摘要
ALOG (Arabidopsis LSH1 and Oryza G1)基因家族成员广泛存在陆生植物中,并在其生长发育的过程中起到关键作用。在水稻ALOG基因家族中总共有10个成员(G1, G1L1～G1L9),G1、G1L5和G1L6已经被证明是影响水稻花发育的关键基因,然而其余7个基因成员的功能还未知。本研究使用CRISPR/Cas9基因编辑系统对水稻ALOG基因家族功能未知的7个成员进行基因敲除,得到它们的突变株系,借此研究它们的基因功能并观察植株表型。在得到的所有T0代突变体植株中,G1L1、G1L2、G1L3、G1L8和G1L9经过测序和解码已经检测到各自纯合的突变株,经过对纯合突变株基因型和氨基酸序列的分析发现,它们各自的序列都发生移码并且编码的氨基酸序列都存在不同程度的突变,说明基因敲除成功。通过对T0代纯合植株的表型观测,我们在G1L1和G1L2的纯合突变株中发现了穗发育缺陷的表型,即主穗轴变长,且上面的小穗变少。本研究利用CRISPR/Cas9基因编辑技术对水稻ALOG基因家族成员进行敲除,为进一步研究水稻发育尤其是水稻花发育调控分子机制提供了重要的遗传材料。
The ALOG (Arabidopsis LSH1 and Oryza G1)gene family are widely found and play a key role in growth and development of terrestrial plants.There are 10 members ofALOG gene family (G1,GIL1,GIL2-GIL9) in rice.In previous research,G1,GIL5 and GIL6 have been proved to be a key gene that affecting rice flora development,while the function of remaining 7 members is still unknown.In this study,the remaining 7 members of rice ALOG gene family were knocked out by the CRISPR/Cas9 gene editing system to obtain their mutant lines,so as to study their gene function and observe their phenotypes.Among all To mutant plants obtained,the target site of GIL1,GIL2,GIL3,GIL8 and GIL9 have been sequenced and decoded.Therefore,their homozygous mutants had been identified.By analyzing the genotypes and amino acid sequences of homozygous mutants,it was found that their respective sequences were frameshift and the amino acid sequences encoded by them were mutated to different degrees,which indicated that the gene knockout was successful.Based on the phenotypic observation of the homozygous plants of To generation,we found the phenotype of the ear of grain developmental defects in the homozygous mutant plants of GIL1 and GIL2,that is,the main spike axis became longer and the spikelet on the top became less.In this study,CRISPR/Cas9 gene editing thchnique was used to knockout ALOG gene family members in rice,which might provide genetic materials for further research on the molecular mechanism of rice development,especially the regulation of rice flower development.
作者
张强
罗能杰
韦圣博
金健
Zhang Qiang;Luo Nengjie;Wei Shengbo;Jin Jian(State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,Guangxi University,Nanning,530004)
出处
《分子植物育种》
CAS
CSCD
北大核心
2019年第3期834-840,共7页
Molecular Plant Breeding
基金
广西高等学校高水平创新团队及卓越学者计划(2016
第三批)资助
