Analysis On Different Gene Expression And Molecular Mechanism For Creeping-rooted Type Medicago Varia

Alfalfa is the highest value of legume forages for cultivation and use in the world. The creeping-rootedness of alfalfa has important significance to improving the ability of resistance and nurture new cultivar.In this paper we use the creeping-rooted alfalfa "BL-101" for the experimental material and construct the Transcriptomic of library creeping-rooted(CR) and noncreeping-rooted(NCR) separately. Analyzing the library transcriptome with next generation sequences(RNA-seq). Reasearching the expression the differential gene through cDNA-SRAP technology in order to search the molecular mechanism of creeping-rootedness of alfalfa and finding the relevant excellent gene. The results as follow:1. Separate the RNA from CR and NCR root tissue using Trizol method and construct the library. Through the analysis of the library which sequenced with RNA-seq technology that we obtain 15978 differently expressed genes totally. The highest multiples differentially expressed genes is GA3 related proteins, so we determine that the happening of creeping-rootedness induced by GA3. According to the FDR methord we get 960 up-regulated unigenes, and 997 down-regulated unigenes. The result of GO analyse is we can classify the Unigene from three aspect, there are 237 kinds of gene in molecular function,114 kinds of gene in cellular component and 584 kinds of gene in biological process.2. Find 5 Unigenes are related with the root development from the biological process, two of the five genes is up regulated unigene and making sequence alignment through NCBI, we draw a conclusion that one gene is RAV subtribe of AP2/EREBP transcription factor family and the other is transferrin receptor protein.3. Using cDNA-SRAP technology screen the differently expressed genes, and spraying GA3 outside as induction.we choose 15 primers for PCR augmentation get 134 strips,76 of them are diversity.42 strips are recyled which are stable repeatability and difference to sequencing. Sequence Alignment show that 9 genes are unknown genes,26 are functional genes.the most of these functional genes are zinc finger protein thioredoxin h7,MYB transcription factor,AP-2 complex subunit beta-1,xyloglucan galactosyltransferase.4.we determine that the occurrence of creeping-rooted is induced by GA, through signal transduction makes the RAV transcription factor was expressed, protein kinases and enzymes participate participate in root metabolism, acetyl-CoA and pyruvate dehydrogenase participate in root respiration, than the root node developed. Golgi membrane secreted the signal substances, and ion exchange proteins on plasma membrane controled the material exchange, WRKY transcription factors and ubiquitin ligases promote the performance of creeping-rooted organization.