[1]王亚乐,李雪君,孙焕,等.抗烟草黑胫病基因NtR1A-10的克隆及其响应病原侵染的分子机制[J].江苏农业学报,2025,(09):1696-1703.[doi:doi:10.3969/j.issn.1000-4440.2025.09.004]
　WANG Yale,LI Xuejun,SUN Huan,et al.Cloning of the tobacco black shank resistance gene NtR1A-10 and its molecular mechanism in response to pathogen infection[J].,2025,(09):1696-1703.[doi:doi:10.3969/j.issn.1000-4440.2025.09.004]
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抗烟草黑胫病基因NtR1A-10的克隆及其响应病原侵染的分子机制()

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2025年09期
页码:
1696-1703
栏目:
遗传育种·生理生化
出版日期:
2025-09-30

文章信息/Info

Title:
Cloning of the tobacco black shank resistance gene NtR1A-10 and its molecular mechanism in response to pathogen infection
作者:
王亚乐; 李雪君; 孙焕; 孙计平
(河南省农业科学院烟草研究所,河南许昌461000)
Author(s):
WANG Yale; LI Xuejun; SUN Huan; SUN Jiping
(Tobacco Research Institute, Henan Academy of Agricultural Sciences, Xuchang 461000, China)
关键词:
烟草; 黑胫病; 抗病基因NtR1A-10; 苯丙氨酸解氨酶途径
Keywords:
Nicotiana tabacum; black shank; resistance gene NtR1A-10; phenylalanine ammonia-lyase pathway
分类号:
S572
DOI:
doi:10.3969/j.issn.1000-4440.2025.09.004
文献标志码:
A
摘要:
为解析抗烟草黑胫病基因NtR1A-10的抗病防御机制,本研究基于美洲茄(Solanum americanum) Rpi-amr3基因序列,从抗烟草黑胫病品种664-01中克隆到NtR1A-10基因,并对其进行生物信息学和表达模式分析。结果表明,NtR1A-10蛋白相对分子量为4.01×104,包含富含亮氨酸的重复序列(LRR)结构域,属于不稳定亲水性蛋白,其氨基酸序列共有37个可磷酸化氨基酸位点。茎中NtR1A-10的相对表达量极显著高于根和叶片(P＜0.01)。以感病品种红花大金元为遗传背景构建NtR1A-10过表达株系（OE2、OE5、OE13、OE20）,通过接种烟草疫霉(Phytophthora nicotianae)评估其抗病能力。接种烟草疫霉14 d后,接种烟草疫霉的OE2株系和OE5株系发病率和病情指数均极显著低于野生型红花大金元(P＜0.01),接种烟草疫霉的OE5株系中NtPR1和NtNPR的相对表达量极显著高于野生型(P＜0.01),表明NtR1A-10过表达株系中水杨酸的合成被激活。接种烟草疫霉的OE5株系中NtPAL相对表达量极显著高于野生型(P＜0.01),而接种烟草疫霉的OE5株系中NtEPS1、NtEDS5、NtICS相对表达量极显著低于野生型(P＜0.01),表明烟草疫霉侵染后,NtR1A-10可能通过调控苯丙氨酸解氨酶(PAL)途径促进烟草水杨酸(SA)的生物合成,进而激活水杨酸(SA)信号通路下游基因NtPR1的响应,增强烟草对烟草黑胫病的抗性。本研究揭示了NtR1A-10抗烟草黑胫病的分子机制,为培育高效抗病烟草新品种提供理论依据。
Abstract:
To elucidate the disease resistance defense mechanism of the tobacco black shank resistance gene NtR1A-10, based on the sequence of Rpi-amr3 gene from Solanum americanum, this study cloned the NtR1A-10 gene from the tobacco variety 664-01 resistant to tobacco black shank and conducted bioinformatics and expression pattern analyses on it. The results showed that the relative molecular weight of the NtR1A-10 protein was 4.01×104, containing a leucine-rich repeat (LRR) domain. It was classified as an unstable hydrophilic protein, and there were a total of 37 phosphorylatable amino acid sites in its amino acid sequence. The relative expression level of NtR1A-10 in the stems was extremely significantly higher than that in the roots and leaves (P＜0.01). Using the susceptible variety Honghuadajinyuan as the genetic background, overexpression lines of NtR1A-10 (OE2, OE5, OE13, OE20) were constructed, and their disease resistance was evaluated by inoculating with Phytophthora nicotianae. Fourteen days after inoculation with Phytophthora nicotianae, the disease incidence and disease index of the inoculated OE2 and OE5 lines were both extremely significantly lower than those of the wild-type Honghuadajinyuan (P＜0.01), and the relative expression levels of NtPR1 and NtNPR in the inoculated OE5 line were extremely significantly higher than those in the wild type (P＜0.01), indicating that the synthesis of salicylic acid was activated in the NtR1A-10 overexpression lines. The relative expression level of NtPAL in the inoculated OE5 line was extremely significantly higher than that in the wild type (P＜0.01), while the relative expression levels of NtEPS1, NtEDS5, and NtICS in the inoculated OE5 line were extremely significantly lower than those in the wild type (P＜0.01), indicating that after infection with Phytophthora nicotianae, NtR1A-10 may regulate the phenylalanine ammonia-lyase (PAL) pathway to promote the biosynthesis of salicylic acid (SA) in Nicotiana tabacum, thereby activating the response of downstream gene NtPR1 in the salicylic acid signaling pathway, and enhancing the resistance of Nicotiana tabacum to tobacco black shank. This study reveals the molecular mechanism of NtR1A-10 against tobacco black shank, providing a theoretical basis for the breeding of highly disease-resistant new tobacco varieties.

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备注/Memo

备注/Memo:
收稿日期：2024-11-07基金项目：河南省烟草公司洛阳市公司科技项目(2022410300200053);河南省农业科学院基础性科研项目(2024JC09)作者简介：王亚乐(1993-),女,河南许昌人,硕士,研究实习员,主要从事烟草遗传育种研究。(E-mail) yms_0905@126.com通讯作者：孙计平,(E-mail)sunjiping2002@126.com
更新日期/Last Update: 2025-10-27