Cloning and Expression Analysis of Gibberellin Receptor Gene DoGID1A in Dioscorea opposita
LONG Wen-Hong1, MENG Jin-Gui1, XU Sheng-Sheng1, *, ZHANG Xue-Mei2, DUAN Yan-Bi3, YANG Rong-Ping1
1 College of Horticulture and Landscape, Yunnan Agricultural University, Kunming 650201, China;
2 College of Agronomy and Biotechnology, Yunnan Agricultural University, Kunming 650201, China;
3 Experiment Teaching Center for Agriculture Specialty, Yunnan Agricultural University, Kunming 650201, China
Abstract：Gibberellin receptor GID1 (gibberellin insentive dwarf 1), one of important elements in gibberellin (GA) signal transduction pathway, directly affects the effect of gibberellin on plant. A cDNA sequence encoding gibberellin receptor GID1 was cloned from epidermis of bulbils of 'Niuwei Yam' (Dioscorea opposita) with reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) techniques. The expression of the cloned yam GID1 gene at different stages after bulbils were harvested and the response of the gene expression to paclobutrazole were tested by qRT-PCR. The results showed that the obtained gene had a full length of 1 123 bp with 996 bp opening reading fram encoding 331 amino acids, and named DoGID1A (GenBank No.MK268684). Bioinformation analysis showed that the molecular weight and theoretic isoelectric piont of the protein was 3.72×104 D and 8.25, respectively. The DoGID1A protein was unstable protein with an abhydrolase domain (93~304 aa) but without transmembrance domain or signal peptide. Homologous alignment showed that DoGID1A protein shared 73.56% sequence similarity with 13 specieces including Musa acuminata, and shared conserved many domains including motifs linking hormone GA and protein DELLA. Phylogenetic tree showed all dicotyledons were cluster in a group, monocotyledonous plants except rice (Oryza sativa) and yam shared another group. Yam was clustered out alone, so was rice. qRT-PCR analysis showed the expression of DoGID1A gene increased during yam bulbil sprouting and was enhanced by paclobutrazol treatment. These results indicated that DoGID1A gene could be associated with yam bulbil sprouting. This study provides basic information for further research on expression pattern and function of this gene.
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