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| 含粘质沙雷氏菌几丁质酶SchiA基因的植物转化质粒pBG1112构建和水稻遗传转化 | |
| 引用本文： | 何迎春,李小湘,高必达.含粘质沙雷氏菌几丁质酶SchiA基因的植物转化质粒pBG1112构建和水稻遗传转化[J].农业生物技术学报,2003,11(2):121-126. |
| 作者姓名： | 何迎春 李小湘 高必达 |
| 作者单位： | 1. 湖南中医学院基础课部生物化学教研室,长沙,410007 2. 湖南省水稻研究所,长沙,410125 3. 湖南农业大学植物保护学院,长沙410128 |
| 基金项目： | 湖南省教委高校科研项目 |
| 摘 要： | 摘要:将一个2.8 kb的CaMV35S启动子/SchiA编码区/Nos终止区融合基因插入到植物转化载体pCAMBIA1301的多克隆酶切位点，得到1个14.6 kb的新植物转化质粒pBG1112,用花器介导法转化水稻（Oryza sativa ），经PCR检测，初步证实已将目的基因整合到受体植物的基因组中。一部分转基因T3代潮霉素抗性阳性植株对水稻纹枯病(Rhizoctonia solani )和稻瘟病（Pyricularia oryzae）的抗性较非转化对照增强。RT-PCR表明抗病性植株为阳性，而不抗病的植株为阴性。将RT-PCR产物测序后，用BLAST软件分析序列可知，该序列为细菌几丁质酶基因核苷酸序列而不是植物几丁质酶基因的核苷酸序列。T4代转基因水稻的几丁质酶活力高于对照未转基因植株，表明转入的外源几丁质酶基因能正常表达。 |
| 关 键 词： | 关键词： 细菌几丁质酶基因 转化 水稻 纹枯病 稻瘟病 |
Construction of a New Plant Transformation Vector pBG1112 with a Chitinase Gene SchiA from Serratia marcescens and Its Genetic Transformation in Rice |
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| Abstract: | Abstract: A 2.8 kb CaMV 35S promoter/Schi A coding region/Nos terminator fusion gene was inserted into the polycloning site of a 11.8 kb binary vector pCAMBIA1301 to produce a new 14.6 kb plasmid pBG1112, which was used in rice (Oryza sativa L.) transformation by floral organ-mediated method. T3 rice plants were screened by hygromycin solution and the positive plants were examined by PCR for presence of transgene and by RT-PCR for matured mRNA. Bioassay for some of the T3 transgenic rice plants which displayed both hygromycin-resistance and RT-PCR positive showed the increased resistance to rice sheath blight (Rhizoctonia solani ) and rice blast（Pyricularia oryzae）. The sequencing result showed that the nucleotide acid sequences of RT-PCR products were the same as that of transgene analyzed by BLAST software. Chitinase activity of T4 transgenic rice was higher than that of non-transgenic rice,which showed that the transferred exogenous chitinase gene could be expressed normally. |
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