烟草烟碱相关miRNA筛选及验证  

Identification and validation of nicotine related miRNAs in tobacco

作　　者：许亚龙[1] 陈千思[1] 谢小东[1] 刘萍萍[1] 翟妞[1] 金静静[1] 王晨[1] 曹培健[1] XU Yalong;CHEN Qiansi;XIE Xiaodong;LIU Pingping;ZHAI Niu;JIN Jingjing;WANG Chen;CAO Peijian(Zhengzhou Tobacco Research Institute of CNTC,Zhengzhou 450001,China)

机构地区：[1]中国烟草总公司郑州烟草研究院,郑州450001

出　　处：《烟草科技》2021年第2期10-17,共8页Tobacco Science & Technology

基　　金：国家烟草专卖局烟草基因组计划重大专项“烟草基因功能元件全景图构建及关键基因挖掘与调控节点研究”[110201601033(JY-07)];郑州烟草研究院院长科技发展基金项目“烟碱合成相关miRNA功能研究”(902018CA0240)。

摘　　要：为挖掘烟草中可能参与烟碱调控相关的miRNA,利用生物信息学方法,进行了烟草全基因组水平的miRNA鉴定,筛选出烟碱相关的miRNA,并利用qPCR和GC-MS技术对相关过表达miRNA材料开展了功能验证。结果显示,利用红花大金元3个时期5个不同组织(根、茎、叶、花和腋芽)共39个样品的miRNA测序数据,共鉴定出了2 925个miRNA,其中1 245个miRNA是已知的,包括43个烟草中已经报道的miRNA。过表达miR2635的材料靶基因NtQPT2表达量下降约48%~67%,过表达miRNA203材料的靶基因NtMATE1表达量下降达98%,两种过表达材料中平均烟碱含量分别降低了约69%和62%,表明miRNA2635和miRNA203可分别通过降低靶基因NtQPT2和NtMATE1的表达量进而降低烟草中的烟碱含量。In order to identify miRNAs involving nicotine biosynthesis in tobacco,whole-genome miRNA identification was conducted and nicotine related miRNAs were predicted by bioinformatics.The functional verification of miRNA overexpressed plants was performed by qPCR and GC-MS technologies.The results showed that from 39 samples of cv.Honghuadajinyuan,including 5 tissues(root,stem,leaf,flower and axillary bud),at 3 developmental stages,a total of 2925 potential miRNAs were identified,and 1245 of which were conserved among other plants,and 43 candidates had already been reported in tobacco by other studies.The expression of NtQPT2 decreased by 48%-67%in miRNA2635 overexpressed plants,the expression of NtMATE1 decreased by 98%in miRNA203 overexpressed plants,and the average nicotine contents of two types of overexpressed plants decreased by about 69%and 62%respectively;which indicated that overexpressions of miRNA2635 and miRNA203 could significantly decrease the accumulation of nicotine by downregulating the expressions of NtQPT2 and NtMATE1.

关 键 词：烟草 烟碱 MIRNA 靶基因 NtQPT2 NtMATE1 

分 类 号：TS413[农业科学—烟草工业]