Comparison on the Regeneration System and Genetic Transformation Condition in Four Ground Cover Chrysanthemum Cultivars

doi:10.11924/j.issn.1000-6850.2013-1863

Abstract

Abstract: In order to establish the regeneration system and definite the transformation condition of the four ground cover Chrysanthemum cultivars, ‘Shenyun’, ‘Fanxingfen’, ‘Dushiliren’ and ‘Fenmeigui’, the leaves of the aseptic seedlings were used as explants to analyze the effect of plant growth regulator hormone combination on regeneration buds induced from callus, and then the genetic transformation condition of these four Chrysanthemum morifolium was compared through the mediation of Agrobacterium tumefaciens. This study will lay the research foundation for cultivating new Chrysanthemum morifolium using the transgenic means. The result indicated that the optimum condition for regeneration buds inducing on MS medium was NAA 1.0 mg/L+ BA 1.0 mg/L, NAA 0.3 mg/L+ BA 1.0 mg/L, NAA 1.5 mg/L+ BA 0.5 mg/L and NAA 0.5 mg/L+ BA 1.5 mg/L for ‘Shenyun’, ‘Fanxingfen’, ‘Dushiliren’ and ‘Fenmeigui’ respectively. The genetic transformation conditions of the leaves of aseptic seedlings were pre-cultured for 24 hours, infected with A. tumefaciens for 10 minutes which OD600 was 0.4, 0.5, 0.3 and 0.4 respectively, co-cultivated for 48 hours, later-cultivated for 2 days, the mannose concentration was 10 g/L and the cefsulodin concentration was 300 mg/L. The target bands of genes could be obtained in some resistant plants of these four Chrysanthemum morifolium by PCR detection, which indicated that the target gene was integrated into their genomes.

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