【摘要】 在前期已克隆得到的早酥梨(Pyrus bretschneidericv.Zaosu)抗病基因同源类似物Pb-Zs3(GenBank登录号:EU939783)的基础上,通过RT-PCR获得Pb-Zs3的cDNA序列,并利用RACE技术获得该基因的全长cDNA序列,命名为早酥梨抗黑星病相关基因(Vnp1),提交至GenBank(GenBank登录号:FJ763188)。生物信息学分析表明,早酥梨Vnp1基因的全长cDNA序列为1200bp,包含一个完整的921bp的开放读码框架(ORF),编码306个氨基酸残基,预测其分子质量为34.6kD。同时,该基因含有与抗病相关的功能结构域核酸结合位点(NB-ARC),同源性比对显示其与苹果黑星病菌(Venturia inaequalis)诱导后的苹果EST序列ABEA004598(GenBank登录号:EB150292)同源性达71%。对早酥梨Vnp1基因进行半定量RT-PCR研究的结果表明,该基因在梨黑星病菌(Venturia nashicola)诱导后216h内的叶片中的表达水平呈明显上升趋势。更多还原

【Abstract】 Based on the disease resistance gene analog Pb-Zs3(GenBank accession No.EU939783) which was cloned from Zaosu pear(Pyrus bretschneideri cv.Zaosu) in previous study, its expressed cDNA sequence was acquired with RT-PCR.The technique of rapid amplification of cDNA end(RACE) was used to clone its full-length cDNA sequence, and this gene was named pear scab related disease resistance gene(Vnp1) which has been submitted to GenBank(GenBank accession No.FJ763188).Bioinformatic Analysis revealed that Vnp1 gene of Zaosu pear was a novel gene, its cDNA was 1 200 bp.The complete open reading frame(ORF) was 921 bp in length, encoding 306-residue protein with a calculated molecular mass of 34.6 kD.Meanwhile, a disease resistance related domain nucleotide binding adaptor shared by APAF-1, R proteins, and CED-4(NB-ARC) was also detected in its encoding protein.What’s more, a EST sequence of ABEA004598(GenBank accession No.EB150292) of apple EST project induced by Venturia inaequalis showed 71% identity with Vnp1 gene of Zaosu pear by homology comparison.Semi-Quantitative RT-PCR revealed that the transcript of Vnp1 gene of Zaosu pear was significantly enhanced in the leaves of pear under the inducing of Venturia nashicola for 216 h.更多还原