Construction Of The Expression Vector With VP Gene And Analysis Of The Transformation

Wheat(Triticum aestivum Linn)is China’s important crop,and is one of the world’s earliest and most widely planded grain crops.It belongs to gramineae,monocotyledon,angiosperms.The caryopsis of wheat contains starch,fat,proteins,vitamins and enzymes,which has a very high nutritional value and medicinal value.However,the wheat planting has been affected by the drought and soil salinization,which hinders the process of agricultural modernization in our country.According to statistics,ten percent of the available land area in our country has different level of salinization,and our country has more than four hundred million acres of saline land and salinization of soil,accounting for twenty-five percent of the total arable land in the national.At the same time,the water resources of our country is poor.Especially in recent decades,with the deterioration of ecological environment as well as the decrease of cultivated land,the shortage of water resource has become the biggest obstacle to agricultural production.In view of more and more serious drought,saline and alkaline land and lacking of high quality of resources in wheat breeding,this experiment intended to design primers according to H~+-PPase genes of arabidopsis,rice,barley,corn,and amplificated in wheat,and at last acquired the Wheat tonoplast’s H~+-PPase gene by using the methods of molecular biology.Then this experiment designed primers from pCAMBIA 1380 vector to acquair CAMV35S gene,and designed primers from pEGFPN1 to acquair GFP gene.And then the experiment constructed the expression vector after cnstructing the three cloning vectors.The experiment transformed the entire pCAMBIA 1380-35S-TVP-GFP vector into arabidopsis by agrobacterium conversion,and screened the homozygous strain.Through the germination experiment,the statistics of green seedlings,the proline determination of seedlings,the proline determination of plants and the determination of the conductivity this experiment can analysis the salt tolerance and the drought resistant of arabidopsis.So that this experiment can determine the relationship between H~+-PPase gene and the resistance of plants,also lay a foundation for cultivating high quality new strains of wheat.