亲环素AtCYP1 RNA干涉载体的构建及对拟南芥发育的影响

Vector Construction of Ubi::AtCYP1i and Functional Analysis of CYP1 in Arabidopsis thaliana

根据拟南芥AtCYP1基因序列设计特异引物,以拟南芥总DNA为模板,扩增AtCYP1基因中344 bp转录本,插入表达载体PTCK303,构建目的基因RNA干扰载体Ubi::AtCYP1i。利用改良的农杆菌浸染技术获得拟南芥RNAi转基因株系,RT-PCR分析结果表明转基因株系中AtCYP1基因的表达量低于野生型,表型观察结果表明RNAi转基因纯合株系抽苔时间比野生型晚3.32 d,抽苔叶片数较野生型多2.49片,其开花时间、结出第一个种荚的时间、株高等方面也与野生型存在明显差异。此结果说明AtCYP1可能参与了拟南芥的早花发育过程,为进一步研究其在植物生长发育中的功能奠定了基础。

According to gene sequence of Arabidopsis AtCYP1,specific primers designed and Arabidopsis genomic DNA as template,the 344 bp AtCYP1 gene transcripts was amplified and transformed into the expression vector PTCK303,to build the target gene RNAi vector Ubi：：AtCYP1i.Using RNAi transgenic Arabidopsis lines,RT-PCR analysis showed that transgenic expression of genes in AtCYP1 lower than wild-type.Phenotype observed results showed that RNAi transgenic homozygous lines bolting time 3.32 days later than wild-type,bolting a few more leaves than wild-type 2.49.Its flowering time,seed pod bear the first time,plant height,it is also obvious differences between the wild type.This result showed AtCYP1 may be involved in early Arabidopsis flower development process.