title: Cloning and functional validation of molybdenum cofactor sulfurase gene from Ammopiptanthus nanus.
abstract: The molybdenum cofactor sulfurase gene ( AnMCSU ) was cloned from xerophytic desert plant Ammopiptanthus nanus and validated for its function of tolerance toward abiotic stresses by heterologous expression in Arabidopsis thaliana. Molybdenum cofactor sulfurase participates in catalyzing biosynthesis of abscisic acid, which plays a crucial role in the response of plants to abiotic stresses. In this study, we cloned molybdenum cofactor sulfurase gene (AnMCSU) from a super-xerophytic desert plant, Ammopiptanthus nanus, by using rapid amplification of cDNA ends method. This gene has a total length of 2544 bp, with a 5'- and a 3'-untranslated region of 167 and 88 bp, and an open reading frame of 2289 bp, which encodes an 84.85 kDa protein of 762 amino acids. The putative amino acid sequence shares high homology and conserved amino acid residues crucial for the function of molybdenum cofactor sulfurases with other leguminous species. The encoded protein of the AnMCSU gene was located in the cytoplasm by transient expression in Nicotiana benthamiana. The result of real-time quantitative PCR showed that the expression of the AnMCSU gene was induced by heat, dehydration, high salt stresses, and ABA induction, and inhibited by cold stress. The heterologous expression of the AnMCSU gene significantly enhanced the tolerance of Arabidopsis thaliana to high salt, cold, osmotic stresses, and abscisic acid induction. All these results suggest that the AnMCSU gene might play a crucial role in the adaptation of A. nanus to abiotic stress and has potential to be applied to transgenic improvement of commercial crops.
authors.affiliation: Maize Research Institute, Sichuan Agricultural University, Chengdu, 611130, Sichuan, People's Republic of China.
authors.forename: Hao Qiang
authors.initials: HQ
authors.lastname: Yu
authors.forename: Yuan Yuan
authors.initials: YY
authors.lastname: Zhang
authors.forename: Tai Ming
authors.initials: TM
authors.lastname: Yong
authors.forename: Yan Ping
authors.initials: YP
authors.lastname: Liu
authors.forename: Shu Feng
authors.initials: SF
authors.lastname: Zhou
authors.forename: Feng Ling
authors.initials: FL
authors.lastname: Fu
authors.forename: Wan Chen
authors.initials: WC
authors.lastname: Li
mesh_terms: D000040:Abscisic Acid / Q000378:metabolism / Q000494:pharmacology; D000222:Adaptation, Physiological / Q000187:drug effects / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003067:Coenzymes / Q000235:genetics / Q000378:metabolism*; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics; D007887:Fabaceae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics* / Q000502:physiology; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant; D018525:Germination / Q000187:drug effects; D006720:Homozygote; D008353:Mannitol / Q000494:pharmacology; D008667:Metalloproteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D000090006:Molybdenum Cofactors; D010641:Phenotype; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011392:Proline / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011621:Pteridines / Q000378:metabolism*; D015203:Reproducibility of Results; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012965:Sodium Chloride / Q000494:pharmacology; D013347:Subcellular Fractions / Q000187:drug effects / Q000201:enzymology; D013466:Sulfurtransferases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D000040:Abscisic Acid; D003067:Coenzymes; D018076:DNA, Complementary; D008353:Mannitol; D008667:Metalloproteins; D000090006:Molybdenum Cofactors; D010940:Plant Proteins; D011392:Proline; D011621:Pteridines; D012965:Sodium Chloride; D013466:Sulfurtransferases; C022261:molybdopterin