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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

Southern blot analysis of OsARG gene copy number in the genomes of transgenic cotton plants.The probe was labeled with the radioactive isotope [α-32P] dCTP. (A) Two copies of OsARG were integrated into the ARG-26 genome. (B) One copy of OsARG was integrated into the ARG-38 genome. The under labels indicate the following samples: M, marker. 1, positive control. 2, negative control. 3, genomic DNA digested using EcoRI. 4, genomic DNA digested using HindIII.
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pone.0141530.g003: Southern blot analysis of OsARG gene copy number in the genomes of transgenic cotton plants.The probe was labeled with the radioactive isotope [α-32P] dCTP. (A) Two copies of OsARG were integrated into the ARG-26 genome. (B) One copy of OsARG was integrated into the ARG-38 genome. The under labels indicate the following samples: M, marker. 1, positive control. 2, negative control. 3, genomic DNA digested using EcoRI. 4, genomic DNA digested using HindIII.

Mentions: Two transgenic cotton plants were obtained by Agrobacterium-mediated genetic transformation; they each exhibited normal growth characteristics, designated as ARG-26 and ARG-38. The offspring of these plants were screened using PCR amplification of the OSARG gene in kanamycin resistant cotton plants; the size of the PCR product for the positive plants was shown as Fig 2. Southern blot analysis was used to investigate the number of OsARG copies integrated into the genomes of the T1generation cotton plants. Genomic DNA from transformed cotton plants was digested using EcoRI and HindIII. The probe length was 966 bp No EcoRI or HindIII restriction sites were present in the probe region. The hybridization results indicated that there were two strong bands detected in the EcoRI and HindIII lanes of the ARG-26 genomic DNA (Fig 3A).One band was detected in the EcoRI and HindIII lanes of the ARG-38 genomic DNA (Fig 3B). These results suggested that two copies of the OsARG gene were integrated into the ARG-26 genome, and that one copy was integrated into the ARG-38 genome. T3 generation homozygous offspring originating from ARG-26 and ARG-38 were bred by self-pollination.


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)

Southern blot analysis of OsARG gene copy number in the genomes of transgenic cotton plants.The probe was labeled with the radioactive isotope [α-32P] dCTP. (A) Two copies of OsARG were integrated into the ARG-26 genome. (B) One copy of OsARG was integrated into the ARG-38 genome. The under labels indicate the following samples: M, marker. 1, positive control. 2, negative control. 3, genomic DNA digested using EcoRI. 4, genomic DNA digested using HindIII.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0141530.g003: Southern blot analysis of OsARG gene copy number in the genomes of transgenic cotton plants.The probe was labeled with the radioactive isotope [α-32P] dCTP. (A) Two copies of OsARG were integrated into the ARG-26 genome. (B) One copy of OsARG was integrated into the ARG-38 genome. The under labels indicate the following samples: M, marker. 1, positive control. 2, negative control. 3, genomic DNA digested using EcoRI. 4, genomic DNA digested using HindIII.
Mentions: Two transgenic cotton plants were obtained by Agrobacterium-mediated genetic transformation; they each exhibited normal growth characteristics, designated as ARG-26 and ARG-38. The offspring of these plants were screened using PCR amplification of the OSARG gene in kanamycin resistant cotton plants; the size of the PCR product for the positive plants was shown as Fig 2. Southern blot analysis was used to investigate the number of OsARG copies integrated into the genomes of the T1generation cotton plants. Genomic DNA from transformed cotton plants was digested using EcoRI and HindIII. The probe length was 966 bp No EcoRI or HindIII restriction sites were present in the probe region. The hybridization results indicated that there were two strong bands detected in the EcoRI and HindIII lanes of the ARG-26 genomic DNA (Fig 3A).One band was detected in the EcoRI and HindIII lanes of the ARG-38 genomic DNA (Fig 3B). These results suggested that two copies of the OsARG gene were integrated into the ARG-26 genome, and that one copy was integrated into the ARG-38 genome. T3 generation homozygous offspring originating from ARG-26 and ARG-38 were bred by self-pollination.

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