleotide polymorphism markers in 2 years. And analysis of genetic diversity in QTL segment was carried, with the resequencing data of 1 308 cultivated soybeans and 203 wild soybeans. Result Through GWAS, three QTL loci that explained greater than 10% of the observed phenotypic variance were detected, namely qPdh-Chr08 (Gm08: 3048312), qPdh-Chr15 (Gm15: 312814), and qPdh-Chr16 (Gm16: 29951529). Among them, qPdh-Chr16 was known gene pdh1 (Pod dehiscence habit 1). Combining the resequencing data of wild and cultivated soybeans, it was found that there were population differentiation sequences caused by artificial selection in regions of qPdh-Chr08 and qPdh-Chr15. And two important candidate genes Glyma.08G038600 and Glyma.15G003600 were identified based on population differentiation sequences analysis. Through functional annotation, it was found that these two genes were involved in the metabolism of auxin and lignin, respectively. Conclusion Two new QTL loci qPdh-Chr08 and qPdh-Chr15 of soybean pod dehiscence habit were detected using two-year’s data. Differential genes between wild and cultivated soybean are identified within the QTL region, and two important candidate genes are ultimately identified. Table and Figures | Reference | Related Articles | Metrics

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Advances in the Biosynthesis of 5-aminolevulinic Acid （5-ALA） LYU Huan-huan, ZHANG Gao-yang, WANG Sai-di, SUN Zhong-ke, LI Cheng-wei, LUO De-ping Biotechnology Bulletin    2025, 41 (11): 75-88.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0153 Abstract （1772）   HTML （6）    PDF（pc） （2287KB）（133）       Knowledge map    Save 5-aminolevulinic acid (5-ALA) is a non-protein amino acid naturally occurring in living organisms, and has been widely used in the fields of medicine, health care, agriculture and animal additives, etc. 5-ALA is mainly synthesized by chemical and biological methods, but the complexity of the chemical synthesis process and cost problems have limited its large-scale development, and biosynthesis method has highlighted its great potential for industrial development due to its advantages of environmental protection and high efficiency. The biosynthesis method, with its advantages of environmental protection and high efficiency, has shown its great potential for industrial development. In recent years, the biosynthesis of 5-ALA has gradually become a research hotspot due to the rapid development of interdisciplinary disciplines such as synthetic biology and metabolic engineering. In this paper, we reviewed the development of 5-ALA biosynthesis in the past decades, summarized the progress of various metabolic engineering strategies based on the C4 and C5 pathways, cofactor regeneration pathway, tricarboxylic acid cycle and other pathways for the construction of high-efficiency cell factories, and highlighted the application and significance of the dynamic regulation based on biosensors and the establishment of high-throughput screening methods in the metabolic engineering modification of the bacterial strains. We also put forward several strategies and prospects for further improving the synthetic yield of 5-ALA and breaking the bottleneck of its industrial application, with a view to providing certain references and research bases for the efficient synthesis and industrial manufacturing of 5-ALA. Table and Figures | Reference | Related Articles | Metrics

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Differential Accumulation of Carotenoids in Ludisia discolor under Different Light Qualities Based on Multiomics ZHANG Yu-xuan, ZHANG Shi-yi, CHEN Hui-fang, CAI Kun-xiu, LI Chen-ye, YANG Jun-jie, ZHENG Tao, QIU Ming-yue, YANG You-si-yuan, CHEN Ying Biotechnology Bulletin    2025, 41 (10): 98-109.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0379 Abstract （1739）   HTML （5）    PDF（pc） （2829KB）（88）       Knowledge map    Save Objective This study is aimed to investigate the effects of different light qualities on the accumulation of carotenoid compounds in the leaves of Ludisia discolor and their molecular mechanisms, which may provide a theoretical reference for the standardized cultivation of this medicinal plant. Method This study used Ludisia discolor ‘Minre Yuanshuai’ to analyze the metabolic and transcriptional regulatory mechanisms under white light (W), blue light (B), and yellow light (Y) treatments. Liquid chromatography-mass spectrometry (LC-MS/MS) and high-throughput transcriptome sequencing (RNA-seq) technologies were utilized to obtain metabolomic and transcriptomic data, respectively. The white light group was used as a control to analyze the effects of blue and yellow light on carotenoid content, related metabolites and gene expression. RT-qPCR was adapted to verify the expression patterns of eight key genes closely related to carotenoid synthesis. Result The blue light treatment significantly increased total carotenoid levels in L. discolor leaves, whereas yellow light treatment caused no substantial changes. The metabolomic results implied that there were 23 differential metabolites related to carotenoid synthesis, including xanthoxic acid, abscisic alcohol, strigolactone ABC-rings, and neurosporaxanthin. Transcriptome analysis revealed nine differentially expressed metabolic enzyme genes (such as CrtZ, Z-ISO, PSY) and six key transcription factors (ERF002, ERF059, ERF066), which may regulate carotenoid synthesis in response to gibberellin, methyl jasmonate, auxin, salicylic acid, and abscisic acid signals. RT-qPCR confirmed that the eight key genes play a potential role in the regulation of carotenoid metabolism. Conclusion Under blue light treatment, ERF and bZIP transcription factors in L. discolor leaves regulate the expressions of downstream enzyme genes by binding to cis-acting elements related to gibberellins, methyl jasmonate, growth hormone, salicylic acid, and abscisic acid, leading to significant accumulation of carotenoid-related metabolites. Table and Figures | Reference | Related Articles | Metrics

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Cloning and Functional Characterization of 6-OMT Gene Related to Isocorydine Biosynthesis in Dactylicapnos scandens LI Ming, LIU Xiang-yu, WANG Yi-na, HE Si-mei, SHA Ben-cai Biotechnology Bulletin    2025, 41 (2): 309-320.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0544 Abstract （1736）   HTML （14）    PDF（pc） （4284KB）（106）       Knowledge map    Save Objective (S)-norcoclaurine O-methyltransferase (6-OMT) is the key rate-limiting enzyme in the biosynthesis of isocorydine. This study is aimed to verify the function of the 6-OMT gene from Dactylicapnos scandens through cloning and in vitro enzyme activity assays, thereby laying the foundation for elucidating the isocorydine biosynthetic pathway in D. scandens. Method The DsOMT gene was mined from the transcriptome data of D. scandens. The full-length cDNA sequence was obtained via PCR amplification, and the protein structure of DsOMT was analyzed using bioinformatics. The expressions of the DsOMT gene in different tissues were examined. A pET-28a-DsOMT prokaryotic expression vector was constructed and transferred into Escherichia coli BL21(DE3) for induced expression. The protein was then purified and subjected to in vitro enzyme assays to characterize its function. Result Four DsOMT candidate genes, named DsOMT07, DsOMT08, DsOMT010, and DsOMT012, were identified from the transcriptome data, and their full-length cDNA sequences were successfully amplified. All four DsOMT proteins lacked transmembrane domains and signal peptides, classifying them as extramembrane proteins. Phylogenetic analysis indicated that these genes were closely related to the 6-OMT subfamily. Amino acid sequence analysis suggested potential catalytic activity at the 6-OH site of the upstream pathway for (S)-norcoclaurine. Expression profiling revealed that these four DsOMT genes were highly expressed in the roots. SDS-PAGE results showed that DsOMT proteins were highly soluble and efficiently expressed in Escherichia coli. In vitro enzyme assays demonstrated that DsOMT010 catalyzed the O-methylation of the C6 position of (S)-norcoclaurine, forming (S)-coclaurine. Conclusion Four DsOMT genes are successfully cloned and identified as extramembrane proteins closely related to the 6-OMT subfamily. These genes show high expressions in the roots. Additionally, heterologous expression of DsOMT is achieved in E. coli, and the purified proteins are characterized for their in vitro enzyme functions. Among them, DsOMT010 is identified as a (S)-norcoclaurine C-6 O-methyltransferase. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in the Biosynthesis of Polylactic Acid LU Tian-yi, LI Ai-peng, FEI Qiang Biotechnology Bulletin    2025, 41 (4): 47-60.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0982 Abstract （1715）   HTML （34）    PDF（pc） （2620KB）（148）       Knowledge map    Save Polylactic acid (PLA) is a non-natural biodegradable plastic polymerized from lactic acid. It demonstrates remarkable biodegradability and serves as a significant alternative to traditional petroleum-based plastics, particularly in the background of achieving carbon peak and carbon neutrality. As a typical carbon-neutral material, PLA is increasingly recognized as a crucial raw material for fostering national economic and social development. Currently, PLA is primarily produced through a combination of biological fermentation and chemical polymerization. However, this systhesis method is fraught with complexities, high costs, and potential risks of toxic residue accumulation. Consequently, the exploration of more environmentally sustainable and efficient production methods has become a central focus in the field of PLA synthesis. With the rapid advancements in synthetic biology, protein engineering and metabolic engineering, the key enzymes of PLA biosynthesis have been gradually identified and modified, the PLA biosynthesis pathways have been designed and assembled, leading to the establishment of cell factories for one-step synthesis of PLA utilizing microorganisms with industrial properties, thereby offering a new solution for the green synthesis of PLA. Nonetheless, the biosynthesis method is confronted with challenges, including low PLA yield and suboptimal product performance, which hinder its alignment with industrial standard. Therefore, enhancing the efficiency of PLA biosynthesis and improving product performance have emerged as critical objectives in the development of PLA biosynthesis technologies. In this paper, PLA and its synthesis methods were introduced briefly, and the advantages and disadvantages of chemical synthesis and biosynthesis were systematically analyzed. Subsequently, the pathways and key enzymes of PLA biosynthesis were summarized, and the regulatory strategies of PLA biosynthesis were summarized from the aspects of protein engineering and metabolic engineering. Finally, the key challenges and future research trends in the upgrading and development of PLA biosynthesis technology were systematically analyzed and prospected, aiming to provide useful reference for the design and development of more efficient and greener PLA biosynthesis system. Table and Figures | Reference | Related Articles | Metrics

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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 Biotechnology Bulletin    2025, 41 (7): 117-127.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-1266 Abstract （1689）   HTML （8）    PDF（pc） （5458KB）（130）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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Cloning of Gene CtbHLH128 in Safflower and Response Function Regulating Drought Stress LI Kai-jie, WU Yao, LI Dan-dan Biotechnology Bulletin    2025, 41 (8): 234-241.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0059 Abstract （1615）   HTML （14）    PDF（pc） （2621KB）（71）       Knowledge map    Save Objective Basic helix-loop-helix (bHLH) transcription factors represent one of the largest families of transcription factors in eukaryotes, and their members play crucial roles in plant growth, development, and stress responses. This study enriches the research on the function of the bHLH transcription factor family genes and lays an important foundation for analyzing the molecular regulatory mechanism of safflower (Carthamus tinctorius L.) seed development. Method Based on the complete CDS sequence of CtbHLH128 in the genome, primers were designed using Primer 6.0 software to clone and obtain the correct sequence of the gene, and bioinformatics was used to analyze the number of amino acids encoded by the gene, conserved domains, and phylogenetic tree. The CtbHLH128 fragment was constructed into a plant expression vector and transformed into Arabidopsis thaliana by Agrobacterium infection. Herbicide screening and PCR were used to identify positive strains. Drought stress was applied to the positive strains at the seedling and germination stages to explore the role of this gene in drought stress. Result The CtbHLH128 consists of 948 bp nucleotides, encoding 315 amino acids. The molecular weight of its encoded protein is 33 742.32 Da, and the theoretical isoelectric point is 9.02. CD-Search analysis results revealed that its C-terminus contained a typical HLH conserved domain located at amino acid residues 249-301. Phylogenetic tree revealed high similarity with bHLH128 gene in sunflower and lettuce. The CtbHLH128 gene was overexpressed in Arabidopsis, and three T3 positive strains were obtained through resistance screening and PCR identification. The positive strains were subjected to drought stress-related phenotypic analysis. The results revealed that transgenic Arabidopsis plants showed greater sensitivity to drought conditions at the seedling stage, but enhanced resistance to drought at the germination stage. Conclusion This suggests that the CtbHLH128 gene may possess potential functions in the responses to drought stress and seed germination. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in RNA Binding Proteins in Plant Disease Resistance LYU Yue, ZHANG Jie-wei, WANG Bo Biotechnology Bulletin    2025, 41 (6): 12-26.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0056 Abstract （1586）   HTML （11）    PDF（pc） （2242KB）（131）       Knowledge map    Save During their growth and development, plants are continuously exposed to complex environmental stresses that severely constrain their growth, agronomic traits, and productivity. To combat biotic stresses such as pathogen infection, plants have evolved multilayered sophisticated regulatory networks. In recent years, post-transcriptional regulation has emerged as a novel research hotspot in plant immunity, demonstrating unique advantages in the resistance to disease through dynamic regulation of messenger RNA (mRNA) metabolism. RNA-binding proteins (RBPs), functioning as core executors in plant resistance-to-resistane networks, act as "molecular switches" in plant-pathogen interactions by recognizing specific RNA motifs to regulate critical processes including pre-mRNA alternative splicing, mRNA stability, alternative polyadenylation (APA), translation efficiency, and RNA modifications. This review systematically elaborates RBP-mediated post-transcriptional regulatory mechanisms and their functions during plant-pathogen interactions. For instance, at the pathogen-recogned stage, RBPs regulate mRNA stability of immune receptors to enable rapid activation of defense signals. During disease resistance responses, RBPs mediate alternative splicing of resistance genes to generate transcript variants with distinct subcellular localization or functional activities. Recent studies also reveal novel pathways in plant immunity where RNA epigenetic modifications (e.g., m6A) regulate RBP recruitment efficiency. This article provides in-depth analysis of the multilayered defense systems constructed through RBPs and their molecular regulatory mechanisms, while proposing future research directions including deciphering RBP-mediated disease resistance mechanisms, modifying RBP regulatory elements through multi-omics integration, and developing novel disease-resistant breeding strategies. Comprehensive understanding of RNA regulatory codes in plant immunity will offer theoretical foundations for creating broad-spectrum resistant germplasm and provide crucial references for developing innovative green control strategies. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in Temperature Regulation of Chloroplast Development in Crops JIANG Yun-bo, CHEN Xue-xue, ZHAO Yu-sheng Biotechnology Bulletin    2025, 41 (10): 20-31.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0756 Abstract （1569）   HTML （45）    PDF（pc） （1040KB）（134）       Knowledge map    Save Chloroplasts are the key organelles responsible for photosynthesis in plants. As semi-autonomous structures unique to plant cells, their development is regulated by both internal growth and developmental signals as well as external environmental cues. Among these factors, temperature plays a critical role in shaping chloroplast development. It influences various aspects, such as the chloroplast membrane system, morphology, plastid division, and differentiation, ultimately impacting their development and function. This review begins with a concise overview of the structure, function, and developmental processes of chloroplasts in response to temperature fluctuations. It then summarizes research progress on plant leaf color mutants, shedding light on the molecular basis of temperature regulation in chloroplast development. Additionally, the review explores the mechanisms of temperature regulation in chloroplasts across three levels: Transcriptional regulation of chloroplast genes, post-transcriptional regulation of chloroplast genes, and protein synthesis and homeostasis in chloroplast. Finally, it discusses future directions, offering insights into how leaf color-related genes can be leveraged to uncover the mechanisms underlying temperature-responsive chloroplast development, with applications in both research and agricultural practices. In conclusion, this review thoroughly examines how environmental temperature changes affect chloroplast development and physiological functions in crops, while also exploring the broader implications for photosynthesis. By addressing these impacts, it aims to provide theoretical insights and practical guidance for developing molecular breeding strategies that enhance crop adaptability and photosynthetic efficiency in the face of global climate change. Table and Figures | Reference | Related Articles | Metrics

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Identification and Expression Analysis of the C3H Gene Family in Grona styracifolia across Different Varieties HUANG Jin-heng, HUANG Xi, ZHANG Jia-yan, ZHOU Xin-yu, LIAO Pei-ran, YANG Quan Biotechnology Bulletin    2025, 41 (4): 243-255.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0883 Abstract （1565）   HTML （15）    PDF（pc） （4367KB）（648）       Knowledge map    Save Objective The C3H family genes play a significant role in regulating plant growth and development. Investigating the characteristics and functions of the C3H gene family in Grona styracifolia will lay a foundation for studying its roles in the physiological and morphological formation of this species. Method Based on the comparison of transcriptome data between ‘Guangyao Da 1’ and wild G. styracifolia, the C3H family genes were identified. Bioinformatics techniques were utilized to analyze the physicochemical properties, chromosomal positions, phylogenetic relationships, conserved motifs, and domains of these genes. real-time quantitative PCR (RT-qPCR) was applied to examine the expression patterns of the C3H family genes in different tissues and varieties of G. styracifolia. The correlation between C3H genes and key genes involved in flavonoid biosynthesis was analyzed. Additionally, the total flavonoid content in the stem tips and bases of different varieties were measured. Result The G. styracifolia C3H gene family consists of 27 members distributed across 9 chromosomes, primarily encoding basic, unstable, and hydrophilic proteins. Phylogenetic analysis shows that the G. styracifoliaC3H family genes are divided into three groups, with members in the same group being relatively conserved. Analysis of conserved motifs and domains indicates that the most frequently occurring motif is related to auxin regulation, and there is cooperative interaction with domains such as ANKYR among the G. styracifolia C3H family members. Expression pattern analysis reveals that the G. styracifoliaC3H family genes mainly function in the stems of it and are highly expressed at the stem tips of the ‘Guangyao Da 1’ variety. Correlation analysis found that the expressions of G. styracifolia C3H family genes are often positively correlated with key genes in flavonoid biosynthesis. Total flavonoid content measurement showed that at the stem tip, the total flavonoid content of ‘Guangyao Da 1’ is significantly higher than that of wild creeping G. styracifolia, while the opposite is true at the stem base. Conclusion The GsC3H family genes may indirectly influence the agronomic traits of ‘Guangyao Da 1’ G. styracifolia by regulating the biosynthesis of flavonoids in the stem segments. Table and Figures | Reference | Related Articles | Metrics

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Strengthening Fundamental Research on Photosynthesis to Support Crop Breeding for High Yield SUN Jing, YANG Yun-long, LIU Rong-zhi, YU Hong, LU Tie-gang Biotechnology Bulletin    2025, 41 (10): 1-5.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0634 Abstract （1564）   HTML （42）    PDF（pc） （497KB）（211）       Knowledge map    Save Strengthening fundamental research on the biology of high photosynthetic efficiency in crops is a critical pathway to breaking the bottleneck of grain yield per unit area and advancing seed industry innovation and agricultural modernization. Significant progress has been made in areas such as the structural analysis of photosynthetic membrane proteins, the discovery of high-photosynthetic-efficiency genes, and the simulation of C4 pathways. Technologies like gene editing and synthetic biology have preliminarily enhanced the solar energy utilization efficiency in crops such as soybeans and rice. However, challenges persist in China’s fundamental research on high photosynthetic efficiency, including insufficient systematicity in research, lagging phenotyping platforms, a shortage of interdisciplinary talent, and weak genetic evaluation systems. Moreover, amid intense global competition, China’s participation in major international collaborative projects remains limited. This review summarizes the progress and challenges in high-photosynthetic-efficiency crop research both domestically and internationally, highlighting breakthroughs in foundational studies, the potential of technological applications, and interdisciplinary trends. It systematically identifies the shortcomings in China’s high-photosynthetic-efficiency research and proposes targeted recommendations. Moving forward, it is recommended to strengthen top-level design and establish interdisciplinary research projects; develop multi-scale phenotyping platforms; accelerate the training of interdisciplinary professionals; advance innovations in gene editing and intelligent design technologies; and promote the creation of high-photosynthetic-efficiency germplasm and the translation of research outcomes into practice, thereby providing scientific and technological support for safeguarding food security. Reference | Related Articles | Metrics

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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 Biotechnology Bulletin    2025, 41 (7): 281-291.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-1245 Abstract （1562）   HTML （18）    PDF（pc） （2347KB）（97）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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A CRISPR-Cas12a-based Detection Method for Respiratory Syncytial Virus YAO Xue-chun, LI Lei, WANG Zhi-xian, SHENG Chang-zhong, ZHOU Zeqi, TAN Cherie S Biotechnology Bulletin    2025, 41 (1): 103-109.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0433 Abstract （1545）   HTML （15）    PDF（pc） （4457KB）（111）       Knowledge map    Save 【Objective】Respiratory syncytial virus（RSV）is the most common cause of acute respiratory infections in infants. This study is aimed to establish a rapid and specific detection method for RSV based on the combination of RT-ETA and CRISPR-Cas12a.【Method】Specific crRNA and ERA primers were designed and synthesized for conserved fragments of the RSV N gene, and the optimal primer-crRNA sequence combination was selected to establish the detection system. The sensitivity and specificity of the method were evaluated by detecting the generated fluorescent signal.【Result】According to the activation principle of Cas12a, RSV-crRNA1 for RSV A and RSV-crRNA2 for RSV B were designed and synthesized, respectively. There was a synergistic interaction when RSV-crRNA1 and RSV-crRNA2 were mixed into the detection system. Meanwhile, RSVA and RSVB could be detected simultaneously by the mixed crRNA. With the highest fluorescence intensity, the forward prime combined with the reverse primer（RSV-F + RSV-R3J）was selected as the optimal primer pair. This method was able to detect RSV at titers as low as 5×102 copies/mL within 35 min under the constant temperature condition of 39℃. And no cross-reaction with other pathogens was observed. Only RSV nucleic acids were detected, indicating it was of high specificity.【Conclusion】Based on the a RT-ERA-CRISPR/Cas12a technology, a method of detecting RSV with high sensitivity, strong speficity, apid, and low cost for RSV detection is successfully established. This method does not demand complicated instrumentation, and it can be applied in any fluorescence laboratory equipment capable of providing a constant temperature. Thus this method is suitable for point-of-care test of RSV, and may also for detecting other pathogen infections. Table and Figures | Reference | Related Articles | Metrics

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Screening, Identification and Biocontrol Potential Analysis of an Antagonistic Strain against Ralstonia solanacearum MU Xue-nan, WU Tong, ZHENG Zi-wei, ZHANG Yue, WANG Zhi-gang, XU Wei-hui Biotechnology Bulletin    2025, 41 (1): 276-286.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0598 Abstract （1534）   HTML （20）    PDF（pc） （6537KB）（298）       Knowledge map    Save 【Objective】Tomato bacterial wilt caused by Ralstonia solanacearum（Rs）is main disease of tomato. In this study, the antagonistic strain against R. solanacearum and with growth promoting was screened from tomato rhizosphere, and its physiological and biochemical characteristic and biocontrol effect were investigated, which may provide a theoretical basis for the further development of biological control agents.【Method】Filter paper slice method was employed to screen the antagonistic strain against Rs. Physiological and biochemical characteristic and 16S rRNA sequences were analyzed to identify the antagonistic strain. Pot experiments were conducted to evaluate the biocontrol and growth promoting effects of the antagonistic strain on bacterial wilt and tomato, respectively. 16S rRNA gene amplicon sequencing and fluorescence quantitative PCR were used to investigate the effect of antagonistic strain on the bacterial community in tomato rhizosphere.【Result】An antagonistic strain A72 was screened out from the tomato rhizosphere. Strain A72 was identified as Bacillus siamensis by 16S rRNA gene sequence, and it had the ability to resolve phosphorus, release potassium, produce IAA, secrete extracellular hydrolases and siderophore, and form biofilms. The pot experiments showed that its control effect against tomato bacterial wilt was 63.80%, and it significantly increased the root length, plant height, dry weight, fresh weight and chlorophyll content of tomato plants. Application of strain A72 significantly decreased the density of R. solanacearum and altered the structure and composition of bacterial community in tomato rhizosphere.【Conclusion】Strain A72 has a good growth promotion effect on tomato seedlings and may effectively control tomato bacterial wilt. Table and Figures | Reference | Related Articles | Metrics

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Advances in RNA Interference Technology for Plant Functional Genomics and Crop Improvement HUANG Wen-jing, REN Si-chao, LIN Li, WANG You-ping, WU Jian Biotechnology Bulletin    2025, 41 (9): 1-21.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0645 Abstract （1532）   HTML （33）    PDF（pc） （3684KB）（280）       Knowledge map    Save RNA interference (RNAi) is a sequence-specific gene silencing mechanism mediated by double-stranded RNA (dsRNA) and conserved in eukaryotes. Its molecular mechanism primarily involves the processing of dsRNA, the generation of small interfering RNA (siRNA) or microRNA (miRNA), and the sequence-specific degradation or translational inhibition of target mRNA mediated by the RNA-induced silencing complex (RISC). Based on this mechanism, RNAi technology has become a powerful tool for plant functional genomics research and demonstrates immense potential in the genetic improvement of crops for resistance to diseases and pests, as well as in green and sustainable pest control. This review systematically summarizes the discovery history and molecular mechanism of RNAi and focuses on its three core applications: Virus-induced gene silencing (VIGS) utilizes engineered viral vectors to achieve transient silencing of host genes, providing an efficient approach for plant gene functional studies. Host-induced gene silencing (HIGS) aims to breed transgenic crops with durable resistance to fungi, viruses, and pests by expressing RNAi molecules within the plant to target and silence key genes in pathogens or pests. Spray-induced gene silencing (SIGS), as a non-transgenic strategy, involves spraying designed target-specific dsRNA. Upon uptake by the target pathogen or pest, this dsRNA inhibits the expressions of their key genes, thereby interfering with their infection/infestation process, offering a novel, environmentally friendly strategy for disease and pest control. In the future, further advancements in RNAi technology are expected to benefit from the deep integration of synthetic biology, artificial intelligence, and nano-delivery systems to optimize target design, enhance silencing efficiency, and reduce off-target risks. With breakthroughs in key technologies and the overcoming of application bottlenecks, RNAi holds exceptionally broad application prospects for advancing plant science research and achieving sustainable agriculture. Table and Figures | Reference | Related Articles | Metrics

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Positive Regulation of Anthocyanin Biosynthesis by PfMYB80 Transcription Factor in Perilla frutescens LI Rui, HU Ting, CHEN Shu-wei, WANG Yao, WANG Ji-ping Biotechnology Bulletin    2025, 41 (6): 243-255.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-1211 Abstract （1528）   HTML （6）    PDF（pc） （3832KB）（106）       Knowledge map    Save Objective The R2R3-MYB transcription factor is mainly involved in regulating the biosynthesis pathways of secondary metabolites such as flavonoids and anthocyanins. To verify its function in the anthocyanin biosynthesis of perilla (Perilla frutescens (L.) Britt.) would lay the foundation for elucidating the role of R2R3-MYB transcription factors in regulating plant anthocyanin synthesis. Method This study employed bioinformatics analysis to identify the R2R3-MYB transcription factors across the entire genome of perilla, and to predict their physicochemical properties, phylogenetic evolution, chromosome localization, and cis-acting elements of the promoter. R2R3-MYB members potentially involved in regulating the biosynthesis ofperilla anthocyanin were screened through correlation analysis, and the highly expressed PfMYB80 gene coding sequence in leaves was cloned to explore the regulatory effect of PfMYB80 on anthocyanin synthesis in perilla and its response mechanism to red and blue light stress. Result A total of 186 R2R3-MYB members were identified. Phylogenetic analysis revealed that PfMYB80 and PfMYB146 from perilla are most closely related to the genes in subgroup S6 of Arabidopsis thaliana, which are known to regulate anthocyanin synthesis in plants, suggesting their potential involvement in the regulation of anthocyanin synthesis. Analysis of promoter cis-acting elements indicated that the promoter regions of Perilla R2R3-MYB genes contain light stress-responsive elements. RT-qPCR results showed that the expression of the PfMYB80 gene gradually increases during different developmental stages of Perilla leaves, consistent with the trend of anthocyanin synthesis, suggesting its possible role in anthocyanin biosynthesis. This protein is localized in the nucleus. Expression profile analysis of PfMYB80 and anthocyanin synthesis-related structural genes revealed that the expression of the LDOX gene in Perilla aligns with that of PfMYB80 and matches the trend of anthocyanin accumulation, indicating that the PfMYB80 transcription factor might regulate anthocyanin synthesis by directly controlling the transcriptional expression of the LDOX gene. Functional analysis through transgenic tobacco demonstrated that PfMYB80 responds to blue light induction and positively regulates anthocyanin synthesis. Conclusion The PfMYB80 transcription factor in Perillapositively regulate the biosynthesis of anthocyanins and respond to light response. Overexpression of PfMYB80 in tobacco under blue light treatment increases anthocyanin accumulation and enhances SOD enzyme activity and reduces POD enzyme activity and MDA content. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in the Function and Their Action Mechanism of Plant PPR Protein WANG Yi-fan, ZHU Hong-liang Biotechnology Bulletin    2025, 41 (6): 27-37.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-1185 Abstract （1516）   HTML （19）    PDF（pc） （1336KB）（131）       Knowledge map    Save RNA editing is a post-transcriptional level process of processing and modification that makes mature RNA different from its DNA template strand by base insertion, deletion, or replacement, which is a universal correction mechanism in plants to restore conserved amino acids that have been altered by DNA mutations. PPR proteins (pentatricopeptide repeat protein) are an important class of RNA editing factors that are widely distributed in plants and constitute one of the largest families of higher plants. At present, many studies have shown that it plays an important role in plant growth and development. Its main function is to affect the expression of organelle genes by participating in the processing of RNA precursors, such as achieving the transformation of RNA C-U, participating in intron splicing, affecting mRNA stability and translation, etc., thus affecting photosynthesis, respiration, plant development and environmental response. In this paper, the classification, localization, function and mechanism of PPR protein reported in recent years are reviewed, the application prospect of PPR protein is also prospected, aiming to provide theoretical basis for the subsequent research and analysis of the function of PPR protein and its application. Table and Figures | Reference | Related Articles | Metrics

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Research Progress in Microalgal Lipid Synthesis and Cultivation of High-lipid Strain ZOU Tao-zhen, LI Peng-fei, LI Xin-dong, WAN Huan, ZHANG Yi Biotechnology Bulletin    2025, 41 (1): 25-38.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0522 Abstract （1515）   HTML （14）    PDF（pc） （4197KB）（179）       Knowledge map    Save Microalgal lipids, as a potential renewable energy source and biofuel resource, hold significant importance in addressing the energy crisis and promoting green development. The characteristics of algal strains affect various stages of microalgal cultivation, lipid extraction and conversion. Selecting suitable original strains for high-lipid breeding is expected to overcome the bottleneck of low overall lipid yield in the production process. Stress cultivation induces physiological and metabolic changes in microalgae by altering external growth conditions, thereby promoting lipid accumulation. This essentially utilizes the stress response of microalgae, requiring an optimal balance between lipid accumulation and growth. Mutagenesis induces random mutations in microalgae cells through physical or chemical means, essentially acting as external stress-induced random mutations, necessitating the selection of mutants with desirable traits. Genetic engineering breeding involves the precise modification of the microalgal genome through molecular biological techniques, characterized by high precision, high cost, and high complexity. Exploring the integration of high-lipid strain cultivation with resource-efficient concepts can achieve a more economical and environmentally-friendly model biomass energy production, driving the development of the biomass energy industry. This paper provides an overview of the mechanisms of microalgal lipid synthesis and its regulatory strategies, summarizes the methods for promoting lipid production in microalgae, including stress induction, mutagenesis, and genetic engineering, as well as the synergy between high-lipid strains and resource-efficient production. The paper emphasizes the importance of cultivating high-lipid strains for sustainable biofuel production. By enumerating the key technologies and mechanisms of various cultivation methods in current high-yield microalgal lipid research, the paper highlights the research directions and bottlenecks in microalgal lipid production. Future research may focus on the exploration and improvement of the lipid metabolism regulatory network in oil-producing microalgae, the innovation of high-throughput breeding methods, and the optimization of resource-efficient cultivation systems. Table and Figures | Reference | Related Articles | Metrics

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Cloning and Functional Analysis of CsWAK8 Gene from Camellia sinensis during Cold Stress JIAO Xiao-yu, WU Qiong, LIU Dan-dan, SUN Ming-hui, RUAN Xu, WANG Lei-gang, WANG Wen-jie Biotechnology Bulletin    2025, 41 (2): 210-220.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0542 Abstract （1509）   HTML （13）    PDF（pc） （6729KB）（118）       Knowledge map    Save Objective The wall-associated kinase (WAK) is a unique class of receptor-like kinase (RLK) that plays an important role in regulating plant growth and responding to both biotic and abiotic stresses. Exploring the function of CsWAK8 in responding to cold stress may provide a theoretical basis for further analysis of the cold resistance mechanisms in Camellia sinensis. Method The CsWAK8 gene was cloned from the leaves of C. sinensis. The quantitative real-time PCR method was used to analyze the expression pattern of the CsWAK8 gene in different tissues and different cold-resistant tea tree varieties during the wintering period. An agrobacterium-mediated method was used to heterologously express the CsWAK8 gene in Arabidopsis thaliana. The cold-treated phenotypic observation, enzyme activity determination, and cold-response-related gene expression detection of transgenic plants were also carried out. Result The CDS of CsWAK8 gene is 2 307 bp, and encodes a protein of 768 amino acids, which contained a conserved domain unique to the WAK family. CsWAK8 was highly expressed in the mature leaves of C. sinensis, and its expression in the leaves and roots of cold-sensitive tea tree varieties was significantly higher than that in cold-tolerant ones during the wintering period. Nine transgenic lines of A. thaliana were acquired through the heterologous over-expressions of the CsWAK8 gene, and three of these lines were analyzed for their cold resistances. Under cold stress, the root lengths and survival rates of the transgenic lines were significantly lower than those of the wild type. Additionally, the degree of wilting in the rosette leaves of potted seedlings was higher in the transgenic lines compared to the wild type, and the MDA content in the transgenic variety L48 was significantly higher than that of the wild type after 6 h of freezing treatment. Furthermore, qPCR analysis revealed that the relative expressions of AtCBFs in the CsWAK8 gene transgenic A. thaliana under cold treatment were significantly lower than those in the wild type. Conclusion The transgenic A. thaliana is more sensitive to cold treatment than wild-type A. thaliana. CsWAK8 may play a negative regulatory role in response and tolerance to cold stress through the CBF-mediated cold-signaling pathway. Table and Figures | Reference | Related Articles | Metrics

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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 Biotechnology Bulletin    2025, 41 (7): 299-311.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0070 Abstract （1503）   HTML （10）    PDF（pc） （4691KB）（91）       Knowledge map    Save 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. Table and Figures | Reference | Related Articles | Metrics

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Genome-wide Identification and Expression Analysis of CCDs Family in Brassia rapa L. WANG Chen, LIU Guo-mei, CHEN Chang, ZHANG Jin-long, YAO Lin, SUN Xuan, DU Chun-fang Biotechnology Bulletin    2025, 41 (3): 161-170.   DOI: 10.13560/j.cnki.biotech.bull.1985.2024-0722 Abstract （1462）   HTML （15）    PDF（pc） （3191KB）（179）       Knowledge map    Save Objective The members of the carotenoid cleavage dioxygenase gene family in Brassia rapa L. were identified to gain insight into the function and tissue expression characteristics of the BrCCD family genes. Method Bioinformatics methods were used to identify the CCDs gene family in B.rapa L., construct a phylogenetic tree, and analyze the chromosome distribution, gene structure, conserved motif, intraspecific collinearity and promoter cis elements. Combined with transcriptome data and RT-qPCR technology, the expression of CCDs genes in different tissues was analyzed. Result Total 16 BrCCD genes were identified in B.rapa L., which were divided into six subgroups and distributed unevenly on eight chromosomes. Colinearity analysis showed that there was a collinearity between seven pairs of CCDs genes. Cis element analysis showed that CCDs family members may respond to various regulatory processes such as growth and development, hormone regulation, and abiotic stress. Transcriptome and quantitative PCR data analysis showed that the expression of BrCCD was tissue-specific, and the expression of BrCCD-L gene was higher in flowers, leaves and seeds, BrCCD1b was higher in flowers, and the rest was higher in seeds and leaves. Result Total 16 CCDs genes were identified in the genome of B.rapa L., which showed different expression patterns in different tissues. The BrCCD-L gene was highly similar to the CCD2 gene with saffron genus, and was significantly expressed in different tissues. Table and Figures | Reference | Related Articles | Metrics

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Progress in the Study of Proteins Related to Photosynthetic Product Transport in Plants LYU Ruo-tong, SUN Jing, LI Xin-ying, WANG Xu-jing, AI Peng-fei, WANG Yan-wei Biotechnology Bulletin    2025, 41 (10): 43-53.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0494 Abstract （1455）   HTML （13）    PDF（pc） （1292KB）（104）       Knowledge map    Save The transport of photosynthetic products is a critical process in plant growth and development, involving multiple specialized transporters that play essential roles in maintaining normal plant metabolism, regulating energy allocation, and facilitating nutrient exchange between source and sink tissues. Recent years have witnessed significant advances in research on sugar transporters. Sucrose transporters (SUT) and sugars will eventually be exported transporters (SWEET) primarily mediate sucrose translocation between different plant tissues and organs, while monosaccharide transporters (MST) facilitate transmembrane monosaccharide transport. Invertases (INV) and sucrose synthases (SUS) participate in sucrose hydrolysis and synthesis, thereby regulating the transport of photosynthetic assimilates within plants. The coordinated action of these proteins is crucial for controlling the allocation and metabolism of photosynthetic products, with profound implications for plant responses to environmental changes and yield optimization. This review summarizes major transporters associated with photosynthetic product transport in plants, highlighting the biological functions and molecular mechanisms of sucrose transporters, SWEET proteins, invertases, and related proteins across diverse plant species. It further explores their potential applications in plant development, stress resistance, and yield regulation. Finally, this review aims to provide new theoretical foundations and practical strategies for enhancing crop productivity and advancing agricultural production. Table and Figures | Reference | Related Articles | Metrics

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Cloning and Expression Analysis of the UV-B Receptor Gene McUVR8 in Mentha canadensis L. KANG Qin, WANG Xia, SHEN Ming-yang, XU Jing-tian, CHEN Shi-lan, LIAO Ping-yang, XU Wen-zhi, WU Wei, XU Dong-bei Biotechnology Bulletin    2025, 41 (8): 255-266.   DOI: 10.13560/j.cnki.biotech.bull.1985.2025-0013 Abstract （1454）   HTML （15）    PDF（pc） （4291KB）（48）       Knowledge map    Save Objective The ultraviolet-B (UV-B) receptor UVR8 plays a key role in regulating plant growth and development in response to adversity stress. The aim is to explore the protein properties and expression characteristics of McUVR8, a UV-B receptor gene in mint (Mentha canadensis L.), and to predict its interacting proteins, thus to provide theoretical references for the subsequent probing of the function of McUVR8. Method The McUVR8 gene was obtained by RT-PCR, and its physicochemical properties, protein structure and subcellular localization, and interacting proteins were analyzed and predicted using a variety of bioinformatics tools. Tissue and leaf sequence expression patterns of McUVR8 and its expression changes under different hormones and stresses were analyzed using RT-qPCR. Result McUVR8 is 1 353 bp in length, encoding 450 amino acids, distributed in both nucleus and cytoplasm, with conserved helix-looped RCC1 core domain and C17 and C27 domains, and there is high homology with SsUVR8 of Salvia splendens, SdBUR8 of Salvia divinorum and ShUVR8 of Salvia hispanica, which are also in the Labiaceae family. McUVR8 was expressed in different tissues and leaf sequences of M. canadensis, with some differences, with the lowest expression in the roots, the highest expression in the flowers, and the highest expression in the first leaf in different leaf sequences. In addition, the McUVR8 gene responded to MeJA, ABA, and drought, NaCl, and two heavy metal and aluminum stress treatments in both leaves and roots. Finally, McUVR8 was screened and verified to interact with McCOP1. Conclusion McUVR8 may have important roles in regulating mint growth and development, responding to hormonal or adversity signals, and it may be involved in regulating mint secondary metabolism, adversity response, and other processes by binding to COP1, WRKYs, and other interacting proteins. Table and Figures | Reference | Related Articles | Metrics