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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。
The full-length cDNA sequence of FaNBS20 was cloned by homologous cloning and RT-PCR using cultivated strawberry (Fragaria × ananassa Duch.) ’Jiuxiang’ as experimental material. The real-time fluorescence quantitative PCR was used to detect the exogenous SA treatment synergistic strawberry Effect of Anthracnose Pathogen Infection on FaNBS20 Expression. The results showed that the open reading frame of FaNBS20 was 3 573 bp, encoding 1 190 amino acids with a predicted protein molecular weight of 134.5861 kD, containing 1 TIR domain, 1 NB-ARC domain and 1 leucine-rich Region and up to 99% homology with Fragaria vesca subsp. Vesca anti-tobacco mosaic virus protein (XP_004292718). Inoculation of strawberry anthracnose had a significant effect on the expression of FaNBS20, an anti-anthracnose variety in strawberry. The expression of FaNBS20, a susceptible variety, was always at a low level, indicating that the gene may be related to the resistance to anthracnose . The resistance of strawberry varieties to strawberry anthracnose increased significantly after SA treatment, and the duration of the two strawberry varieties could reach more than 10 days. Under the synergistic induction of SA and strawberry anthracnose, the expression level of FaNBS20 gene was 38 times and 7.2 times higher than that of the resistant variety ’Sweet Charlie’ and the susceptible variety ’Jiuxiang’ respectively. The above results show that salicylic acid can improve the strawberry resistance to anthrax infection, and the expression level of FaNBS20 gene may be an important member of salicylic acid involved in pathogenic response to anthracnose resistance.