Isolation And Functional Analysis Of GsRNF12Gene In Wild Soybean

Wild soybean has great resistance and cultivated soybean origins from wild soybean. Wild soybean is animportant germplasm resources for cultivated soybean breeding. Therefore, utilization of wild soybeanresources is very important value for cultivated soybean breeding improvement.By using RT-PCR technique, GsRNF12gene was isolated from Wild soybean. The results of sequencefunctionally annotated by Blast in NCBI indicate ORFof GsRNF12is723bp encoding240amino acids,including C3HC4ring domain. The amino acid sequence alignment and phylogenetic tree analysis indicatedthat GsRNF12was related most closely to the GmRNF12in soybean, implying that they may be diversifiedfrom a common ancestor during evolution. Original nuclear expression vector pET-28a-GsRNF12wasconstructed. The results of SDS-PAGE showed that GsRNF12gene was successfully expressed in engineeringbacteria BL21.Real-time PCR technique was applied to clarify the express pattens response to various abiotic stressesabout GsRNF12. GsRNF12expression was induced by drought, salt, low-temperature, SA and ABA.GsRNF12gene had no tissue-specific expression characteristics which expressed in roots, stems, leaves.GsRNF12gene was higher in root than in other tissues.The plant expression vectors for GsRNF12gene was construced and transfornled into Arabidopsis byAgrobacterium-mediated transformation. Identification of low tempeture, drought, high salt resistanceindicated that the T3generation of transgenic Arabidopsis plants showed increasing resistance to lowtemperaturem, drought, and high salt. The primarily results showed that the transgenic plants can enhance thetolerance to low temperature, drought and sal than wild-type.