银中杨叶片离体再生体系的建立及Gt5GT7基因的遗传转化

以银中杨幼嫩枝条为外植体,探究不同预培养方式、激素配比对其叶片离体再生体系的影响.结果表明,枝条在MS营养液中预培养3d为最优方式;不定芽诱导最佳培养基为MS＋TDZ 0.1 mg/L＋NAA 0.02 mg/L,叶片可不经历愈伤组织直接形成丛生芽,最适生根培养基为1/2 MS＋NAA 0.05 mg/L＋IBA 0.01 mg/L＋活性炭0.6 g/L.利用发根农杆菌介导法,建立Gt5GT7基因的银中杨叶片遗传转化体系,叶片GUS染色结果初步证明,Gt5GT7基因成功整合入银中杨叶片,且最适宜侵染的菌液A600值为0.45～0.6....

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Published in广东农业科学 Vol. 41; no. 13; pp. 144 - 146
Main Author 徐洪伟 高峰 杨伟新 王晶 周晓馥
Format Journal Article
LanguageChinese
Published 吉林师范大学植物资源科学与绿色生产吉林省重点实验室,吉林四平,136000 2014
Subjects
Gt5GT7
激素配比
离体再生
遗传转化
银中杨
预培养
离体再生
银中杨
预培养
Gt5GT7
激素配比
遗传转化
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ISSN1004-874X

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Summary:以银中杨幼嫩枝条为外植体,探究不同预培养方式、激素配比对其叶片离体再生体系的影响.结果表明,枝条在MS营养液中预培养3d为最优方式;不定芽诱导最佳培养基为MS＋TDZ 0.1 mg/L＋NAA 0.02 mg/L,叶片可不经历愈伤组织直接形成丛生芽,最适生根培养基为1/2 MS＋NAA 0.05 mg/L＋IBA 0.01 mg/L＋活性炭0.6 g/L.利用发根农杆菌介导法,建立Gt5GT7基因的银中杨叶片遗传转化体系,叶片GUS染色结果初步证明,Gt5GT7基因成功整合入银中杨叶片,且最适宜侵染的菌液A600值为0.45～0.6.
Bibliography:44-1267/S
Taking young branches of Populus albaxP, berolinensis as plant material, effects of pre-cultures, cytokinin/ auxin ratio on in vitro regeneration were investigated. Results showed that pre-cultured in MS liquid medium for 3 d was the optimal method. The most suitable bud induction medium was MS＋TDZ 0.1 mg/L＋NAA 0.02 mg/L, in which multiple buds could directly form. The optimal rooting medium was 1/2 MS＋NAA 0.05 mg/L＋IBA 0.01 mg/L＋ active carbon 0.6 g/L. Moreover, Gt5GT7 gene was transformed into regenerative leaves mediated by Agrobacterium rhizogenesis. GUS staining illustrated that transformation system was successfully established. Moreover, the optimal A600 was 0.45-0.6.
Populus alba × P. berolinensis; pre-cuhure; cytokinin/auxin ratio; in vitro regeneration; GtSGTT; genetic transformation
XU Hong-wei, GAO Feng, YANG Wei-xin, WANG Jing, ZHOU Xiao-fu （Key Laboratory of Jilin Province for Plant Resources Science and Green Production, Jilin Normal University, Siping 136000, China）
ISSN:1004-874X