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

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。

Abstract: 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.