Cloning of galactinol synthase gene from Ammopiptanthus mongolicus and its expression in transgenic Photinia serrulata plants

• Published in: Gene Vol. 513; no. 1; pp. 118 - 127
• Main Authors: Song, Jian, Liu, Jing, Weng, Manli, Huang, Yanyan, Luo, Lei, Cao, Pengxiu, Sun, Haiwei, Liu, Jie, Zhao, Jinhong, Feng, Dianqi, Wang, Bin
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15-01-2013
• Subjects: abscisic acid; Abscisic Acid - metabolism; AMG gene; Amino Acid Sequence; amino acids; Ammopiptanthus mongolicus; Base Sequence; Cloning, Molecular; cold; cold stress; Cold Temperature; cold tolerance; Cold-tolerance gene of tree; DNA; Exons; Fabaceae - genetics; Galactinol synthase gene of Ammopiptanthus mongolicus (AmGS); Galactosyltransferases - biosynthesis; Galactosyltransferases - genetics; Gene expression; gene expression regulation; Gene Expression Regulation, Plant - genetics; Gene Expression Regulation, Plant - physiology; genes; genetic transformation; genomics; inositol 3-alpha-galactosyltransferase; Introns; Molecular Sequence Data; Nucleotide sequence; Nucleotides; Open Reading Frames; Photinia; Photinia - genetics; Photinia - metabolism; Plants, Genetically Modified - genetics; Plants, Genetically Modified - metabolism; Promoters; Stop codon; Stress; Transformation; Transgenic ligneous plant; Transgenic plants; Trees; LT50; ORF; Galactinol synthase gene of Ammopiptanthus mongolicus (AmGS); Cold-tolerance gene of tree; GS; TAIL-PCR; REC; AFLP; RACE; TDF; EB; EC; Transgenic ligneous plant
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2012.10.058

A cold induced galactinol synthase gene (AmGS) and its promoter sequence were identified and cloned from the cold-tolerant tree Ammopiptanthus mongolicus by using cDNA-AFLP, RACE-PCR and TAIL-PCR strategies combined with its expression pattern analysis after cold inducing treatment. Accession number of the AmGS gene in GenBank is DQ519361. The open reading frame (ORF) region of the AmGS gene is 987 nucleotides encoding for 328 amino acid residues and a stop codon. The genomic DNA sequence of AmGS gene contains 3 exons and 2 introns. Moreover, a variety of temporal gene expression patterns of AmGS was detected, which revealed the up-regulation of AmGS gene in stresses of cold, ABA and others. Then the AmGS gene was transformed into Photinia serrulata tree by Agrobacterium-mediated transformation, and the transgenic plants exhibited higher cold-tolerance comparing with non-transformed plants.
► We clone the novel galactinol synthase gene (AmGS) in Ammopiptanthus Mongolicus. ► The promoter sequence of AmGS gene possesses cold and ABA responsive element. ► AmGS highly express induced by cold, exogenous ABA treatment and other tolerance. ► Transgenic Photinia serrulata plants overexpressing AmGS enhance cold tolerance. ► The increasing relative electrical conductivity in transgenic plants was lower.