马铃薯泛素结合酶基因StUBC17的克隆与功能分析

doi:10.13560/j.cnki.biotech.bull.1985.2018-0763

摘要/Abstract

摘要： 泛素结合酶（Ubiquitin-conjugating Enzyme E2,UBC）与植物生长发育和抗病反应密切相关。为了解泛素结合酶基因对植物抗晚疫病的贡献,从马铃薯栽培种“合作88”中克隆出一个E2泛素结合酶基因,命名为StUBC17,并对其受晚疫病菌诱导表达特性及功能进行分析。利用病毒诱导的基因沉默（Virus-induced gene silencing,VIGS）技术降低本氏烟（Nicotiana benthamiana）中StUBC17同源基因（NbUBC）的转录水平,再接种晚疫病菌进行抗病性鉴定。进一步在基因沉默植株上瞬时表达马铃薯抗病基因和其相应的无毒基因（R3a+AVR3a、R3b+AVR3b和Rx+CP）及INF1。StUBC17编码区全长447 bp,编码148个氨基酸,其蛋白分子量为16.52 kD,理论等电点为7.72。表达分析结果表明,StUBC17受晚疫病菌诱导表达。抗病鉴定结果显示,与对照植株相比,NbUBC沉默植株的抗病性显著降低。沉默NbUBC不影响过敏反应（Hypersensitive responses,HR）的发生。StUBC17是植物防御晚疫病所需,但该基因的沉默并不影响R3a、R3b、Rx及INF1介导的HR反应。

关键词: 马铃薯, 晚疫病, StUBC17, 本氏烟, 病毒诱导基因沉默

Abstract: Ubiquitin-conjugating enzyme E2 is closely correlated with plant growth and disease resistance. To understand the contribution of ubiquitin-conjugating enzyme gene in plant resistance to late blight（Phytophthora infestans），an E2 ubiquitin-conjugating enzyme gene（designated as StUBC17）was cloned from potato cultivar “C88”，and its induced expressions by P. infestans and functions were analyzed. The transcriptions of NbUBC（homolog of StUBC17）was silenced in Nicotiana benthamiana using virus-induced gene silencing（VIGS）method，and the resistance to disease was validated by inoculating P. infestans. Furthermore，the disease-resistant gene and corresponding non-toxic genes of R3a+AVR3a、R3b+AVR3，Rx+CP，and INF1 were transiently expressed in NbUBC-silenced plants. The 447 bp coding region of StUBC17 encoded 148 amino acid residues with 16.52 kD of an estimated molecular mass and 7.72 of a calculated pI. Quantitative reverse transcriptase PCR analysis showed that the expression of StUBC17 was up-regulated after P. infestans infected it. By inoculating leaves in vitro，the identification of disease resistance showed that NbUBC-silenced plants presented an increased susceptibility to P. infestans；however，silencing of NbUBC demonstrated no effect on the hypersensitive responses. Our results suggest that the StUBC17 is necessary for potato plant to defense P. infestans，but its silencing do not affect the R3a-，R3b-，Rx- and INF- triggered hypersensitive responses

Key words: potato, late blight, StUBC17, Nicotiana benthamiana, virus-induced gene silencing

雷朝霞, 刘晶, 白易平, 唐唯, 王洪洋. 马铃薯泛素结合酶基因StUBC17的克隆与功能分析[J]. 生物技术通报, 2019, 35(1): 35-41.

LEI Zhao-xia, LIU Jing, BAI Yi-ping, TANG Wei, WANG Hong-yang. Cloning and Functional Analysis of a Potato Ubiquitin-conjugating Enzyme Gene StUBC17[J]. Biotechnology Bulletin, 2019, 35(1): 35-41.

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