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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 T 1  generation plants revealed that the best line among the  GmCnx1  transgenic plants accumulated 4.25 μg g -1  h -1  and30 pmol L -1 , respectively (approximately 2.6-fold and 3.9-fold higher than non-transgenic control plants).In addition, overexpression of GmCnx1 boosted 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.

Overexpression of a GmCnx1 Gene Enhanced Activity of Nitrate Reductase and Aldehyde Oxidase, and Boosted Mosaic Virus Resistance in Soybean