Study On Cloning And Function Aanlysis Of ERF1Gene In Potato

Ethylene responsive factor (ERF) as AP2/EREBP transcription factor subfamily,is unique to the plant, and then participate in plant cell growth and development,hormone response, biological and non-biological adversity stress resistance, etc.through the AP2domain structure of ERF combined with cis-function componentsGCC-box/DRE/CRT of the downstream genes promoter region to regulatedownstream gene expression.Previously, the traits of rice, wheat, tomato and otherplants have been acquired comprehensive improvement by introducing ERF toregulate downstream genes, however the research of ERF gene is still relatively littlein potato. In this study, in-depth research of the structure and function of ERF1inpotato has been completed by cloning ERF1gene of potato, constructing ofexpression vector, analyzing of bioinformatics， drought resistance of transgenicplant analysis and downstream gene expression quantity, etc. Obtaining the followingresults:1. The length of573bp cDNA sequences of potato ERF1transcription factor hasbeen cloned by RT-PCR technique, which is100%homology to the publishedsequence (GenBank accession No. JN125860). It encode191amino acid residueswhich27-85is AP2conserved domain so belonging to the ERF type of EREBPsub-tribe of AP2/EREBP family. And encode non-hydrophobic protein which has nosignal peptide.2. The potato ERF1sense expression vectors that were drived by rd29A andCaMV35S promoters respectively were constructed and transformed into potatomediation by Agrobactium tumefacienses L. Obtained44transgenic lines included23lines overexpression of ERF1driven by CaMV35S,21lines overexpression of ERF1driven by rd29A. Quantitative real-time polymerase chain reaction (qRT-PCR)analysis indicated that the ERF1gene expressed in roots, stems and leaves of thetransgenic plants, and especially higher expression in leaves. Its downstream genesP5CS, CDPK, ZEP, SDR, PR2expression levels in transgenic plants was significantlyhigher than non-transgenic plants under drought stress.3. Under drought stress, the growth performance of the transgenic plants wasbetter than non transgenic plants in quantitative estimation of the height of plantlets,the roots length, the thickness of the stem and fresh weight of plantlets. In addition, Over-expression ERF1in Solanum tuberosum L., as results, sense transgenic potatoincrease the proline content, while decrease MDA content, to enhance tolerance todrought stress in the transgenic potato plants.In summary, overexpression of ERF1transgenic plants has improved droughtstress resistance to some extent, ERF1may positively regulate the expression of stress-related genes involved in the physiological regulation and then adjust the abioticstress responses in potato plants.