宝山堇菜Pb耐性相关基因筛选及表达特性  

The screening and expression pattern analysis of lead tolerant related gene in Viola baoshannensis

作　　者：邓冬梅[1] 艾红霞 邓金川[2] 廖斌[2] 

机构地区：[1]广西科技大学生物与化学工程学院∥广西糖资源绿色加工重点实验室,广西柳州545006 [2]中山大学生态进化学院∥生物防治国家重点实验室∥广东植物资源重点实验室,广东广州510275

出　　处：《中山大学学报（自然科学版）》2018年第1期29-35,共7页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：国家自然科学基金(31570506;31700470);广西自然科学基金(2017GXNSFBA198099);广西重点实验室基金(YB2014201;2016TZYKF10)

摘　　要：Cd超富集植物宝山堇菜Viola baoshannensis也有一定Pb耐性/富集能力,为了解宝山堇菜对Pb的耐性/富集分子机制,采用抑制消减杂交技术(suppression subtractive hybridization,SSH),筛选了宝山堇菜根在Pb诱导下差异表达的基因片段,进而对与Pb耐性与富集相关的基因克隆片段(expressed sequence tags,ESTs)进行半定量RT-PCR和Northorn杂交验证。利用SSH方法,从宝山堇菜根部获得27个在Pb胁迫下特异表达的无重复的有意义EST克隆片段。半定量RT-PCR和Northern杂交进一步证实和Lycopersicon esculentum木葡聚糖内源转糖基酶(xyloglucan endotransglycosylase,XTH)基因、Arabidopsis thaliana GTP结合蛋白基因(GTP binding mRNA)基因相似的2个克隆片段在宝山堇菜受Pb胁迫时,为上调表达基因。可初步假设,Pb胁迫下,伴随有细胞壁的变化,GTP结合蛋白可能参与宝山堇菜体内Pb胁迫信号的传导。Viola baoshanensis, an identified Cd hyperaccumulator can also tolerate and accumulate a certain amount of Pb. To understand the resistance/enrichment molecular mechanism of Viola baoshansis to Pb, suppression subtractive hybridization （SSH） was performed to screen gene segments induced by Pb, and semi-quantitive RT PCR along with Northorn blotting was then used to verify the relationship between the screened expressed sequence tags （ESTs） and Pb tolerant/accumulation in V. baoshanensis. Twenty-seven different cDNA fragments were obtained via sequences analysis of cDNA inserts obtained from SSH subtractive cDNA library of V. baoshanensis roots. The results of both RT PCR and Northern blot analysis showed that two cDNA fragments with high homology with xyloglucan endotransglycosylase （XTH） and GTP binding gene were up-regulated under Pb stress. It could be speculated that, in response to Pb stress, cell wall might have some changes and GTP binding protein might play a role in signal transduction.

关 键 词：超富集植物 宝山堇菜 PB SSH 

分 类 号：X53[环境科学与工程—环境工程]