AGROBACTERIUM-MEDIATED TRANSFORMATION OF WHITE CLOVER USING DIRECT SHOOT ORGANOGENESIS

被引:50
作者
VOISEY, CR [1 ]
WHITE, DWR [1 ]
DUDAS, B [1 ]
APPLEBY, RD [1 ]
EALING, PM [1 ]
SCOTT, AG [1 ]
机构
[1] AGRESEARCH,GRASSLANDS RES CTR,PLANT MOLEC GENET LAB,PRIVATE BAG 11008,PALMERSTON NORTH,NEW ZEALAND
关键词
TRIFOLIUM-REPENS L; AGROBACTERIUM-TUMEFACIENS; TRANSFORMATION; DIRECT REGENERATION;
D O I
10.1007/BF00232627
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
White clover (Trifolium repens L.) plants from the cultivars Grasslands Huia and Grasslands Tahora have been transformed using Agrobacterium-mediated T-DNA transfer. Transgenic plants regenerated directly from cells of the cotyledonary axil. To transform white clover, shoot tips from 3 day old seedlings were co-cultivated with A. tumefaciens strain LBA4404 carrying the plasmid vector pPE64. This vector contains the neomycin phosphotransferase II gene (nptII) and beta-glucuronidase reporter gene (gus) both under the control of the CaMV 35S promoter. Kanamycin-resistant plants regenerated within 42 days after transfer onto selective media. Integration of the nptII and gus genes into the white clover genome was confirmed using Southern blotting, and histochemical analysis indicated that the gus gene was expressed in a variety of tissues. In reciprocal crosses between a primary transformant and a non-transformed plant the introduced gus gene segregated as a single dominant Mendelian trait.
引用
收藏
页码:309 / 314
页数:6
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