Study On The Tissue Culture And Genetic Transformation With Hypocotyl Of Sunflower

The hypocotyl of sunflower with different genotypes were used as materials in this research. Effects of genotype, the culture medium, the type and concentration of endocrine on tissue culture and regeneration were alse investigated, Using Agrobacterium tumefaciens, the affecting factors of transformation of sunflower were also tested with hypocotyl of different genotype sunflower as receptor.The regeneration frequency of induced bud with hypocotyl in different culture medium was further researched by taking different genotypes sunflower as material.The result showed that, shoot regeneration were better achieved from the medium of MS + 0.03mg/LIAA +1.2mg/L6-BA. The shoot s could grow up well from the medium of 1/2MS+1.0mg/LIBA without addition of hormone.The hypocotyl of three different genotypes of sunflower(NC208, HA300 and Nuoza212) were transformed with Agrobracterium tumefaciens EHA101 carrying the target Psy gene. Meanwhile the factors affecting transformation was studied. Transgenic sunflower's plants were examined by PCR and PCR-Southern analysis. The results as follows:The fit concentration of Hyg is 10mg/L. The time of co-culture in 3 days might be suitable. 10 minutes is suitable for infection. The fit range of OD₆₀₀ is 0.6. PCR and PCR-Southern analysis showed that the target Psy gene integrated into the genome of sunflower.