Transgenic Methods Of Chinese Cabbage:Agrobacterium-mediated Transformation By In Vitro Tissue Culture And In Planta Infiltration

Leaf disks hom five Chinese cabbage(Brascia camPestris L.ssppebnensis )were perfOnned tO eight sortS of plant hormone experimentS. The resultS revealed thatfOur genetyPes could acquire higher regeneration frequency except the No.3 typewhen 1mg/l 6-BA' 1mg/l NAA and 3mgh AgNO3were added to the MS medium.forng fOur genotypes the regeneration abi1ity had higher diversffication. PriInaryexpsrAnentS were carried out to stUdy the factors that could affect the regenerationability. The results showed that the genetype was the primary factor. Although addingAgNOa could Anprove the regeneration frequency signifcantly it was not the oulyfactOr. Because thc regeneration frequency was simi1ar when the density of AgNO3rWd from 0.5 to 9 mg/l. Also the site and the tAne of acquire explants did littleeffect on regrnendion.Explants from No.10, which had the highest regeneration frequency, were treatcdwith nopaline apobacterium C58 which carried gni gcne. By histochemicalidentdication, the best suitable treatment was that precultured the explants there days,co-cultured the exPlantS and agrobacterium fOr tWo or there days and adjusted the pHof the co-cultured medium to 5.2. Using the above treatheent procedure, TuMV-NIaand LMV-CP genes which could lead to higher resistance to TuMV virus wereintroduced into the Chinese cabbage after agrObacterium affected. PCR andPCR-Southem blotting revealed that the target gene had been integrated to thegenome of Chinese cabbage. Although the temPOrary exPression frequency was ashigh as 60% the transformation frequency at last was only 0.67% when the explantsapplied to the target agobacterium. Possible reason was given and the advice waspresented also.Floral cut sites and tbe whole floral of Brassica camPetris L.ssp. chinensiS wereaffected by agrobacterium in planta, and one herbicide resistant plant was aCquired byscreened thc Tl generation seeds which collected from 8is sortS treatheent. PCR andPCR-Southem blotting showed that the two target gene were also inttgrated tO thecabbage genome. The procedure became sforplifier by the omission of vacuumtreatment, but the efficiency of transfOnnation required twrovement.