Abstract:

GAI protein belongs to the DELLA subfamily of the GRAS family, and is involved in plant growth processes such as development, photosynthesis and stress tolerance. In the early research of the authors’ laboratory, it was found that the GAI gene of sugar beet M14 line (BvM14-GAI) was up-regulated under salt stress. To investigate the functions of BvM14-GAI, BvM14 was selected as experimental material, and the full-length of BvM14-GAI gene was amplified by reverse transcription polymerase chain reaction (RT-PCR). In addition, the bioinformatics analysis and subcellular localization were conducted. The results showed that the full length cDNA of BvM14-GAI gene was obtained, and the results of bioinformatics analysis revealed that the full-length of BvM14-GAI gene was 1824 bp which encoded 607 amino acids with the predicted molecular mass of 66 kDa and the pI of 5.37. Multiple sequence alignment of BvM14-GAI and other GAI proteins showed that BvM14-GAI protein was highly homologous with the GAI proteins in spinach (Spinacia oleracea) and quinoa (Chenopodium quinoa). Subcellular localization prediction showed that BvM14-GAI protein was localized in the nucleus. Subsequently, the BvM14-GAI gene was ligated with the pCAMIA2300-eYFP vector, and the recombinant plasmids were transformed into Agrobacterium EHA105 and injected into tobacco. Finally the results exhibited that the BvM14-GAI protein was localized in the nucleus. In conclusion, the full-length cDNA of the BvM14-GAI gene is successfully obtained in this experiment, and subcellular location analysis confirms that the BvM14-GAI is localized in the nucleus. This work lays an important foundation for studying the function of BvM14-GAI gene.

Key words: sugar beet M14 line, BvM14-GAI gene, bioinformatics analysis, subcellular localization