摘要
利用同源克隆结合RACE技术从久香草莓茎尖cDNA中分离到一个抗病相关TIR-NBS-LRR类基因FaNBS1（GenBank登录号：HQ845018），解析了该基因及其编码蛋白的组织结构和同源特征；并采用半定量RT-PCR技术分析了该基因在不同组织中的表达差异和外源植物生长物质处理下的表达变化。该基因在基因组上长为2434bp，由3个外显子组成的转录本包含长1893bp的读码框。所编码蛋白含630个氨基酸残基，在15～146是保守的TIR结构域，191～492是NB-ARC结构域。FaNBS1与大豆TIR-NBS-LRR抗性蛋白ADF78118的全序列一致性为50．20％，在NB-ARC保守结构域的一致性为52．06％。半定量RT-PCR分析显示，FaNBS1在检测的所有组织中都表汰佃存摹套分年组织中表达水平最高,日存叶中的表达水平明显受到外源水杨酸和脱落酸处理的影响.
Homologous cloning combined with the RACE （rapid amplification of cDNA ends） technique was used to isolate the disease resistance related gene FaNBS1 （Accession No.： HQ845018） from shoot apical meristems of Fragaria × ananassa Duch. cv. Jiuxiang. After characterizing the exon-intron organization of FaNBS1, modular structure and homology of its deduced protein, semi-quantitative RT-PCR was performed to study its expression pattern in different tissues and responses to external plant growth regulators. The genomic sequence for FaNBS1 is 2 434-bp-long, consisting of 3 exons. This gene harbors a 1 893 bp open reading frame （ORF） encoding a 630 amino acid peptide, with a TIR domain （amino acids 15 to 146） at the N terminal and a NB-ARC domain in the middle region （amino acids 191 to 492）. The deduced amino acid sequence showed the highest similarity to TIR-NBS-LRR disease resistance protein ADF78118 from Glycine max, with 50.20% identity in whole sequence and 52.06% identity in NB-ARC domain. Semi-quantitative RT-PCR analysis showed that this gene expressed in all tissues tested, but the expression level was the highest in shoot apical meristem and was significantly affected by salicylic acid and abscisic acid in leaves.
出处
《果树学报》
CAS
CSCD
北大核心
2011年第6期1025-1031,共7页
Journal of Fruit Science
基金
上海农科院科技发展基金项目(农科发2008(02))
上海市科技兴农重点攻关项目(沪农科攻字(2008)第1-4号)
上海市科委自然科学基金项目(10ZR1426700),上海市科委青年科技启明星计划(09QA1405300)
关键词
草莓
NBS—LRR基因
同源克隆
基因结构
进化树
RT—PCR
Fragaria× ananassa Duch. cv. Jiuxiang
NBS-LRR disease resistance gene
Homologouscloning
Gene organization
Phylogenetic tree
RT-PCR
