利用CRISPR/Cas9系统编辑日本结缕草ZjSGR基因技术研究

日本结缕草(Zoysia japonica)是一种常见的具有众多优良性状的暖季型草种,秋冬季节在我国北方地区会较早出现叶片脱绿现象,在一定程度上限制了日本结缕草在北方的大面积使用。ZjSGR是从结缕草中分离出来的一种衰老诱导基因,负调控叶片滞绿性状。本研究利用CRISPR/Cas9系统,通过基因枪介导的遗传转化方法实现对日本结缕草ZjSGR基因的编辑,经测序验证获得1株单碱基缺失突变植株。突变植株叶片深绿,在正常生长温度(28～30℃)条件下,和对照相比,叶绿素含量更高(P <0.05)。本研究为进一步阐述ZjSGR基因在日本结缕草滞绿性状调控中的作用机理提供了基础研究材料,也为培育日本结缕草滞绿品种育种工作奠定了基础。

ZjSGR gene editing of Zoysia japonica using the CRISPR/Cas9 system

Zoysia grass(Zoysia japonica)is a common warm-season turfgrass and forage grass species with many excellent characteristics.However,early leaf de-greening occurs in northern parts of China in late autumn and early winter.This limiting factor restricts the wide usage of Zoysia japonica.ZjSGR,a senescence inducing gene isolated from Zoysia grass,plays a negative role in the Zoysia grass leaf color retention process.In this study,the CRISPR/Cas9 system was used to edit the ZjSGR gene of Zoysia japonica by particle bombardment transformation.One ZjSGR mutant plant was obtained after sequencing verification.The mutant plants which appeared dark green had a higher concentration of chlorophyll under natural conditions(28~30℃)compared to wild-type plants(P<0.05).This study provides plant materials for further functional studies of the ZjSGR gene.The mutant plants are a valuable germplasm for the stay-green type Zoysia grass breeding program.