Establishment Of Regeneration System Of Perennial Ryegrass,Creeping Bentgrass And Agrobacterium Tumefaciens-mediated Transformation With Cry8C

Perennial ryegvass(Lolium perenne L. ) and creeping bentgrass(Agrostis stolonifera L.) are one of the major cool-season turfgrass widely utilized at home and abroad. Improving insect-resistant by plant genetic engineering is of momentous significance to control the destroy of the underground pest, reduce the usage of pesticide, protecting environments, save the maintenance of turfgrass. In this dissertation, perennial ryegrass and creeping bentgrass regeneration system have been eatablished and Agrobacterium-mediated transformation factors were systematically studied.and obtained trandgenic plants by PCR assay. The main results are as follows:(1) The mature seeds of Cutter , Imagine (Lolium perenne L.) was as explant, when the medium contain 9 mg/L 2,4-D, the best callus induction rate of Cutter was produced up to 41.3%; when the medium contain 7 mg/L 2,4-D, the best callus induction of Imagine up to 33.3%; the embryogenic calli induction rate of Cutter up to 24.7% and Imagine up to 18.7% when the medium contain 7 mg/L 2,4-D. It is useful to facilitate the callus induction rate and improve the callus quality when supplementation of TDZ; When the hypocotyls of perennial ryegrass as explant, the embryogenic calli induction ratio was low than mature seeds as explant. It could improve the embryogenic calli induction ratio when supplementation of Proline and casein hydrolysate. The mature seeds of Southshore (Agrostis stolonifera L.) as explant, callus induction rate of Southshore up to 68.1 % when supplementation of 6mg/L 2,4-D and 0.2 mg/L TDZ; embryogenic calli induction ratio up to 45.6% supplementation with 5mg/L 2,4-D and 0.1 mg/L TDZ.After the embryogenic callus was subculture for two months ,the highest differentiation rate of Cutter up to 31.7% with combination of 0.3mg/L TDZ and 0.7mg/L 6-BA; the highest differentiation rate of Imagine up to 26 % with combination of 0.3mg/L TDZ,0.5mg/L 6-BA,and the highest differentiation rate of Southshore up to 52.7% with combination 0.3mg/L TDZ and 0.5mg/L 6-BA.After the embryogenic calli was subculture for five months, the differentiation rate of Cutter drop to 20.3%, Imagine drop to Imagine up to 12.3%,and Southshore drop to 41.2...