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Overexpression of a GmCnx1 gene enhanced activity of nitrate reductase and aldehyde oxidase, and boosted mosaic virus resistance in soybean.

Zhou Z, He H, Ma L, Yu X, Mi Q, Pang J, Tang G, Liu B - PLoS ONE (2015)

Bottom Line: 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).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.

View Article: PubMed Central - PubMed

Affiliation: Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China.

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.

No MeSH data available.


Related in: MedlinePlus

NR (upper) and AO (bottom) activities in leaf of both WT and T1 GmCnx1transgenic soybean plants.Data were calculated by significance analysis method, and the significant differences were compared with the control. WT, negative control, non-transgenic plants. L1, L2, L3, L4, L7, L10, L17, L18, L22 and L26 represent the GmCnx1 transgenic line numbers. Values are the mean ± SD (n = 3). Error bars represent standard errors of means from three replicates. The same letter at each bar means no significant difference at P< 0.05.
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pone.0124273.g004: NR (upper) and AO (bottom) activities in leaf of both WT and T1 GmCnx1transgenic soybean plants.Data were calculated by significance analysis method, and the significant differences were compared with the control. WT, negative control, non-transgenic plants. L1, L2, L3, L4, L7, L10, L17, L18, L22 and L26 represent the GmCnx1 transgenic line numbers. Values are the mean ± SD (n = 3). Error bars represent standard errors of means from three replicates. The same letter at each bar means no significant difference at P< 0.05.

Mentions: As NR and AO activity was causally linked with the biosynthesis of Moco, they were measured in leaves of WT and T1 transgenic plants. The results showed that, NR and AO activities increased in GmCnx1 transgenic plants compared with WT (Fig 4). In particular, for NR activity, the GmCnx1 transgenic soybean plants produced high level (4.25 μg g-1 h-1) in line 10, which were 2.6-fold higher than the non-transgenic plants (1.62 μg g-1 h-1), and for AO activity, the level in the leaf was 3.9-fold higher in GmCnx1 transgenic soybean plants (30.00 pmol L-1) in line 10 compared with the non-transgenic plants (7.71 pmol L-1). That was consistent with the expression of GmCnx1 gene in leaf and root. Taken together, it can be concluded that the differentGmCnx1 levels among the different transgenic lines correspond to similar differences at NR and AO activities in leaf.


Overexpression of a GmCnx1 gene enhanced activity of nitrate reductase and aldehyde oxidase, and boosted mosaic virus resistance in soybean.

Zhou Z, He H, Ma L, Yu X, Mi Q, Pang J, Tang G, Liu B - PLoS ONE (2015)

NR (upper) and AO (bottom) activities in leaf of both WT and T1 GmCnx1transgenic soybean plants.Data were calculated by significance analysis method, and the significant differences were compared with the control. WT, negative control, non-transgenic plants. L1, L2, L3, L4, L7, L10, L17, L18, L22 and L26 represent the GmCnx1 transgenic line numbers. Values are the mean ± SD (n = 3). Error bars represent standard errors of means from three replicates. The same letter at each bar means no significant difference at P< 0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401665&req=5

pone.0124273.g004: NR (upper) and AO (bottom) activities in leaf of both WT and T1 GmCnx1transgenic soybean plants.Data were calculated by significance analysis method, and the significant differences were compared with the control. WT, negative control, non-transgenic plants. L1, L2, L3, L4, L7, L10, L17, L18, L22 and L26 represent the GmCnx1 transgenic line numbers. Values are the mean ± SD (n = 3). Error bars represent standard errors of means from three replicates. The same letter at each bar means no significant difference at P< 0.05.
Mentions: As NR and AO activity was causally linked with the biosynthesis of Moco, they were measured in leaves of WT and T1 transgenic plants. The results showed that, NR and AO activities increased in GmCnx1 transgenic plants compared with WT (Fig 4). In particular, for NR activity, the GmCnx1 transgenic soybean plants produced high level (4.25 μg g-1 h-1) in line 10, which were 2.6-fold higher than the non-transgenic plants (1.62 μg g-1 h-1), and for AO activity, the level in the leaf was 3.9-fold higher in GmCnx1 transgenic soybean plants (30.00 pmol L-1) in line 10 compared with the non-transgenic plants (7.71 pmol L-1). That was consistent with the expression of GmCnx1 gene in leaf and root. Taken together, it can be concluded that the differentGmCnx1 levels among the different transgenic lines correspond to similar differences at NR and AO activities in leaf.

Bottom Line: 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).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.

View Article: PubMed Central - PubMed

Affiliation: Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China.

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.

No MeSH data available.


Related in: MedlinePlus