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目的克隆三七Panax notoginseng(Burk)F.H.Chen多聚半乳糖醛酸酶抑制蛋白(polygalacturonase-inhibiting protein,PGIP)基因(Pn PGIP)的全长c DNA序列,分析该基因的表达特性。方法根据三七中编码PGIP的EST(expressed sequence tag)序列设计引物,采用c DNA末端快速扩增技术克隆Pn PGIP基因的全长c DNA序列;用q RT-PCR分析Pn PGIP基因的表达水平。结果 PnPGIP基因的cDNA全长为1 171 bp,含有981 bp的开放阅读框,13 bp的5’-非翻译区以及177 bp的3’-非翻译区,编码含326个氨基酸的蛋白质,PnPGIP蛋白的相对分子质量约为36 770,等电点约为5.83;q RT-PCR分析结果显示,Pn PGIP基因的表达量分别在三七接种茄腐镰刀菌和人参链格孢后4 h和2 h内迅速上升;此外,信号分子茉莉酸甲酯(methyl jasmonate,Me JA)、乙烯利(ethylene,ETH)、H2O2、水杨酸(salicylic acid,SA)处理均能不同程度地诱导Pn PGIP基因的表达水平。结论 PnPGIP基因在转录水平响应茄腐镰刀菌和人参链格孢的侵染,并受几种逆境胁迫相关信号分子的诱导,Pn PGIP基因可能参与三七对茄腐镰刀菌和人参链格孢的防卫反应。
Objective To clone the full length cDNA of PGH gene of Panax notoginseng (Burk) F.H.Chen polygalacturonase-inhibiting protein (PGIP) and analyze its expression characteristics. Methods According to the sequence of expressed sequence tag (EST) encoding PGIP in Panax notoginseng, primers were designed and the full-length c DNA sequence of Pn PGIP gene was cloned by c-terminal rapid amplification. The expression of Pn PGIP gene was analyzed by q RT-PCR. Results The full-length cDNA of PnPGIP gene was 1 171 bp in length and contained 981 bp of open reading frame, 13 bp of 5’-untranslated region and 177 bp of 3’-untranslated region, encoding a 326 amino acid protein, PnPGIP protein The relative molecular mass was about 36 770 and the isoelectric point was about 5.83. Q RT-PCR analysis showed that the expression of Pn PGIP gene was detected at 4 h and 2 h after inoculation with Fusarium solani and ginseng In addition, the signal molecules such as methyl jasmonate (Me JA), ethylene (ETH), H2O2 and salicylic acid (SA) could induce different degrees of Pn PGIP gene The expression level. Conclusions The PnPGIP gene is responsive to the infection of Fusarium solani and Alternaria solani at the transcriptional level and is induced by several signal transducers related to stress. Pn PGIP gene may be involved in the pathogenesis of Fusarium solani and Alternaria solani. Defense response.