Biotechnology Bulletin

Research Progress in Plant SUMOylation

SANG Tian, WANG Peng-cheng

2023, 39(3): 1-12. doi:10.13560/j.cnki.biotech.bull.1985.2022-0928

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SUMOylation is a highly-conserved post-translational modification in eukaryotes. Under the synergistic work of the SUMOylase system, mature SUMO can covalently target to the lysine residue in substrate proteins in the form of iso-peptide bonds. The SUMOylation is highly dynamic and can regulate various biological processes by affecting target proteins'stability, activity, localization, etc. The conjugated SUMO molecular can be removed from its targets by SUMO proteases and re-enter the SUMO catalytic cycle. Reports indicate that SUMOylation is involved in stress response, growth, and flowering. In this review, we briefly summarize the current processes of the plant SUMOylation pathway and discusses the identification of SUMO tragets by biochemistry and proteomics strategies.

Advances in the Development and Regulation of Vascular Cambium

GE Yan-rui, ZHAO Ran, XU Jing, LI Ruo-fan, HU Yun-tao, LI Rui-li

2023, 39(3): 13-25. doi:10.13560/j.cnki.biotech.bull.1985.2022-0865

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The vascular cambium, part of the secondary lateral meristem, contributes to the lateral growth of a plant. In recent years, many studies have strengthened our understanding of the vascular cambium. However, we still know far less about the vascular cambium than the apical meristem. Genetic and molecular studies have revealed that the proliferation and differentiation of cambium is regulated by many factors, including long-distance hormone signals, short-range peptide signals, and interactions between them. In addition, a large number of transcription factors and microRNAs also play a crucial role in the regulation of the vascular cambium activities. This review focuses on the novel discoveries on the development of vascular cambium and its regulation of proliferation and differentiation. The review also summarizes the current research and prospects for future research in this field.

Multifaceted Roles of bHLH Phosphorylation in Regulation of Plant Physiological Functions

LIU Cheng-xia, SUN Zong-yan, LUO Yun-bo, ZHU Hong-liang, QU Gui-qin

2023, 39(3): 26-34. doi:10.13560/j.cnki.biotech.bull.1985.2022-0775

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bHLH（basic Helix-Loop-Helix）is the second largest class of transcription factors in plants and plays an important role in the transcriptional regulatory network of plant growth and development and stress responses. Protein phosphorylation as a quick and flexible posttranslational modification affects the transcriptional activity, localization, protein interaction and stability of transcription factors. To understand the multifaceted roles of bHLH phosphorylation, here we review recent advances in the phosphorylation of bHLH family members, including the structure, classification, and function of the bHLH transcription factors, as well as the changing of their physiological and biochemical function caused by mutations at the phosphorylation sites, aiming to provide theoretical basis for improving agronomic traits such as nutrient use efficiency, quality and stress tolerance from the perspective of phosphorylation regulation.

PIP/PIPL: A Kind of Endogenous Plant Peptide Regulating Plant Stress Response and Development

CUI Jun-mei, WEI Jia-ping, DONG Xiao-yun, WANG Ying, ZHENG Guo-qiang, LIU Zi-gang

2023, 39(3): 35-42. doi:10.13560/j.cnki.biotech.bull.1985.2022-0816

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Plant endogenous polypeptides are composed of natural amino acids in different composition and arrangement from the precursor proteins cut into mature polypeptides, which play biological function in development and stress response after secreted outside the cells and recognized by their receptors. The plant endogenous polypeptide family PIP（PAMP induced secret peptide）/PIP-Like（PIPL）contains SGPS and GxGH motifs recognized by receptor like kinase 7（RLK7）, and plays important roles in pathogen resistance, virus resistance, salt resistance and development. Here, we review the signal transduction pathways of PIP/PIPL family polypeptides in the process of resistance to stress and development, discuss important unsolved scientific issues, and prospect possible applications, aiming to provide the references on further researches on PIP/PIPL.

Research Progress in Plant Endophytic Fungi for Root-knot Nematode Control

YI Xi, LIAO Hong-dong, ZHENG Jing-yuan

2023, 39(3): 43-51. doi:10.13560/j.cnki.biotech.bull.1985.2022-1010

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The diseases caused from root-knot nematode is the one damaging crops seriously but quite difficult to be controlled, and is becoming more and more serious with the development of facility agriculture in China. Conventional chemical control methods are not suitable for sustainable development of agriculture due to high toxicity and ecological damage. As a biological control fungus that can be stably parasitized in crops, endophytic fungi achieve stable, efficient and safe control of root-knot nematode diseases by inhibiting egg hatching, reducing the viability of J2 stage nematode larvae, inhibiting nematode invasion, delaying female development, reducing the number of laid eggs, and decreasing the number of root knots and egg masses in crop roots. In recent years, the action mechanism of endophytic fungi has received extensive attention and research, and significant progress has been earned. This paper reviews the research progress of biological control mechanism of endophytic fungi for root knot-nematode in recent years, summarizes four main mechanisms such as direct attack, resource competition, metabolite stress and defense activation of endophytic fungi, and discusses the existing issues, aiming to provide help for further development and application of plant endophytic fungi control technology.

Roles of Phosphate-solubilizing Microorganisms in the Passivation and Phytoremediation of Heavy Metal Contaminated Soil

ZHANG Hua-xiang, XU Xiao-ting, ZHENG Yun-ting, XIAO Chun-qiao

2023, 39(3): 52-58. doi:10.13560/j.cnki.biotech.bull.1985.2022-0784

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Passivation and phytoremediation are primary means for the ecologsical remediation of heavy metal contaminated soil, and microorganisms can further strengthen the passivation and phytoremediation of heavy metal contaminated soil. We introduced the basic principles of passivation and phytoremediation of heavy metal contaminated soil. Then we reviewed the study progresses on the solubilization of insoluble phosphates in soil by phosphate-solubilizing microorganisms, passivation remediation of heavy metal contaminated soil by phosphates, enhancement of phosphate passivation and phytoremediation of heavy metal contaminated soil by phosphate-solubilizing microorganisms. Further we discuss the heavy metals resistance of phosphate-solubilizing microorganisms and the phosphate-solubilizing mechanism. We also explored the enhancement mechanism of phosphate passivation and phytoremediation of heavy metal contaminated soil by phosphate-solubilizing microorganisms. The aim of this study is to provide a certain theoretical and technical references for bioremediation of heavy metal contaminated soil.

Research Progress in Metabolites Produced by Bacillus Against Three Common Plant Pathogenic Fungi

WANG Wei-chen, ZHAO Jin, HUANG Wei-yi, GUO Xin-zhu, LI Wan-ying, ZHANG Zhuo

2023, 39(3): 59-68. doi:10.13560/j.cnki.biotech.bull.1985.2022-1315

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Plant pathogenic fungi are one of the major threats to agricultural production. Applying biological agents to control pathogenic fungi is widely considered to be a safer and more sustainable strategy. Bacillus species can produce a variety of antifungal active substances（lipopeptides, bacteriocins and enzymes, etc.）, which is the most widely used biocontrol bacteria at present. Biocontrol agents based on Bacillus spp. and its metabolites can effectively control plant pathogenic fungi and play an important role in agricultural production. This paper focuses on the biological control potential of Bacillus metabolites and their antagonistic properties and mechanisms against three common plant pathogenic fungi（Magnaporthe oryzae, Fusarium oxysporum, and Botrytis cinerea）. Several important Bacillus metabolites were introduced by investigating the related literatures published in recent years on the antifungal activities of Bacillus metabolites, and the antifungal effects and mechanisms of Bacillus metabolites on important plant pathogenic fungi were summarized. And meanwhile the research methods and effects of Bacillus metabolites on the damages of cell wall and cell membrane of pathogenic fungi, inhibition of fungal spore germination and mycelial growth, and competitive binding with fungal DNA were summarized. The aim of this review is to provide guidance for the preparation and application of Bacillus biocontrol agents in the future.

Application of Lipidomics in Toxicological Studies

WANG Xin-lu, WANG Meng, ZHAI Wen-lei

2023, 39(3): 69-80. doi:10.13560/j.cnki.biotech.bull.1985.2022-0793

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As major components of cells, lipids play important roles in a variety of life activities, such as intracellular material transport, signal transduction and apoptosis, etc. Lipidomics provides a systematic analysis of lipids in living organisms to understand their interactions and the interactions with other biomolecules. The authors mainly introduced the analysis methods of lipidomics, including the pretreatment methods of samples, detection methods and instruments, and data analysis methods. Besides that, the authors reviewed the research progress of lipidomics in neurotoxicity, hepatotoxicity, immunotoxicity and endocrine disrupting toxicity, aiming to provide ideas and reference for studying the mechanism of compound toxicity. Finally, the authors suggested that lipidomics technology should be used to study the combined toxic effects of different compounds in the future, to screen relevant lipid biomarkers based on the lipidomics, as well as to provide the scientific basis for risk warning of combined toxicity and relevant toxic mechanism study.

Development and Genetic Analysis of Two Nullisomic Lines（NC1 and NC2）in Natural Brassica napus

CAI Meng-xian, GAO Zuo-min, HU Li-juan, FENG Qun, WANG Hong-cheng, ZHU Bin

2023, 39(3): 81-88. doi:10.13560/j.cnki.biotech.bull.1985.2022-0577

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The nullisomic（2（n-1））line losing a pair of chromosome from the normal organism chromosome set has been recognized as a rare and important genetic resources for revealing the missing chromosome. Herein, after screening the offsprings from the cross and backcross between Brassica napus and the restituted B. rapa（RBR, 2n =20, AⁿAⁿ）, we used the integration of specific C chromosome molecular markers and fluorescence in situ hybridization technology to screen two nullisomic lines of chromosome C1（NC1）and chromosome C2（NC2）in natural B. napus（2n =38, AⁿAⁿCⁿCⁿ）for the first time. Both two nullisomic lines had 18 bivalents at diakinesis and a dominant segregation of 18:18 at meiosis anaphase I（MA I）in the pollen mother cells. However, some lagging chromosomes were observed at MA I and MA II in both two lines. Compared to their parental B. napus, the two lines presented significantly smaller phenotype at biomass, pollen stainability, and seed setting, also showed some specific features. For example, the NC1 had the leaves with obvious burrs, and NC2 showed significantly early flowering time of two months earlier than its parental B. napus, indicating some genes inhibited such characters were locating on the missing chromosome. The two nullisomic lines effectively reduce the complexity of B. napus genome, which can be used to map the target traits, or to verify the gene function, and thus to provide convenience for future genetic analysis of B. napus.

Development Core SNP Markers for Tobacco Germplasm Genotyping

YU Shi-zhou, CAO Ling-gai, WANG Shi-ze, LIU Yong, BIAN Wen-jie, REN Xue-liang

2023, 39(3): 89-100. doi:10.13560/j.cnki.biotech.bull.1985.2022-0810

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Based on kompetitive allele specific PCR（KASP）technology platform, the tobacco SNP（single nucleotide polymorphism）markers and corresponding primers were developed and validated，which could assist tobacco germplasm evaluation, genetic diversity analysis, and core germplasm screening. Using Python and Perl tools, 1 179 154 SNPs covering tobacco genome were designed and screened for KASP primers, and their accuracy and applicability were verified by experiment. As results, 217 621 SNPs were enough for designing corresponding KASP primers. Then 1 378 SNPs were selected for experimental validation, and 732 were approved for SNP marker. Further 48 SNP markers, with an average PIC of 0.36 and an average MAF of 0.39, were finally identified as core markers, and they were evenly distributed on 24 chromosomes of the tobacco genome. The various germplasm, especially the genotypes of current major tobacco cultivars could be distinguished by the 48 core SNPs, and the marking is of extremely high reliability.

Selection and Validation of Reference Genes for RT-qPCR in Allium tuberosum Infected by Botrytis squamosa

SONG Hai-na, WU Xin-tong, YANG Lu-yu, GENG Xi-ning, ZHANG Hua-min, SONG Xiao-long

2023, 39(3): 101-115. doi:10.13560/j.cnki.biotech.bull.1985.2022-0851

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Chinese chive gray mold caused by Botrytis squamosa is one of the major factors affecting the production and quality of Chinese chive（Allium tuberosum）. In order to screen stable reference genes in Chinese chive leaves infected with gray mold for quantitative gene expression analysis, Chinese chive leaves mock-inoculated and inoculated with B. squamosa for 24, 48 and 72 h were taken as materials. Thirteen genes including UBC1, UBC2, UBQ1, UBQ2, GAPDH3, GAPDH4, TUB, EF-1α, 40S RP, DDX, eIF-1A, PABP and DnaJ were selected as candidate reference genes based on previous RNA-Seq data of Chinese chive. The expressions of the candidate reference genes were detected by real-time quantitative PCR（RT-qPCR）and evaluated by geNorm, NormFinder, BestKeeper and RefFinder statistical algorithms. The results showed that among the thirteen candidate reference genes, UBQ1 had the smallest variation range of Ct value and the most stable expression. The most suitable reference genes evaluated by geNorm, NormFinder and BestKeeper software were different. Comprehensive evaluation of RefFinder website showed that UBC2 and UBQ1 had the better stability in Chinese chive leaves after inoculation with B. squamosa, and DDX displayed the lower stability. In order to verify the reliability of the selected reference genes, six candidate genes with different stability were used as internal controls for quantitative analysis, and normalized the expression levels of GST676 and PRP902 in Chinese chive leaves at different time points after inoculation with B. squamosa. The expression trends of GST676 and PRP902 corrected by UBC2 or UBQ1 with fine stability were consistent with that in RNA-Seq data, while the expression trends of GST676 and PRP902 corrected by 40S RP, eIF-1A, GAPDH4 or DDX with relative poor stability had some difference with that in RNA-Seq data. These results of software analysis and experimental verification showed that UBC2 and UBQ1 were the most suitable reference genes in Chinese chive leaves infected by B. squamosa for quantitative expression analysis. The results lay the foundation for future research on the expression and function of key genes in Chinese chives leaves during infection with B. squamosa.

Efficient Generation of Secondary Libraries During Functional Nucleic Acids Screening

LI Tian-shun, LI Chen-wei, WANG Jia, ZHU Long-Jiao, XU Wen-tao

2023, 39(3): 116-122. doi:10.13560/j.cnki.biotech.bull.1985.2022-0743

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In vitro screening of functional nucleic acids（FNAs）by SELEX is an essential prerequisite for their various functions. In the SELEX process, the preparation of high-quality single-strand secondary library is one of the most basic and critical technologies, which largely determines the success or failure of screening. Here, we comprehensively summarize the generation strategies of single-strand DNA for FNAs selection, such as amplification-based methods, enzymatic digestion techniques, magnetic separation with streptavidin-coated beads, and secondary library regeneration strategies based on mobility shift. The advantages, disadvantages and key details of various methods are elaborated with aiming to lay a foundation for the further development of efficient FNAs screening methods.

Isolation and Expression Analysis of SiMAPK3 in Setaria italica L.

WANG Hai-long, LI Yu-qian, WANG Bo, XING Guo-fang, ZHANG Jie-wei

2023, 39(3): 123-132. doi:10.13560/j.cnki.biotech.bull.1985.2022-0819

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Mitogen-activated protein kinases（MAPKs）are a class of serine/threonine protein kinases that play important roles in plant growth and development, response to stress, and hormone signal transduction. In this study, the SiMAPK3 gene, which was the highest protein homology to AtMAPK3 in Arabidopsis, was cloned from the foxtail millet variety Yugu 1. The physicochemical properties, structure and function of SiMAPK3 protein were systematically analyzed by bioinformatics methods. The expressions of SiMAPK3 gene in different tissues and different abiotic stress in the early shooting stage of the foxtail millet was analyzed by RT-qPCR. The results showed that the open reading frame of SiMAPK3 gene was 1 128 bp, encoding a deduced protein of 376 amino acids, with a predicted molecular weight of 43 427.85 Da and an isoelectric point of 5.46. The SiMAPK3 in the foxtail millet was a hydrophilic extramembrane protein without signal peptide. Its secondary structure included 44.27% α helix, 14.67% β sheet, 5.07% extended chain and 36.00% random coil. SiMAPK3 amino acid 44-328 contained a Pkinase conserved domain, belonging to the MAPK protein kinase family. The tertiary structure of SiMAPK3 had high similarity with Arabidopsis MAPK protein, SiMAPK3 contained 11 serines, 8 threonines, 4 tyrosines and a large number of potential phosphorylation sites. Protein structural analysis showed that it had a conserved PKinase domain. RT-qPCR analysis showed that SiMAPK3 was expressed in the roots, stems and leaves of foxtail millet in the early shooting stage, and its expression level was the highest in leaves, which was about 25 times that in roots. SiMAPK3 responded to low temperature（4℃）, high salt（300 mmol/L NaCl）, drought and ABA（200 μmol/L）and JA（200 μmol/L）and other stresses. After 12 h of low temperature, the expression of SiMAPK3 was up-regulated 16-fold, and after 3 h of high-salt, the expression of SiMAPK3 was up-regulated 8-fold. The cloning and functional analysis of the SiMAPK3 gene provide an important basis for further analysis of the leaves development and the signal transduction in response to low temperature and high salt in foxtail millet.

Characterization of Soybean VAS1 Gene Family and Its Involvement in Lateral Root Development

YANG Chun-hong, DONG Lu, CHEN Lin, SONG Li

2023, 39(3): 133-142. doi:10.13560/j.cnki.biotech.bull.1985.2022-0690

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Arabidopsis VAS1 gene encodes a pyridoxal-phosphate-dependent aminotransferase, which regulates the auxin amount through converting the indole pyruvic acid to tryptophan. Genome-wide characterization and expression pattern analysis of soybean aminotransferase VAS1 was conducted to explore their roles in root development, for laying foundation in further mining biological functions of VAS1 in soybean plant（Glycine max）. Bioinformatics approaches were used to identify and analyze the gene members of the soybean VAS1 family, and then fluorescence quantitative analysis was used to analyze the expression pattern of this gene family under different stress treatments. In addition, GmVAS1-1 gene was cloned for further functional analysis. The interaction factors of GmVAS1-1 were screened through yeast two-hybrid method. The results showed that there were two members in the soybean VAS1 gene family, and were named as GmVAS1-1 and GmVAS1-2. Phylogenetic tree，gene structure and conserved motifs analysis indicated that soybean VAS1 was more closely related to legumes（Phaseolus vulgaris，Medicago truncatula，Vigna unguiculata，Lotus corniculatus and Cicer arietinum）. There were multiple stress or light response elements in the promoter region. Expression pattern analysis showed that soybean VAS1 genes were highly expressed in the endosperm at the different seed development stages. Fluorescent quantitative PCR analysis revealed that the expression of GmVAS1 was significantly up-regulated in soybean roots under drought or salt stress condition. The number of lateral roots in overexpressed GmVAS1-1 Arabidopsis was significantly less than that of wild type, suggesting that GmVAS1-1 was involved in regulating lateral root development in plants. A total of 17 proteins interacting with GmVAS1-1 were screened by yeast two-hybrid assays and network prediction, which laid a molecular theoretical basis for further exploring the function and mechanism of GmVAS1-1 gene. In summary, soybean VAS1 gene family are highly expressed in the endosperm and respond to various abiotic and nutritional stresses, among which GmVAS1-1 is involved in the development of lateral root.

Genome Wide Association Analysis of Thousand Seed Weight in Brassica napus L.

XIAO Xiao-jun, CHEN Ming, HAN De-peng, YU Pao-lan, ZHENG Wei, XIAO Guo-bin, ZHOU Qing-hong, ZHOU Hui-wen

2023, 39(3): 143-151. doi:10.13560/j.cnki.biotech.bull.1985.2022-0824

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This study aims to excavate the SNP loci and candidate genes significantly associated with the seed number per silique（SPS）in Brassica napus L. Having 300 B. napus inbreeding lines as experimental materials, we investigated the phenotypes of SPS under two locations（Jiangxi Agricultural University（JXAU）in Nanchang and Jiangxi Institute of Red Soil（JXIRS）in Jinxian in a year. Then we conducted the genome-wide association study via general linear model（GLM）and mixed linear model（MLM）combining 201 817 SNPs markers determined previously. And finally we predicted the functions of relevant candidate genes within the 100 kb region flanking trait significantly associated SNP loci. The results showed that the broad phenotypic variations of SPS in the two locations among the 300 300 B. napus samples existed, and two germplasm with more SNP were screened. Total 39 SNPs loci remarkably associated with SPS were excavated by general linear model（GLM）. Furthermore, 3 SNPs of SPS were detected severally using the mixed linear model（MLM）, and they all were detected using GLM. And 19 candidate orthologous genes to documented Arabidopsis the seed number per silique, like CIK, ERF022 and EDE1, were found near our eight SNPs loci. The results are conducive to analyzing the genetic basis of SPS of rapeseed and would lay a foundation for studying the regulation mechanism and guiding the genetic improvement of SPS.

Transcriptome Analysis of Diploid and Autotetraploid in Cucumber Fruit

XIE Yang, XING Yu-meng, ZHOU Guo-yan, LIU Mei-yan, YIN Shan-shan, YAN Li-ying

2023, 39(3): 152-162. doi:10.13560/j.cnki.biotech.bull.1985.2022-0974

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Fruit shape and sugar content are important quality traits closely related to the appearance, nutrition and flavor of melon vegetables, respectively. To investigate the molecular mechanism of fruit shape and sugar content in cucumber caused by ploidy differences, the diploid（long fruit/low sugar）and autotetraploid（short fruit/high sugar）cucumber with significant differences of shape and sugar content were selected as the study materials, and RNA-seq analysis on the fruits at 0 and 9 d after flowering were performed. The results showed that a total of 1 874 differentially expressed genes（DEGs）were identified from diploid and autotetraploid at 9 d after flowering, including 818 up-regulated genes and 1 056 down-regulated genes. GO analysis demonstrated that the DEGs were significantly enriched in DNA replication（GO:0006260）, DNA metabolic process（GO:0006259）, protein-DNA complex（GO:0032993）, chromosome（GO:0005694）, and protein dimerization activity（GO:0046983）. KEGG analysis revealed that the DEGs were enriched in “plant hormone signal transduction”（csv04075）, “starch and sucrose metabolism”（csv00500）and other metabolic pathways. The up-regulated expressions of AUX1, DELLA, B-ARR and TCH4 and down-regulated expression of GH3, GID1, PP2C and CYCD3 related to “plant hormone signal transduction”, as well as the up-regulated expression of SPS and SUSY related to “starch and sucrose metabolism”, may correlate with fruit shape index decreasing shortening and sugar content increasing in the cucumber. Quantitative reverse transcription-PCR（RT-qPCR）analysis showed that the expression pattern of selected DEGs was consistent with the expression trend of RNA-Seq except for ARF4, among which SUSY was highly differentially expressed between diploid and autotetraploid cucumber fruits, suggesting that SUSY plays a critical role in regulating fruit shape and sugar content in cucumber autotetraploid.

Regulation of Burkholderia sp. GD17 on the Drought Tolerance of Cucumber Seedlings

WANG Qi, HU Zhe, FU Wei, LI Guang-zhe, HAO Lin

2023, 39(3): 163-175. doi:10.13560/j.cnki.biotech.bull.1985.2022-0753

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Drought is one of the major constraints on agricultural productivity. Plant growth promoting rhizobacteria（PGPR）could effectively alleviate drought-induced damages to plants. However, the involved mechanisms still need to be explored. This study emphasized the regulatory roles of Burkholderia sp. GD17 in cucumber seedling responses to drought stress. The experiment consisted of GD17-bacterized and non-bacterized plants, with or without drought treatment, then the physiological and biochemical parameters, and gene expression were analyzed at 7 d after withdrawing water, and the mechanisms of GD17 promoting growth and antagonizing were evaluated. As results, on the 5th day of inoculation, the number of GD17 in the roots reached 6.2×10⁶ CFU/g fresh weight, and remained at this level until the 15th day when drought treatment was implemented. Under regular irrigation, fresh and dry weight of aerial part of GD17-bacterized plants were 39% and 36% higher than those of non-bacterized plants, respectively. Following drought stress, the former was 38% and 32% higher than the latter, respectively, and the relative water content was 8.5% higher in the former than the latter. Inoculation of GD17 efficiently alleviated drought-induced oxidation damage as indicated by lower malondialdehyde content（45%）and electrolyte leakage（26%）in GD17-bacterized leaves than in non-bacterized ones following drought stress. The activities of superoxide dismutase, peroxidase and catalase in the GD17-bacterized leaves were significantly lower than those in non-bacterized ones under regular irrigation, while they were higher in the former than the latter under drought stress. Leaf proline content increased significantly under drought stress, with a greater degree in the bacterized plants. The net photosynthetic rate was higher in bacterized plants than in non-bacterized ones under regular irrigation, but the stomatal conductance, intercellular carbon dioxide concentration and transpiration rate did not change significantly. However, the values of these parameters in the bacterized plants were significantly higher than those in non-bacterized plants under drought conditions. Chlorophyll fluorescence imaging further showed that inoculation with GD17 effectively alleviated the drought-induced damage to photosynthetic apparatus and the impairment of photosynthetic efficiency. The expressions of antioxidation-, proline synthesis- and transcription factor-related genes were up-regulated following drought stress, especially in GD17-bacterized plants. In conclusion, inoculation with GD17 efficiently promotes cucumber seedling growth and tolerance to drought. The possible mechanisms might be associated with improving antioxidant defense, alleviating photosynthetic damage, enhancing osmotic substance synthesis and cell water retention, and up-regulating expression of transcription factors. GD17 strain presents potential application in cucumber agricultural production.

Identification and Analysis of LBD Gene Family and Expression Analysis of Fruit Development in Cucumis melo

CHEN Qiang, ZHOU Ming-kang, SONG Jia-min, ZHANG Chong, WU Long-kun

2023, 39(3): 176-183. doi:10.13560/j.cnki.biotech.bull.1985.2022-1264

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The lateral organ boundaries domain（LBD）genes are widely found in higher plants, and can participate in the regulation of plant growth, development and stress response. Identification and analysis of the LBD transcription factor in melon（Cucumis melo）is aimed to unravel its regulatory role in the process of fruit development. Bioinformatics was used to identify the members of LBD transcription factor in melon genome and named as CmLBD1-CmLBD32. Multiple-sequence alignment was used to analyze the structure characteristics of LBD-domain in the melon. Maximum-likelihood and neighbor-joining were to have phylogenetic tree analyzed. The expressions of LBD gene family during fruit development were deteted by RT-qPCR using extracted RNAs from the fruits（five developing and mature fruits were harvested at 20, 25, 30, 35 and 40 d after pollination）. Total 32 LBD genes were identified from melon genome, and they can be divided into 7 categories and were distributed on 12 chromosomes. The multiple-sequence alignment analysis demonstrated that there were typical LOB domains in 32 LBD genes. LBD family genes expressed in different stages of fruits, the expressions of CmLBD2 and CmLBD28 gradually increased with the processes of fruit development, while relatively low for rest family members. Subcellular localization suggested that CmLBD2 protein was located in the cell membrane. The correlation analysis indicated that there was a negative correlation between the CmLBD2 expression and the fruit firmness during fruit ripening. CmLBD2 expression was inhibited by 1-Methylcyclopropene（1-MCP）after the fruit was processed with 1-MCP, indicating that CmLBD2 gene expression may be regulated by ethylene. In sum, LBD genes in the melon may play different roles in fruit development process, and CmLBD2 may be involved in ethylene regulation related to fruit ripening and softening after harvest.

Analyses of Transcription and Metabolic Differential in the Flower Development Processes of ‘Rose rugosa cv. Plena’

ZHAO Yan-xia, ZHANG Jing-ying, SUN Jun-fei, WANG Jiang-hui, SUN Jia-bo, LV Xiao-hui

2023, 39(3): 184-195. doi:10.13560/j.cnki.biotech.bull.1985.2022-0729

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In order to explore the key genes and metabolic pathways regulating floral aroma during the development of ‘Rose rugosa cv. Plena’, transcriptome and metabolome sequencing were performed on the samples at different development stages of flowers. The 4435, 2444, 7021 and 4123 differentially expressed genes in the comparison of flower bud stage, early opening stage, half opening stage, full opening stage and falling stage respectively were detected via transcriptomics analysis, of which 214 were differentially expressed genes in five stages. A total of 508 metabolites and 230 differential metabolites were detected by metabonomics. There were significant differences in metabolites at different developmental stages, and the metabolites were divided into 4 clusters. The 58 differentially expressed genes and 11 differentially expressed metabolites were found in the analysis of synthetic pathways of phenylpropanoid and terpene in ‘Rosa rugosa’. Combined with transcriptome and metabolome analysis, the genes and volatile substances changed significantly during flower development. Monoterpenoids were formed in flower bud stage, volatiles gradually accumulated in half opening stage, and the release of metabolites decreased in falling stage. Typical rose aroma substances were mainly formed in half opening stage. Therefore, choosing the appropriate harvest time is conducive to the formation and retention of rose fragrance quality.

Cloning and Functional Analysis of LiCMK Gene in Lilium ‘Siberia’

LIU Si-jia, WANG Hao-nan, FU Yu-chen, YAN Wen-xin, HU Zeng-hui, LENG Ping-sheng

2023, 39(3): 196-205. doi:10.13560/j.cnki.biotech.bull.1985.2022-0714

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4-diphosphocytidyl-2-C-methyl-D-erythritol kinase（CMK）is a crucial enzyme for terpenoid synthesis in the methylerythritol phosphate（MEP）pathway. To uncover its regulating effect on the aroma of the lilies, gene LiCMK was cloned from Lilium ‘Siberia’ and analyzed by bioinformatics. Specifically, the protein was located by subcellular localization and spatial-temporal expression patterns of LiCMK gene were explored by means of fluorescence quantitative PCR. And the validation was performed by instantaneously silenced LiCMK through viral induced gene silencing（VIGS）. As a result, LiCMK was 1 200 bp in length and encoded 399 amino acids, belonging to IspE family, which was relatively similar to CMK in Elaeis guineensis. The expression of LiCMK rose and fell corresponding to the flowering and reached the peak when it came to full flowering stage. Moreover, the expression in flower was significantly higher than that in stems and leaves. In addition, LiCMK protein was located in chloroplast. LiCMK expression dropped by about 84% after LiCMK was silenced instantaneously in Lilium ‘Siberia’, leading to the expressions of major monoterpene synthesis genes decreased, i.e., myrcene synthase gene（MYS）, ocimene synthase gene（OCS）and linalool synthase gene（LIS）plunged by 57%, 59% and 63%, respectively, and the release of major monoterpenes, myrcene, ocimene and linalool significantly decreased. To conclude, LiCMK gene is indispensable to the synthesis of monoterpenoid compounds and floral release in Lilium ‘Siberia’, which lays the foundation for future researches about the synthesis and regulatory mechanism of monoterpenoid compounds and the floral scent release.

Cloning and Expression of PdANS in Paeonia delavayi and Correlation with Anthocyanin Content

PING Huai-lei, GUO Xue, YU Xiao, SONG Jing, DU Chun, WANG Juan, ZHANG Huai-bi

2023, 39(3): 206-217. doi:10.13560/j.cnki.biotech.bull.1985.2022-0532

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Flower color is one of the most important quality traits of ornamental plants and plays an important role in the process of plant growth and development. Exploring the mechanism of flower colors regulation in Paeonia delavayi lays the theoretical foundation for improving the ornamental value. The petals were used as the material to clone PdANS, using bioinformatics ally to analyze its characteristics, and combined with real-time fluorescence quantitative PCR and HPLC to investigate the correlation between the expressions of PdANS in different tissues, different developmental periods and the anthocyanin content of each tissue. The results showed that the full length of PdANS was 1 121 bp, including an open reading frame（ORF）of 1 064 bp, encoding 354 amino acids, the relative molecular mass was 40.41 kD, the theoretical isoelectric point（pI）was 5.48, the molecular formula was C₁₈₁₉H₂₈₇₆N₄₇₄O₅₄₀S₁₂, the aliphatic index was 88.64, the instability index（II）was 55.32, and the average hydrophilicity was -0.435. It was presumed that it was an unstable hydrophilic protein. And there was no signal peptide sequence and transmembrane helix region, which was a non-transmembrane protein without secretory function. The phylogenetic analysis showed that PdANS of P. delavayi was most closely related to the ANS proteins of P. suffruticosa, Paeonia lactiflora and other plants. The results of the RT-qPCR revealed that PdANS was expressed in the petals, anthers, sepals, bracts and pedicels, with the highest expression in petals. The results of HPLC showed that Cy3G5G, Pn3G5G, Cy3G and Pn3G were detected in the extracts of different tissues, with the content of the anthocyanin of petals > anthers > sepals > pedicels > bracts. The correlation analysis showed that the anthocyanin contents were highly significantly correlated with the expression of PdANS. It is speculated that PdANS plays an important role in formation of flower color in P. delavayi and is involved in the biosynthesis of anthocyanin substances.

Expression Analysis and Interaction Protein Validation of CsPPR and CsCPN60-like in Albino Tea Plant（Camellia sinensis）

WANG Tao, QI Si-yu, WEI Chao-ling, WANG Yi-qing, DAI Hao-min, ZHOU Zhe, CAO Shi-xian, ZENG Wen, SUN Wei-jiang

2023, 39(3): 218-231. doi:10.13560/j.cnki.biotech.bull.1985.2022-0802

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Our group screened out two genes（CSS0013384 and CSS0036305）related to the albino leaf color of Camellia sinensis Baijiguan through the transcriptome, in order to explore the expression patterns of CSS0013384 and CSS0036305 in albino tea plants interacting proteins. Using cultivar Baijiguan as material, the full-length cDNA sequences of CSS0013384 and CSS0036305 were cloned. Bioinformatics, yeast one-hybrid and yeast two-hybrid were used to analyze the protein physicochemical properties, the phylogenetic tree, the chromosome location, the gene structure, the protein structure, the protein regulation and interaction network, and gene expression pattern. Bioinformatics analysis showed that CSS0013384 and CSS0036305 belonged to the pentatricopeptide repeat protein（PPR）and chaperone（CPN60-like）families, respectively. The lengths of their protein coding sequence（CDS）was 1 893 bp and 1 752 bp, the number of encoded amino acids was 631 and 575, the protein mass was 71.87 kD and 60.79 kD, the isoelectric points were 8.93 and 6.21, the subcellular localization prediction results indicated that CSS0013384 was localized in chloroplast, and CSS0036305 was localized in mitochondria. The RT-qPCR results of the second leaf of Baijiguan under shading and restored light treatment and tea plant varieties with different leaf colors revealed that CSS0013384 and CSS0036305 were highly expressed in albino leaves. CSS0002807 belonged to the PIF transcription factor family, and the yeast one-hybrid results indicated that CSS0002807 bound to the CSS0013384 promoter. CSS0013384 and CSS0036305 may be involved in the development of chloroplast and mitochondria in albino tea leaves, and play an important role in the process of leaf albino. The results may provide a reference for further research on the mechanism of tea leaf albino.

Semi-lethal Low Temperature and Taxane Content of Taxus Under Low Temperature Stress

JIANG Lu-yuan, FENG Mei-jing, DU Yu-qing, DI Bao, CHEN Duan-fen, QIU De-you, YANG Yan-fang

2023, 39(3): 232-242. doi:10.13560/j.cnki.biotech.bull.1985.2022-0762

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To determine the electrical impedance parameters suitable for the determination of semi-lethal low temperature of Taxus and explore the effects of low temperature on the taxane biosynthesis in Taxus, the electrical impedance parameters and taxane content changes of Taxus under low temperature stress were examined. It will lay a theoretical foundation for expanding the introduction range of Taxus and using low temperature conditions to increase the contents of taxane content in Taxus. Using T. yunnanensis, T. chinensis var. mairei （Changzhi, Shanxi）, T. chinensis, T. cuspidata and T. × media as materials, the semi-lethal low temperature of these trees were determined by conductivity method and electrical impedance method respectively. Moreover, the taxane contents and abscisic acid contents in the leaves of Taxus under low-temperature stress were determined by LC-MS method. The results showed that T. chinensis, T. chinensis var. mairei（Changzhi, Shanxi）, T. cuspidata and T. × media all tolerated low temperature of -14.668- -9.106℃. The T. yunnanensis tolerated -8.802- -3.521℃, which was higher than the ons of other four Taxus species. The semi-lethal low temperature of one-year-old branches of five Taxus measured by electrical impedance τ_m parameter was significantly correlated with that measured by conductivity method, and the correlation coefficient was 0.912. The taxol content in the leaves of one-year-old Taxus was higher than that in the current-year under low temperature stress. The taxol content in the leaves of one-year-old T. yunnanensis and T. cuspidata significantly increased with the decrease of treatment temperature, while the baccatin III content and 10-deacetyltaxol content in the leaves of one-year-old T. chinensis significantly increased. Moreover, ABA content in the leaves of Taxus increased with the decrease of temperature, and ABA content in the current-year leaves was higher than that in the one-year-old leaves. Compared with T. yunnanensis, T. chinensis, T. chinensis var. mairei（Changzhi, Shanxi）, T. cuspidata and T. × media tolerated lower temperature. Low temperature is beneficial to the synthesis of taxol in the leaves of one-year-old T. yunnanensis and T. cuspidata, and the synthesis of baccatin III and 10-deacetyltaxol in the leaves of current-year T. chinensis.

Identification of Phytate Phosphorus-solubilizing PGPB in Avena sativa Rhizosphere from Alpine Grassland and Functional Characteristics of Dominant Genus Pseudomonas sp.

LI Qi, YANG Xiao-lei, LI Xiao-lin, SHEN You-lei, LI Jian-hong, YAO Tuo

2023, 39(3): 243-253. doi:10.13560/j.cnki.biotech.bull.1985.2022-0686

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This work aims to directly screen the microbial resources dissolving phytate phosphorus in Avena sativa rhizosphere from alpine grassland and plant growth promoting bacteria（PGPB）and to analyze the genes encodingphytase. The strains were isolated and screened using national botanical research institute's phosphate growth medium（NBRIP）and the 16S rRNA gene sequencing were used to identify the strains. The dominant strains were used to determine phytase activity and growth-promoting characteristics. The complete sequence of phytase was amplified by degenerate PCR and high-efficiency thermal asymmetric interleaved PCR（hiTAIL-PCR）, and bioinformatics analysis and heterologous expression of new phytase. A total of 107 strains were obtained, of which 51 strains formed a clear phosphate solubilizing circle on NBRIP medium, and the strains belonged to two phyla, 10 families and 11 genera, which Pseudomonas（52.94%）were the predominant bacteria. Fourteen Pseudomonas sp. were detected to have phytase activity, and to have the growth-promoting ability of dissolving organic/inorganic phosphorus, secreting 3-indoleacetic acid（IAA）, fixing nitrogen and antagonizing phytopathogenic fungi. Three BPPhy（PHY65, PHY101 and PHY131）sequences of strains were cloned from fourteen Pseudomonas sp., which were predicted to be β-propeller phytases（BPPhy）family proteins, and the specific activity of recombinant PHY65 was 28.2 U/mg. The results may provide excellent strain resources for the development and utilization of phosphate solubilizing agents, and provide a theoretical basis for the production and application of phytase.

Transcriptome and Candidate Effectors Analysis of Pratylenchus brachyurus

HU Li-li, LIN Bo-rong, WANG Hong-hong, CHEN Jian-song, LIAO Jin-ling, ZHUO Kan

2023, 39(3): 254-266. doi:10.13560/j.cnki.biotech.bull.1985.2022-0773

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Pratylenchus brachyurus is an important migratory endoparasitic phytopathogenic nematode. Analysis of the transcriptome is an important basis for further research on its parasitic characteristics and pathogenic mechanism. In the transcriptome of P. brachyurus, 72 516 unigenes were generated and 38 266（52.76%）unigenes were annotated against seven protein databases. In the carbohydrate-active enzymes analysis, 541 proteins were predicted to have plant/fungal cell wall degrading activities, and belonged to the families of GH5, GH30, GH53, GH28, PL3, GH16, GH18 and GH20 respectively. There were 56 protein sequences in the transcriptome demonstrating similarities with orthologues involved in RNAi pathway of Caenorhabditis elegans. Further analysis showed that 51 protein sequences with homology to known plant-parasitic nematode effector proteins were identified. Nine candidate effectors were selected and their potential ability in suppressing plant defense was tested. Seven of them suppressed programmed cell death（PCD）triggered by Gpa2/Rbp-1. The results revealed that the gene distribution characteristics of P. brachyurus were similar to of the same genus of Pratylenchus spp. The candidate effectors may play a critical role in suppressing plant defense.

Isolation, Identification and Bio-activity of Three Bacillus Strains with Biocontrol Function

SHEN Yun-xin, SHI Zhu-feng, ZHOU Xu-dong, LI Ming-gang, ZHANG Qing, FENG Lu-yao, CHEN Qi-bin, YANG Pei-wen

2023, 39(3): 267-277. doi:10.13560/j.cnki.biotech.bull.1985.2022-0722

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Based on healthy tobacco rhizosphere soil, bacterial strains with diverse functions and high activity were isolated and screened to provide efficient and diversified biocontrol resources for the biological control of crop diseases. The plate confrontation method was used to screen the strains with high activity and wide antibacterial spectrum, and the inhibition rate of the strains on Fusarium oxysporum, mycelia growth and spore germination rate were determined. PCR was adapted to detect the antibiotic synthesis genes of the strains, and the control effects of the functional strains on tomato fusarium wilt were detected by indoor pot experiments. The enzyme production, phosphorus and potassium solubilization, nitrogen fixation and siderophorer production abilities of the functional strains were measured in vitro. The functional strains were identified by combining morphological, physiological and biochemical and 16S rDNA universal primers. A total of 127 strains were isolated from the rhizosphere soil of tobacco, and 24 strains presented the inhibitory effects on Fusarium oxysporum sp. of tomato. The inhibitory rates of 3 top-activity strain SH-1471, SH-1464 and SH-1439 against F. oxysporum sp. of tomato were 82.0%, 74.0% and 75.0%, respectively. It caused the mycelia distortion of F. oxysporum sp. of tomato and vesicle structure was formed, and thus inhibited the spore germination of F. oxysporum sp. of tomato by 62.7%, 50.0% and 37.0%, respectively. The three functional strains had genes for antibiotic synthesis, including srfA, fenB, ituA, ituD and bymA. The results of pot experiment showed that SH-1471, SH-1464 and SH-1439 had 83.7%, 60.7% and 59.0% control effect against tomato fusarium wilt. In addition, the three strains all had the ability of producing protease and cellulase, solubilizing phosphorus, fixing nitrogen, and secreting siderophores. The SH-1471 was identified as Bacillus amyloliquefaciens（Gen-bank ON417363）, and SH-1464 as Bacillus paramycoides（Gen-bank ON417365）and SH-1439 as Bacillus siamensis（Gen-bank ON417364）. Strains SH-1471, SH-1464 and SH-1439 have high-efficiency and broad-spectrum antibacterial ability and diversified functions, and they may be used as efficient biocontrol resources for a variety of diseases and had great development potential.

Identification and Diversity Analysis of Mycoviruses from the Phytopathogenic Fungus Colletotrichum spp. of Walnut

XU Xiao-wen, LI Jin-cang, HAI Du, ZHA Yu-ping, SONG Fei, WANG Yi-xun

2023, 39(3): 278-289. doi:10.13560/j.cnki.biotech.bull.1985.2022-0732

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Colletotrichum spp. are the pathogens of walnut anthracnose, a major disease of walnuts, but little has been known about the types and their genomic sequence information of viruses carried by phytopathogenic fungus Colletotrichum spp. of walnut. In this study, 25 strains isolated from walnut fruits suffering walnut anthracnose in three different provinces of China were mined，identified and classified via meta-transcriptomic approach. After homologous alignment analysis, 22 kinds of viral genome sequences were obtained, 19 of them were novel. Among the 22 mycoviruses, 21 were positive single-stranded RNA viruses and one was dsRNA virus. The positive single-stranded RNAs belonged to family Narnaviridae, Mitoviridae and Botourmiaviridae, respectively, while the dsRNA belonged to Alternaviridae family. The RT-PCR validation results showed that all 22 mycoviruses were detected in our Colletotrichum spp. strains, and the virus carriage rate of 25 strains was 100%, with each strain being infected by at least 1 to 11 viruses. The results enriched the genomic information of viruses carried by Colletotrichum spp. and laid the foundation for the subsequent in-depth analysis of the diversity and molecular characteristics of mycoviruses from Colletotrichum.

Genome Sequencing and Bioinformatics Analysis of Gelidibacter sp. PG-2

ZHANG Zhi-xia, LI Tian-pei, ZENG Hong, ZHU Xi-xian, YANG Tian-xiong, MA Si-nan, HUANG Lei

2023, 39(3): 290-300. doi:10.13560/j.cnki.biotech.bull.1985.2022-0919

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Gelidibacter sp. PG-2 is a carotenoid-producing strain, and genome-wide sequencing was conducted in order to further study its carotenoid-producing mechanisms. After the carotenoids were extracted from PG-2, qualitative analysis was performed by LC-MS/MS, the content was calculated by liquid UV full wavelength scanning, and the metabolic pathways of carotenoid-producing in this strain were predicted based on the results of whole genome sequencing. The results showed that the carotenoids produced by PG-2 were zeaxanthin with a content of 185.81 μg/g of dry weight, the whole genome size of PG-2 was 3 850 413 bp, the GC content was 37.91%, the total rRNA was 5, the total tRNA was 37, and the sRNA was 19, which were annotated in COG, GO and KEGG databases to genes 3 401, 1 797 and 1 495, respectively. The strain PG-2 contained three core genes associated with carotenoid production, crtI, crtZ, and lcyB, and their metabolic pathways were predicted. GenBank accession number JAMJTV000000000 was obtained after Genome Framework Sequencing Data submitted to NCBI. The above results demonstrate that the strain PG-2 has the ability to produce carotenoids, and it is speculated that its carotenoid production is related to the genes crtI, crtZ and lcyB.

Isolation of a Novel Heterotrophic Nitrification-Aerobic Denitrification Bacterium Paracoccus sp. QD-19 and Its Characterization of Removing Nitrogen

ZHANG Yu-hong, DONG Xian-bo, LIU Xiang-yu, XU Jia-qi, XU Zi-ling

2023, 39(3): 301-310. doi:10.13560/j.cnki.biotech.bull.1985.2022-0834

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The study is aimed to isolate a novel strain with simultaneous heterotrophic nitrification and aerobic denitrification ability from the activated sludge of the south sewage treatment plant in Shenyang, and to evaluate the nitrogen-removing characteristics, which may lay a foundation for improving nitrogen removal process in wastewater facility. The strain Paracoccus sp. QD-19 was observed by morphological and identified by 16S rRNA gene sequencing. The nitrogen-removing capabilities of the strain were explored by NH₄Cl,NaNO₂, and KNO₃as sole nitrogen source. The effects of carbon source types, C/N ratio, pH value, temperature, shaking speed and inoculation amount（v:v）on the nitrogen removal were explored. A novel strain with heterotrophic nitrification-aerobic denitrification was obtained, identified as Paracoccus by 16S rRNA gene sequencing, and designated as that Paracoccus sp. QD-19. Results showed that 100% of ammonia nitrogen（below 300 mg/L）in wastewater was removed by the strain, and the removal rates achieved 8.707 mg/（L·h）. There was no nitrite accumulation in the nitrogen removal process. The stain Paracoccus sp. QD-19 removed 99% of nitrite or nitrate nitrogen in wastewater in 36 h, and the removal rates achieved 4.944 and 5.666 mg/（L·h）separately. The optimal conditions for ammonia nitrogen removal were: sodium succinate as carbon resource, 10（C/N ratio）, 7（pH value）, 1%（inoculation amount）, 30℃（temperature）, and 140 r/min（shaking speed）. These findings indicate that the strain Paracoccus sp. QD-19 possesses heterotrophic nitrification-aerobic denitrification capability, and the characterization without the accumulation of nitrite and nitrate nitrogen, which provides the practical basis for the improvement of nitrogen-removing process in the wastewater treatment plant.

Cloning, Tissue Expression Profile and Function Prediction of GPX3 Gene in Tibetan Chicken

CHEN Chu-wen, LI Jie, ZHAO Rui-peng, LIU Yuan, WU Jin-bo, LI Zhi-xiong

2023, 39(3): 311-320. doi:10.13560/j.cnki.biotech.bull.1985.2022-0967

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The aim is to clone and have bioinformatics analysis of GPX3 gene in Tibetan chickens, to detect its expressions in different tissues, and to identify the mRNA expressions of GPX3 interacting proteins and analyse their correlations for laying the foundation for further exploring the effect of GPX3 gene on immune function in Tibetan chickens. In this experiment, having 70-day-old healthy Tibetan chickens as study material, RT-PCR was applied to clone the CDS sequence of GPX3 gene and to perform the biological analysis. Real-time fluorescent quantitative PCR（qPCR）was used to detect the expression differences of GPX3 gene in the hearts, livers, spleens, lungs and kidneys of Tibetan chickens, and to analyze the mRNA expressions of 10 related proteins that may interact with GPX3 protein in spleen. A full length of 799 bp GPX3 gene sequence was cloned successfully, including 78 bp of 5' UTR, 660 bp of CDS, and 61 bp of 3' UTR, encoding 219 amino acids. GPX3 mRNA was expressed in 5 tissues of Tibetan chickens, and the highest in the spleen, predicting that the result of the highest expression of GPX3 in the spleen might be related to this protein immune function. GPX3 protein showed a highly and significantly positive correlation with the predicted reciprocal protein SOD2, suggesting that GPX3 might play an important role in drug resistance and reproductive regulation. A 660 bp CDS of GPX3 gene was cloned successfully in this study, predicting that it might be highly significantly and positively correlated with SOD2. This study may provide a theoretical basis for further understanding the role of the antioxidant function of GPX3 gene in the immune mechanism of Tibetan chickens.

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2023, 39(3): 321-321.

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