Table of Content

26 April 2026, Volume 42 Issue 4

Previous Issue    Next Issue

Global Strategies， Technologies， and Policy Recommendations for Crop Intelligent Design and Breeding

LIN Qiao, XIAN Guo-jian, LI Hui-hui, ZHANG Xue-fu, SUN Tan

2026, 42(4):  1-16.  doi:10.13560/j.cnki.biotech.bull.1985.2025-1157

Asbtract ( 1228 )   HTML ( 12)   PDF (2175KB) ( 984 )  

Figures and Tables | References | Related Articles | Metrics

Seed is a crucial, irreplaceable component in agriculture. Intelligent design and breeding employs technologies such as big data, artificial intelligence, and gene editing to model and simulate the entire crop growth process from breeding to harvest. This marks a substantial shift from traditional “experimental optimization” to “computational optimization”. This paper traces the evolution of crop breeding from its early domestication stages to the era of intelligent design and breeding. Using VOSViewer software and the LDA topic model, we analyze scientific literature to identify global research themes and technological trends inintelligent design and breeding. Additionally, we further discuss the layout and its characteristics of the breeding policies in agriculturally advanced countries, such as the United States. We also address the issues and challenges faced by China in this field and offer development recommendations. The study reveals that: 1) Global research in intelligent design and breeding focuses on three main themes and nine major technological directions, particularly AI phenotyping technology systems, intelligent computational breeding technology systems, and genotype-environment-phenotype fitness associating technology systems. 2) International policies emphasize the importance of the Breeding 4.0 concept, integrating high technologies such as gene editing, big data, and artificial intelligence. In contrast, China’s policies are predominantly centered on the molecular breeding stage, with a strong focus on germplasm resource protection, key technological breakthroughs in seed sources, and enhancements of traditional breeding methods. 3) To address the challenges in policy formulation, core technology development, and the application of research in intelligent design and breeding, we propose that China should strengthen top-level design, accelerate technology-driven innovation, and improve the innovation ecosystem.

Advances in the Studies of Plant C2 Domain Abscisic Acid-related Protein

YAN Qi-qi, BU Yu-fen, ZHANG Xiao-xin, MA Xiao-cen, JING Yan-ping

2026, 42(4):  17-25.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0590

Asbtract ( 370 )   HTML ( 5)   PDF (1565KB) ( 531 )  

Figures and Tables | References | Related Articles | Metrics

C2 domain proteins (C2DPs) are a class of proteins widely present in eukaryotes, characterized typically by containing a C2 domain composed of approximately 130 amino acid residues. Plant C2-domain abscisic acid (ABA)-related proteins (CARs), which belong to subfamily I of C2DPs, consist of one C2 domain and a plant-specific “sig” insertion domain. The C2 domain binds to phospholipids in a Ca²⁺- dependent manner to mediate the membrane localization of proteins. The “sig domain” enables CAR proteins to bind to complexes of different types of signaling proteins, thereby participating in the regulation of plant growth and development as well as responses to stress conditions. This review systematically summarizes the structural characteristics, tissue expression patterns, and subcellular localizations of CAR proteins, with particular emphasis on their functions and mechanisms in ABA signal transduction, phototropic and gravitropic growth, iron nutrient uptake, and responses to biotic and abiotic stresses. Additionally, future research directions are proposed, aiming to provide a theoretical reference for in-depth analysis of the regulatory network of plant CAR proteins and their application for developing stress-resilient crops.

Advances in Coordinated Tolerance Mechanisms to Abiotic Stresses in Rice

YIN Ya-long, ZHANG Ming-yang, WANG Jie-min, MIAO Xue-xue, CHEN Jin, WANG Wei-ping

2026, 42(4):  26-37.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0791

Asbtract ( 182 )   HTML ( 4)   PDF (1023KB) ( 509 )  

Figures and Tables | References | Related Articles | Metrics

Global climate change is intensifying the occurrence of compound abiotic stresses, such as salinity-drought, drought-heat, salinity-heat, and salinity-drought-heat in rice (Oryza sativa) production, constituting a major threat to global food security. Although targeted genetic improvement has successfully enhanced crop tolerance to individual abiotic stresses, the inherent genetic complexity and nonlinear interactions among multiple stresses have impeded systematic increase of crop's resistance to compound abiotic stress. Understanding the coordinated tolerance mechanisms in plants under multifaceted stress conditions is therefore key to advancing crop performance under such challenges. This review systematically summarizes the principal response mechanisms of rice to individual abiotic stresses (salinity, drought, and heat) as well as to compound stresses (salinity-drought, drought-heat, salinity-heat, and salinity-drought-heat). This review outlines fundamental adaptation strategies to single stresses, such as ion homeostasis and osmotic adjustment and elucidates integrated response mechanisms involving signal transduction, transcriptional regulation, metabolic reprogramming, and epigenetic modifications under compound stress conditions. Furthermore, it identifies critical limitations in current compound stress research, including poor field relevance, the absence of standardized multi-omics data integration, and insufficient mechanistic insight into nonlinear stress interactions. To address these gaps, we propose future strategies such as enhancing cross-scale validation linking field phenotyping to molecular mechanisms, establishing unified platforms for multi-omics data standardization, and optimizing genetic solutions that balance stress adaptation with growth and productivity. This review provides a conceptual framework for breeding rice with enhanced compound stress tolerance and offers valuable insights for developing resilient cultivars suited to increasingly variable and stressful climates.

Advances in Flexible Wearable Biosensors for Early Warning of Crop Stress and Disease-pest Infestation： A Comprehensive Review

HE Jun-jie, HE Yang, YU Wei, GUAN Hua-nan

2026, 42(4):  38-52.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0911

Asbtract ( 111 )   HTML ( 9)   PDF (3060KB) ( 71 )  

Figures and Tables | References | Related Articles | Metrics

Flexible wearable biosensors have high sensitivity, stability, flexibility, and stretchability, enabling conformal adhesion to crop epidermis and real-time conversion of physiological signals into electrical signals for dynamic monitoring of plant physiological changes. In recent years, such sensors have rapidly advanced in the field of crop health monitoring and stress early-warning, providing a novel paradigm for real-time diagnosis of growth status, diseases, pests, and abiotic stresses. However, their large-scale application has been limited by key factors including long-term stable adhesion, environmental tolerance, multi-signal integration capability, and cost-effectiveness. This review summarized recent advances in plant wearable flexible sensors for crop health monitoring and stress early-warning, with a focus on their core characteristics, primary classifications, and sensing performance. Furthermore, it analyzed the critical bottlenecks in the translation of flexible wearable sensors into agricultural practices from multiple dimensions: early identification of diseases and pests, water status, metabolic changes, pesticide residues, and morphological strain. It also prospected the future pathways of their integration with artificial intelligence, nanotechnology, and advanced materials, which may provide important references for theoretical breakthroughs and technological implementation in crop health management within the context of smart agriculture.

Research Progress in Drug Resistance Mechanisms and Novel Therapeutic Strategies of Staphylococcus aureus

WANG Yu, HE Yun-jia, YIN Hai-xin, MA Yue, GUO Hai-yong

2026, 42(4):  53-64.  doi:10.13560/j.cnki.biotech.bull.1985.2025-1032

Asbtract ( 157 )   HTML ( 5)   PDF (922KB) ( 97 )  

Figures and Tables | References | Related Articles | Metrics

Staphylococcus aureus and its methicillin-resistant strains (MRSA) have become one of the central challenges in the global fight against antibiotic resistance due to their strong multidrug resistance and high pathogenicity. The resistance of S. aureus is mediated by multiple mechanisms, including aberrant expression of PBP2a encoded by the mecA/mecC genes, horizontal transfer of resistance determinants, biofilm formation, and the generation of persister cells. Given the limited efficacy of conventional antibiotics against resistant strains, developing novel strategies to combat S. aureus infections has become a key focus in both clinical and research fields. Nanotechnology-based antimicrobial therapies have demonstrated great potential in overcoming bacterial resistance through targeted delivery and multi-target bactericidal mechanisms. Antimicrobial peptides, with their broad-spectrum activity and low risk of resistance induction, have attracted increasing attention, while alternative approaches such as bacteriophage therapy, phage-derived lysins, and immunotherapy have also achieved significant progress. However, the clinical translation of these emerging strategies remains challenged by issues of toxicity, stability, and the potential risk of resistance dissemination. Future research should focus on optimizing drug delivery systems, enhancing targeting precision, and minimizing adverse effects, while further elucidating the dynamic mechanisms underlying resistance evolution, to promote the safe and effective clinical translation of anti-infective strategies and ultimately achieve effective control of S. aureus infections.

Application Status of Genome-wide Association Study (GWAS) in the Study of Reproductive Traits in Sow

OU Qi, FENG Yao, WEI Liu-ting, ZHUANG Zhan-wei, ZHAO Yun-xiang, CHEN Fu-mei

2026, 42(4):  65-71.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0382

Asbtract ( 95 )   HTML ( 4)   PDF (980KB) ( 104 )  

Figures and Tables | References | Related Articles | Metrics

The reproductive traits of sows play a critically important role in modern swine production. These traits encompass a series of key performance indicators, including litter size, number of live births, number of weaned piglets, litter weight at birth, litter weight at weaning, and sow longevity. They not only directly determine production efficiency but also profoundly influence the profitability and competitiveness of the entire swine industry chain.However, sow reproductive performance is a typical complex trait characterized by generally low heritability. Its genetic mechanisms are involved in numerous genes and their interactions, including additive effects, epistasis, and potential epigenetic regulation. Concurrenlty, these traits are significantly influenced by non-genetic factors such as environmental conditions, management practices, and feeding regimens (such as including nutrition, temperature, humidity, stocking density, stress levels, and health management). Consequently, the genetic improvement of these traits has remained a major challenge in animal breeding research. With the continuous development of Genome-Wide Association Study (GWAS) methodology, researchers can now utilize high-throughput genotyping platforms to screen for genetic loci associated with target phenotypes across the entire genome, thereby enabling precise dissection of complex traits. This review aims to systematically summarize the developmental overview of GWAS technology, including its statistical principles, methodological evolution, and improvements in detection power. It further emphasizes the application progress of this technology in genetic analysis of key reproductive traits in sows, synthesizing important genetic signals that have been identified and their biological functions. Finally, the review discusses challenges faced by GWAS in studying sow reproductive trait and provides perspectives on future research directions, such as integrating multi-omics data to gain deeper insights into causal mutations and regulatory networks, and leveraging big data and artificial intelligence to optimize breeding value estimation, thereby providing references for the breeding of highly prolific sows.

Research Advances in Enhancing the Soluble Expression of Recombinant Heterologous Proteins

PANG Xin-li, ZHANG Hong-bing, LIU Xiao-qing, WANG Yuan, WU Ning-feng, TIAN Jian, GUAN Fei-fei

2026, 42(4):  72-82.  doi:10.13560/j.cnki.biotech.bull.1985.2025-1013

Asbtract ( 192 )   HTML ( 5)   PDF (857KB) ( 250 )  

Figures and Tables | References | Related Articles | Metrics

Enhancing the soluble expression of recombinant heterologous proteins is a key challenge in bioengineering, pharmaceutical development, and industrial biotechnology, directly impacting protein functionality and large-scale production applications. This review systematically summarizes multi-level optimization strategies, from gene sequence to expression host, to improve the expression level, solubility, and stability of target proteins. At the codon level, strategies encompass host-preference-based codon adaptation, GC content adjustment, and the use of deep learning models to optimize mRNA structure and predict translation efficiency, thereby significantly increasing protein yield. At the amino acid level, rational design (e.g., hydrophobic core engineering, surface charge optimization), application of solubility-enhancing tags, and incorporation of non-standard amino acids can effectively improve protein folding, stability, and solubility. At the expression host level, the optimization of transcription and translation can be achieved through the selection and engineering of microbial strains (such as Escherichia coli, Bacillus subtilis, and yeast) and the precise design of various regulatory elements (e.g., promoters, ribosome binding sites, terminators) using artificial intelligence. The deep integration of artificial intelligence and biotechnology is driving the field of soluble protein expression from an empirical-driven approach towards a precise and predictable new “design-build-test” paradigm. This review aims to provide a comprehensive theoretical reference and technical outlook for the multidimensional and intelligent enhancement of soluble protein expression.

Optimization of a High-performance and Low-cost Fluorescence Detection Buffer with Broad Compatibility across Cas12a Orthologs

LI Ya-qi, SUN Meng, LI Xiu-li, WEI Jing-na, ZHAO Lin-lin, ZHAO Yun-ping, LIU Zheng-hui, SU Fan

2026, 42(4):  83-91.  doi:10.13560/j.cnki.biotech.bull.1985.2025-1073

Asbtract ( 99 )   HTML ( 2)   PDF (1833KB) ( 41 )  

Figures and Tables | References | Related Articles | Metrics

Objective Current CRISPR/Cas12a nucleic acid detection systems commonly use conventional restriction enzyme buffers, which are not specifically optimized for Cas12a-mediated fluorescence activation. This limitation reduces detection sensitivity and increases system cost. The study aimed to construct a broadly compatible, cost-effective, and fluorescence-optimized reaction system for multiple Cas12a proteins to improve nucleic acid detection performance. Method Fluorescence quantification using a fluorescence detector and visual readout were employed to systematically evaluate the effects of pH (7.3–7.9, 25 ℃), Tris-HCl concentration (5–50 mmol/L), calcium ion (Ca²⁺, 0.1–1 mmol/L), and magnesium ion (Mg²⁺, 10–30 mmol/L) on Cas12a fluorescence signal. Based on these results, a simplified reaction buffer (CasRB), free of antioxidants and protein stabilizers, was developed. CasRB performance was compared with commercial NEB buffers, and its compatibility was tested in three Cas12a orthologs: Francisella novicida Cas12a (FnCas12a), Acidaminococcus sp. Cas12a (AsCas12a), and Lachnospiraceae bacterium Cas12a (LbCas12a). Result The optimized CasRB reduced buffer cost by over 99.9% compared with commercial buffers by eliminating high-cost components such as dithiothreitol (DTT) and protein stabilizers. Fluorescence signal-to-noise ratio increased more than tenfold, significantly enhancing naked-eye visualization. CasRB showed strong cross-ortholog compatibility, providing comparable fluorescence performance in FnCas12a, AsCas12a, and LbCas12a systems. Conclusion Systematic optimization of reaction conditions produce a CasRB buffer that combined cost reduction and enhanced fluorescence sensitivity. The buffer addresses the compatibility limitations of conventional buffers in Cas12a-based nucleic acid detection systems, offering a versatile platform for multiple Cas12a proteins.

Expression Analysis and Interaction Protein Screening of GmRLK19 in Soybean

XU Meng-ge, SONG Huo-yan, LUO Jia, SU Yi, ZHOU Hui-wen, WANG Can, KONG Ke-ke

2026, 42(4):  92-100.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0903

Asbtract ( 77 )   HTML ( 10)   PDF (4268KB) ( 282 )  

Figures and Tables | References | Related Articles | Metrics

Objective GmRLK19, an important candidate gene for soybean 100-seed weight, was identified in our previous study. This study aims to analyze the expression pattern and screen for interacting proteins of GmRLK19, providing a basis for further investigation into its biological function. Method The bioinformatics was employed to analyze the structural characteristics of the GmRLK19 gene. The tissue expression pattern of GmRLK19 was analyzed using quantitative real-time PCR (RT-qPCR) technology. Subcellular localization was determined by fusing GmRLK19 with a fluorescent tag and observing with confocal microscopy. Proteins interacting with GmRLK19 were screened using yeast two-hybrid technology, and their potential functions were predicted using database information. Result The secondary structure of GmRLK19 protein is predominantly composed of random coils and α-helices, with a total of 145 predicted phosphorylation sites. Gene GmRLK19 is expressed ubiquitously across all tested tissues, showing the highest transcript levels in seeds at 30 d after flowering. Subcellular localization experiments confirmed that the GmRLK19 protein is primarily localized to the plasma membrane. Through yeast two-hybrid screening, a total of 187 positive clones were identified, among which 148 candidate interacting proteins were screened by PCR amplification of the inserted fragments after transformation. Functional analysis revealed that the interacting proteins function primarily in regulating plant growth and development, signal transduction, and metabolic processes. The GO enrichment analysis revealed significant enrichment for interacting proteins involved in photosynthesis and protein translation. Conclusion GmRLK19 is expressed in various tissues, and its encoded protein localizes primarily to the cell membrane. The 148 candidate interacting proteins are obtained via screening, suggesting that GmRLK19 influences soybean seed weight likely through multiple pathways, such as plant hormone signaling and photosynthesis.

Identification and Expression Analysis of CAD Gene Family in Soybean(Glycine max (L.) Merr.)

SU Yan-zhu, LI Da, ZHANG Ai-ai, LIU Yong-guang, ZHANG Xiu-rong, XUE Qi-qin

2026, 42(4):  101-113.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0928

Asbtract ( 147 )   HTML ( 7)   PDF (5112KB) ( 79 )  

Figures and Tables | References | Related Articles | Metrics

Objective The characteristics and expression analysis under different stresses of the Cinnamyl Alcohol Dehydrogenase (CAD) gene family (GmCAD) in soybean were studied, providing a theoretical basis for further research on the biological function of the GmCADs gene family. Method Based on the soybean genome data, the CAD gene family members were screened and identified using bioinformatics methods. The characteristics of the encoded proteins, phylogenetic relationships, gene structure and conserved motifs were analyzed. Result A total of 18 GmCADs (GmCAD1-GmCAD18) were identified in the whole soybean genome, which were unevenly distributed on 13 chromosomes, encoding 219 to 364 amino acids. Phylogenetic analysis revealed that the GmCADs gene family can be divided into four subfamilies, and the genes within the same subfamily share similar gene structures and conserved motifs. The collinearity analysis indicates that fragment duplication was the main form of expansion in the GmCADs gene family. Selection pressure analysis showed that the GmCADs gene was under purifying selection. The promoters of the GmCADs gene family containing abundant cis-acting elements were mainly associated with the light response, hormone response, stress response and growth development processes. The protein interaction network prediction results showed a complex protein interaction network, which was formed by all GmCADs genes through five indirect proteins, and GO functional analysis significantly enriched in terms such as CAD activity and lignin metabolism process. The GmCAD members demonstratecertain selectivity in expression under different tissues and stress conditions (such as drought, salt, cold, shade, and high temperature stress). RT-qPCR validation indicates the expressions of nine GmCAD significantly changed at different time points under salt and drought treatments, mainly showing upregulation. Conclusion The distribution, structure and function of the 18 soybean CAD members are diverse, and the GmCADs gene may respond to abiotic stress during the growth and development of soybean plants.

Identification of ZF-HD Gene Family in Arachis hypogaea and Analysis in Response to Abiotic Stress

CHEN Deng-ke, LAN Gang, XIA Zhi, HOU Bao-guo, YANG Liu-liu, CAO Cai-rong, LI Peng-bo, WU Cui-cui

2026, 42(4):  114-128.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0806

Asbtract ( 111 )   HTML ( 7)   PDF (14841KB) ( 42 )  

Figures and Tables | References | Related Articles | Metrics

Objective To analyze the basic characteristics of the ZF-HD gene family in peanut (Arachishypogaea) and its expression patterns under various abiotic stresses, laying a foundation for revealing the role of ZF-HD in regulating abiotic stress in peanut. Method Bioinformatics methods were used to screen the peanut ZF-HD gene family members and analyze 1) their physicochemical properties, phylogenetic relationships, conserved motifs, conserved structural domains, gene structures, promoter cis-acting elements, chromosomal localization and gene covariance; 2) the expression patterns of the AhZHDs genes in different tissues and organs of peanut and in different abiotic stresses based on the transcriptome data; 3) the expressions of some members of this family in peanut following different abiotic stress treatments using RT-qPCR. The expression patterns of AhZHDs genes in different tissues and organs of peanut under different abiotic stresses were analyzed based on transcriptome data, and the expression of some members of this family in different abiotic stresses was analyzed by real-time fluorescence PCR. Result A total of 40 peanut ZF-HD genes were identified in the peanut Tifrunner genome, which were classified into six subfamilies, ZHD Ⅰ, ZHD Ⅱ, ZHD Ⅲ, ZHD Ⅳ, ZHD Ⅴ and MIF, according to their phylogenetic relationships. AhZHDs genes in the same subfamily had similar structural features, and most of them had no introns. Cis-acting element analysis indicated that the members of the peanut ZF-HD gene family were widely involved in hormone response, growth and development response, abiotic stress and light response. Chromosomal localization and covariance analysis showed that the 40 AhZHDs genes were unevenly distributed on 16 chromosomes, and that the whole genome duplication (WGD) event or fragment duplication might be the main driving force for the evolution of the AhZHDs genes. Protein interactions analysis showed that most AhZHD proteins had complex interactions with each other and might be involved in peanut development and stress response through co-regulation. Transcriptome analysis and qRT-PCR validation indicated that nine AhZHD genes (AhZHD5/9/10/17/23/29/30/32/40) showed diverse expression patterns across various peanut tissues and under stress conditions, with the majority showing significantly altered expression levels in response to abiotic stress, subcellular localization experiments showed that AhZHD5/17/29/32 function in the nucleus. Conclusion The 40 AhZHD genes differ in structure and characterization, and AhZHDs are widely involved not only in peanut growth and development and hormone signaling, but also play important regulatory roles in abiotic stresses.

Integrated Analysis of Transcriptome and Lipid Metabolome Reveals the Differences in α-Linolenic Acid Synthesis Regulation in Different Perilla frutescens

WANG Yu-kun, YUAN Yuan, WANG Bin, ZHU Yun-na, REN Xiao-qiang, REN Fei, YE Hong

2026, 42(4):  129-140.  doi:10.13560/j.cnki.biotech.bull.1985.2025-1008

Asbtract ( 112 )   HTML ( 2)   PDF (11572KB) ( 15 )  

Figures and Tables | References | Related Articles | Metrics

Objective This study aims to analyze the differences in α-linolenic acid (ALA) content between QO10 and QS5 seeds of Perilla frutescens, and find out genes related to ALA synthesis and regulation. The results will provide genetic resources for the further creation of new perilla varieties with high ALA content. Method Using morphological index determination, transcriptomics, and lipid metabolomics techniques, a systematic analysis was performed on the seed morphology, differentially expressed genes (DEGs), and lipid metabolite profiles of perilla QO10 and QS5. Result The 1 000-seed weight of QS5 seeds was significantly higher than that of QO10. Seed coat color difference analysis indicated that the seed coat color of QS5 was brighter than that of QO10, and the two were easily distinguishable in terms of seed coat color. Results of lipid metabolomics analysis demonstrated that there were significant differences in the types and relative contents of lipid metabolites between QO10 and QS5 seeds; the contents of stearic acid (SA) and ALA in QS5 seeds were higher than those in QO10. Transcriptomic analysis results showed that 188 differentially expressed genes (DEGs) were enriched in 6 GO terms and 5 KEGG pathways related to fatty acid (FA) metabolism. Based on the results of transcriptome-lipid metabolome integrated analysis, 15 key enzyme and protein-coding genes were finally screened out. These included ACSL, FABP, FAD2, ENR, KAR, and KAS as well as lipase (lipase)-coding gene, LCAT3, FAR, SCL, and HMGCR. Furthermore, transcriptomic analysis revealed that 38 WRKY transcription factors (TFs) and 26 MYB TFs were differentially expressed; 2 WRKY TFs and 2 MYB TFs with the largest fold changes in differential expression were selected as candidate TFs. Finally, 10 candidate genes related to the regulation of ALA biosynthesis were verified by RT-qPCR, and the quantitative results were consistent with those of transcriptome sequencing. Conclusion There are significant morphological differences between the seeds of QO10 and QS5, and QS5 contained higher levels of SA and ALA. Nineteen genes related to the regulation of ALA biosynthesis are screened out, whose differential expression in QO10 and QS5 seeds is a potential cause of the difference in ALA content.

Screening of Differentially Expressed Genes in Tomato Defense against Meloidogyne incognita Based on RNA-seq and Expression Characteristics Analysis of RPP13 Gene

FENG Cheng-hao, DANG Yu-le, WANG Zhi-ze, NIE Wei-dan, YANG Zhong-min, DU Chong

2026, 42(4):  141-152.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0657

Asbtract ( 79 )   HTML ( 3)   PDF (22241KB) ( 56 )  

Figures and Tables | References | Related Articles | Metrics

Objective By conducting RNA-seq and weighted gene co-expression network analysis on resistant and susceptible tomato materials inoculated with root-knot nematodes (RKNs) ( Meloidogyne incognita), we aim to focus on the signaling pathways and genes related to disease resistance, verify the root expression patterns of target genes, and provide theoretical basis for subsequent molecular function research on these genes. Method The resistant lines ‘18060’ and susceptible lines ‘Moneymaker’ were used for inoculation, and root samples were collected for transcriptome sequencing and WGCNA at the time points of no inoculation, 2 d and 4 d after inoculation The main pathways and regulatory genes that might be involved in disease resistance were screened, and the determination of the tissue-specific expression and root expression of these target genes were completed. Result As the inoculation time prolonged, the number of differentially expressed genes (DEGs) gradually increased. In the comparison between the resistant and susceptible groups at 2 dpi, 1 900 DEGs were screened out, and at 4 dpi the number of DEGs increased to 3 012. KEGG analysis showed that the “plant-pathogen interaction” was significantly enriched in both the within-group and between-group comparisons. In the between-group comparison, the DEGs enriched in this pathway mainly encoded receptor-like proteins/kinase, calcium signaling-related proteins, heat shock proteins, and transcription factors, etc. Among which 5 R genes encoding RPP13 protein were also screened out. Focusing on the DEGs during the inoculation period, the significant module MEbrown was screened out via the WGCNA, and the top 10 nodes based on the in degree and out degree were selected as hub genes based on the in degree and out degree. Two DEGs encoding RPP13 were found among them. The tissue-specific expression and root expression pattern identification further indicated that Solyc07g039410.3 might play a key role in tomato defense against the invasion of M. incognita. Conclusion Six RPP13 genes potentially involved in the process of resisting M. incognita are screened out. Among them, Solyc07g039420.1 and Solyc07g039410.3 demonstrate specific expression characteristics in the roots. At 4 dpi, the hub gene Solyc07g039410.3 is significantly induced and its expression increases by 6.23 times, indicating this gene plays a critical role in the defense of tomato against M. incognita.

Cloning and Functional Analysis of AeF3H Gene in Okra

SUN Ting, ZHANG Yan, LIU Yu-shan, FENG Yuan-yuan, QIN Heng-shan, ZHANG Jun, HE Xiao-gang, ZHANG Jing-rong

2026, 42(4):  153-160.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0837

Asbtract ( 95 )   HTML ( 7)   PDF (3813KB) ( 151 )  

Figures and Tables | References | Related Articles | Metrics

Objective The AeF3H was cloned from okra (Abelmoschus esculentus), and its biological function was characterized via transformation of Arabidopsis thaliana, aiming to provide a theoretical basis for further elucidating the flavonoid accumulation in okra. Method The primers were designed based on the A. esculentus transcriptome data, and the seed’s cDNA was used as template to amplify the AeF3H gene by PCR. The sequence features were analyzed using bioinformatics. The expression characteristics of AeF3H gene in different tissues of okra were analyzed using real-time fluorescence quantitative PCR (RT-qPCR). A 35S::AeF3H::YFP overexpressing vector was constructed and transformed into Arabidopsis via the floral dip method. Transgenic plants were screened, and the total flavonoids contents in the T₃ lines were determined by Al(NO₃)₃ colorimetric method. The subcellular localization of AeF3H was analyzed by transient expression in Arabidopsis protoplasts. Result The open reading frame (ORF) of AeF3H was 1 101 bp in length and encoded a 366-amino acid protein. The genomic DNA spanned 1 304 bp, containing 3 exons and 2 introns. The deduced protein molecular weight was 41.08 kD and its theoretical isoelectric point was 5.32. Phylogenetic tree analysis revealed that AeF3H demonstrated the closest homology to the F3H from Hibiscus trionum. RT-qPCR results showed that AeF3H had a tissue-specific expression profile, with particularly high expressions detected in young seeds and buds, and the lowest levels detected in the stem and pod. Subcellular localization showed that the AeF3H protein was located in both nucleus and cytosol. Arabidopsis plants overexpressing AeF3H presented a 4.75-fold increase in total flavonoid content compared to wild-type plants. Conclusion AeF3H has tissue-specific expression, and its protein is localized to both the nucleus and cytoplasm. AeF3H plays a crucial role in the flavonoid biosynthetic pathway in okra. The overexpression of AeF3H may significantly increase total flavonoid content in transgenic Arabidopsis.

Functional Analysis of VvHSP18.2 Overexpression in Regulating Salt-alkali Resistance in Grapevines

XU Yu-jiao, SUN Yu-shuai, LIU Dao-qi, ZHANG Li, ZHANG Zhi-chang, YAO Yu-xin

2026, 42(4):  161-169.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0692

Asbtract ( 667 )   HTML ( 1)   PDF (3653KB) ( 36 )  

Figures and Tables | References | Related Articles | Metrics

Objective Saline-alkali stress severely affects grape growth, development, and quality formation. This study aims to evaluate the function of VvHSP18.2 in regulating grape saline-alkali resistance, identify key genes involved in grape salt-alkali tolerance, and establish a theoretical foundation for the genetic improvement of stress resistance in grapes. Method Phylogenetic analysis of VvHSP18.2 was conducted using bioinformatics tools, and its promoter was analyzed. RT-qPCR was used to assess the expression of VvHSP18.2 under various stress treatments, including NaHCO₃. VvHSP18.2 was overexpressed in Arabidopsis and grape, and its function was evaluated through phenotypic observation and physiological indicators. Result VvHSP18.2 belonged to the heat shock protein CI subfamily and is mainly expressed in grape callus, fruit, and roots. Its promoter region contained multiple cis-acting elements responsive to drought, salt, and high-temperature stresses. VvHSP18.2 rapidly responded to multiple abiotic stresses: its expression was significantly induced by NaCl, especially NaHCO₃, at 6 and 12 h post-treatment; it was significantly induced by PEG6000 at 1 hour post-treatment; and its expression significantly decreased at 3 and 6 h after AlCl₃ treatment. The overexpression of VvHSP18.2 in grape roots and Arabidopsis alleviated phenotypic damage caused by NaHCO₃ treatment, reduced electrolyte leakage and MDA content, and enhanced resistance to NaHCO₃ stress. Furthermore, VvHSP18.2 overexpression increased the activities of SOD, CAT, and POD enzymes in Arabidopsis, reduced the accumulation of O^2- and H₂O₂, indicating that it enhanced stress resistance by improving reactive oxygen species (ROS) scavenging capacity. In contrast, VvHSP18.2 overexpression had no significant effect on NaCl resistance in Arabidopsis. Conclusion VvHSP18.2 is involved in regulating the stress response in grapes. The overexpression of VvHSP18.2 enhances saline-alkali stress resistance in both grape and Arabidopsis by increasing antioxidant enzyme activities and improving ROS scavenging capacity, thereby regulating the balance of ROS metabolism.

Identification of the MYB Gene Family in Pomelo (Citrus grandis) and Functional Analysis in Chlorosis of Graft Incompatibility

GUO Dao-xiang, SU Quan, LI Ao-ting, WANG Yan, CHEN Qing, WANG Xiao-rong, HE Wen

2026, 42(4):  170-181.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0873

Asbtract ( 91 )   HTML ( 2)   PDF (5253KB) ( 61 )  

Figures and Tables | References | Related Articles | Metrics

Objective Grafting is commonly used in citrus propagation, while scion-rootstock incompatibility often occurs in orchards and leads to economic losses. This study is aimed to investigate the role of MYB family genes in leaf chlorosis associated with scion-rootstock incompatibility, and to elucidate the molecular mechanisms underlying scion-rootstock interaction in Citrus. Method We performed a genome-wide identification and systematic analysis of the MYB gene family in pummelo (Citrus grandis). Based on transcriptome data and RT-qPCR from leaves at different yellowing stages of compatible/incompatible graft combinations, key MYB genes were screened. By employing transient overexpression in tobacco leaves, the function of the candidate gene CgMYB53 in leaf chlorosis associated with scion-rootstock incompatibility was preliminarily validated. Result A total of 143 MYB family members were identified in the pomelo genome, classified into four subfamilies, R1-MYB (35 members), R2R3-MYB (104 members), R3-MYB (3 members), and R4-MYB (1 member). Promoter sequences of MYB members primarily contained cis-acting elements responsive to light (53.23%), hormones (27.94%), and stress (16.07%). Comparative transcriptome analysis of leaves from compatible and incompatible graft combinations, 10 differentially expressed MYB genes were identified. RT-qPCR results showed that the relative expression of CgMYB53 was relatively low in the green leaves, but relatively high in the yellow leaves. Transient overexpression of CgMYB53 in tobacco leaves resulted in significant decrease in chlorophyll a and chlorophyll b content, accompanied by significant increase in amylose, amylopectin, and total starch content,starch synthesis-related genes CgGBSS2、CgISA2 and CgBE3 were highly expressed. Conclusion This study identified 143 MYB family members in pomelo and screened 10 MYB TFs potentially involved in leaf chlorosis associated with scion-rootstock incompatibility. Preliminary functional validation indicates that CgMYB53 may participates in starch synthesis related genes expression, inducing leaf chlorosis by regulating starch accumulation.

Overexpression of Mulberry MnERF2 Enhances Resistance to Drought in Arabidopsis thaliana

DONG Ya-ru, NIE Yu-xia, ZHU Hong, WANG Zhao-hong, LIU Hui-fen, GUO Guang

2026, 42(4):  182-189.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0448

Asbtract ( 95 )   HTML ( 0)   PDF (4866KB) ( 39 )  

Figures and Tables | References | Related Articles | Metrics

Objective AP2/ERF transcription factors are plant-specific regulators that play a critical role in plant responses to abiotic stresses, such as low temperature, drought, waterlogging, and high salinity. Elucidating the molecular function of the mulberry ERF transcription factor MnERF2 in drought stress response may provide a theoretical foundation and genetic resources for stress-resistant mulberry breeding. Method MnERF2-overexpressing Arabidopsis thaliana lines were obtained via the floral dip method. Both transgenic seedlings and mature plants were subjected to drought stress, followed by phenotypic observation and physiological/biochemical analysis. Result The water loss rate of MnERF2-overexpressing A. thaliana lines is significantly lower than that of the wild-type. The overexpressingseedlings presented superior root development and mature transgenic plants showed significant increase in biomass accumulation and chlorophyll content under drought stress. The overexpressing lines demonstrated enhanced activities of antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and glutathione S-transferase (GST). Levels of ascorbic acid (ASA), glutathione (GSH), and proline significantly elevated, meanwhile the relative electrical conductivity, malondialdehyde (MDA) content, and the levels of reactive oxygen species (ROS; O₂^•-, H₂O₂, •OH) markedly reduced under drought stress. Conclusion MnERF2 positively regulates the responses to drought stress. Overexpressing MnERF2 increase the tolerance to drought in transgenic A. thaliana by improving water retention and biomass accumulation, promoting proline biosynthesis, boosting antioxidant enzyme activity, and increasing antioxidant content.

Analysis of Phenotypic Characteristics and Root Transcriptomics of Sugarcane with Different Genotypes

YANG Ting, YANG Zong-tao, AI Jing, WANG Yu-tong, LI Yan-ye, DENG jun, LIU Jia-yong, ZHAO Yong, ZHANG Yue-bin

2026, 42(4):  190-201.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0966

Asbtract ( 80 )   HTML ( 2)   PDF (37993KB) ( 11 )  

Figures and Tables | References | Related Articles | Metrics

Objective To clarify the phenotypic variation pattern of sugarcane germplasm resources and the regulatory mechanism of key genes, and provide molecular targets for the directional breeding of nitrogen-efficient varieties. Method Seventeen sugarcane genotype materials with clear genetic backgrounds were used as the research objects. Seven agronomic traits such as plant height, stem diameter, and single-stem weight were systematically measured to evaluate their genetic diversity. Phenotypic groups were divided by cluster analysis, and a total of six extreme materials from two groups were selected for root transcriptome sequencing. The DESeq2 software was used to screen differentially expressed genes (DEGs), and GO and KEGG functional enrichment analyses were performed. Weighted gene co-expression network analysis (WGCNA) was used to mine core genes related to group differentiation, and their expression patterns were verified by RT-qPCR. Furthermore, the differences in nitrogen absorption and utilization abilities of the two types of materials were verified through a ¹⁵N isotope absorption experiment. Result The phenotypic variation coefficients of 17 materials ranged from 0.10 to 0.84, and the genetic diversity indices were between 2.55 and 2.83. By cluster analysis they were divided into Group Ι (7 materials) and Group Ⅱ (10 materials). The two groups showed significant differences in traits such as stem diameter (1.12 cm vs. 1.39 cm), number of effective stems (28.0 vs 15.1), and brix (11.1 vs 14.9). Transcriptome analysis identified a total of 9 724 differentially expressed genes (DEGs), which were significantly enriched in pathways such as linoleic acid metabolism (ko00591), flavonoid biosynthesis (ko00941), α-linolenic acid metabolism (ko00592), glutathione metabolism (ko00480), glycolysis/gluconeogenesis (ko00010), glyoxylate and dicarboxylate metabolism (ko00630), starch and sucrose metabolism (ko00500), and ether lipid metabolism (ko00565). Via WGCNA the 10 hub genes were identified significantly associated with phenotypes, including Sspon.02G0013210-1A (AP2 transcription factor), Sspon.02G0008140-1T (GTP-binding protein), etc. The ¹⁵N absorption experiment showed that the materials in Group II had a stronger nitrogen absorption capacity, while the materials in Group I presented higher nitrogen accumulation characteristics. Conclusion Wild species materials adapt to low-nitrogen environments by increasing the number of effective stems, while hybrid materials achieve biomass and sugar accumulation by enhancing nitrogen absorption and utilization efficiency. The coordinated regulation of carbon and nitrogen metabolism and redox balance by key genes is an important molecular basis for the formation of nitrogen efficiency in sugarcane.

Genetic Diversity Analysis of Citri Fructus Germplasm Resources Based on Phenotypic Traits and SSR Markers

WEI Li-na, JIANG Jing-long, DENG Xi-xi, LI Yan, LI Li, DENG Jia-rui, DING De-kuan

2026, 42(4):  202-215.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0846

Asbtract ( 108 )   HTML ( 0)   PDF (3751KB) ( 110 )  

Figures and Tables | References | Related Articles | Metrics

Objective Citri fructus germplasm resources in China were investigated to analyze phenotypic variation of fruit traits, genetic diversity, population structure and genetic differentiation characteristics, providing a scientific basis for germplasm conservation and superior cultivar breeding. Method The total of 40 batches of fruits and 90 leaves samples of plants from 16 provinces (37 cities) of China were used as the test materials, and then the genetic diversity analysis was conducted using 15 phenotypic traits and screened 10 pairs of SSR markers. Result The Shannon-Wiener index H and Simpson’s diversity index D of 7 quality traits of citri fructus from different regions varied from 0.526 to 1.365 and 0.349 to 0.711, respectively. Among the 8 quantitative traits coefficient of variation ranged from 21% (number of segments) to 99% (single-fruit weight). According to phenotypic traits, the tested materials could be divided into three groups: Ⅰ (Xiangyuan and Zhique), Ⅱ (Xiangpao) and Ⅲ (Juyuan). The 10 pairs of primers with good polymorphism and stable bands were screened for PCR of SSR primers. The results showed that tthe number of alleles (Na), the number of effective alleles (Ne), Shannon’s diversity index (I), observed heterozygosity (Ho), and expected heterozygosity (He) at the locus level were 1.832, 1.354, 0.331, 0.824, and 0.210, respectively. By clustering, population structure and PCoA analysis, the 90 germplasm were divided into four groups. Xiangyuan and Zhique were further separated. The genetic diversity of the four groups was different with the descending order of Zhique (Ho=0.682, He=0.199), Juyuan (Ho=0.659, He=0.175), Xiangpao (Ho=0.436, He=0.133) and Xiangyuan (Ho=0.391, He=0.127). The Mantel test showed that the genetic distance of the citri fructus and geographical distance of the provenances were significantly correlated (r=0.558, P=0.010). The molecular variance analysis of xiang yuan population exhibited 73% of the variation within the population. Conclusion Citri fructus germplasm resources in China have rich phenotypic genetic variation and high diversity.

Identification and Functional Analysis of the MpPP2A-C Gene in Marchantia polymorpha

JIANG Xin-hua, FANG Tian-yu, ZHANG Jing-jing, LI Xiang-yuan, ZHANG Bang-yue, LIAO Xiao-shan, RONG Duo-yan

2026, 42(4):  216-226.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0820

Asbtract ( 93 )   HTML ( 10)   PDF (4202KB) ( 254 )  

Figures and Tables | References | Related Articles | Metrics

Objective To identify the MpPP2A-C gene of Marchantia polymorpha, analyze its expressions and study its interaction with the MpPP2A-A protein, so as to provide a theoretical basis for clarifying the function of the MpPP2A-C gene. Method The phylogenetic trees of PP2A-C of Marchantia polymorpha and other plants were constructed, and homology alignment was performed using the PP2A-C gene sequence of Arabidopsis thaliana to identify the PP2A-C gene of M. polymorpha, and bioinformatics analysis was conducted on it. The expression pattern of the MpPP2A-C gene in tissue specificity and response to abscisic acid was explored by RT-qPCR. The interaction relationship between the MpPP2A-C subunit and the MpPP2A-A subunit was explored by using yeast two-hybrid experiments. Result Three MpPP2A-C genes (MpPP2A-C1, MpPP2A-C2, and MpPP2A-C3) were identified in the M. polymorpha genome. These genes were highly conserved in bryophytes, and their encoded proteins had conserved structural features, though they differed in physicochemical properties and subcellular localization. The MpPP2A-C gene was expressed in the apical notch, gemma cup and the rest of thallus. Among them, the expressions of MpPP2A-C1 and MpPP2A-C3 were the highest at the apical notch, while there was no significant difference in the expressions of MpPP2A-C2 among various tissues. ABA treatment significantly inhibited the growth of gemma. The expressions of MpPP2A-C1 increased with the rise of ABA concentration, there was no significant difference in the expression of MpPP2A-C2, while the expression of MpPP2A-C3 first increased and then decreased. The yeast two-hybrid experiment inferred that MpPP2A-A interacted with MpPP2A-C1 and MpPP2A-C3. Conclusion The MpPP2A-C is highly conserved, can respond to abscisic acid, and interacts with the A subunit. These results lay the foundation for future research on the function of the PP2A-C gene.

Identification of Litsea cubebaTCP Genes and Their Roles in the Regulation of Terpenoid Biosynthesis

NI Fei-fei, CHEN Yi-cun, GAO Ming, ZHANG Sheng-jiao, PENG Fang-you, CHEN Tao-mei, ZHAO Yun-xiao, WANG Yang-dong

2026, 42(4):  227-238.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0740

Asbtract ( 60 )   HTML ( 2)   PDF (4468KB) ( 178 )  

Figures and Tables | References | Related Articles | Metrics

Objective To screen and validate TCP transcription factors potentially involved in regulating terpenoid biosynthesis in Litsea cubeba. Method Based on the whole-genome sequence of L. cubeba and transcriptome data from fruits at different developmental stages, bioinformatics approaches were employed to systematically analyze the physicochemical properties, chromosomal distribution, gene structures, conserved motifs, cis-acting elements, and collinearity of TCP transcription factors. RT-qPCR was applied to examine the expression patterns of CIN subfamily members in different tissues and fruit developmental stages. Subcellular localization of key transcription factors was performed, and their functions were further validated through yeast one-hybrid and transient expression assays. Result A total of 30 TCP gene family members (LcTCPs) were identified, unevenly distributed across nine chromosomes and designated LcTCP1 to LcTCP30. Their open reading frames ranged from 504 to 1 755 bp, with predicted molecular weights of 17 711.98–63 421.45 Da. These genes were classified into three subfamilies via phylogenetic analysis: PCF (17 members), CIN (7 members), and CYC/TB1 (6 members), consistent with the conserved motif patterns. Gene family expansion was mainly driven by whole-genome duplication events, and members within each subfamily had high conservation. RT-qPCR analysis revealed that LcTCP6 and LcTCP11 of the CIN subfamily showed relatively high expression in the fruits and during the peak stage of essential oil biosynthesis, suggesting their potential involvement in terpenoid metabolism. Notably, LcTCP11 showed an expression pattern similar to that of the key terpenoid biosynthetic gene LcDXS5 during fruit development and was localized in the nucleus. Functional assays confirmed that LcTCP11 regulated the expression of LcDXS5. Conclusion Thirty TCP transcription factors were identified from the L. cubeba genome. Among them, LcTCP11 showed co-expression with the key terpenoid biosynthetic gene LcDXS5 and was highly expressed in fruits compared with other tissues, suggesting that LcTCP11 may participate in regulating terpenoid biosynthesis in L. cubeba through regulating LcDXS5.

Function of Transcription Factor NtMYB96a in Regulating the Tolerance of Tobacco to Drought

LIU Qing-yuan, WU Hong-qi, CHEN Xiu-e, CHEN Jian, JIANG Yuan-ze, HE Yan-zi, YU Qi-wei, LIU Ren-xiang

2026, 42(4):  239-250.  doi:10.13560/j.cnki.biotech.bull.1985.2025-1094

Asbtract ( 146 )   HTML ( 9)   PDF (26916KB) ( 30 )  

Figures and Tables | References | Related Articles | Metrics

Objective This study investigated the role of the tobacco NtMYB96a gene in drought-stress responses using transcriptome sequencing, with the aim of elucidating its regulatory mechanisms and providing candidate genetic resources for developing drought-resistant tobacco germplasm. Method The NtMYB96a coding sequence was cloned, and an overexpression vector was constructed and introduced into tobacco. Subcellular localization and tissue-specific expression analyses were performed. Drought-related physiological traits of NtMYB96a-overexpressing lines were evaluated at the seedling stage by assessing in vivo antioxidant activity. RNA-seq analysis was conducted on confirmed overexpression lines, and differentially expressed genes were subjected to GO and KEGG functional annotation and enrichment analyses. Result Subcellular localization showed that NtMYB96a is localized in the nucleus. RT-qPCR analysis revealed that there are expression in the roots, stems, and leaves, with the highest levels in the leaves across seedling, rosette, and vigorous growth stages. Under drought stress, NtMYB96a overexpressing lines had enhanced drought tolerance relative to wild-type plants, including reduced wilting, significantly increased CAT, POD, and SOD activities, and markedly decreased MDA content. Transcriptome analysis indicated that NtMYB96a alters the expression of genes involved in photosynthesis, carbon fixation, porphyrin metabolism, and photosynthetic antenna proteins, as well as several drought-responsive transcription factor families such as bHLH, MYB-related, and WRKY. Conclusion NtMYB96a positively regulates tobacco drought-stress responses by enhancing antioxidant enzyme activity and modulating key drought-related metabolic pathways and transcription factor networks, thereby improving plant drought tolerance.

Functional Analysis of the Tobacco NtPPO5 Gene Using VIGS and CRISPR/Cas9 Technology

PENG Chao-feng, XIA Lin, LIU Rui-xia, YAN Xin-ke, MU Yang-wei, LIU Meng-ru, YANG Jun, WU Ming-zhu

2026, 42(4):  251-262.  doi:10.13560/j.cnki.biotech.bull.1985.2025-1055

Asbtract ( 90 )   HTML ( 5)   PDF (14656KB) ( 209 )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the sequence characteristics, phylogenetic relationships, tissue-specific expression patterns of the polyphenol oxidase gene NtPPO5 in flue-cured tobacco (Nicotiana tabacum L.) K326, and its regulatory effects on PPO activity and total phenol content. Method To elucidate the function of the NtPPO5 gene (Accession number is Nta24g03870.10), it was cloned from the common cultivated tobacco cultivar K326. Subsequently, phylogenetic and amino acid sequence analyses were conducted to clarify the evolutionary relationships between NtPPO5 and its homologs. Furthermore, the expression patterns of NtPPO5 in various tobacco tissues were examined. Using both VIGS and CRISPR/Cas9 systems, loss-of-function mutants of NtPPO5 were generated through gene silencing and knockout, respectively. Based on this, changes in NtPPO5 expression levels, PPO activity, and total phenolic content were analyzed in the leaves of the mutant plants. Result The amino acid sequence encoded by the NtPPO5 gene contained typical conserved domains of the PPO family, including tyrosinase, PPO1_DWL, and PPO1_KFDV. Phylogenetic analysis revealed that NtPPO5 clusters within the same clade as NtomPPO from Nicotiana tomentosiformis and NtPPO1 from N. tabacum, indicating a close evolutionary relationship. Tissue-specific expression analysis showed that NtPPO5 was expressed in the root, stem, leaf, and flower tissues, with the highest expression observed in leaves. In both VIGS-silenced and CRISPR/Cas9 knockout plants, the expression of NtPPO5 and PPO activity significantly decreased, while the total phenolic content markedly increased. Conclusion NtPPO5 is a typical member of the PPO family, closely related to NtomPPO and NtPPO1. This gene is highly expressed in the leaves, with its expression positively correlated with PPO activity and negatively correlated with total phenol content.

Functional Analysis of PsMYB43 Regulating Linalool Synthesis in Tree Peony

WANG Zhen-quan, ZHONG Yi, MA Bo, WU Jing, HU Zeng-hui

2026, 42(4):  263-271.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0832

Asbtract ( 64 )   HTML ( 4)   PDF (4505KB) ( 215 )  

Figures and Tables | References | Related Articles | Metrics

Objective The monoterpene substance linalool is an important component of the floral fragrance in tree peony (Paeonia suffruticosa ‘OuKan’), and MYB-type transcription factors play a crucial role in the synthesis of monoterpenes. Exploring the role of PsMYB43 in the synthesis of linalool in tree peony, aiming to provide genetic resources and theoretical basis for genetic improvement of floral fragrance. Method Using the P. suffruticosa ‘OuKan’ as the experimental material, the CDS sequence of the PsMYB43 gene was first cloned, and its physicochemical properties, conserved structural domains, and phylogenetic relationships were analyzed using bioinformatics software. Secondly, RT-qPCR was used to analyze its expression patterns in different floral development stages and organ tissues. Subcellular localization was performed using tobacco, and finally the function of PsMYB43 was validated through transient overexpression and transient silencing to clarify its role in the synthesis of monoterpene compound linalool. Result The CDS sequence of PsMYB43 is 1 140 bp and encodes 379 amino acids. PsMYB43 is an unstable hydrophilic protein with two incomplete HTH domains, belonging to the R2R3-type MYB transcription factor family. Systematic evolutionary analysis revealed that the PsMYB43 protein in P. suffruticosa is most closely related to the PdMYB12L protein in P. delavayi. With flowering developing, the expressions first increased and then decreased, reaching its peak at the half-opening stage, consistent with the release pattern of linalool. PsMYB43 has the highest expressions in the outer petals during full blooming, while its expressions relatively low in carpels and stamens. Subcellular localization indicated that PsMYB43 is localized in the cell nuclear. Following transient overexpression of PsMYB43 in snapdragons, the levels of monoterpenes such as linalool, myrcene, and linalool significantly increases, rising by 2.0, 1.67, and 3.4-fold, respectively. Following transient silencing of PsMYB43 in tree peony, linalool release decreases significantly by 38%. Conclusion PsMYB43 positively regulates the synthesis of monoterpenoid linalool in tree peony petals.

Screening and Identification of Antagonistic Bacterium against Banana Fusarium wilt and Its Biocontrol Effects

LIN Bao-mei, LI Shan-shan, LI Hai-ming, HONG Jia-min, ZHANG Shuai, WU Miao-hong, WU Wei-jian, WU Shui-jin

2026, 42(4):  272-286.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0891

Asbtract ( 84 )   HTML ( 8)   PDF (91274KB) ( 228 )  

Figures and Tables | References | Related Articles | Metrics

Objective Banana Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense (FOC), is a devastating soil-borne disease that seriously impacts banana production in terms of cultivation area, yield, and fruit quality. This study is aimed to screen effective biocontrol strains against FOC to facilitate the green and sustainable management of this disease. Method Antagonistic bacteria were isolated from the rhizosphere soil of banana plants. A potent isolate, GX-13, was selected and identified based on its morphological, physiological and biochemical characteristics, 16S rRNA gene sequence, and whole genome sequence, with subsequent annotation for potential biocontrol-related genes. Additionally, we assessed its plant-growth-promoting traits (phosphate solubilization, ammonia production, indole-3-acetic acid (IAA) synthesis) and antifungal properties (production of siderophores and cell wall-degrading enzymes). The efficacy of its sterile fermentation supernatant, volatile and non-volatile metabolites against FOC were evaluated in vitro, along with its inhibitory spectrum against other plant pathogenic fungi. Finally, the biocontrol efficacy and plant-growth-promoting ability of GX-13 were validated in a pot experiment. Result A bacterial strain, GX-13, showing antagonistic activity against FOC, was identified as Bacillus subtilis. GX-13 presented plant growth-promoting traits, including phosphate solubilization, ammonia production, indole-3-acetic acid synthesis, and antifungal functional characteristics such as secretion of fungal cell wall-degrading enzymes (protease, β-1,3-glucanase, cellulase) and siderophore production. The non-volatile metabolites of GX-13 caused structural damage to FOC hyphae, resulting in an inhibition rate of 76.85% and significantly reducing hyphal invasiveness. In contrast, its volatile metabolites and sterile fermentation supernatant showed lower inhibition rates (20.01% and 16.48%, respectively). GX-13 also demonstrated a broad antifungal spectrum, effectively inhibiting eight other phytopathogenic fungi, including Fusarium oxysporum f. sp. niveum, Fusarium oxysporum f. sp. lycopersici, Magnaporthe oryzae, Colletotrichum acutatum, Aspergillus carbonarius, Colletotrichum musae, Phyllosticta graminicola and Pseudopestalotiopsis theae. In pot experiments, GX-13 achieved control efficacies of 68.42 % against leaf yellowing and 50.00% against corm browning. Furthermore, it significantly promoted plant growth, with a highly significant enhancement in root development (P<0.01). Gene function annotation and secondary metabolite biosynthetic gene cluster analysis revealed that GX-13 harbored numerous genes involved in the synthesis fungal cell wall-degrading enzymes, antimicrobial compounds, and pathways responsible for plant growth promotion and pathogen antagonism. Conclusion The Bacillus subtilis strain GX-13 effectively controls banana Fusarium wilt through a synergistic mechanisms, including direct antifungal activity, broad-spectrum antagonism, and plant growth promotion.

Physicochemical Properties and Bacterial Community Characteristics of Rhizosphere Soil in Olive Orchards under Different Soil Conditions

ZHANG Jian-xia, JIANG Cheng-ying, WU Wen-jun, JIN Gao-ming, ZHANG Cong-cong, YAO Yu-fang, ZHANG Rong, QI Jian-li

2026, 42(4):  287-296.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0755

Asbtract ( 784 )   HTML ( 4)   PDF (4699KB) ( 95 )  

Figures and Tables | References | Related Articles | Metrics

Objective To provide a basis for optimizing cultivation techniques, this study explored the effects of different soil conditions (loess, phyllite soil, and debris flow alluvial fan soil) in the non-traditional olive-growing area of Longnan, Gansu on the rhizosphere microorganisms of olive trees. Method The study focused on 15-year-old ‘Lei Xing’ olive trees in Wudu district, Longnan. Soil physical and chemical properties were measured, and the bacterial community structure and function were analyzed through metagenomic sequencing. Result All three soil types showed slightly alkaline pH (7.87-7.93). Debris flow alluvial fan soil (Cs) possessed significantly higher levels of key nutrients compared to loess (As) and schist (Bs) soils, including available nitrogen (AN: 186.03 mg/kg), total phosphorus (TP: 2.23 g/kg), and soil organic matter (SOM: 64.58 g/kg).Actinobacteria and Proteobacteria were the dominant bacterial phyla in both the rhizosphere and root endosphere compartments across all soil types. The relative abundance of Actinobacteria was significantly elevated in the Cs rhizosphere soil. PCA revealed that root endosphere microbial communities were relatively similar regardless of soil type. In contrast, rhizosphere microbial communities from Bs and Cs soils clustered together, showing significant divergence from those in As soil. RDA indicated that soil pH, total nitrogen (TN), total phosphorus (TP), and organic matter (OM) were the primary environmental factors driving variation in microbial community composition, collectively explaining 99.28% of the observed variance. Notably, the relative abundance of Actinobacteria showed a significantly positive correlation with TN, TP, and OM contents. KEGG annotation demonstrated that metabolic functions dominated the functional profiles across all samples. Metagenomic analysis further revealed a higher abundance of genes associated with nitrogen cycling pathways in the Cs rhizosphere soil compared to the other soil types. Conclusion This study demonstrates that soil type significantly shapes the structure and functional potential of olive rhizosphere microbial communities in the non-traditional Longnan region, primarily through its influence on nutrient availability and pH. Debris flow alluvial fan soil (Cs), characterized by higher nutrient content and a suitable pH, fostered a distinct and potentially beneficial microbial community enriched in Actinobacteria. Consequently, Cs soil should be prioritized as a reference soil type for new olive orchard site selection in similar non-traditional regions. These findings provide a scientific basis for enhancing olive cultivation efficiency through optimized soil management and site selection.

Optimization of the Fermentation Process and Plant Growth-Promoting Effects of a Salt-tolerant Microbial Consortium from the Rhizosphere of Elaeagnus angustifolia

DONG Yu-tao, BAO Hui-fang, He Miao, HOU Peng, LI Feng, WANG Zhi-xi, QIN Yao, LUO Qing-hong

2026, 42(4):  297-309.  doi:10.13560/j.cnki.biotech.bull.1985.2025-1104

Asbtract ( 119 )   HTML ( 4)   PDF (4143KB) ( 39 )  

Figures and Tables | References | Related Articles | Metrics

Objective To investigate the fermentation process of salt-tolerant and plant growth-promoting microbial consortia and evaluate their growth-promoting effects on plants under salt stress, providing theoretical basis for plant cultivation and microbial product development in saline-alkali regions of Xinjiang. Method Two salt-tolerant PGPR (plant growth-promoting rhizobacteria) strains isolated from the rhizosphere of Elaeagnus angustifolia in saline-alkali soil were selected to construct a bacterial consortium. The fermentation medium and culture conditions were optimized using single-factor experiments and Box-Behnken response surface methodology. Biological traits (viable cell count, biomass, and growth kinetics) and plant growth-promoting traits (solubilization of organic, inorganic phosphorus, nitrogen fixation and indole-3-acetic acid (IAA) production) were analyzed before and after optimization. A pot experiment was conducted to assess the consortium's effects on plant height, root length, leaf number, stem diameter, and fresh weight under salt stress. Result The optimal fermentation medium consisted of 14 g/L molasses, 11 g/L peptone, 5 g/L yeast extract, and 5 g/L Na₂HPO₄. Optimal culture conditions were pH 7.0, inoculation volume 10%, incubation temperature 37 ℃, shaking speed 180 r/min, and filling volume 40 mL per 150 mL flask. Under these conditions, viable cell count reached 7.28×10⁹ CFU/mL, representing a 36.04-fold increase over the unoptimized medium, with a shorter lag phase (6 h). The optimized consortium showed enhanced plant growth-promoting activity, with organic phosphorus solubilization, inorganic phosphorus solubilization, nitrogen fixation, and IAA production increased by 56.21%, 44.38%, 69.70% and 64.57%, respectively. Under salt stress, compared with the sterile water control group (CK), the optimized microbial consortium treatment group (FYA) showed significant increases in growth parameters of Elaeagnus angustifolia, with plant height, root length, leaf number, stem diameter, and fresh weight increasing by 63.76%, 34.17%, 36.24%, 71.15%, and 73.68%, respectively. Compared to the unoptimized compound bacterial treatment group (FYB), the FYA group showed improvements of 19.32%, 5.96%, 15.58%, and 17.86% in plant height, root length, stem diameter, and fresh weight, respectively. Conclusion The optimized fermentation process significantly enhanced both the viability and growth-promoting traits of the salt-tolerant microbial consortium, contributing to improved plant growth under salt stress and offering potential for application in saline-affected agricultural systems.

Identification and Control Characteristics of Antagonistic Bacteria against Tobacco Target Spot Disease

ZHU Zhi-yan, SONG Zhi-wei, HE Shan, ZHANG De-qing, HE Yong, TIAN Zhi-hong

2026, 42(4):  310-320.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0678

Asbtract ( 78 )   HTML ( 7)   PDF (2369KB) ( 206 )  

Figures and Tables | References | Related Articles | Metrics

Objective To isolate and screen biocontrol bacteria against tobacco target spot disease. Method The antagonists were isolated and screened by plate coating method and plate confrontation method from Rhododendron simsii Planch. rhizosphere. After identification by physiological and biochemical analysis and 16S rDNA gene sequence analysis, the phylogenetic tree was constructed, and the antibacterial activities of the fermented broth and the crude extract of ethyl acetate were determined concurrently. Pot culture method was used to determine the control effect of antagonists on tobacco target spot disease, as well as the effects on the activities of malondialdehyde (MDA), antioxidant enzymes and the regulation and expression of pathogenesis related protein genes. Result A strain of Burkholderia gladioli QLDJ-1 with antagonistic effect against Rhizoctonia solani was obtained. The relative inhibition rate of QLDJ-1 against R. solani was (72.107 ± 2.788) %. Compared with the control group, the disease index of target spot of tobacco plants treated with QLDJ-1 decreased by 32.02%. The activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) increased by 33.72%, 51.45% and 17.49% respectively. The content of malondialdehyde (MDA) decreased by 62.44%. The expressions of pathogenesis-related genes such as PR1b, NmIMSP, P450-1, GST and hrs203J in the tobacco plants treated with QLDJ-1 were different from those of the control. Conclusion QLDJ-1 can effectively control tobacco target spot disease and has the potential to be developed as a biocontrol agent.

Growth-promoting Effect of Arcopilus aureus XBAa1 on Tobacco and Its Colonization in Tobacco Root

GENG Ming-yan, FENG Hui, HE Lei, ZHANG Guo-zheng, WANG Jing

2026, 42(4):  321-331.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0732

Asbtract ( 58 )   HTML ( 7)   PDF (16855KB) ( 304 )  

Figures and Tables | References | Related Articles | Metrics

Objective This work is to investigate the growth-promoting effects of Arcopilus aureus strain XBAa1 on tobacco and the colonization dynamics in roots. Method By reconstruction of the tobacco-XBAa1 fungal symbiosis, its endophytic performance and plant growth promotion was evaluated. Based on the PEG-mediated protoplast transformation method, GFP gene carried by the P3641 plasmid was integrated into A. aureus XBAa1 genome, the transformants stably expressing GFP were obtained. Transformants were used to infect tobacco roots. The infection sites and colonization levels of strain XBAa1 within tobacco roots were detected using laser confocal microscopy (LCM) and quantitative absolute real-time PCR assays. Result A. aureus XBAa1 endophytically colonized in tobacco roots, which significantly promoted tobacco growth. The biomasses of the shoots and roots increased remarkably, with the chlorophyll a+b contents in the leaves enhanced by 63.17% compared with CK. It regulated the changes in hormones of tobacco, with cytokinin content increased by 143.65% and abscisic acid content decreased by 46.56% relative to CK, respectively. Meanwhile, it reduced the degree of oxidative damage in tobacco, and the malondialdehyde content decreased by 38.02% compared with CK. The optimal cell wall lysis conditions for protoplast preparation of XBAa1 were enzymatic hydrolysis with 12.5 mg/mL megalyase solution treated for 3 h. For the obtained XBAa1-Y3 transformant, laser confocal microscopy observations revealed that it colonized stably in the intercellular spaces of tobacco root tissue cells. Under the condition of sterile substrate in the greenhouse, the colonization biomasses of XBAa1-GFP strain in tobacco roots showed a “first increasing and then decreasing” trend within 30 d post-inoculation (dpi), reaching a peak of 1.23×10⁷ copies/mL at 15 dpi; at 30 dpi, the colonization density decreased to 8.12×10⁶ copies/mL. Conclusion A. aureus XBAa1 had stable colonization in tobacco roots and remarkable growth-promoting effects on host plants, indicating considerable application potential in agriculture. The successfully obtained GFP-tagged XBAa1 transformant verified that protoplasts are feasible materials for subsequent genetic transformation, establishing a critical basis for an in-depth clarifying the migration traits of XBAa1 in tobacco plants and its biocontrol functions.

Fermentation Optimization of a Surfactin-producing Bacillus Strain and Its Application in Prevention and Control

SU Chang, WEN Feng, WANG Ye-lin, SHAO Rui-ying, TANG Jia-jie, XIA Zhan-feng

2026, 42(4):  332-344.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0996

Asbtract ( 119 )   HTML ( 7)   PDF (36058KB) ( 38 )  

Figures and Tables | References | Related Articles | Metrics

Objective This study aimed to optimize the fermentation conditions and evaluate the insecticidal activity of a surfactin-producing strain, Bacillus australimaris TRM82479, isolated from desert soil. Method Key influencing factors were screened using the Plackett-Burman design, followed by optimization of fermentation conditions via the Box-Behnken response surface methodology. The effects of various nutrients (including carbon and nitrogen sources), cultivation time, temperature, shaking speed, initial pH, inoculation volume, and medium volume on surfactin production were determined using the oil displacement method and HPLC-MS. The insecticidal spectrum of the bacterial suspension was determined using the feed poisoning method and leaf dipping method; and the field control effects of the bacterial suspension at different concentrations were evaluated via the five-point sampling method. Result The optimal liquid medium composition and fermentation parameters for surfactin production were as follows: Maltose 8.912 g/L, yeast extract powder 7.135 g/L, sodium chloride 7 g/L, magnesium sulfate 0.2 g/L, fermentation temperature 28 ℃, fermentation time 75 h, shaking speed 160 r/min, medium volume 150 mL, and vaccination rate of 4%.Under these optimized conditions, surfactin yield increased approximately 1.88-fold, reaching a concentration of about 1 355.78 mg/L. The bacterial culture had insecticidal activity against seven agricultural pests, demonstrating a broad insecticidal spectrum. The highest mortality was observed against aphids, with a corrected mortality rate of 91.23% at 48 h. In field trials, the supernatants at concentrations of 10⁸, 10⁷, and 10⁶ CFU/mL provided control efficacies against aphids of 77.33%, 70.48%, and 66.26%, respectively, after seven days, all of which were higher than the 56.95% efficacy achieved by the chemical pesticide flonicamid (25%). Conclusion After optimizing the fermentation process of strain TRM82479, the yield reaches 1 355.78 mg/L, it exerts a good control effect on multiple agricultural pests, and its field control effect against aphids after 7 d is as high as 77.33%. This is significantly higher than that of the chemical pesticide 25% flonicamid, indicating great potential for development as a microbial insecticide.

Inositol Biosynthesis and Fermentation Optimization of an Inositol Biosynthesis Pathway Using Citrus sinensis Inositol Phosphate Synthase Gene Csino3

YAN Xin, WU Xiao, HE Si-yu, DUAN Yu-huan, QIU Wu-xia, YUAN Xiao-qin, MAO Xin-fang, LIU Zhong-yuan

2026, 42(4):  345-356.  doi:10.13560/j.cnki.biotech.bull.1985.2025-0879

Asbtract ( 92 )   HTML ( 8)   PDF (19042KB) ( 101 )  

Figures and Tables | References | Related Articles | Metrics

Objective To establish an efficient myo-inositol biosynthetic pathway through prokaryotic co-expression of Citrus sinensis myo-inositol phosphate synthase gene Csino3 and Escherichia coli MG1655 inositol-1-monophosphatase gene suhB. To increase the inositol yield by optimizing fermentation conditions and knocking out pgi and gldA genes in E. coli. Method The prokaryotic co-expression vector pETDuet-1-Csino3-suhB was constructed and transformed into E. coli BL21(DE3) for heterologous expression. Protein expression was induced with IPTG and the expression of soluble protein was identified via SDS-PAGE analysis. Four fermenting parameters, glucose concentration, IPTG concentration, inoculum size, and initial pH were optimized through single-factor experiments. Further fermentation condition was optimized using response surface methodology. Then myo-inositol yield was enhanced by knocking out the pgi gene (encoding glucose-6-phosphate isomerase) and the gene gldA (glycerol dehydrogenase) to modify the metabolic pathway of E. coli . Result Successful heterologous co-expression of the recombinant plasmid pETDuet-1-Csino3-suhB in E. coli BL21(DE3) confirmed that both Csino3 and SuhB proteins were expressed in soluble forms under 25 ℃ induction. The fermentation conditions for the target strain were optimized using single-factor experiments and response surface methodology (RSM). The optimal conditions were determined as follows: Glucose concentration 9.7 g/L, IPTG concentration 0.5 mmol/L, inoculum size 8%, and initial pH 8.1. Under these conditions, myo-inositol production reached 309 mg/L. Based on the above mentioned fermentation conditions, further optimization was performed for the knockout strain △gldAΔpgi-pETDuet-1-CSino3-suhB/E.coli BL21(DE3). Under an initial glycerol concentration of 20 g/L, the myo-inositol production reached 3.97 g/L, representing an approximately 12.8-fold increase compared to the pre-knockout strain. Conclusion The recombinant strain pETDuet-1-CSino3-suhB /E.coli BL21(DE3) can efficiently produce inositol. Knocking out the pgi and gldA gene to redirect the metabolic pathway of E. coli significantly enhances inositol yield.

Content

2026, 42(4):  346. 

Asbtract ( 4 )   PDF (1297KB) ( 1 )  

Related Articles | Metrics

copyright

2026, 42(4):  347. 

Asbtract ( 4 )  

Related Articles | Metrics

Cover

2026, 42(4):  348. 

Asbtract ( 4 )  

Related Articles | Metrics