发根农杆菌介导的星油藤毛状根遗传转化体系建立

Establishment of an Agrobacterium rhizogenes-mediated genetic transformation system of hairy roots in sacha inchi(Plukenetia volubilis L.)

木本油料植物星油藤(Plukenetia volubilis)的种子油富含α-亚麻酸,是一种新型健康食用油料植物,然而其遗传转化体系的缺乏极大地限制了该植物的基因功能和分子育种研究。本研究基于裸眼可视报告基因RUBY系统,分析了发根农杆菌(Agrobacterium rhizogenes)菌株类型、侵染方式以及抽真空处理对星油藤毛状根诱导和遗传转化效率的影响,初步建立了发根农杆菌介导的星油藤毛状根遗传转化体系。研究结果表明:利用发根农杆菌K599菌株进行幼苗切根和下胚轴基部裹菌处理是最优的诱导星油藤产生毛状根的方法,可获得含有超量表达RUBY毛状根的嵌合植株,并且转基因毛状根可进一步诱导形成愈伤组织。该毛状根遗传转化体系的建立为进一步开展星油藤的遗传转化和基因功能研究提供参考。

Sacha inchi(Plukenetia volubilis)is a promising woody oil plant,the seed oil of which is rich in α-linolenic acid.However,the lack of genetic transformation method greatly limited the investigation of gene function and molecular breeding in sacha inchi.In this study,based on the naked-eye visual reporter system RUBY,the effects of different Agrobacterium rhizogenes strains,infection methods,and vacuum treatment on the induction of hairy roots and genetic transformation efficiency were invesitgated.The results show that coating hypocotyle bases of seedlings with primary roots removed with A.rhizogenes K599 showed the highest effciency in inducing the hairy roots.Abundant composite plants with red hairy roots overexpressing RUBY were obtained by using this method.The calli were further induced in the callus-inducing media by directly using the transgenic red hairy roots as explants.Thus,in this study,we report the first A.rhizogenes-mediated hairy root transformation system in sacha inchi.This protocol would lay a foundation for further establishment of the efficient genetic transformation method in sacha inchi,and also facilitate the functional characterization of novel genes in the sacha inchi root system.