受PVY诱导的烟草NtERD1的基因分离与表达分析

Gene Isolation and Expression Analysis of NtERD1 Induced by PVY in Tobacco (Nicotiana tabacum L.)

通过抑制差减杂交和cDNA芯片法从受PVY诱导的烟草抑制差减杂交文库中筛选得到一段长为745 bp的基因EST片段,经Blast同源性比对分析,该序列与植物脱水诱导早期应答基因(ERD)在核苷酸和氨基酸水平上高度同源。ORfinder分析表明,该序列包含一个492 bp的开放读码框,编码163个氨基酸。氨基酸Blastp比对显示,该氨基酸序列与其他植物脱水诱导早期应答蛋白具有很高的同源性。所以筛选得到的这个新基因属于烟草脱水诱导早期应答基因,被命名为NtERD1(GenBank登录号为GU144573)。实时荧光定量PCR分析表明,NtERD1在PVY接种早期上调表达,因此NtERD1受PVY侵染诱导,可能对烟草抗病毒具有重要作用。

A 745 bp EST was obtained from the tobacco（Nicotiana tabacum L.） suppression subtractive hybridization（SSH） library induced by PVY（potato virus Y）,using SSH and cDNA chip technologies.Sequence alignments with Blast indicated that the EST revealed a high degree of similarity with other members of the plant early responsive to dehydration（ERD） genes in both nucleic and amino acid levels.The cDNA contained an open reading frame（ORF） of 492 bp consisting of 163 amino acids.Amino acid Blastp indicated that the predicted protein had a high degree of homology with other ERD proteins.Thus the new gene screened belongs to tobacco ERD genes and was named as NtERD1（GenBank accession no.GU144573）.The results of real-time PCR showed that NtERD1 was up-regulated in leaves infected with PVY in early stage.Therefore NtERD1 was induced by PVY infection and might play important roles in tobacco resistance to viruses.