title: Overexpression of a GmCnx1 gene enhanced activity of nitrate reductase and aldehyde oxidase, and boosted mosaic virus resistance in soybean.
abstract: Molybdenum cofactor (Moco) is required for the activities of Moco-dependant enzymes. Cofactor for nitrate reductase and xanthine dehydrogenase (Cnx1) is known to be involved in the biosynthesis of Moco in plants. In this work, a soybean (Glycine max L.) Cnx1 gene (GmCnx1) was transferred into soybean using Agrobacterium tumefaciens-mediated transformation method. Twenty seven positive transgenic soybean plants were identified by coating leaves with phosphinothricin, bar protein quick dip stick and PCR analysis. Moreover, Southern blot analysis was carried out to confirm the insertion of GmCnx1 gene. Furthermore, expression of GmCnx1 gene in leaf and root of all transgenic lines increased 1.04-2.12 and 1.55-3.89 folds, respectively, as compared to wild type with GmCnx1 gene and in line 10 , 22 showing the highest expression. The activities of Moco-related enzymes viz nitrate reductase (NR) and aldehydeoxidase (AO) of T1 generation plants revealed that the best line among the GmCnx1 transgenic plants accumulated 4.25 μg g(-1) h(-1) and 30 pmol L(-1), respectively (approximately 2.6-fold and 3.9-fold higher than non-transgenic control plants).In addition, overexpression ofGmCnx1boosted the resistance to various strains of soybean mosaic virus (SMV). DAS-ELISA analysis further revealed that infection rate of GmCnx1 transgenic plants were generally lower than those of non-transgenic plants among two different virus strains tested. Taken together, this study showed that overexpression of a GmCnx1 gene enhanced NR and AO activities and SMV resistance, suggesting its important role in soybean genetic improvement.
authors.affiliation: Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: Molecular Epigenetices of the Ministry of Education (MOE), Northeast Normal University, Changchun, China.
authors.forename: Hongli
authors.initials: H
authors.lastname: He
authors.affiliation: Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China.
authors.forename: Luping
authors.initials: L
authors.lastname: Ma
authors.affiliation: Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China.
authors.forename: Xiaoqian
authors.initials: X
authors.lastname: Yu
authors.affiliation: Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Mi
authors.affiliation: Molecular Epigenetices of the Ministry of Education (MOE), Northeast Normal University, Changchun, China.
authors.forename: Jingsong
authors.initials: J
authors.lastname: Pang
authors.affiliation: Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China.
authors.forename: Guixiang
authors.initials: G
authors.lastname: Tang
authors.affiliation: Molecular Epigenetices of the Ministry of Education (MOE), Northeast Normal University, Changchun, China.
authors.forename: Bao
authors.initials: B
authors.lastname: Liu
mesh_terms: D016960:Agrobacterium tumefaciens; D042931:Aldehyde Oxidase / Q000378:metabolism*; D003067:Coenzymes / Q000096:biosynthesis; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D060467:Disease Resistance; D005822:Genetic Vectors; D008667:Metalloproteins / Q000096:biosynthesis; D008969:Molecular Sequence Data; D000090006:Molybdenum Cofactors; D009029:Mosaic Viruses / Q000502:physiology*; D050901:Nitrate Reductase / Q000378:metabolism*; D010802:Phylogeny; D010935:Plant Diseases / Q000235:genetics* / Q000821:virology; D018515:Plant Leaves / Q000378:metabolism / Q000821:virology; D010940:Plant Proteins / Q000235:genetics / Q000502:physiology*; D018517:Plant Roots / Q000378:metabolism; D030821:Plants, Genetically Modified; D017434:Protein Structure, Tertiary; D011621:Pteridines; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D030262:Soybean Proteins / Q000235:genetics / Q000502:physiology*; D013025:Glycine max / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D015854:Up-Regulation; D014968:Xanthine Dehydrogenase / Q000235:genetics / Q000502:physiology*
chemical_list: D042931:Aldehyde Oxidase; D003067:Coenzymes; D018076:DNA, Complementary; D018744:DNA, Plant; D008667:Metalloproteins; D000090006:Molybdenum Cofactors; D050901:Nitrate Reductase; D010940:Plant Proteins; D011621:Pteridines; D011993:Recombinant Fusion Proteins; D030262:Soybean Proteins; D014968:Xanthine Dehydrogenase; C022261:molybdopterin