Opitimisition Of Tissue Culture System And Transformation Of Herbicide Genes To Elite Rice Varieties Of JiLin Province

The tissue culture system for elite rice (Japonica) varieties (Fengyou 20l andChaochan 2, high yield, good quality, but blast disease sensitive) cultivated in Jilinprovince was optimized. A herbicide gene was delivered into those elite varieties.Herbicide transgenic rice plants was obtained. Results are as follows f1 .The tissue cultural system of mature embryos of elite cultivated rice varietiesin Jilin province was optimized.l. l The callus induction and differentiation efficiency of different genotypes isquite different. The callus induction efficiency of Ji 89-48, Chaochan 2, Fengyou501, and Fengyou 307 could be as high as 86.4, 88.7, 80.8 and 8I.6 percentrespectively. And the differentiation efficiency of Ji 89-45, Chaochan 2 and Fengyou501 could be as high as 50. 1, 45.8 and 49.5 percent respectivelyl.2Reducing 2.4-D concentration to 1 rng/L, adding NAA (1 mg/L) and KT (0.5mgth) in the subculture medium can improve the growth of caIlus. And rnoreyellowish, grain-shape embryogenic ca1li was obtained.l .3Drying treatment of calli for l6h before differentiation could increase p1antregenera1ion efficiency by 20.6-37.2 percent.2.Parameters of Gene Gun (PDS-l000) transformation of the cal1us derived frommature el11bryo of elite rice varieties were studied by Gus gene transcient expression.2.l When the distance between DNA-gold pellet and the target cells was 9 cm,bombard for 2 times, the transformation efficiency was the highest. The Gus genetranscient expression efficiency was as high as 88.2 percent.2.2When the diStance was fixed, the Gus gene expression efficiency wasincreased with the increase of bombarded times. When the diStance was 9 cm, andbombarded for l, 2 and 3 times, Gus gene expression efficiency was 30.6, 88.3 and90.0 percent respectively.3. A Herbicide resistant gene (bar) has been delivered into rice. And herbicideresistant transgenic rice p1antS were obtained.Using the embryogenic caIli derived from mature embryo of elite rice varieties(high yield and early maturing) as the expIant, several transgenic rice plants wereobtained via Biolistic gene delivering method. bar gene had been inserted into ricelllNovember 2002 Quan Dongxinggenome demonstrated by PCR and Southern hybridization. Three transgenic rice plants (Do-1, D0-2 and D0-3) showed herbicide resistance. Gene silence was observed in one transgenic rice plant (D0-5).