Jube Sandro, Borthakur Dulal
Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, 1955 East-West Road, Honolulu 96822, HI, USA.
Plant Cell Tissue Organ Cult. 2009;96(3):325-333. doi: 10.1007/s11240-008-9490-x.
The tree-legume Leucaena leucocephala (leucaena) is used as a perennial fodder because of its fast-growing foliage, which is high in protein content. The use of leucaena as a fodder is however restricted due to the presence of the toxin mimosine. Improvements in the nutritional contents as well as other agronomic traits of leucaena can be accomplished through genetic transformation. The objective of this research was to develop a transformation protocol for leucaena using phosphinothricin resistance as the plant selectable marker. Explants obtained from immature zygotic embryos infected with the Agrobacterium tumefaciens strain C58C1 containing the binary plasmid pCAMBIA3201 produced four putative transformed leucaena plants. Transformation was con- firmed by PCR, RT-PCR, Southern blot, Western analyses, GUS-specific enzyme activity and herbicide leaf spraying assay. A transformation efficiency of 2% was established using this protocol.
豆科树木银合欢因其叶片生长迅速且蛋白质含量高，而被用作多年生饲料。然而，由于含有毒素含羞草素，银合欢作为饲料的使用受到限制。通过基因转化可以实现银合欢营养成分以及其他农艺性状的改良。本研究的目的是利用草丁膦抗性作为植物选择标记，开发一种银合欢的转化方案。从未成熟合子胚获得的外植体，用含有二元质粒pCAMBIA3201的根癌农杆菌菌株C58C1进行感染，产生了四株推定的转化银合欢植株。通过PCR、RT-PCR、Southern杂交、Western分析、GUS特异性酶活性和除草剂叶片喷洒试验对转化进行了确认。使用该方案确定的转化效率为2%。
