发根农杆菌介导的杨梅遗传转化研究初报

为了建立发根农杆菌Agrobacterium rhizogenes介导的杨梅Myrica rubra遗传转化体系,利用携带拟南芥Arabidopsis thaliana LEAFYcDNA片段的发根农杆菌R15834感染东魁杨梅M. rubra‘Dongkui’和荸荠杨梅M. rubra‘Boji’的子叶、叶片和茎段。结果表明,只从东魁杨梅子叶上诱导出毛状根,而且产生毛状根的东魁杨梅子叶数在10%以下。杨梅子叶产生毛状根受多种因素影响,新种子比旧种子容易产生毛状根,在1/2 MS(Murashige and Skoog)培养基培养较在MS培养基上培养好。东魁杨梅子叶上的毛状根经聚合酶链反应(polymerase chain reaction,PCR)检测和Southern杂交检测,证实已将拟南芥的LEAFYcDNA片段成功导入并整合到东魁杨梅子叶中。图2参10

Genetic transformation of Myrica rubra mediated by Agrobacterium rhizogenes

Myrica rubra(China bayberry)is a particular fruit tree to China.It is also a ideal tree species of afforestation.The purpose is to establish a genetic transformation system of Myrica rubra mediated with Agrobacterium rhizogenes.Cotyledons,leaves,and stem segments of M.rubra 'Dongkui'and M.rubra 'Boji' were infected with A.rhizogenes R15834 containing a pDC202 vector carriing the LEAFY gene,which controls floral meristem,cultured in 1/2 Murashige and Skoog(MS) or MS medium.Results showed that only less than 10%of the cotyledons of M.rubra'Dongkui'induced hairy roots.Hairy roots were produced from new seeds of M.rubra'Dongkui'more easily than old seeds,and the cotyledons grew hairy roots on the 1/2 MS media more readily than the MS media.By PCR and Southern hybridization test,it was proved that the LEAFY gene of Arabidopsis thaliana was successfully transferred and integrated into a nuclear genome through cotyledons of M.rubra 'Dongkui'.Ch,2 fig.10 ref.]