摘要
开花时间是作物重要发育性状,对作物产量有显著影响。miRNA是一类由生物体内源MIR基因转录后加工而形成的小分子RNA,可通过切割靶基因影响靶基因的表达。前期研究基础上在甘蓝型油菜中发现一个未知生物学功能的新的小分子RNA,novel-Bna-miR1040,本研究过表达Bna-miR1040的甘蓝型油菜较野生型花期延迟。降解组测序表明BnaEIF3A是Bna-miR1040的潜在靶基因,5'RACE和烟草瞬时转化试验表明,Bna-miR1040可以切割BnaEIF3A,使BnaEIF3A的表达量降低;进化分析结果表明,BnaEIF3A在不同物种中具有保守性;亚细胞定位结果显示BnaEIF3A定位在细胞核上;通过CRISPR/Cas9创建BnaEIF3A的功能缺失突变体,突变体较野生型也表现为花期延迟。本研究结果为甘蓝型油菜miRNA调控开花时间提供了数据,为甘蓝型油菜不同花期品种选育提供了理论参考。
Flowering time is a critical developmental trait in crops that significantly influences yield.MicroRNAs(miRNAs),a class of small endogenous RNAs processed from MIR gene transcripts,regulate gene expression by mediating transcript cleavage.In this study,we identified a novel miRNA—novel-Bna-miR1040—in rapeseed(Brassica napus),whose biological function had not been previously characterized.Transgenic rapeseed plants overexpressing Bna-miR1040 exhibited delayed flowering compared to wild-type plants.Degradome sequencing identified BnaEIF3A as a potential target of Bna-miR1040.This interaction was validated by 5′RACE and transient expression assays in tobacco leaves,which confirmed that Bna-miR1040 cleaves BnaEIF3A,significantly reducing its transcript levels.Evolutionary analysis showed that BnaEIF3A is highly conserved across diverse plant species.Subcellular localization analysis revealed that the BnaEIF3A protein is exclusively localized in the nucleus.To further investigate its function,we generated BnaEIF3A loss-of-function mutants using CRISPR/Cas9 genome editing.These mutants also exhibited delayed flowering phenotypes compared to wild-type controls.Together,our findings demonstrate that Bna-miR1040 regulates flowering time in rapeseed by suppressing BnaEIF3A expression,providing novel insights for molecular breeding strategies aimed at optimizing flowering time in rapeseed.
作者
马毅娜
吴晓明玉
李藕琪
王圆
陈丽
张盈川
赵伦
文静
傅廷栋
沈金雄
Ma Yi-Na;Wu Xiao-Ming-Yu;Li Ou-Qi;Wang Yuan;Chen Li;Zhang Ying-Chuan;Zhao Lun;Wen Jing;Fu Ting-Dong;Shen Jin-Xiong(National Key Laboratory of Crop Genetic Improvement,Huazhong Agricultural University/National Engineering Research Center of Rapeseed,Wuhan 430070,Hubei,China)
出处
《作物学报》
北大核心
2026年第2期349-362,共14页
Acta Agronomica Sinica
基金
国家自然科学基金项目(31930032)
财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-12)资助。
作者简介
通信作者:沈金雄,E-mail:jxshen@mail.hzau.edu.cn;第一作者:马毅娜,E-mail:mayina202202@163.com。
