华北农学报 ›› 2019, Vol. 34 ›› Issue (2): 66-71. doi: 10.7668/hbnxb.201751433

所属专题: 薯类作物 生物技术

• 作物遗传育种·种质资源·生物技术 • 上一篇    下一篇

马铃薯StPR1基因克隆及表达特异性分析

李秀钰, 贺付蒙, 韩瑛琪, 赵潇璨, 武佳文, 朱元芳, 周磊, 石奇海, 冯哲, 李凤兰   

  1. 东北农业大学 生命科学学院, 黑龙江 哈尔滨 150030
  • 收稿日期:2019-01-04 出版日期:2019-04-28
  • 通讯作者: 李凤兰(1973-),女,黑龙江哈尔滨人,副教授,博士,主要从事植物病理学研究。
  • 作者简介:李秀钰(1993-),女,内蒙古兴安盟人,在读硕士,主要从事植物病理学研究。
  • 基金资助:
    国家自然基金青年科学基金项目(31201470);国家自然科学基金项目(J1210069);哈尔滨市应用科技研究与开发项目(2015RAQXJ021)

Cloning and Expression Characteristics of Solanum tuberosum StPR1 Gene

LI Xiuyu, HE Fumeng, HAN Yingqi, ZHAO Xiaocan, WU Jiawen, ZHU Yuanfang, ZHOU Lei, SHI Qihai, FENG Zhe, LI Fenglan   

  1. College of Life Science, Northeast Agricultural University, Harbin 150030, China
  • Received:2019-01-04 Published:2019-04-28

摘要: 为了探究病程相关蛋白(Pathogenesis related protein,PR蛋白)与马铃薯抗病的相关性,检验其在马铃薯中抗软腐病、青枯病、干腐病的能力。在前期的马铃薯抗病性研究中对PRs的17个家族基因结合转录组数据进行分析及基因的筛选,最终确定PR1为试验的研究对象。选用马铃薯大西洋品种为材料,由黑龙江省农科院提供,主要进行StPR1基因的克隆,并对其进行生物信息学分析;将真菌接骨木镰孢、燕麦镰孢以及导致软腐病和青枯病等细菌为胡萝卜软腐欧文氏菌、菊欧氏菌、胡萝卜软腐欧文氏菌马铃薯黑胫亚种、茄科雷尔氏菌接种到马铃薯块茎后进行StPR1基因表达特异性分析。结果显示,StPR1基因长度为540 bp,编码179个氨基酸,蛋白质疏水性及亲水性预测PR1蛋白序列的GRAVY值在+4~-3之间,含有亲水区及疏水区,蛋白质二级结构预测其属于混合型蛋白,蛋白质三级结构预测发现其为紧密复杂的螺旋结构,具有SCP_PR-1_like保守结构域,其与辣椒PR1基因在一个进化分支上,相似性较高。qRT-PCR结果显示,StPR1基因的表达量表现为抗真菌胁迫高于细菌胁迫,并预测其抗干腐病的能力强于抗软腐病及青枯病的能力。综上所述,当马铃薯受到外界病原菌侵害时,StPR1在马铃薯的抗病过程中发挥着重要的作用。

关键词: 马铃薯, StPR1, 基因克隆, 表达特异性, 抗病

Abstract: In order to explore the correlation between pathogenesis related protein (PR protein) and potato disease resistance, the ability to resist soft rot, bacterial wilt and dry rot in potato was tested. In the previous potato disease resistance study, 17 family gene-binding transcriptome data of PRs were analyzed and the genes were screened, and PR1 was finally determined. The potato Atlantic variety was selected as the material, which was provided by Heilongjiang Academy of Agricultural Sciences. The StPR1 gene was cloned and analyzed by bioinformatics. F.sambucinum and F.avenaceum were used. And E.carotovora subsp. Carotovora Borgey (Ecc), E.chrysanthemi Burkholder. Atroseptica Dye (Ech), can cause soft rot and bacterial wilt. The specific analysis of StPR1 gene expression was carried out after inoculation of potato tuber with E. carotovora subsp. Mc Fadden et Dimock (Eca) and Ralstonia solanacearum (RS). The results showed that the StPR1 gene was 540 bp in length and encoded 179 amino acids. The hydrophobicity and hydrophilicity of the protein are predicted.The GRAV value of the PR1 protein sequence was between +4 and-3, containing hydrophilic and hydrophobic regions, and protein secondary structure. It is predicted to be a mixed protein. The tertiary structure of the protein is predicted to be a tightly complex helical structure with a SCP_PR-1_like conserved domain, which is highly similar to the pepper PR1 gene on an evolutionary branch. The results of qRT-PCR showed that the expression level of StPR1 gene was higher than that of bacterial stress in antifungal stress, and its ability to resist dry rot was better than that against soft rot and bacterial wilt. In summary, when the potato is attacked by external pathogens, StPR1 plays an important role in the disease resistance of the potato.In summary, when the potato is attacked by external pathogens, StPR1 plays an important role in the disease resistance of the potato.

Key words: Solanum tuberosum, StPR1, Gene clone, Expression characteristics, Disease resistance

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引用本文

李秀钰, 贺付蒙, 韩瑛琪, 赵潇璨, 武佳文, 朱元芳, 周磊, 石奇海, 冯哲, 李凤兰. 马铃薯StPR1基因克隆及表达特异性分析[J]. 华北农学报, 2019, 34(2): 66-71. doi: 10.7668/hbnxb.201751433.

LI Xiuyu, HE Fumeng, HAN Yingqi, ZHAO Xiaocan, WU Jiawen, ZHU Yuanfang, ZHOU Lei, SHI Qihai, FENG Zhe, LI Fenglan. Cloning and Expression Characteristics of Solanum tuberosum StPR1 Gene[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(2): 66-71. doi: 10.7668/hbnxb.201751433.

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