转EPSPS基因抗草甘膦棉花的遗传分析

Genetic Analysis of Transgenic Glyphosate-resistance Cotton with EPSPS Gene

为分析转基因抗草甘膦棉花早代遗传情况,以花粉管通道法获得的26个转5-烯醇式丙酮酰莽草酸-3-磷酸合酶基因(EPSPS)抗草甘膦棉花转化事件为材料,以其背景亲本中棉所49为对照,喷施草甘膦后对转基因棉T1、T2分离比例进行考察。T1田间抗性鉴定结果表明,经卡方检测20个转化事件T1分离符合3∶1的分离规律,即外源基因插入1个位点;6个转化事件不符合1对基因的分离规律,出现了偏分离。T2田间抗性鉴定结果表明,通过花粉通管法共获得152个纯合株系,分别来源于25个转化事件;对T2不纯合株系继续进行分离比例的考察,发现来源于15个转化事件的57个株系符合3∶1的分离规律;此外卡方检测结果表明,每个转化事件都有不符合3∶1分离规律的株系,且其中10个转化事件没有符合3∶1分离规律的株系。表明通过花粉管通道法获得的转基因植株中外源基因的整合和遗传均较复杂。

The purpose of this study was to analysis the genetic characters of transgenic glyphosate-resistance cotton with EPSPS gene,using the glyphosate-resistant cotton transformation events G6-1-G6-26 as the materials,and their non-transgenic genetic background cultivar CCRI-49 as control. Field resistance test in T1 plants showed that the 20 of 26 transformation events were consistent with the ratio of 3 ∶ 1 and the other transformation events were inconsistent with single-gene Mendelian inheritance according to χ2test. Field resistance test in T2 plants showed152 homozygous resistance lines were obtained derived from 25 transformation events,respectively. Fifty-seven lines derived from 15 transformation events were consistent with the ratio 3 ∶ 1 through the analysis of heterozygous resistance lines. Furthermore,every transformation event had lines which were inconsistent with the ratio of 3 ∶ 1,and 10 transformation events did not have lines which were consistent with the ratio of 3 ∶ 1. These suggested the integration and genetic mode of exogenous gene transformed by the pollen tube pathway method were complex.