The Expression Of Populus Euphratica Large Fragment Gene Into Arabidopsis And Salt Tolerance Analysis About Transgenic Strains

Mediated with Agrobacterium tumefaciens,Arabidopsis thaliana were taken as the receptor plant and transferred large gene fragments of PopuLus euphratica to it. The main resuLts of the study were as follows:1.At present,The activation experiment of 1884 agrobacterium tumefaciens was carried out, including 1570 agrobacterium activation was successful.The value of OD600 was between 0.8-1, the activation rate was 83.33%.2.1570 BIBAC clones of P. euphratica were transferred in arabidopsis thaliana,sntibooti cs plate screening transgenic T1 generation,and 590 from these clones got the Arabidopsi s thaliana transformants by screening on antibiotic containing plate.3.Then sowing 590 strains of T2 generation, gathering seedling leaves and using Tissue Lyser which combined with method of CTAB to extract DNA,and using micro uLtraviolet spectrophotometer to test its purity.then statistics strains whose ratio of A260/280 (excluding the pollution impact on DNA purity.such as phenolics. RNA. and carbohydrat, peptides,) that between 1.8-2.0 was totally 521.4.Take the NPTII gene as a primer to PCR identification for target band amplification, target band is 540 bp, a total of 403 positive strains were plants.5.Sowing the seed of T2 generation in medium with kar, part of the resuLting transformation totally 119 homozygous for screening experiment, isolated from T2 homozygous 49, the proportion is 41.18%.6.49 strains was obtained by separation of salt tolerance at the sieve experiment (150 mM) NaCl concentration, received salt resistance performance of 11 strains; Further validation tests confirmed (NaCl concentration 150 mM and 175 mM),2 strains obtained the stable salt resistant performance.7.The salt resistance of 11 strains in NaCl aqueous solution treatment when sowing,the RWC about 3 strains attained to 90%,when the concentration of NaCl was 50,100,150 mM; Medium showed that the resistance to salt strain largest salt concentration is 150 mM.8.There were 2 mutants obtained which phenotype changed in the gradient of salt processing.This research mainly to collect egina popuLus euphratica resources to salt tolerance with BIBCA library technology.we made the large fragment gene of Euphrates poplar expressed in arabidopsis thaliana and using its advantages to screening,identiflcation the function of popuLus euphratica gene, It is of great significanc to analysis Euphrates poplar’s salt-tolerant mechanism and mining salt-tolerant genetic resourcese. This is not only enriched the contents of the p. euphratica salt tolerance studies,but also provided the evidence the study about other plant salt resistance.