一种快速有效的741杨离体叶片再生芽方法

在对杂交品种741杨(Populus alba(P.davidiana×P.simonii)×P.tomentosa)进行农杆菌介导法转基因的试验中发现了一种快速有效的叶片再生芽的方法.首先叶片外植体在培养基Ⅰ(MS培养基添加0.5 mg/L BA和1.0 mg/L 2,4-D)上培养2～3 d,再转移到培养基SH(MSmedium containing 2.0 mg/L of BA and 0.1 mg/L of NAA)上培养10 d,然后再转移到培养基Ⅱ(MSmedium with 0.5 mg/L of BA)上,培养大约5 d之后86.7%的叶片外植体产生的芽,每片叶片外植体(1 cm×1 cm)可产生40～50个芽.但是,如果叶片外植体在培养基Ⅰ上培养的时间长于5 d,再依次转移到培养基SH和Ⅱ上,则叶片会产生大量根.

A new efficient method for rapid in-vitro shoot regeneration on leaf explants of Poplar 741

In the experiments of Agrobacterium tumefaciens-mediated transformation of the hybrid poplar 741, Populus alba ( P. davidiana × P. simonii) × P. tomentosa, we developed a rapid and efficient method of shoot regeneration from the leaf explants in-vitro on Murashige and Skoog (MS) medium. The leaf explant segments was first cultured on medium Ⅰ ( MS medium supplemented with 0.5 mg/L of BA and 1.0 mg/L of 2,4-D ) for 2 ～ 3 days and then transferred to medium SH ( MS medium containing 2.0mg/L of BA and 0. 1 mg/L of NAA) to culture for 10 days. 86.7% of leaf segments produced shoots after the leaf segments were transferred from medium SH to medium Ⅱ (MS medium with 0.5 mg/L of BA) for 1 week. The average shooting number was 40 ～ 50 per 1 cm × 1 cm of leaf segment. However, if the leaf explants incubated on medium Ⅰ longer than 5 days, they produced more roots instead of shoots.