华北农学报 ›› 2006, Vol. 21 ›› Issue (5): 99-104. doi: 10.3321/j.issn:1000-7091.2006.05.024

所属专题: 玉米 生物技术

• 论文 • 上一篇    下一篇

抗玉米丝黑穗病转基因株系的分子检测及抗病株系选育

杜建中1, 孙毅1, 郝曜山1, 刘俊玲2   

  1. 1. 山西省农业生物技术研究中心, 山西太原 030031;
    2. 清徐县农业局, 山西清徐 030400
  • 收稿日期:2006-05-16 出版日期:2006-10-28
  • 通讯作者: 孙毅
  • 作者简介:杜建中(1960- ),男,山西芮城人,副研究员,主要从事分子生物学研究工作。
  • 基金资助:
    国家“863”计划项目(2001AA212131)

Molecular Check and Measure of Transgenic Maize Lines Resistancing to Maize Head Smut and Breeding Selection for Transgenic Lines Resistancing to Disease

DU Jian-zhong1, SUN Yi1, HAO Yao-shan1, LIU Jun-ling2   

  1. 1. The Agri-Biotechnology Research Center of Shanxi Province, Taiyuan 030031, China;
    2. Bureau of Agriculture of Qing-xu County, Qingxu 030400, China
  • Received:2006-05-16 Published:2006-10-28

摘要: 采用花粉介导方法将质粒pGL IIRC1导入玉米自交系海921中,获得了T0种子128粒,种子经潮霉素抗性筛选得到抗潮霉素植株16株。对抗性植株分离的统计分析表明,多数情况下几丁质酶基因以单拷贝形式导入转基因植株,后代分离比约为3:1。抗性苗移栽到大田后,56叶期取样进行PCR扩增、PCR Southern blot杂交分析,对T1的分析结果表明,几丁质酶基因已导入转基因植物细胞中。对T2,T3植株的检测结果显示,目的基因已整合到转化植株基因组中并可随植株世代稳定遗传。田间抗病鉴定结果表明,转基因植株及后代的玉米丝黑穗病发病率明显比对照降低。根据分子检测及田间抗病鉴定结果,得到GH05024,GH05028 2个抗丝黑穗病的纯合转基因株系。

关键词: 玉米, 几丁质酶基因, 转基因, 丝黑穗病, 选育

Abstract: In this study 128 transgenic maize seeds that were obtained from transgenic plants transformed by pollen-mediated approach on maize(Zea mays L.) inbred line Hai 92-1,the vector was plasmid pGL II-RC-1.The result of seeds selected by hygromycin-screening had proved that 16 plants was hygromycin-resistant plants.Statistics analysis for the hygromycin-resistant plants suggested that single copy of the chintinase gene was transferred into the cell of majority of T-(0) transgenic plants,segregation ratio of the progeny of transgenic plants for hygromycin resistant and susceptible was about 3:1,P<0.01.The results of detecting to T-1 transgenic plants by PCR amplification and PCR Southern blot hybridization showed that chitinase gene was transferred into the cells of transgenic plants.The same methods detected to T2 and T3,their results indicated that chitinase gene was integrated into the genome of transgenic plants and stable inheritance in transgenic plants and their progeny.The results of Bio-assay for head smut-resistance activity indicated that the disease incidence of transgenic plants and their progeny was lower than that of control plants.Based on the results above,GH05024 and GH05028.2 transgenic maize lines that resisted to maize head smut were obtanined by breeding selection.

Key words: Maize, Chitinase gene, Transgene, Maize head smut, Breeding selection

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引用本文

杜建中, 孙毅, 郝曜山, 刘俊玲. 抗玉米丝黑穗病转基因株系的分子检测及抗病株系选育[J]. 华北农学报, 2006, 21(5): 99-104. doi: 10.3321/j.issn:1000-7091.2006.05.024.

DU Jian-zhong, SUN Yi, HAO Yao-shan, LIU Jun-ling. Molecular Check and Measure of Transgenic Maize Lines Resistancing to Maize Head Smut and Breeding Selection for Transgenic Lines Resistancing to Disease[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2006, 21(5): 99-104. doi: 10.3321/j.issn:1000-7091.2006.05.024.

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