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Expression of the Rice Arginase Gene OsARG in Cotton Influences the Morphology and Nitrogen Transition of Seedlings.

Meng Z, Meng Z, Zhang R, Liang C, Wan J, Wang Y, Zhai H, Guo S - PLoS ONE (2015)

Bottom Line: Field experiments indicated that the polyamine and nitrogen content increased by more than two-fold in the T3 generation plants of the transgenic cotton lines ARG-26-2, ARG-26-7, ARG-38-8, and ARG-38-11, as compared with the control plants.After harvesting cotton fibers grown in field conditions, we analyzed the quality of fiber and found that the fiber length was increased in the transgenic lines.Our results indicate that the OsARG gene could potentially be used to improve cotton fiber length traits.

View Article: PubMed Central - PubMed

Affiliation: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.

ABSTRACT
Arginase is the only enzyme capable of producing urea in plants. This enzyme also contributes to many important biological functions during plant growth and development, such as seed development, root development and plant nitrogen using. The unique rice arginase gene OsARG is known to affect nitrogen use efficiency and is also associated with higher yields in rice. In this study, we transformed OsARG into upland cotton R18 by Agrobacterium-mediated genetic transformation and analyzed the function of OsARG in transgenic cotton. Two independent OsARG expression transgenic cotton lines, ARG-26 and ARG-38, were obtained via transformation. Southern blot analysis indicated that two copies and one copy of the OsARG gene were integrated into the ARG-26 and ARG-38 genomes, respectively. Enzyme activity and RNA transcription analysis revealed that the OsARG gene is highly expressed in cotton. The nitric oxide content and the morphology of ARG-26 and ARG-38 seedlings were both affected by expression of the OsARG gene. Field experiments indicated that the polyamine and nitrogen content increased by more than two-fold in the T3 generation plants of the transgenic cotton lines ARG-26-2, ARG-26-7, ARG-38-8, and ARG-38-11, as compared with the control plants. After harvesting cotton fibers grown in field conditions, we analyzed the quality of fiber and found that the fiber length was increased in the transgenic lines. The average cotton fiber length for all of the transgenic cotton lines was two millimeters longer than the fibers of the control plants; the average cotton fiber lengths were 31.94 mm, 32.00 mm, 32.68 mm and 32.84 mm in the ARG-26ARG-26-2, ARG-26-7, ARG-38-8 and ARG-38-11 lines, respectively, but the average fiber length of the control plants was 29.36mm. Our results indicate that the OsARG gene could potentially be used to improve cotton fiber length traits.

No MeSH data available.


Related in: MedlinePlus

Map of the plant expression vector.
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pone.0141530.g001: Map of the plant expression vector.

Mentions: Rice arginase has a transit peptide and is known to be localized to mitochondria. Given the existence of certain differences in subcellular localization mechanisms between monocots and dicots, we hypothesized that the rice arginase gene could be expressed in cotton and targeted into mitochondria if a rice arginase construct replacing its native transit peptide with a dicot mitochondrial transit peptide was introduced into cotton transgenically. A new ORF was constructed that replaced the transit peptide nucleotide sequence (60bp, 20AA) of the OsARG CDS (HM369061, 1023bp, 340AA) with the mitochondrion transit peptide (Mtp) nucleotide sequence (63 bp, 21AA) of the Arabidopsis thaliana aldehyde dehydrogenase gene (AtALDH, ALDH2B7).The new OsARG ORF was inserted into the PUC19 plasmid using the PstI and XhoI sites. Promoter and terminator sequences were inserted into PUC19 OsARG via enzyme digestion. The CaMv35S promoter was digested using PstI and HindIII, and the NOS terminator was digested using PstI and EcoRI. The transformed plasmid pBIMtpOsARG was constructed by inserting CaMv35S::OsARG:: NOS into pBI121 using HindIII and EcoRI (Fig 1).


Expression of the Rice Arginase Gene OsARG in Cotton Influences the Morphology and Nitrogen Transition of Seedlings.

Meng Z, Meng Z, Zhang R, Liang C, Wan J, Wang Y, Zhai H, Guo S - PLoS ONE (2015)

Map of the plant expression vector.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4631492&req=5

pone.0141530.g001: Map of the plant expression vector.
Mentions: Rice arginase has a transit peptide and is known to be localized to mitochondria. Given the existence of certain differences in subcellular localization mechanisms between monocots and dicots, we hypothesized that the rice arginase gene could be expressed in cotton and targeted into mitochondria if a rice arginase construct replacing its native transit peptide with a dicot mitochondrial transit peptide was introduced into cotton transgenically. A new ORF was constructed that replaced the transit peptide nucleotide sequence (60bp, 20AA) of the OsARG CDS (HM369061, 1023bp, 340AA) with the mitochondrion transit peptide (Mtp) nucleotide sequence (63 bp, 21AA) of the Arabidopsis thaliana aldehyde dehydrogenase gene (AtALDH, ALDH2B7).The new OsARG ORF was inserted into the PUC19 plasmid using the PstI and XhoI sites. Promoter and terminator sequences were inserted into PUC19 OsARG via enzyme digestion. The CaMv35S promoter was digested using PstI and HindIII, and the NOS terminator was digested using PstI and EcoRI. The transformed plasmid pBIMtpOsARG was constructed by inserting CaMv35S::OsARG:: NOS into pBI121 using HindIII and EcoRI (Fig 1).

Bottom Line: Field experiments indicated that the polyamine and nitrogen content increased by more than two-fold in the T3 generation plants of the transgenic cotton lines ARG-26-2, ARG-26-7, ARG-38-8, and ARG-38-11, as compared with the control plants.After harvesting cotton fibers grown in field conditions, we analyzed the quality of fiber and found that the fiber length was increased in the transgenic lines.Our results indicate that the OsARG gene could potentially be used to improve cotton fiber length traits.

View Article: PubMed Central - PubMed

Affiliation: Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, China.

ABSTRACT
Arginase is the only enzyme capable of producing urea in plants. This enzyme also contributes to many important biological functions during plant growth and development, such as seed development, root development and plant nitrogen using. The unique rice arginase gene OsARG is known to affect nitrogen use efficiency and is also associated with higher yields in rice. In this study, we transformed OsARG into upland cotton R18 by Agrobacterium-mediated genetic transformation and analyzed the function of OsARG in transgenic cotton. Two independent OsARG expression transgenic cotton lines, ARG-26 and ARG-38, were obtained via transformation. Southern blot analysis indicated that two copies and one copy of the OsARG gene were integrated into the ARG-26 and ARG-38 genomes, respectively. Enzyme activity and RNA transcription analysis revealed that the OsARG gene is highly expressed in cotton. The nitric oxide content and the morphology of ARG-26 and ARG-38 seedlings were both affected by expression of the OsARG gene. Field experiments indicated that the polyamine and nitrogen content increased by more than two-fold in the T3 generation plants of the transgenic cotton lines ARG-26-2, ARG-26-7, ARG-38-8, and ARG-38-11, as compared with the control plants. After harvesting cotton fibers grown in field conditions, we analyzed the quality of fiber and found that the fiber length was increased in the transgenic lines. The average cotton fiber length for all of the transgenic cotton lines was two millimeters longer than the fibers of the control plants; the average cotton fiber lengths were 31.94 mm, 32.00 mm, 32.68 mm and 32.84 mm in the ARG-26ARG-26-2, ARG-26-7, ARG-38-8 and ARG-38-11 lines, respectively, but the average fiber length of the control plants was 29.36mm. Our results indicate that the OsARG gene could potentially be used to improve cotton fiber length traits.

No MeSH data available.


Related in: MedlinePlus