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Csu-miR260转基因水稻插入序列的表达检测
引用本文:罗彪,周池,陈彦超,周霆,柳初微,周镇中,陈浩,饶力群,汪启明.Csu-miR260转基因水稻插入序列的表达检测[J].植物保护,2021,47(6):201-207.
作者姓名:罗彪  周池  陈彦超  周霆  柳初微  周镇中  陈浩  饶力群  汪启明
作者单位:1. 湖南农业大学生物科学技术学院, 长沙410128; 2. 华中农业大学作物遗传改良国家重点实验室, 武汉430070
基金项目:转基因生物新品种培育重大专项(2018ZX08011-01B); 国家自然科学基金(31772178)
摘    要:本文以具有抗二化螟性状的Csu-miR260转基因水稻为试验材料, 采用TaqMan探针实时荧光定量PCR (TaqMan RT-qPCR)和茎环实时荧光定量PCR(stem-loop RT-qPCR)分别对Csu-miR260转基因抗虫水稻中Csu-miR260插入序列和剪切形成的amiR260s的表达情况进行了定量检测。发现Csu-miR260插入序列和剪切形成的20 nt和21 nt两种长度amiR260s在转基因抗虫水稻苗期的根、茎、叶中表达, 且无组织表达特异性。该试验解决了人工miRNA作物中amiRNA及前体检测引物的特异性问题, 为人工miRNA植物干扰序列的表达分析提供技术参考, 并为miRNA转基因作物遗传表达相关安全评价提供了数据参考。

关 键 词:人工miRNA    抗虫水稻    TaqMan实时定量PCR    茎环实时定量PCR    转基因安全评价
收稿时间:2020/8/9 0:00:00
修稿时间:2020/9/14 0:00:00

Detection of expression of inserted sequence in Csu-miR260 transgenic rice
LUO Biao,ZHOU Chi,CHEN Yanchao,ZHOU Ting,LIU Chuwei,ZHOU Zhenzhong,CHEN Hao,RAO Liqun,WANG Qiming.Detection of expression of inserted sequence in Csu-miR260 transgenic rice[J].Plant Protection,2021,47(6):201-207.
Authors:LUO Biao  ZHOU Chi  CHEN Yanchao  ZHOU Ting  LIU Chuwei  ZHOU Zhenzhong  CHEN Hao  RAO Liqun  WANG Qiming
Affiliation:1. College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China; 2. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, China
Abstract:In this article, the expression of insert sequence Csu-miR260 from transgenic rice with resistance to Chilo suppressalis and two length(20 nt and 21 nt) of amiRNA formed by shearing Csu-miR260 were quantitatively determined by using TaqMan probe real-time fluorescent quantitative PCR (TaqMan RT-qPCR) and stem-loop real-time fluorescent quantitative PCR (stem-loop RT-qPCR). It was found that the expression of the insert sequence Csu-miR260 and amiRNA formed by shearing Csu-miR260 were no tissue specificity in the roots, stems and leaves of the transgenic insect-resistant rice seedlings. This test solves the problem of specificity of amiRNA and precursor detection primers in artificial miRNA crops, provides technical reference for the expression analysis of artificial miRNA plant interference sequences. It also provides a data reference for the safety evaluation of genetic expression in miRNA transgenic crops.
Keywords:artificial miRNA  insect-resistant rice  TaqMan RT-qPCR  stem-loop RT-qPCR  safety evaluation of transgenic plants
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