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26 April 2026, Volume 42 Issue 4

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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

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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 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 (Oryza sativa) 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.

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

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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. Methods Fluorescence quantification using a fluorescence detector and visual readout were employed to evaluate systematically 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). Results 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 produced a CasRB buffer that combined cost reduction and enhanced fluorescence sensitivity. The buffer addressed the compatibility limitations of conventional buffers in Cas12a-based nucleic acid detection systems, offering a versatile platform for multiple Cas12a proteins.

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

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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

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Objective To analyze the basic characteristics of the ZF-HD gene family in peanut (A. hypogaea) 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 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

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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 were 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 were screened out, whose differential expression in QO10 and QS5 seeds is a potential cause of the difference in ALA 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

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Objective To clarify the phenotypic variation pattern of sugarcane germplasm resources and theregulatory 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 weighted gene co-expression network analysis (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.

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

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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.

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

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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.

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

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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.