OsBTF3过表达转基因水稻株系的创制及其分子鉴定

doi:10.3969/j.issn.2095-2341.2012.01.07

生物技术进展 ›› 2012, Vol. 2 ›› Issue (1): 39-43.DOI: 10.3969/j.issn.2095-2341.2012.01.07

OsBTF3过表达转基因水稻株系的创制及其分子鉴定

李广旭1,2,陈华民1,吴静1,吴茂森1,何晨阳1*

1.中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193;
2.辽宁省农业科学院果树研究所, 辽宁熊岳 115009

收稿日期:2011-11-04 出版日期:2012-01-25 发布日期:2011-12-14
通讯作者: 何晨阳,研究员,博士生导师,主要从事分子植物病理学研究。Tel:010-62894147; E-mail:cyhe@caas.net.cn
作者简介:李广旭,博士研究生,主要从事分子植物病理学研究。
基金资助:
国家转基因生物新品种培育重大专项(2009ZX08009-044B,2011ZX08001-002)资助。

Generation and Molecular Identification of Transgenic Rice Lines with Over-expressed OsBTF3 Gene

LI Guang-xu, CHEN Hua-min, WU Jing, WU Mao-sen, HE Chen-yang

1.State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
2.Institute of Pomology, Liaoning Academy of Agricultural Sciences, Liaoning Xiongyue 115009, China

Received:2011-11-04 Online:2012-01-25 Published:2011-12-14

摘要/Abstract

摘要： 本研究旨在创制OsBTF3过表达转基因水稻株系,为验证OsBTF3基因在水稻抗性和生长发育中的功能、评价其在水稻农艺性状遗传改良中的应用价值提供试验材料。通过基因过表达载体构建、水稻愈伤组织诱导、农杆菌介导愈伤组织转化、植株再生、潮霉素抗性(Hyg^R)筛选及PCR验证、基因过表达RT-Q-PCR检测等方法,成功地获得了97个T₀代和20个T₁代过表达转基因株系,并分别得到分子验证。与野生型对照株相比,5个T₁代过表达株系中的OsBTF3基因表达水平显著提高,平均高达3.58倍。因此,由组成型表达的35S启动子驱动的OsBTF3基因在转基因水稻株系中成功地得到了增量表达,并对水稻生长发育、抗病性和抗逆性具有调控作用。

关键词: OsBTF3, 过表达, 分子鉴定,生物学功能

Abstract: The transgenic rice lines were generated with over-expressed OsBTF3, which can further be used not only for functional analysis of OsBTF3 gene in biotic and abiotic stress resistance and growth and development, but also for evaluation of its potential use in genetic improvement of agronomic trails of rice. The generation of OsBTF3 gene overexpression construct, Agriobacterium-mediated transformation of callus tissues, Hyg^R screening, regeneration of transgenic plants and PCR validation were conducted. Through Agrobacterium-mediated rice transformation method, 97 transgenic lines of T₀ generation and 20 transgenic lines of T₁ generation were successfully generated and validated respectively. The transcription of OsBTF3 in 5 transgenic lines of T₁ generation is significantly higher than the control plants with average of 3.58 times. OsBTF3 gene was successfully overexpressed under the control of 35S promoter resulting in significant changes in growth and development, biotic and abiotic stress resistance in the transgenic rice lines that were generated in this study.

Key words: OsBTF3, overexpression, molecular identification, biological functions

李广旭,陈华民,吴静,吴茂森,何晨阳. OsBTF3过表达转基因水稻株系的创制及其分子鉴定[J]. 生物技术进展, 2012, 2(1): 39-43.

LI Guang-xu1,2, CHEN Hua-min1, WU Jing1, WU Mao-sen1, HE Chen-yang1*. Generation and Molecular Identification of Transgenic Rice Lines with Over-expressed OsBTF3 Gene [J]. journal1, 2012, 2(1): 39-43.

OsBTF3过表达转基因水稻株系的创制及其分子鉴定

Generation and Molecular Identification of Transgenic Rice Lines with Over-expressed OsBTF3 Gene

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