| 3种筛选NPT-Ⅱ标记转基因油菜方法研究 |
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| 引用本文： | 韦献雅,牛应泽,张其坤.3种筛选NPT-Ⅱ标记转基因油菜方法研究[J].中国农学通报,2009,25(14):37-41. |
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| 作者姓名： | 韦献雅 牛应泽 张其坤 |
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| 作者单位： | 1. 四川省乐山市农牧科学研究所,四川乐山,614216
2. 四川农业大学油菜研究中心,四川农业大学作物基因资源与遗传改良教育部重点实验室,四川雅安,625014 |
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| 基金项目： | 四川省科技厅应用基础研究项目"拟南芥CRY1基因导入甘蓝型油菜的遗传表达及分子鉴定" |
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| 摘 要： | 利用卡那霉素溶液叶片涂抹法、种子浸泡法以及卡那霉素MS培养基筛选法三种方法对含有NPT-Ⅱ标记基因的转基因油菜种子（T0）进行筛选。结果表明,三种筛选方法的最佳卡那霉素选择压为200mg/L；涂抹叶片法4-5d可以明显识别植株对卡那霉素抗性与否，此法筛选到的4株植株PCR检测全部呈阳性，可靠性达100%；浸泡种子法2-3d可以明显识别植株对卡那霉素抗性与否，此法筛选到的5株植株PCR检测全部呈阳性，可靠性100%；MS培养基筛选法一般10d可以明显植株识别对卡那霉素抗性与否，此法筛选到20株植株，移栽后存活5株，PCR检测3株呈阳性，2株是假阳性植株，可靠性为60%。因此可以认为卡那霉素涂抹叶片和浸泡种子两种方法是转基因油菜进行大规模、快速的筛选及后代的遗传分析的理想方法。
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| 关 键 词： | 油菜 转基因植株 NPT-II 筛选方法 |
| 收稿时间： | 2009-02-23 |
| 修稿时间： | 2009-05-07 |
Study on Three Methods Screening Transgenic Rapeseed Tagging NPT-II |
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| WEI Xian-Y.Study on Three Methods Screening Transgenic Rapeseed Tagging NPT-II[J].Chinese Agricultural Science Bulletin,2009,25(14):37-41. |
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| Authors: | WEI Xian-Y |
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| Institution: | Wei Xianya, Niu Yingze, Zhang Qikun (1 Institue of Agricultural Science Le Shan of Si Chuan, Leshan Sichuan 614216; 2Rapeseed Research Center of Sichuan Agricultural University, Key Laboratory of Crop Genetic Resources and Improvement, Ministry of Education, Siohuan Agricultural University, Ya＇an Sichuan 625014) |
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| Abstract: | Abstract: In order to obtain the simple method of sieving the seeds of transgenic rapeseed tagging NPT-II. Using three methods such as Kanamycin daubing vane, dipping seeds and MS culture medium containing Kanamycin to study. The results indicated that the best Kanamycin choice press was 200 mg/L. The method of Kanamycin daubing vane can distinguish to Km- resistance or not for 4-5 days. Four plants were chosen in this method and its PCR detection was positive. The reliability reached 100%. The method of dipping seeds can distinguish to Km- resistance or not for 2-3 days. Five plants were chosen in this method and its PCR was positive. The reliability reached 100%. The method of Kanamycin MS culture medium can distinguish to Km- resistance or not commonly for 2-3 days. Twenty plants were chosen in this method but only five saved in transplanting. The result of PCR detection is that there plants were positive and two were negative. The reliability reached 60%. So, it could be thought Kanamycin daubing vane and dipping seeds were ideal methods for fast screening transgenic rapeseed and descendant’s heredity analysis. |
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| Keywords: | Key words: rapeseedzz transgenic-plantzz NPT-IIzz screenzz |
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