Transformation Of Gene Of GmDREB3and BYCV-GPV-CP Into Immature Embryos Of Wheat By Particle Bombardment

Wheat transgenic study began in the early1990s,gene gun transformation is the mainmethod that get transgenic wheat seedings.However,wheat gene gun transformation researchis rather backward, transformation frequency is low,those phenomenon exists generally,thetransformation system also needs further perfection. So it is necessary to study transformationprocess of factors to improve the wheat transformation frequency and accelerate theapplication of transgenic wheat and industrialization process.GmDREB3is a transcriptionfactor gene isolated from soybeans,product of this gene control resistance gene throughinteraction with DRE.BYDV-GPV-CP gene improve the crops resistance through produce coatprotein in plants. To obtain the best optimum conditions of wheat genetic transformationsystem, study on the effect of genotypes of wheat,Induction time of immature embryocallus,dosage Of gold dust per gun,and culture medium of screening and differentiation onregeneration plant rate. The main research results are as follows:1. Different genotypes of wheat on regeneration rate has remarkable effect.Five differentgenotypes of wheat of regeneration rate is significant differences. yang wheat18ofregeneration rate is the highest, achieve7.45%. The Second is yang wheat12, regenerationplant rate is6.25%. And xi nong979of regeneration plant rate is the lowest.2. wheat immature embryo callus by nine days induction and recover two weeks is the beststate. The callus of physiological activity and regeneration ability is higher.these callusrelatively easy to accept and integrate the exogenous gene.3. When dosage Of gold dust per gun is60μg,regenerative plants rate is higher. Too higerof dosage Of gold dust will make callus bear irreversible damage, these injured callus almostlose the regeneration ability.While low dosage Of gold dust will indirectly reduce dosage ofexogenous DNA dosage, thereby reducing gene gun transformation efficiency.4. Screening and differentiation culture medium of type on plantlet regeneration frequencyhas very great effect. At the same condition,useing screening and differentiation medium(1/2MS+NAA10mg/L+KT1.0mg/L+Bialaphos2mg/L) of differentiation frequency issignificantly higher than the other four culture mediums. Regeneration plant differentiationrate reached3.72%.The concentration of lowering the inorganic salt in1/2MS.Application of1/2MS.medium produced some better effect, make the regeneration plant rate improved alot.5. Adding PP333(4mg/L) in culture medium can promote the transgenic seedlings to plantand increase tillering. These regeneration seedlings is stonger.By contrast, addingPP333(2mg/L)of regeneration seedlings is slender and emaciated,these plants grow too fast totransplant successfully. And adding PP333(6mg/L) of regeneration seedlings is the moststrong, but plant growth is too slow.6.The results of specific PCR analysis of the transgenic seedlings show that GmDREB3genes and BYDV-GPV-CP genes are already integrated into wheat genomes.