PLA3基因生物信息学分析及在赤霉素调控水稻种子萌发中的作用

Bioinformatics analysis of PLA3 gene and its involvement in gibberellin regulation of rice seed germination

水稻(Oryza sativa) PLA3基因编码谷氨酸羧肽酶,影响种子萌发等生长发育过程。本研究运用CRISPRCas9技术获得‘日本晴’水稻pla3突变体,该突变体具有胚变大、易萌发的表型。通过对PLA3蛋白进行多物种同源比对和进化分析,发现其具有较高的保守性。对PLA3基因的时空表达和响应不同激素的表达进行了分析,发现其在胚中表达较高,并受赤霉素(GA)等诱导。同时对PLA3蛋白的亚细胞定位、种子萌发过程中GA的含量及转录组学差异等进行分析,结果表明:PLA3定位于内质网;野生型与pla3种子萌发中不同阶段(12和30 h)胚内GA含量差异较大;转录组测序筛选获得与GA信号途径相关的41个差异基因;pla3中信号途径基因如GID2表达出现明显上调。综上所述, PLA3基因与种子萌发相关并参与GA调控种子萌发的过程。

Gene PLA3(PLANTOCHRON3) in rice(Oryza sativa) encodes glutamate carboxypeptidase, affecting the growth and development process, such as seed germination. In this study, CRISPR-Cas9 technology was used to obtain the mutant pla3 in rice ‘Nipponbare’, which has a phenotype with larger embryo and easy germination. Through homologous alignment and phylogenetic analysis of multiple species, it was found that PLA3 has a high conservatism. In addition, the spatial-temporal expression analysis and the responses to different hormones of gene PLA3 showed that it was relatively highly expressed in the embryo and induced by gibberellin(GA). In addition, the subcellular localization of PLA3 protein, GA content and transcriptomic differences during seed germination were analyzed, which indicates that PLA3 was located in the endoplasmic reticulum, and the embryo GA content of wild-type and pla3 during different stages(12 and 30 h) of seed germination were quite different;41 differential genes related to GA signaling pathway were obtained via transcriptome sequencing screening;the expression of pla3 signal pathway genes, like GID2, was significantly up-regulated. In summary, gene PLA3 is related to seed germination and participates in the process of GA regulating seed germination.