| 大花蕙兰转齿兰环斑病毒外壳蛋白基因及检测 |
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| 引用本文： | 彭博,魏莉,杨凯,李潞滨.大花蕙兰转齿兰环斑病毒外壳蛋白基因及检测[J].林业科学研究,2016,29(2):234-237. |
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| 作者姓名： | 彭博 魏莉 杨凯 李潞滨 |
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| 作者单位： | 中国林业科学研究院林业研究所, 林木遗传育种国家重点实验室, 北京 100091;邯郸市环境监测中心站, 河北 邯郸 056002;北京农学院农业应用新技术北京市重点实验室, 北京 102206;中国林业科学研究院林业研究所, 林木遗传育种国家重点实验室, 北京 100091 |
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| 基金项目： | 中国特色花卉种业关键技术研究(2012BAD01B07) |
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| 摘 要： | 目的]通过植物转基因技术获得抗病毒大花蕙兰种质资源,优化转化体系和鉴定方法.方法]本研究克隆了齿兰环斑病毒外壳蛋白基因,并构建了该基因的pBI121表达载体,用根癌农杆菌介导法转化大花蕙兰,尝试以巢式PCR方法检测转基因再生植株.结果]优化了大花蕙兰遗传转化体系,建立了利用巢式PCR技术检测转基因大花蕙兰植株的方法,获得了32株转基因株(系).结论]优化了以类原球茎为外植体的农杆菌介导转化大花蕙兰的方法,确定以5%~10%类原球茎存活时的抗生素(卡那霉素)浓度为筛选浓度;获得了转ORSV CP基因大花蕙兰植株;对大花蕙兰转基因植株检测时,巢式PCR较普通PCR更灵敏、准确.
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| 关 键 词： | 大花蕙兰 齿兰环斑病毒 外壳蛋白基因 转基因 巢式PCR |
| 收稿时间： | 2/2/2015 12:00:00 AM |
Genetic Transformation and Detection of the Cymbidium hybridum Modified by Coat Protein Gene of ORSV |
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| PENG Bo,WEI Li,YANG Kai and LI Lu-bin.Genetic Transformation and Detection of the Cymbidium hybridum Modified by Coat Protein Gene of ORSV[J].Forest Research,2016,29(2):234-237. |
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| Authors: | PENG Bo WEI Li YANG Kai and LI Lu-bin |
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| Institution: | State Key Laboratory of Tree Genetics and Breeding, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China;Environmental Monitoring Center of Handan, Handan 056002, Hebei, China;Beijing Key laboratory for Agricultural Application and New Technique, Beijing University of Agriculture, Beijing 102206, China;State Key Laboratory of Tree Genetics and Breeding, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China |
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| Abstract: | Objective]To optimize the transformation system and identification methods for obtaining the germplasm resources of anti-virus Cymbidium hybridum by plant transgenic technology. Method]The coat protein gene (CP) of ORSV was cloned from the cDNA of infected C. hybridum. After sequencing, the gene was constructed into pBI121 expression vector and transformed to the protocorm-like bodies (PLBs) of C. hybridum by Agrobacterium tumefaciens-mediated method. Result]The genetic transformation system was optimized and the identification method of the transgenic plants was established by nest polymerase-chain-reaction (Nest-PCR). There were 32 transgenic plants of C. hybridum detected by Nest-PCR. Conclusion]The Agrobacterium tumefaciens-mediated transformation method of C.hybridum was optimized using PLBs as explants. The antibiotics(kanamycin)screening concentration was determined as 5% to 10% PLBs survival rate. Using the Nest-PCR to detected the transgenic plants was more sensitive and accurate than conventional PCR. |
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| Keywords: | Cymbidium hybridum Odontoglossum ringspot virus coat protein gene transgenic Nest-PCR |
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