Isolation And Characterization Of StERF Transcription Factor Genes From Potato (Solanum Tuberosum L.)

Transcription factor, it can bind with the eukaryotic gene promoter regions of cis element, and with other related protein to activate or inhibit transcription or to participate in regulation of gene response the external environment factors. AP2/ERF transcription factors are a group of large family of transcription factors, in plant. Play a role in plant growth, cell differentiation, flower and seed development, plant hormone response and biological and abiotic stress response. ERF transcription factor bind the target sequence with GCC- box cis element to regulate the expression of related genes. In this paper, The potato(Solanum tuberosum L.) cultivar ‘Zihuabai’(ZHB) was used for all experiments. According the gene sequence to design specific primers, Using RT-PCR acquired StERF1-4 �'� Pti4 genes. Obtain the following results:1. According to the NCBI registed sequence data(StERF1,GenBank accession JN125860、StERF2,GenBank accession JN711505、StERF3,Gen Bank accession JN125857 、StERF4, GenBank accession JN125859 、 Pti4, GenBank accession EU851735) design a pair of oligonucleotide primers, Use RT-PCR method acquire StERF1-4 and Pti4 genes. Bioinformatics analysis results showed that the 5 St ERFs are typical of the ERF transcription factors.2. Use RT-PCR and qRT-PCR methods StERF1/2/4/ and Pti4 have very little expression in the tuber, StERF3 don’t expressed. StERF4 expressed more in the leaves, flowers and roots, StERF1 expressed higher in flower, and StERF3 not expressed. StERFs response to drought, high salt, ET, SA and MeJA, the 5 kinds of stress treatment, there are different levels of response. Pti4 of which can be more strongly affected by high salt stress induced expression, StERF3 can strongly induced by drought stress.3. Construction yeast expression vectors pBD-StERF1/2/3/4 and pBD-Pti4, respectively. pBD-St ERFs transformed yeast Y187, is used to validate StERFs transcription activation, the results indicate that: 5 St ERF proteins have transcriptional activation activity in the yeast.4. Construction yeast expression vectors p AD-StERF1/2/3/4 and p AD-Pti4, respectively. And Construction yeast one-hybrid report vectors pGCC-His2.1 and pmGCC-His2.1. The p AD-StERFs vectors were transformed into the pGCC-His2 or pmGCC-His2.1 yeast strain, the results indicate that: St ERFs proteins can bind with GCC-box, not with mGCC-box, so its involved in plant growth regulation and stress response regulation by combining the GCC-box.