抗软腐病基因在花魔芋块茎组织中特异表达的研究

Tissue Specific Expression of Soft Rot Disease Resistant Gene in Amorphophallus konjac

本研究利用PCR技术从马铃薯块茎基因组DNA中克隆块茎特异性启动子patatin,并从苏云金芽孢杆菌218中克隆aii A(高丝氨酸环内酯酶)基因,构建含aii A基因的抗软腐病植物表达载体p BI121-patatinaii A。并利用农杆菌介导法将aii A基因导入花魔芋,以研究aii A基因在魔芋块茎组织中的特异表达。结果表明,转化植株经GUS染色,仅在球茎部位出现蓝色斑点,经PCR检测,RT-PCR及Southern杂交检测证实aii A基因已整合到花魔芋基因组,初步表明aii A基因可在魔芋球茎内特异性表达。本研究对今后魔芋通过分子遗传工程改良软腐病抗性具有一定的应用价值。

In this study, a plant expression vector pBI121-patatin-aiiA was constructed by carrying a aliA （acyl- homoserine lactonase） gene which was obtained from Bacillus thurirtgiensis and driven by a tuber-specific promoter of patatin gene cloned from Solarium tuberosurn genomic DNA,and then transferred into A morphophallus konjac mediated by Agrobacteriurn tumefaciens. PCR, RT-PCR and Southern blot analysis confirmed that aliA gene was integrated into A. konjac genome. The results of GUS staining showed that the blue spots could be only observed on the corm of transformed A. konjac plants, which preliminarily indicated that aiiA gene could be specifically expressed in the corm of transformed A. konjac plants. This study provided a certain applicable value on resistance improvement of soft rot disease through molecular genetic engineering.