Establishment Of Regeneration And Genetictransformation By Agrobacterium Tumefaciens With Gly Gene Of Perennial Ryegrass And Zoysia

Perennial ryegrass(Lolium perenne L.) and Zoysia grass(Zoysia japonica Steud.) are important forage and turf grasses,and are representatives of cool season and warm season grass as well.The two species were used as materials to study plant regeneration system in diverse cultivars of perennial ryegrassand zoysia grass genetictransferring system of the two plant species by Agrobacterium tumefaciens with Gly gene.The results are as follows:1. The regeneration system of perennial ryegrass and zoysia grass have been estabilished.The three varieties as lvbaoshi ,bainimu,xuanfeng were selected out from eleven ones of perennial ryegrassand were used as acceptors for transformation.MS was used as basic medium aiding with 0.05mg/l 6-BA and 5 mg/l 2,4-D to induce embryogenic callus and subculture as well.Meanwhile,in the selecting of sugar and arga,sucrose and phytagel are the best compared with others by infected zoysia callus,the induced rate is high.At the same time,1 mg/lNAA,1 mg/l KT and 3 mg/l 6-BA are preferable concentration of plants hormone during in differentiation medium.In addition,80 days are appropriate time for getting higher differentiation frequency.2.Genetic transformation has been established.By Agrobacterium tumefaciens mediated,genetic transformation is optimized,using the Agrobacterium strain LBA4404:Embryogenic callus subcultured 80 days,which precultivated 3 days ;Immersed in the bacterial suspension for 15 minute and co-cultivated on medium for 2 days;300 mg/l Cef appending 0.1 mol/l AS is better than Carb in controlling bacterium .3.Transgenic resistant GLY callus are detected by GUS detecting.