The Construction Of An Efficient Tissue Culture System For Paspalum Atratum & The Clone And Construction Of AtGolS2 Drought-resistance Related Gene And PVKH-35S-AtGolS2 Expressing Vector

An efficient system of tissue culture and plant regeneration is essential for genetic transformation. In this research, we used the seedling segment,leaf base,mature embryo and immature male inflorescences of Paspalua atratum cv as the candidate explants .Calli of Paspalua atratum cv were induced on the modified MS medium (MSM), which MS salt + 9.9mg/L vitaminB1+ 9.5mg/L vitamin B6+ 4.5 mg/L nicotinic acid+ 1mg/L hydrolyze casine+ 30g/L sucrose+ 8g/L agar as basic components with different concentration 2.4-D. 2.0mg/L 2.4-D,pH 5.8 was able to induce callus formation from all the four explants in dark conditions for 20 days. The differentiation of calli was obtained on the modified MS medium (MSM) with 6 mg/L 6-BA. The root induction was on the MSM with 0.5 mg/L NAA. Rooted plantlets could be grown well in sand after one week seedling adaptation.In this research, we used mature embryo as explants to construct the systems of tissue culture and regeneration for Paspalua atratum cv. Number of factors affected the system were studied also. The results were shown as below:(1) Mature embryo of Paspalua atratum cv was used as explant , modified MSM consisted of MS salt with 3% agar, pH 5.8. Callus induction on the MSM with 2.0mg/L 2.4-D in dark condition for 20 days and callus regeneration on the MS medium with 2.0mg/L 2.4-D+ 0.1mg/L KT.The differentiation of the calli was on the MSM medium with 6 mg/L 6-BA , 25±1℃,16h light/8h dark, the differentiation ratio of calli reached 65%. The root induction was on MSM+0.5 mg/L NAA, 25±1℃,16h light/8h dark. The 95% seedlings were survive on tiny sand. The whole period of...