Study On Culture And Regeneration Of Zoysia Japonica Steud.and Its Genetic Transformation

Zoysiagrass(Zoysia japonica stued.) commonly is used as warm-season turfgrass. Because of its great advantages in strong adaptability, drought-resistance, extension ability, wear-tolerance elasticity and et al., it is widely used in the following fields: turf planting, ecologica restoration, water & soil conservation, and so on.. However, the green period is relatively short and endure cold isn't strong. The intergration of conventional and modern breeding technique, especially modern biology technique, will be able to decrease the period of breeding greatly. Application of biotechnology in turfgrass breeding improved agronomic characteristic is a developing research. The conditions of establishing the high regeneration system of Zoysiagrass were studied, the genes of IPT and CBF1 were transformed into the calli using microprojectile bombardment and Agrobacterium tumefaciens transformation.Resulted in the production of transgenic plants with improved tolerance to cold and extended green period, which offered a new germplasm to breed new Zoysiagrass with improving the cold tolerance. The obtained transgenic plants can provide the new material resource for Zoysiagrass genetically improvement.The results were as follows: 1.The feasible medium for callus induction from mature was improved MS + 2, 4-D (3.0mg/L)+VB1(4.0mg/L), the induction frequency was up to 43.20%. 2.The feasible medium for callus of subculture:MS+2, 4-D(2mg/L)+6-BA(0.2mg/L)+ABA(0.2mg/L),Induction frequency of Embryo genie callus 28.4%; 3.The differentiation medium employed:MS+KT(2.0mg/L)+1.0(mg/L)+NAA(0.1 mg/L), the differentiation ratio was about 56%. 4. The rooting medium was 1/2 MS+ NAA(0.1mg/L): Rooting ratio was 100% 5.The transgenic plants were obtained by microprojectile bombardment andAgrobacterium tumefaciens of callus induced from mature . 6.The results of PCR analysis displayed that the exogenous CBF1 gene had integrated intothe genome of transgenic Zoysaigrass.7. The assay of RT-PCR showed the transgenic CBF1 had expressed at transcript level. 8.The transgenic frequencies of CBF1 gene were 0.26%...