| In the present studies, the chitinase and j3-l,3-glucanase genes were transformed by the Agrobacteriuni lutnefac lens LBA4404 in which the plasmid pCG II is the binary vector for the two gene expression. Also, the optimum transformation system and the regeneration system of leaf discs and stem segments were studied. The two genes were cloned from the Nicotiana tabacum and could express extracellularly in plant. The results showed: In the optimized regeneration system, the callus induction media for the two kinds of explants were MS+NAAO.Smg/L+6-BA2.Omg/L and MS+NAA l.Omg/L+6-BA 2.Omg/L. The regeneration media for leaflet discs were MS+GA35.Omg/L+6-BA2.2rng/L and MS+GA35 .Omg/L+KT 2.Omg/L-i-6-BA 2.2mgIL. And the regeneration media for stein segment was MS+GA35.Omg/L+6-BA 2.2mg/L. The selective pressure during the callus induction and regeneration was kanamycin 100 mg/L. In the rooting phase it was was kanarnycin 50 mgIL. In the optimum regeneration system, the optimum bacterium concentration in value of 0D600 was 0.5. It had a best promoting effect on the transformation when the acetosyringone concentration was 50 ii mol/L. To leaflet discs, the optimal time of preculture, infection and co-culture were 3 days, l52O minutes and 4 days respectively. To stem segment, they were 2 days, 10S minutes and 4 days. 33 In the transformation of potato using chitinase and β-1,3-g1ucanase genes, transgenic plants were obtained from the callus derived from the leaflet discs and stem segments of an 11?cultivar. The transformation rate of leaflet discs and stem segments were 86.3% and 8 1.5% respectively, and the regeneration rates were 32.1% and 11.1% respectively. The opine assay showed that the regenerated plants had positive reaction, thus it could be proved that they are transformed plants. The primary detection of disease resistance with Fusariurn spp. And Rizoclonia solani Showed that transgenic plants were more resistant to those pathogenic fungi. |
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