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2025年8月5日 星期二
农业生物技术学报  2023, Vol. 31 Issue (12): 2477-2489    DOI: 10.3969/j.issn.1674-7968.2023.12.004
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
番茄病程相关蛋白SlPR1b基因的克隆、表达及其在青枯菌胁迫下的功能分析
陈娜1, 李晓鹏1, 刘进法2, 邵勤1,*
1 宜春学院 生命科学与资源环境学院,宜春 336000;
2 新余市现代农业科技园管理委员会,新余 338000
Cloning, Expression, and Function Analysis Under Ralstonia solanacearum Stress of Pathogenesis-related Protein SlPR1b Gene in Tomato (Solanum lycopersicum)
CHEN Na1, LI Xiao-Peng1, LIU Jin-Fa2, SHAO Qin1,*
1 College of Life Science and Resources and Environment, Yichun University, Yichun 336000, China;
2 Management Committee of Modern Agricultural Science Park of Xinyu, Xinyu 338000, China
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摘要 病程相关蛋白1 (pathogenesis-related protein 1, PR1)基因在植物抗生物和非生物胁迫中起着重要作用。基于本课题组前期青枯菌(Ralstonia solanacearum)胁迫的转录组数据,本研究利用反转录PCR (reverse transcription-PCR, RT-PCR)技术从番茄(Solanum lycopersicum)中克隆到1个青枯菌胁迫相关的PR1基因,并将其命名为SlPR1b (Solyc00g174340.2);利用qPCR方法,研究了SlPR1b基因的组织表达特异性和在青枯菌侵染、水杨酸(salicylic acid, SA)和茉莉酸甲酯(methyl jasmonic, MeJA)处理条件下的表达特性,并构建了SlPR1b基因病毒诱导的基因沉默(virus induced gene silencing, VIGS)载体,并将其转化番茄抗病材料,分析沉默SlPR1b基因番茄在青枯菌胁迫条件下的抗病能力。结果显示,克隆的SlPR1b基因cDNA全长序列为807 bp,其ORF为480 bp,编码159个氨基酸,包含1个保守的CAP-PR-1结构域(cd05381),属于CAP超家族。SlPR1b分子量为17 519.73 D,等电点为8.86,属于具有跨膜结构的分泌蛋白。同源序列比对和系统发育分析表明,SlPR1b与潘那利番茄(Solanum pennellii) SpPR4蛋白的亲缘关系最近,其次为马铃薯(Solanum tuberosum)和辣椒(Capsicum baccatum)。组织特异性表达结果表明,SlPR1b基因在番茄叶片中的表达量最高。此外,SlPR1b基因的表达可被青枯菌、SA和MeJA所诱导。沉默SlPR1b基因会降低番茄植株对青枯病的抗性,该结果表明SlPR1b在番茄抗青枯病中起着正调控作用。本研究为后续进一步研究SlPR1b基因在番茄抗青枯病响应中的调控功能提供理论依据。
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陈娜
李晓鹏
刘进法
邵勤
关键词 番茄青枯病病程相关蛋白(PR)基因克隆表达分析病毒诱导的基因沉默(VIGS)    
Abstract:Pathogenesis-related protein 1 (PR1) gene plays an important role in plant resistance to biotic and abiotic stresses. Based on the transcriptome data of Ralstonia solanacearum stress previously conducted by our research group, in this study, a PR1 gene, named SlPR1b (Solyc00g174340.2) was cloned from tomato (Solanum lycopersicum) by reverse transcription PCR (RT PCR) technology. qPCR was used to reveal SlPR1b gene tissue expression specificity and expression characteristic under the conditions including infection with Ralstonia solanacearum, and treatment with salicylic acid (SA) and methyl jasmonate acid (MeJA). SlPR1b gene virus induced gene silencing (VIGS) vector was constructed and then transformed into tomato resistance material to analyze the resistance ability of SlPR1b gene silenced tomato in R. solanacearum stress condition. The results showed that the full-length cDNA sequence of SlPR1b gene was 807 bp, its ORF was 480 bp, encoding 159 amino acids, including a conserved CAP-PR-1 domain (cd05381), belonging to the CAP superfamily. The predicted molecular weight of SlPR1b was 17 519.73 D, the isoelectric point was 8.86, and the protein was found to be a secreted protein with a transmembrane structure. Homologous sequence alignment and phylogenetic analysis indicated that SlPR1b was highly homologous with a S. pennellii SpPR4 protein, followed by S. tuberosum StPR1b protein. The results of tissue specific expression showed that SlPR1b gene expression level was the highest in tomato leaves. Furthermore, SlPR1b gene expression could be induced by R. solanacearum, SA, and MeJA. Silencing SlPR1b decreased plant resistance to bacterial wilt, these results suggested that SlPR1b played a positive role in tomato resistance to bacterial wilt. This study provided a reference for further exploring the role of SlPR1b gene in the response of tomato to bacterial wilt.
Key wordsTomato bacterial wilt    Pathogenesis-related (PR) protein    Gene cloning    Expression analysis    Virus induced gene silencing (VIGS)
收稿日期: 2022-12-09     
ZTFLH:  S641.2  
基金资助:国家自然科学基金(32260776); 江西省教育厅科学技术研究项目(GJJ190863); 江西省科技厅重点研发计划项目(20202BBFL63002)
通讯作者: *shaoqin2013@126.com   
引用本文:   
陈娜, 李晓鹏, 刘进法, 邵勤. 番茄病程相关蛋白SlPR1b基因的克隆、表达及其在青枯菌胁迫下的功能分析[J]. 农业生物技术学报, 2023, 31(12): 2477-2489.
CHEN Na, LI Xiao-Peng, LIU Jin-Fa, SHAO Qin. Cloning, Expression, and Function Analysis Under Ralstonia solanacearum Stress of Pathogenesis-related Protein SlPR1b Gene in Tomato (Solanum lycopersicum). 农业生物技术学报, 2023, 31(12): 2477-2489.
链接本文:  
https://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2023.12.004     或     https://journal05.magtech.org.cn/Jwk_ny/CN/Y2023/V31/I12/2477
 
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