Limits...
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

The infection rate of SMV-1 (upper) and SMV-7 (bottom) in both non-transgenic and GmCnx1 transgenic soybean plants by DAS-ELISA.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
getmorefigures.php?uid=PMC4401665&req=5

pone.0124273.g006: The infection rate of SMV-1 (upper) and SMV-7 (bottom) in both non-transgenic and GmCnx1 transgenic soybean plants by DAS-ELISA.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: After being infected by SMV-1 and SMV-7 mosaic virus strains, non-transgenic soybean plants showed dwarfing (Fig 5a), mosaic leaves, browning veins and crimple leaves (Fig 5b). In contrast, leaves of T1 transgenic soybean plants remained green, showing no mosaic, shrinkage leaves and other symptoms. Resistance analysis by DAS-ELISA (Fig 6) showed that infection rate of GmCnx1 transgenic plants were generally lower than those of non-transgenic plants. Specifically, for SMV-1, the infection rates of non-transgenic and10 lines of GmCnx1 transgenic plants were 87%, 47%, 40%, 33%, 53%, 33%, 33%, 47%, 40%, 33% and 47%, respectively. For SMV-7, the infection rates of non-transgenic and 10 lines of GmCnx1 transgenic plants were 93%, 40%, 53%, 47%, 40%, 33%, 40%, 47%, 33%, 33% and 40%, respectively. The results above indicated that the GmCnx1 transgenic soybean lines we generated showed higher degrees of resistance to two SMV strains tested.


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)

The infection rate of SMV-1 (upper) and SMV-7 (bottom) in both non-transgenic and GmCnx1 transgenic soybean plants by DAS-ELISA.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.g006: The infection rate of SMV-1 (upper) and SMV-7 (bottom) in both non-transgenic and GmCnx1 transgenic soybean plants by DAS-ELISA.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: After being infected by SMV-1 and SMV-7 mosaic virus strains, non-transgenic soybean plants showed dwarfing (Fig 5a), mosaic leaves, browning veins and crimple leaves (Fig 5b). In contrast, leaves of T1 transgenic soybean plants remained green, showing no mosaic, shrinkage leaves and other symptoms. Resistance analysis by DAS-ELISA (Fig 6) showed that infection rate of GmCnx1 transgenic plants were generally lower than those of non-transgenic plants. Specifically, for SMV-1, the infection rates of non-transgenic and10 lines of GmCnx1 transgenic plants were 87%, 47%, 40%, 33%, 53%, 33%, 33%, 47%, 40%, 33% and 47%, respectively. For SMV-7, the infection rates of non-transgenic and 10 lines of GmCnx1 transgenic plants were 93%, 40%, 53%, 47%, 40%, 33%, 40%, 47%, 33%, 33% and 40%, respectively. The results above indicated that the GmCnx1 transgenic soybean lines we generated showed higher degrees of resistance to two SMV strains tested.

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