Transgenic Research Of Wheat And Plant Regeneration Of Two Desert Plants

Immature embryos of wheat Xinong88, Baofeng7228, Xinongl376, 80101 and Ganmai were used as materials. After precultured for 3-4 days, rice chitinase gene was transferred into scutellum by microprojectile bombardment. Two green plants were finally regenerated from 863 immature embryos bombarded with plasmid pYAO24 containing rice chitinase gene (RC24) and selective marker gene (npt II) on the medium added with 50mg/L kanamycin. Four green plants were finally regenerated from 388 immature embryos bombarded with plasmid pARN6 containing rice chitinase gene (RC24) and pD containing reporter gene (gus) as well as selective marker gene (bar) on the medium supplemented with 2mg/L Basta. These six green plants were identified with PCR and Southern blotting analysis. Plant height, ear length, spikelet numbers, effective tillers, grain numbers per plant and TKW of T0 transgenic plants were examined. Resistance of T1 transgenic lines and CK to rust fungi in the field was identified. The results are as follows:(1) By means of PCR and Southern blotting analysis, seven plants from seven regenerated plants were identified to be transgenic lines containing the target genes. The transformation frequency was 0.40%. There was distinct difference among the 5 cultivars of wheat. Transformation frequency of Baofeng7228 reached 1.04%. Transformation frequency of Xinong88 was 0.50%. No transgenic plant was obtained from Xinong1 376, 80101 and Ganmai.(2) Spikelet numbers, effective tillers, grain numbers per plant and TKW of TO plants, especially TKW decreased in comparison with CK. However, plant height and ear length were not observed in obvious differences.(3) There was no distinct difference resistant to rust fungi between Tl transgenic plants bombarded with plasmid pYAO24 and CK. But the resistance to rust fungi in Tl transgenic plants bombarded with plasmid pARN6 and pD was stronger than that in CK.By means of pollen tube pathway, plasmid pYAO24 was transferred into four cultivars of wheat Xinong88, Baofeng7228, Xinongl 376 and Xinong8727.Kanamycin-resistant screening and PCR analysis of genetic transformed plants showed that three of green plants were PCR positive with the positive rate of 0.16%. There was distinct difference among the four wheat cultivars. Transformation frequency of Baofeng7228 reached 0.41%. Transformation frequency of Xinong88 was 0.19%. No transgenic plant was obtained from Xinongl376 and Xinong8727.