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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

Showing the relationship among GmCnx1 gene, molybdenum enzymes and SMV.Cnx1, Cnx2, Cnx3, Cnx6 and Cnx7 are involved in Moco biosynthesis. Solid arrows represent functions of GmCnx1 observed in this present work while dotted arrows represent future research prospective and open arrows show Moco-dependent enzymes that have been published in other research work [2,3].
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pone.0124273.g007: Showing the relationship among GmCnx1 gene, molybdenum enzymes and SMV.Cnx1, Cnx2, Cnx3, Cnx6 and Cnx7 are involved in Moco biosynthesis. Solid arrows represent functions of GmCnx1 observed in this present work while dotted arrows represent future research prospective and open arrows show Moco-dependent enzymes that have been published in other research work [2,3].

Mentions: SMV seriously affects yield and quality of soybean globally. Due to its wide distribution, great harmfulness, and difficulties in chemical control, cultivating and planting resistant varieties is an economical, safe and effective way to prevent SMV. In the present study we aimed at checking the relationship between GmCnx1 gene and its function in plants with regards to SMV resistance. Based on a microarray analysis of gene expression differences between two contrasting genotypes (SMV resistant and SMV susceptible) upon SMV inoculation, GmCnx1 was selected as a candidate SMV resistant gene [17]. We therefore deduced that the GmCnx1 gene expression in soybean is probably related to the positive regulation on the resistance to SMV. In this study, SMV-1 and SMV-7 were used to test for plant resistance to SMV. Consistent with the results from the microarray experiments, our results showed that transgenic plants indeed have higher SMV resistance compared with non-transgenic plants after the virus infection (Fig 6). We suspect that resistance to SMV might be due to the increase in NR and AO activities which are enhanced by the overexpression of GmCnx1 gene in soybean (Fig 7). In any case, this is the first report about the novel function of GmCnx1 against SMV, which has apparent breeding implications in soybean.


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)

Showing the relationship among GmCnx1 gene, molybdenum enzymes and SMV.Cnx1, Cnx2, Cnx3, Cnx6 and Cnx7 are involved in Moco biosynthesis. Solid arrows represent functions of GmCnx1 observed in this present work while dotted arrows represent future research prospective and open arrows show Moco-dependent enzymes that have been published in other research work [2,3].
© Copyright Policy
Related In: Results  -  Collection

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

pone.0124273.g007: Showing the relationship among GmCnx1 gene, molybdenum enzymes and SMV.Cnx1, Cnx2, Cnx3, Cnx6 and Cnx7 are involved in Moco biosynthesis. Solid arrows represent functions of GmCnx1 observed in this present work while dotted arrows represent future research prospective and open arrows show Moco-dependent enzymes that have been published in other research work [2,3].
Mentions: SMV seriously affects yield and quality of soybean globally. Due to its wide distribution, great harmfulness, and difficulties in chemical control, cultivating and planting resistant varieties is an economical, safe and effective way to prevent SMV. In the present study we aimed at checking the relationship between GmCnx1 gene and its function in plants with regards to SMV resistance. Based on a microarray analysis of gene expression differences between two contrasting genotypes (SMV resistant and SMV susceptible) upon SMV inoculation, GmCnx1 was selected as a candidate SMV resistant gene [17]. We therefore deduced that the GmCnx1 gene expression in soybean is probably related to the positive regulation on the resistance to SMV. In this study, SMV-1 and SMV-7 were used to test for plant resistance to SMV. Consistent with the results from the microarray experiments, our results showed that transgenic plants indeed have higher SMV resistance compared with non-transgenic plants after the virus infection (Fig 6). We suspect that resistance to SMV might be due to the increase in NR and AO activities which are enhanced by the overexpression of GmCnx1 gene in soybean (Fig 7). In any case, this is the first report about the novel function of GmCnx1 against SMV, which has apparent breeding implications in soybean.

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