Cloning And Functional Analysis Of Drought Stress Related Genes From Alfalfa

This experiment used Medicago sativa L.CV.Gongnong NO.3 crepping-rooted alfalfa as experiment materials.After the greenhouse pot culture and man-made drought stress,analyzed the gene expression differences clips between alfalfa under the drought stress and normal supply of water by mRNA differential display.Then sequenced after the Reverse Northern Dot Blotting,as well as analyzed with the results as follows:1,Because the alfalfa has the characteristic which the cuttage easy to survive,the asexual cuttage alfalfa's survival rate may achieve 90%in the greenhouse and in the moisture content and temperature control suitable situation.When particularly stress processing requires a long time or the experimental plant life cycle quite short situation,the negative different clips may effectively reduce which creates as a result of the growing stage changes by using pot culture and the vegetative reproduction to analyze gene expression rather than take the identical plant as an experiment material.2,By DDRT-PCR,the result showed that some special genes were expressed in leaves of alfalfa under drought stress.For half a month with wilting-recovery stress,Then extract RNA from alfalfa and by mRNA differential display technology,we has been selected 32 clips alfalfa differentially expressed genes.3,The establishment of differentially expressed genes in alfalfa research system.Theoretically,by 3 combining Oligo d(t) primers and eight random primers,which can amplified all of the mRNA.This experiment used three Oligo d(t) primers and 10 random primer selected the real and fault different clips by DDRT-PCR Reverse Northern blotting.4,Through DDRT-PCR and Reverse Northern blotting technologies,we selected three positive alfalfa differentially expressed gene fragment affected by drought stress.After cloned and gene sequenced,contrast to the Internet gene database,one is an important drought resistant gene which had significant identity to Arabidopsis thaliana gene encoding water channel proteins,and the other two gene fragments had no similarity to known genes.May be there are new drought resistant genes and need further study.