银中杨;预培养;激素配比;离体再生;Gt5GT7;遗传转化.银中杨叶片离体再生体系的建立及Gt5GT7基因的遗传转化[J].广东农业科学,2014,41(13):144-146 |
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银中杨叶片离体再生体系的建立及Gt5GT7基因的遗传转化 |
Leaf in vitro regeneration and Gt5GT7 genetictransformation in Populus alba伊P. berolinensis |
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DOI: |
中文关键词: 银中杨 预培养 激素配比 离体再生 Gt5GT7 遗传转化 |
英文关键词: Populus alba×P. berolinensis pre-culture cytokinin/auxin ratio in vitro regeneration Gt5GT7 genetic transformation |
基金项目:国家自然科学基金(31070224) |
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中文摘要: |
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1/2MS+NAA0.05mg/L+IBA0.01mg/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。 |
英文摘要: |
Taking young branches of Populus alba伊P. 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. |
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