Induction Study Of Alfalfa Regeneration System By GSTP1-L105V And CYP2A6Gene Transformation

With the extreme expansion of extensive manufacturing,heavy metals andorganic compounds pollution in soil are becoming prominent problems.As the dailyfood safety and health of people are closely related to metals and organic matterpollution in soil,properly handling the pollution problem can be a very importantthing.Glutathione S transferase gene GSTP1-l105V and cytochrome anti oxidase geneCYP2A6were confirmed to be related with heavy metals and organiccompounds.Although Wild Alfalfa embraces heavy metal accumulation and organicmatter decomposition in a certain role, but the effect is not enough to be used fortreatment of soil pollution.This paper aims to insert the two genes above into Alfalfawith agrobacterium mediated method to improve its resistance to heavy metal andcapability for organic.Firstly, we construct the expression vector pBI121-GSTP1-l105V containing onesingle gene in the study.After putting the expression vector into agrobacteriumtumefaciens,then CYP2A6gene was transferred into agrobacterium by freeze thawingmethod.Take the golden empress alfalfa hypocotyls as receptor,after pre-incubation,impregnation,coculture,the first bacteria removal,the second bacterialremoval,subculture,differentiation culture and rooting culture,we obtained100transgenic plants with heavy metals and organic compounds resistance eventually.By PCR and RT-PCR detection and tolerance test on CdSO4and TCE oftransgenic plants,we confirm that the exogenous gene of GSTP1-l105V and CYP2A6were successfully integrated into the genome of Alfalfa on a total of27strains andtolerance to heavy metals and organic matter of transgenic plants increasedsignificantly.