木薯淀粉合成相关酶基因的克隆及表达分析  

Cloning and Expression of Starch Synthesis Related Enzyme Gene in Common Cassava

作　　者：许娟[1] 罗兴录[1] 赵德征[1,2] 

机构地区：[1]广西大学农学院,南宁530004 [2]广西作物遗传改良与生物技术重点实验室,南宁530007

出　　处：《生物技术通报》2012年第11期101-109,共9页Biotechnology Bulletin

基　　金：国家"973"计划课题(2010CB126601);广西自然科学基金重点项目(2010GXNSFD013025);南宁市科技攻关项目(201109044B)

摘　　要：根据核苷酸同源性设计简并引物,克隆了一段大小为665 bp的木薯腺苷二磷酸葡萄糖焦磷酸化酶小亚基基因(sAGP)片段,并根据已知序列设计特异引物,克隆分离得到GBSS基因和SBE基因的cDNA片段。半定量RT-PCR分析结果表明,在木薯不同生长时期,3种淀粉合成相关酶基因的时空表达规律不同,GBSS基因表达有时期特异性和组织特异性,只在块根形成期和块根成熟期表达,在组织部位茎中表达最多,并且在这3种基因中表达水平相对较低。而sAGP基因在这3种基因中表达水平最高,在木薯块根成熟期竟达超量表达水平,并且在华南124和辐选01中有品种差异,差异明显,在辐选01中的表达要优于华南124的表达。SBE基因表达较平稳,在各个时期表达水平都处于中间水平,在4个时期中,块根成熟期时的表达水平相对较高,块根形成期和苗期其次,收获期最低,也具有品种间差异,在辐选01中的表达要优于华南124的表达。In this research,the 665 bp gene fragment were received from cassava adenosine diphosphate glucose pyrophosphorylase small subunit gene,named sAGP.According to the known sequence that design specific primers,we got GBSS and cDNA fragment of SBE gene through cloning separation.Semi-quantitative RT-PCR assay was used to detect the expression of genes in cassava,which showed the results that in different growth periods,three kinds of starch synthesis gene expression pattern has a different rules in time and space.GBSS gene expression has specific period and tissue,only expressed in the mature and formatting root.What’s more,Most of the expression is the organization’s stem,and it has a relatively low level in these three genes.However,the expression of sAGP gene is the highest levels in these three genes that has an over-expression level in maturiture roots.The expression in FX01 stems is superior to SC124.SBE has a relatively stable gene expression.All the expression is in an intermediate level in each period.Descending order of gene expression is maturity,formation,seedling and harvest in the four periods.What’s more,the expression in FX01 stems is much stronger than that in SC124.

关 键 词：木薯 淀粉合成相关酶基因 克隆 半定量RT-PCR 基因表达 

分 类 号：S533[农业科学—作物学]