Studies On Callus Induction And The Establishment Of Agrobacterium Mediated Genetic Transformation System Of Hosta Ventricosa

Hosta ventricosa,also known as calyx purple, purple host. it is Liliaceae host perennial perennial root herb, H. purple flowers and leaves not only have the ornamental value, also have the very strong adaptability and management culture is more easily, So in recent years at home and abroad, it has become a ubiquitous groundcover plants, It is the green to be bestowed favor on newly. In addition, Hosta ventricosa also has certain edible and medicinal value, color is only limited to white flowers and purple flowers. and there are defects such as the lack of aroma、low propagation coefficient、long cycle and so on. People have used the traditional cross breeding of Hosta ventricosa new varieties. But because of long breeding period and species limits many traits can be difficult to obtain through traditional breeding way. For example, it can only change plantaginea leaf traits. and the color of Hosta didn’t develop into other color except white and purple,genetically modified(gm) technology can be very good to overcome these shortcomings for the improvement and modification flower traits provide a shortcut. Now reports mostly are the study of Host tissue culture propagation, But genetically modified by Hosta as explant method research has not been reported. This study selects the Hosta ventricosa as test material, and through the tissue culture technology and agrobacterium mediated method to do the research of the callus induction、proliferation and genetic transformation,and get the required for different explant callus induction and proliferation of different hormone types and appropriate concentration.By various aspects to explore such as different receptors, concentration of agrobacterium infection, infection time, acetyl clove ketone(AS) concentration, and infection way this study selected the optimum transformation conditions, set up the genetic transformation system, for genetic engineering breeding of the directional breeding new varieties of hosta lay a good foundation.1.Different explant callus induction and appreciationLeaves, petioles, pedicel, axillary bud as explants for callus induction, pedicel is the best explant callus induction, the highest callus induction rate is 91%. the best medium is MS+6-BA2.0+NAA0.3(unit mg/L), Callus value-added best culture medium is MS+6-BA3.0+NAA0.4+2,4-D0.2(unit mg/L),the increment rate of up to 95%.2.The study of factors affecting genetic transformation, optimization of genetically modified(gm) system.With callus, peduncle, germ as transformation receptor research influence factors of Agrobacterium mediated genetic transformation Hosta ventricosa in adding the infection way and the way of soaking infect drops. Results show that the transformation receptor material is best callus in add mode of infection, the best concentration of Acetyl clove ketone(AS) is150μmol/L, the best bacteria liquid concentration is OD600=0.6;The best transformation receptor is callus in the soaking infection way, the best time to infect is 10 min, the best concentration of Acetyl clove ketone(AS) is 100μmol/L, the best bacteria liquid concentration is OD600=0.4. The conversion rate comparison of Two types of infection: Add bacteria drops way is the best way of infection.3.Tests of GUS staining and PCR analysis of the resistance of plants prove that carrier has been successfully integrated into the Hosta ventricosa genome.