水稻成花素家族OsDTH11基因的CRISPR/Cas9编辑突变体的创制

Construction of Rice Mutants of the Florigen Family Gene OsDTH11 Based on CRISPR/Cas9 System

水稻成花素家族基因在水稻的开花结实过程中发挥重要作用。水稻基因组中共有19个成花素相关基因,其中Hd3a和RFT1基因已被证实能够促进水稻抽穗开花,其余成花素成员功能未知。本文针对水稻成花素家族成员OsDTH11基因,基于CRISPR/Cas9技术构建了基因编辑载体,并通过农杆菌介导的遗传转化方法成功获得了该基因的敲除突变体。经测序验证发现了2种类型的突变体,一种是纯合的62 bp缺失突变体,另一种是31 bp缺失的杂合突变体。通过自交分离分别获得两种类型稳定遗传的纯合突变体材料,为OsDTH11基因的生物学功能与水稻开花分子机制研究奠定了基础。

The florigen family genes play an important role for the rice flowering.There are 19 members predicted in the rice genome.In these members,Hd3a and RFT1 were identified and characterized as florigen to promote rice flowering.The function of the rest of members remained unknown.In this study,we constructed the CRISPR/Cas9 editing vector of OsDTH11,a member of the florigen family gene,and introduced it into rice callus to produce the editing mutants by Agrobacterium-mediated transformation method.Further sequencing analysis confirmed that the obtained mutants had two types of mutation.One type of mutation was a homozygous 62 bp deletion in the editing mutant.The other was a heterozygous 31 bp deletion in the editing mutant.Finally,we obtained two kinds of homozygous mutants by self-fertilization,and these two kinds of mutation could stablely transmit to the next generation.These mutants laid a foundation for the functional study of OsDTH11 gene.