木薯碱性/中性转化酶MeNINV1基因启动子的克隆及激素应答表达分析

Cloning and Hormone Responsive Expression Analysis of an Alkaline/Neutral Invertase Gene MeNINV1 Promoter from Cassava

为了解木薯碱性/中性转化酶基因MeNINV1对激素的应答模式及调控机制本研究采用PCR技术,从木薯基因组中分离出MeNINV1基因启动子序列。分析结果显示该启动子长度1 714 bp,含有TATA box和CAAT box保守元件,以及四个激素响应元件:ERE(乙烯应答)、ABRE(ABA应答)、TAC-element(SA应答)和P-box(GA应答)。根据启动子上存在的参与激素应答相关的顺式作用元件信息,选用30μmol/L GA3、30μmol/L ABA、100μmol/L SA和1%乙烯利四种外源激素胁迫处理木薯SC8组培苗,利用实时荧光定量PCR技术,研究MeNINV1基因在不同激素处理下的表达模式。结果表明,MeNINV1基因的表达受激素的调控,因此推断碱性/中性转化酶基因MeNINV1启动子上的激素应答元件响应激素诱导,调控基因的表达。研究结果为进一步研究激素信号如何调控木薯块根蔗糖的分解代谢提供理论基础。

In order to study the inducement pattern of hormone and regulating mechanism of MeNINV1 in cassava, we isolated an 1 714 bp promoter region upstream of the gene from cassava genomic DNA using PCR methods. Sequence analysis indicated that the promoter contains typical TATA box and CAAT box, and four Cis-acting elements that relate to hormone responses, such as ERE（ethylene responsive）, ABRE（abscisic acid responsive）, TAC-element（salicylic acid responsive） and P-box（gibberellin responsive）. Online bio-informatic analyses of the promoter sequence find some cis-acting elements that relate to hormone responses in the promoter of MeNINV1. The expression mode to different stresses of MeNINV1 including four hormone treatments： 30 μmol/L GA3, 30 μmol/L ABA, 100 μmol/L SA, and 1% ethephon were studied in tissue culture seedling of cassava plants by Real-time PCR. The results showed that MeNINV1 gene expressions were regulated by hormone, which inferred that the alkaline/neutral invertase promoter of MeNINV1 may have cis-element related to hormone response. The research results provide a theoretical basis for further research on hormone signal how to regulate the sucrose metabolism of cassava tuberous root.