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Studies on Regeneration System and Agrobacterium-mediated Transformation of Carnation

Author: SuXiaoZuo
Tutor: LiMingYang
School: Southwestern University
Course: Floriculture
Keywords: Carnation A . tumefaciens Gus gene genetic transformation
CLC: S682.19
Type: Master's thesis
Year: 2007
Downloads: 162
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Abstract


As one of the major contributors and a commercial leader to the cut-flower market,carnation (Dianthus caryophyllus L.) is an important target for the breeding of new cultivars with novel characteristics.The aim of carnation breeing are not only the character of view and admire but also the character of horticulture.The Septori asp. Caused by alternaria is very rifeness at now,So breeing of fastness to disease become the aim of carnation.So it is very significance to solve problem during breeing by using genetic engineering technology.This experiment based on regeneration system of carnation and using Gus enzyme activity test,studies on agrobacterium-mediated transformation of carnation by LBA4404 to establish A.tumefaciences-mediated transformation system of carnation.The differentiation medium of carnation、containing AS whether or not、the time of pre-cultivation、consistency of the A.tumefaceins cells and the time of dip、the time of co-cultivation and consistency of Km,all of these affect to transformation.The result was positive by Gus enzyme activity te:st,which the foreign gene had been integrated in carnation.Regeneration systems of carnation. The regeneration system from leavies of carnation was established by using different cytokinins combining with other phytohormones. The results showed that the kinds of cytokinin effected the differentiation rate of adventitious buds from leaf.NAA (0.1 mg/L)and TDZ(1.0 mg/L) in relatively low concentration can induce differentiation of adventitious buds from leaf. So the best differentiation medium was: MS+1.0mg/LTDZ+0.1mg/LNAA;The best proliferation medium was: MS+0.5mg/LBA+0.05mg/LNAA.Transformation system of carnation.It was studied that several factors affected genetic transformation of carnation by LBA4404.The optimized condition was that: leaf explants were pre-cultured on MS+1.0mg/LTDZ+0.1mg/LNAA for 2 days;the co-cultivation medium was MS+0.1mg/LNAA+1.0mg/LTDZ+100um/LAS,3days;selection differentiation medium was: MS+0.1mg/LNAA+1.0mg/LTDZ+60mg/LKm+400mg/Lcef;selection and proliferation medium was 1/2MS+0.1mg/LNAA +25mg/LKm.Selection of transgenic plants. Regenerated shoots were inoculated into selection and proliferation medium for rooting selection,and average rootage frequency was 10.7%.

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CLC: > Agricultural Sciences > Gardening > Ornamental Horticulture ( flowers and ornamental trees) > Perennial Flowers > Perennial Flowers Class > Other
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