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水杨酸对草莓炭疽病响应基因FaNBS20 表达的影响
引用本文:张庆雨,刘芳春,段 可,王 飞,王延秀,高清华.水杨酸对草莓炭疽病响应基因FaNBS20 表达的影响[J].园艺学报,2014,41(1):53-60.
作者姓名:张庆雨  刘芳春  段 可  王 飞  王延秀  高清华
作者单位:1 西北农林科技大学林学院,陕西杨凌 712100;2 西北农林科技大学园艺学院,陕西杨凌 712100;3 上海市农业科
学院林木果树研究所,上海 201403;4 甘肃农业大学农学院,兰州 730000
基金项目:上海市科技兴农重点技攻关项目[(2012)第1–3 号];上海市科委现代农业领域重点科技攻关项目(12391901400);农业部公益性行业(农业)科研专项(201003064-5)
摘    要: 以栽培草莓(Fragaria × ananassa Duch.)‘久香’叶片为试材,采用同源克隆和RT-PCR 方 法,克隆FaNBS20 全长cDNA 序列,并利用实时荧光定量PCR 技术检测外源SA 处理协同草莓炭疽病病 原菌侵染对FaNBS20 表达的影响。结果表明,FaNBS20 的开放阅读框为3 573 bp,编码1 190 个氨基酸, 预测的蛋白质分子量为134.5861 kD,含有1 个TIR 结构域、1 个NB-ARC 结构域和1 个亮氨酸的富集区, 与森林草莓(Fragaria vesca subsp. vesca)抗烟草花叶病毒蛋白(XP_004292718)同源性高达99%。接种 草莓炭疽病菌对草莓抗炭疽病品种‘甜查理’FaNBS20 的表达影响显著,易感病品种‘久香’FaNBS20 的表达则一直处于较低水平,表明该基因可能与对炭疽病的抗性相关。两个草莓品种在SA 处理后对草莓 炭疽病的抗性均明显增强,且持续期可以达到10 d 以上。在SA 和草莓炭疽菌协同诱导下,FaNBS20 基 因的表达量在抗性品种‘甜查理’和易感品种‘久香’中分别达到接种前的38 倍和7.2 倍。以上结果表 明,水杨酸能提高草莓对炭疽菌浸染的抗性,表达水平和炭疽病抗性水平相关的FaNBS20 基因可能是水 杨酸参与的对炭疽病抗性响应途径中一个重要的成员。

关 键 词:草莓  水杨酸  胶孢炭疽菌  克隆  表达分析

Effects of Salicylic Acid on the Expression of FaNBS20 Gene Responsive to Colletotrichum gloeosporioides Infection in Fragaria × ananassa
ZHANG Qing-yu,,LIU Fang-chun,DUAN Ke,WANG Fei,WANG Yan-xiu,GAO Qing-hua,.Effects of Salicylic Acid on the Expression of FaNBS20 Gene Responsive to Colletotrichum gloeosporioides Infection in Fragaria × ananassa[J].Acta Horticulturae Sinica,2014,41(1):53-60.
Authors:ZHANG Qing-yu      LIU Fang-chun  DUAN Ke  WANG Fei    WANG Yan-xiu  GAO Qing-hua  
Institution: 1.Department of Medical Oncology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310016, China; 2.Materia Medica Institute of Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China; 3.Hangzhou Sanatorium of Nanjing Military Region, Hangzhou 310007, China.
Abstract:A full-length cDNA of FaNBS20 was isolated by homologous cloning and RT-PCR from Fragaria × ananassa Duch.‘Jiuxiang’. Sequence analysis indicated that FaNBS20 holds a 3 573 bp open reading frame encoding 1 190 amino acids with an estimated molecular mass of 134.5861 kD. Bioinformatics analysis revealed that FaNBS20 contains typical domains of TIR,NB-ARC,and LRR type. FaNBS20 shares a 99% sequence similarity with the TMV resistance protein(GenBank accession No. NC_020492)from Fragaria vesca subsp. vesca. Expression of FaNBS20 was obviously changed by the inoculation of Colletotrichum gloeosporioides,a causal agent of strawberry anthracnose. Opposite changes were observed in two cultivars with different susceptibility to anthracnose. In both the resistant variety ‘Sweet Charlie’and the susceptible‘Jiuxiang’,resistance to C. gloeosporioides infection was clearly improved by spraying salicylic acid and this enhancement lasted for more than 10 d. Following the combined treatment of SA and C. gloeosporioides,expression of FaNBS20 showed a 38-fold and 7.2-fold increase in‘Sweet Charlie’and‘Jiuxiang’,respectively. These results indicate that the expression of FaNBS20 may be positively correlated to strawberry resistance,and external SA can increase strawberry resistance to anthracnose,probably,through a signaling network with FaNBS20 as an important component.
Keywords:Fragaria ×  ananassa  salicylic acid  Colletotrichum gloeosporioides  cloning  expression analysis
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