The Study On ALOG Gene Family Transgenic Knockout Lines Based On CRISPR/Cas9 And The Regulatory Network Of G1L6

ALOG(Arabidopsis LSH1 and Oryza G1)gene family are widely found and play a key role in growth and development in terrestrial plants.There are 10 members of the ALOG gene family(G1,G1L1?G1L9)in rice.In previous research,G1,G1L5 and G1L6 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 and G1L6 of rice ALOG gene family were knocked out by the CRISPR/Cas9 gene editing system to generate the mutant lines.Furthermore,overexpression lines of the 8 ALOG genes were also produced to investigate their function in rice.Our results indicated that all overexpression lines showed no obvious developmental defect,while most of the CRISPR/Cas9-induced lines were also normal,except G1L1 and G1L2.The CRISPR/Cas9-induced transgenic lines showed no difference with WT maybe caused by gene redundancy in ALOG gene family.The phenotype of "beak-like" spikelet of the CRISPR/Cas9-induced G1L6 transgenic line was similar to previous research,and the KO-g1l6 plants exhibited sterility.G1L1 and G1L2 gene knockout lines displayed elongated floral axis with less spikelet.In the analysis of KEGG enrichment pathway,8 starch and sucrose metabolism related DEGs were found which may be related to the sterility of KO-g1l6;15 plant hormone signal transduction related DEGs were also found which revealed that plant hormones may play an important role in the lemma and palea development of KO-gll6.In this study,The RNA-Seq transcriptome analysis technique was performed to explore the regulatory network of G1L6.The ALOG gene family transgenic knockout lines and overexpression lines provided important materials to study ALOG gene members.Our results provide the basis for further analysis of the molecular mechanism of flora development in rice according to the G1L6 regulatory network.