cryIAc基因在转基因玉米中的遗传规律及对抗虫性影响

doi:10.13560/j.cnki.biotech.bull.1985.2015.01.019

摘要/Abstract

摘要： 旨在研究目的基因在转基因植株和后代植株（株系）中的遗传规律及其对转化植株抗虫性的影响。以花粉介导法将cryIAc基因导入玉米自交系‘郑58’和‘昌7-2’，对转化植株及其后代株系进行分子检测和田间抗虫鉴定。结果表明：（1）转化‘郑58’和‘昌7-2’，T₁代分别获得转基因植株24和41个，转化率高达20%以上；（2）转基因T₂代、杂交F₂代及回交1代（B₁）的分子检测结果证明，外源基因的遗传符合孟德尔的3: 1、3: 1和1: 1的遗传分离规律；（3）连续多带的分子检测结果还表明，外源基因可稳定遗传并有效表达，表达水平在9.8-14.3 ng/g叶片鲜重之间；（4）抗虫鉴定结果显示，在阴性对照全部感虫情形下，转基因纯合株系仍表现出较高抗虫活性；（5）此外，回交试验结果还证明外源基因通过杂交可传递给下一代；（6）最终经筛选得到SZ003、SZ005、SC001、SC004和SC007五个高抗虫转基因株系。结果表明,花粉介导法是一种高效、快捷的转化方法，cryIAc基因导入玉米自交系植株后赋予和提高了转基因植株的抗虫活性。

关键词: 转基因玉米, cryIAc基因, 遗传规律, 抗虫性

Abstract: In order to analyse the genetic regularity of target gene cryIAc in transgenic and their descendant plants（lines）, and to study the effect of target gene on insect-resistant activity of transgenic plants at the same time, this research work was conducted. Firstly, cryIAc gene was transformed into maize inbred ‘Zheng 58’ and ‘Chang7-2’ separately by pollen-mediated transformation method. PCR, Southern, ELISA analysis and bioassay in the field of each generation transgenic plants were then conducted according to experimental requirements. Results showed that ⑴ A total of 24 and 41 transgenic plants were obtained through transformed cryIAc gene into maize inbred ‘Zheng 58’ and ‘Chang7-2’, respectively. ⑵ The molecular detection results of transgenic T₂, hybridization F₂ and backcross B₁plants suggested that the segregation of target gene in these lines followed 3: 1, 3: 1 and 1: 1 segregation ratio of the Mendel laws of heredity, respectively. ⑶ The molecular detection results of T₁ to T₄ transgenic plants（lines）showed that the target gene could stably inherited and expressed effectively, the expression amount of the target gene was from 9.8 to 14.3 ng/gleaf fresh weight. ⑷ Moreover, the results of bioassay in the field indicated that in the case of high insect-susceptibility of negative control line, the transgenic lines still showed highly insect-resistant activity. ⑸ In addition, the target gene integrated into genomics of transgenic plants could inherit to the next generation plants by hybridization. ⑹ At last, 5 high insect-resistant transgenic lines, SZ003, SZ005, SC001, SC004 and SC007, was gained by screening. The pollen-mediated transformation method was a effective and shortcut tool used in plant transformation, cryIAc gene could confer and enhance insect-resistant activity of transgenic plants（lines）tansfomated with it.

Key words: transgenic maize, cryIAc gene, genetic regularity, insect-resistance

王建军,杨慧珍,刘佼. cryIAc基因在转基因玉米中的遗传规律及对抗虫性影响[J]. 生物技术通报, 2015, 31(1): 122-130.

Wang Jianjun, Yang Huizhen, Liu Jiao. Genetic Analysis of cryIAC Gene in Transgenic Maize Plants and Its Effects on Insect-resistance[J]. Biotechnology Bulletin, 2015, 31(1): 122-130.

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