| 董福双，高义平，吕孟雨，柴建芳，王海波.利用卡那霉素“浸种法”准确筛选转基因小麦种子的方法研究[J].麦类作物学报,2013,33(6):1105 |
| 利用卡那霉素“浸种法”准确筛选转基因小麦种子的方法研究 |
| Study on Accurately Kanamycin Screening Method of Transgenic Wheat Seed by Seed-immersion |
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| DOI：10.7606/j.issn.1009-1041.2013.06.007 |
| 中文关键词: 卡那霉素 小麦 转基因 抗性筛选 单粒分享量 |
| 英文关键词:Kanamycin Winter wheat Transgene Resistance selection Occupied amount of single grain |
| 基金项目:转基因生物新品种培育重大专项 （2009ZX08010-011B）。 |
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| 摘要点击次数: 1611 |
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| 中文摘要: |
| 为了确定利用卡那霉素“浸种法”筛选转基因小麦后代种子的准确方法，以冬小麦种子为试材，探讨了卡那霉素溶液体积、浓度及种子粒数对筛选结果的影响。结果表明，卡那霉素溶液体积、小麦种子的粒数对筛选结果的影响与卡那霉素浓度同等重要。在卡那霉素浓度为60～180 mg·L-1的范围内，以“单粒分享量”（单粒分享量=卡那霉素浓度×溶液体积/种子粒数）为筛选标准，能有效地对小麦种子进行筛选。利用此方法，我们对小麦金禾9123转基因后代7个株系的412个籽粒进行卡那霉素溶液筛选，获得84株抗性苗，经PCR检测，76株为阳性，卡那霉素筛选正确率达90.5%；同时以7个不同遗传背景的普通小麦品种也验证了此选择方法的可靠性。 |
| 英文摘要: |
| In order to determine accurately Kanamycin screening method of transgenic wheat seed by seed-immersion method , in this paper, winter wheat was used as the experimental material to discuss mainly the influence of knanamycin volume，concentration and grain number on kanamycin screening. The results showed that kanamycin volume and grain number were as importants as kanamycin concentration. Moreover, it’s effective to select resistant seeds according to “occupied kanamycin amount of single grain” within 60～180 mg·L-1 range of kanamycin concentration (occupied kanamycin amount of single grain = kanamycin concentration×volume/grain number). Based on this method we got 84 resistant plants throught knamycin resistances screening of 412 seeds, which came from 7 transgeneic progeny lines. 76 plants were positive by PCR analysis. The correct rate of knamycin resistance screening was 90.5% Meanwhile this method was verified reliability through Kanamycin screening of seven cultivars. We could obtain optimum proportion between security screening effect and reagent amount according the formula of occupied kanamycin amount of single grain. It provides a new way for resistante progeny screening of seed plant. |
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