Bna-novel-miR36421调节拟南芥株型和花器官发育的功能验证  

Functional validation of Bna-novel-miR36421 regulating plant architecture and flower organ development in Arabidopsis thaliana

作　　者：戴丽诗 常玮 张赛[1,2] 钱明超 黎小东 张凯[1,2] 李加纳[1,2,3] 曲存民[1,2,3] 卢坤[1,2,3] DAI Li-Shi;CHANG Wei;ZHANG Sai;QIAN Ming-Chao;LI Xiao-Dong;ZHANG Kai;LI Jia-Na;QU Cun-Min;LU Kun(College of Agronomy and Biotechnology,Southwest University,Chongqing 400715,China;Engineering Research Center of South Upland Agri-culture,Ministry of Education,Chongqing 400715,China;Academy of Agricultural Sciences,Southwest University,Chongqing,400715,China)

机构地区：[1]西南大学农学与生物科技学院,重庆400715 [2]南方山地农业教育部工程研究中心,重庆400715 [3]西南大学农业科学研究院,重庆400715

出　　处：《作物学报》2022年第7期1635-1644,共10页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31871653);国家重点研发计划项目(2018YFD0100500);高等学校学科创新引智基地项目“111项目”(B12006);重庆市自然科学基金重点项目;西南大学种质创制专项资助

摘　　要：MicroRNA参与油菜种子发育、胁迫响应和胚胎发育等多种生物学过程,但其在油菜株型和花器官发育方面的报道还较少。本研究以高/低收获指数油菜中显著差异表达的Bna-novel-miR36421为对象,通过转基因植株表型、靶基因预测、表达量和双荧光素酶报告系统等解析其调控机制。Bna-novel-miR36421与植物miR167家族成员高度同源,可能为油菜新的miR167成员。Bna-novel-miR36421能靶向并抑制Bna.C03ARF6、Bna.C06ARF8、Bna.A09PATL2和Bna.C03DUF581的表达。过表达拟南芥植株中,Bna-novel-miR36421的表达量显著上调,而上述4个靶基因的拟南芥同源基因表达量极显著下降。过表达拟南芥植株株型矮小,茎间缩短,叶片高度卷曲;花器官发育异常,雌蕊膨大,雄蕊花丝缩短,花药不开裂,花粉败育。推测Bna-novel-miR36421可能通过抑制Bna.C03ARF6、Bna.C06ARF8、Bna.A09PATL2和Bna.C03DUF581基因表达,进而对油菜株型和花器官的发育起到重要的调控作用。研究结果对解析miRNA介导的植物株型和花器官发育分子机制,挖掘重要发育性状关键基因奠定了重要基础。MicroRNA is involved in the regulation of various biological processes such as development,stress response,and em-bryonic development in rapeseed,however,only few studies focused on the regulation of miRNA on the plant architecture and floral organ development of rapeseed.In this study,Bna-novel-miR36421 that differentially expressed between the high and low harvest index accessions of rapeseed was identified,whose biological function and regulation mechanism were further character-ized through phenotype analysis in transgenic plants,target gene prediction,expression pattern comparison,and dual luciferase reporter system.The results showed that Bna-novel-miR36421 was highly homologous to miR167 family members,and might be a novel miR167 member in rapeseed.The qRT-PCR and dual luciferase reporter system results indicated that Bna-novel-miR36421 could inhibit the relative expression levels of Bna.C03ARF6,Bna.C06ARF8,Bna.A09PATL2,and Bna.C03DUF581.In the overexpressing Arabidopsis plants,the expression of Bna-novel-miR36421 was significantly increased,while the transcription levels of the Arabidopsis orthologs of its target genes were decreased significantly.Phenotypic observation showed that the plant height of overexpression Arabidopsis plants were reduced,with shortened stems and curled leaves.The development of floral organs was abnormal,with enlarged pistil,shortened stamen filaments,unbreakable anthers,and aborted pollens.Hence,it could be proposed that Bna-novel-miR36421 may regulate plant architecture and floral organ development by repressing Bna.C03ARF6,Bna.C06ARF8,Bna.A09PATL2,and Bna.C03DUF581.The results laid a solid foundation for under-standing the molecular mechanism of miRNA-mediated plant architecture and flower organ development,and mining the key genes involved in plant developmental processes.

关 键 词：甘蓝型油菜 Bna-novel-miR36421 miR167 株型 花器官 

分 类 号：Q943.2[生物学—植物学]