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Salt tolerance and activity of antioxidative enzymes of transgenic finger millet overexpressing a vacuolar H+-pyrophosphatase gene (SbVPPase) from Sorghum bicolor

A vacuolar proton pyrophosphatase cDNA clone was isolated from Sorghum bicolor (SbVPPase) using end-to-end gene-specific primer amplification. It showed 80–90% homology at the nucleotide and 85–95% homology at the amino acid level with other VPPases. The gene was introduced into expression vector pC... Full description

Journal Title: Journal of plant physiology 2014-06-15, Vol.171 (10), p.789-798
Main Author: Anjaneyulu, Ediga
Other Authors: Reddy, Palle Surender , Sunita, Merla Srilakshmi , Kishor, Polavarapu B. Kavi , Meriga, Balaji
Format: Electronic Article Electronic Article
Language: English
Subjects:
Quelle: Alma/SFX Local Collection
Publisher: Germany: Elsevier GmbH
ID: ISSN: 0176-1617
Link: https://www.ncbi.nlm.nih.gov/pubmed/24877670
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title: Salt tolerance and activity of antioxidative enzymes of transgenic finger millet overexpressing a vacuolar H+-pyrophosphatase gene (SbVPPase) from Sorghum bicolor
format: Article
creator:
  • Anjaneyulu, Ediga
  • Reddy, Palle Surender
  • Sunita, Merla Srilakshmi
  • Kishor, Polavarapu B. Kavi
  • Meriga, Balaji
subjects:
  • Antioxidants - metabolism
  • Antioxidative enzymes
  • Base Sequence
  • Chlorophyll - metabolism
  • Eleusine - drug effects
  • Eleusine - enzymology
  • Eleusine - genetics
  • Eleusine - physiology
  • Finger millet
  • Gene Expression
  • Gene Expression Regulation, Plant
  • Germination
  • Inorganic Pyrophosphatase - genetics
  • Inorganic Pyrophosphatase - metabolism
  • Lipid Peroxidation
  • Molecular Sequence Data
  • Na+ content
  • NaCl
  • Plant Leaves - drug effects
  • Plant Leaves - enzymology
  • Plant Leaves - genetics
  • Plant Leaves - physiology
  • Plant Proteins - genetics
  • Plant Proteins - metabolism
  • Plants, Genetically Modified
  • Potassium - metabolism
  • Salt-Tolerance
  • SbVPPase
  • Seedlings - drug effects
  • Seedlings - enzymology
  • Seedlings - genetics
  • Seedlings - physiology
  • Seeds - drug effects
  • Seeds - enzymology
  • Seeds - genetics
  • Seeds - physiology
  • Sequence Analysis, DNA
  • Sodium Chloride - metabolism
  • Sodium Chloride - pharmacology
  • Sorghum - enzymology
  • Sorghum - genetics
  • Stress, Physiological
  • Vacuoles - enzymology
ispartof: Journal of plant physiology, 2014-06-15, Vol.171 (10), p.789-798
description: A vacuolar proton pyrophosphatase cDNA clone was isolated from Sorghum bicolor (SbVPPase) using end-to-end gene-specific primer amplification. It showed 80–90% homology at the nucleotide and 85–95% homology at the amino acid level with other VPPases. The gene was introduced into expression vector pCAMBIA1301 under the control of the cauliflower mosaic virus 35S (CaMV35S) promoter and transformed into Agrobacterium tumifaciens strain LBA4404 to infect embryogenic calli of finger millet (Eleusine coracana). Successful transfer of SbVPPase was confirmed by a GUS histochemical assay and PCR analysis. Both, controls and transgenic plants were subjected to 100 and 200mM NaCl and certain biochemical and physiological parameters were studied. Relative water content (RWC), plant height, leaf expansion, finger length and width and grain weight were severely reduced (50–70%), and the flowering period was delayed by 20% in control plants compared to transgenic plants under salinity stress. With increasing salt stress, the proline and chlorophyll contents as well as the enzyme activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and glutathione reductase (GR) increased by 25–100% in transgenics, while malondialdehyde (MDA) showed a 2–4-fold decrease. The increased activities of antioxidant enzymes and the reduction in the MDA content suggest efficient scavenging of reactive oxygen species (ROS) in transgenics and, as a consequence, probably alleviation of salt stress. Also, the leaf tissues of the transgenics accumulated 1.5–2.5-fold higher Na+ and 0.4–0.8-fold higher K+ levels. Together, these results clearly demonstrate that overexpression of SbVPPase in transgenic finger millet enhances the plant's performance under salt stress.
language: eng
source: Alma/SFX Local Collection
identifier: ISSN: 0176-1617
fulltext: fulltext
issn:
  • 0176-1617
  • 1618-1328
url: Link


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titleSalt tolerance and activity of antioxidative enzymes of transgenic finger millet overexpressing a vacuolar H+-pyrophosphatase gene (SbVPPase) from Sorghum bicolor
sourceAlma/SFX Local Collection
creatorAnjaneyulu, Ediga ; Reddy, Palle Surender ; Sunita, Merla Srilakshmi ; Kishor, Polavarapu B. Kavi ; Meriga, Balaji
creatorcontribAnjaneyulu, Ediga ; Reddy, Palle Surender ; Sunita, Merla Srilakshmi ; Kishor, Polavarapu B. Kavi ; Meriga, Balaji
descriptionA vacuolar proton pyrophosphatase cDNA clone was isolated from Sorghum bicolor (SbVPPase) using end-to-end gene-specific primer amplification. It showed 80–90% homology at the nucleotide and 85–95% homology at the amino acid level with other VPPases. The gene was introduced into expression vector pCAMBIA1301 under the control of the cauliflower mosaic virus 35S (CaMV35S) promoter and transformed into Agrobacterium tumifaciens strain LBA4404 to infect embryogenic calli of finger millet (Eleusine coracana). Successful transfer of SbVPPase was confirmed by a GUS histochemical assay and PCR analysis. Both, controls and transgenic plants were subjected to 100 and 200mM NaCl and certain biochemical and physiological parameters were studied. Relative water content (RWC), plant height, leaf expansion, finger length and width and grain weight were severely reduced (50–70%), and the flowering period was delayed by 20% in control plants compared to transgenic plants under salinity stress. With increasing salt stress, the proline and chlorophyll contents as well as the enzyme activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and glutathione reductase (GR) increased by 25–100% in transgenics, while malondialdehyde (MDA) showed a 2–4-fold decrease. The increased activities of antioxidant enzymes and the reduction in the MDA content suggest efficient scavenging of reactive oxygen species (ROS) in transgenics and, as a consequence, probably alleviation of salt stress. Also, the leaf tissues of the transgenics accumulated 1.5–2.5-fold higher Na+ and 0.4–0.8-fold higher K+ levels. Together, these results clearly demonstrate that overexpression of SbVPPase in transgenic finger millet enhances the plant's performance under salt stress.
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languageeng
publisherGermany: Elsevier GmbH
subjectAntioxidants - metabolism ; Antioxidative enzymes ; Base Sequence ; Chlorophyll - metabolism ; Eleusine - drug effects ; Eleusine - enzymology ; Eleusine - genetics ; Eleusine - physiology ; Finger millet ; Gene Expression ; Gene Expression Regulation, Plant ; Germination ; Inorganic Pyrophosphatase - genetics ; Inorganic Pyrophosphatase - metabolism ; Lipid Peroxidation ; Molecular Sequence Data ; Na+ content ; NaCl ; Plant Leaves - drug effects ; Plant Leaves - enzymology ; Plant Leaves - genetics ; Plant Leaves - physiology ; Plant Proteins - genetics ; Plant Proteins - metabolism ; Plants, Genetically Modified ; Potassium - metabolism ; Salt-Tolerance ; SbVPPase ; Seedlings - drug effects ; Seedlings - enzymology ; Seedlings - genetics ; Seedlings - physiology ; Seeds - drug effects ; Seeds - enzymology ; Seeds - genetics ; Seeds - physiology ; Sequence Analysis, DNA ; Sodium Chloride - metabolism ; Sodium Chloride - pharmacology ; Sorghum - enzymology ; Sorghum - genetics ; Stress, Physiological ; Vacuoles - enzymology
ispartofJournal of plant physiology, 2014-06-15, Vol.171 (10), p.789-798
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02014 Elsevier GmbH
1Copyright © 2014 Elsevier GmbH. All rights reserved.
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1Reddy, Palle Surender
2Sunita, Merla Srilakshmi
3Kishor, Polavarapu B. Kavi
4Meriga, Balaji
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0Salt tolerance and activity of antioxidative enzymes of transgenic finger millet overexpressing a vacuolar H+-pyrophosphatase gene (SbVPPase) from Sorghum bicolor
1Journal of plant physiology
addtitleJ Plant Physiol
descriptionA vacuolar proton pyrophosphatase cDNA clone was isolated from Sorghum bicolor (SbVPPase) using end-to-end gene-specific primer amplification. It showed 80–90% homology at the nucleotide and 85–95% homology at the amino acid level with other VPPases. The gene was introduced into expression vector pCAMBIA1301 under the control of the cauliflower mosaic virus 35S (CaMV35S) promoter and transformed into Agrobacterium tumifaciens strain LBA4404 to infect embryogenic calli of finger millet (Eleusine coracana). Successful transfer of SbVPPase was confirmed by a GUS histochemical assay and PCR analysis. Both, controls and transgenic plants were subjected to 100 and 200mM NaCl and certain biochemical and physiological parameters were studied. Relative water content (RWC), plant height, leaf expansion, finger length and width and grain weight were severely reduced (50–70%), and the flowering period was delayed by 20% in control plants compared to transgenic plants under salinity stress. With increasing salt stress, the proline and chlorophyll contents as well as the enzyme activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and glutathione reductase (GR) increased by 25–100% in transgenics, while malondialdehyde (MDA) showed a 2–4-fold decrease. The increased activities of antioxidant enzymes and the reduction in the MDA content suggest efficient scavenging of reactive oxygen species (ROS) in transgenics and, as a consequence, probably alleviation of salt stress. Also, the leaf tissues of the transgenics accumulated 1.5–2.5-fold higher Na+ and 0.4–0.8-fold higher K+ levels. Together, these results clearly demonstrate that overexpression of SbVPPase in transgenic finger millet enhances the plant's performance under salt stress.
subject
0Antioxidants - metabolism
1Antioxidative enzymes
2Base Sequence
3Chlorophyll - metabolism
4Eleusine - drug effects
5Eleusine - enzymology
6Eleusine - genetics
7Eleusine - physiology
8Finger millet
9Gene Expression
10Gene Expression Regulation, Plant
11Germination
12Inorganic Pyrophosphatase - genetics
13Inorganic Pyrophosphatase - metabolism
14Lipid Peroxidation
15Molecular Sequence Data
16Na+ content
17NaCl
18Plant Leaves - drug effects
19Plant Leaves - enzymology
20Plant Leaves - genetics
21Plant Leaves - physiology
22Plant Proteins - genetics
23Plant Proteins - metabolism
24Plants, Genetically Modified
25Potassium - metabolism
26Salt-Tolerance
27SbVPPase
28Seedlings - drug effects
29Seedlings - enzymology
30Seedlings - genetics
31Seedlings - physiology
32Seeds - drug effects
33Seeds - enzymology
34Seeds - genetics
35Seeds - physiology
36Sequence Analysis, DNA
37Sodium Chloride - metabolism
38Sodium Chloride - pharmacology
39Sorghum - enzymology
40Sorghum - genetics
41Stress, Physiological
42Vacuoles - enzymology
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titleSalt tolerance and activity of antioxidative enzymes of transgenic finger millet overexpressing a vacuolar H+-pyrophosphatase gene (SbVPPase) from Sorghum bicolor
authorAnjaneyulu, Ediga ; Reddy, Palle Surender ; Sunita, Merla Srilakshmi ; Kishor, Polavarapu B. Kavi ; Meriga, Balaji
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1Antioxidative enzymes
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4Eleusine - drug effects
5Eleusine - enzymology
6Eleusine - genetics
7Eleusine - physiology
8Finger millet
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11Germination
12Inorganic Pyrophosphatase - genetics
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16Na+ content
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19Plant Leaves - enzymology
20Plant Leaves - genetics
21Plant Leaves - physiology
22Plant Proteins - genetics
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36Sequence Analysis, DNA
37Sodium Chloride - metabolism
38Sodium Chloride - pharmacology
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40Sorghum - genetics
41Stress, Physiological
42Vacuoles - enzymology
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atitleSalt tolerance and activity of antioxidative enzymes of transgenic finger millet overexpressing a vacuolar H+-pyrophosphatase gene (SbVPPase) from Sorghum bicolor
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abstractA vacuolar proton pyrophosphatase cDNA clone was isolated from Sorghum bicolor (SbVPPase) using end-to-end gene-specific primer amplification. It showed 80–90% homology at the nucleotide and 85–95% homology at the amino acid level with other VPPases. The gene was introduced into expression vector pCAMBIA1301 under the control of the cauliflower mosaic virus 35S (CaMV35S) promoter and transformed into Agrobacterium tumifaciens strain LBA4404 to infect embryogenic calli of finger millet (Eleusine coracana). Successful transfer of SbVPPase was confirmed by a GUS histochemical assay and PCR analysis. Both, controls and transgenic plants were subjected to 100 and 200mM NaCl and certain biochemical and physiological parameters were studied. Relative water content (RWC), plant height, leaf expansion, finger length and width and grain weight were severely reduced (50–70%), and the flowering period was delayed by 20% in control plants compared to transgenic plants under salinity stress. With increasing salt stress, the proline and chlorophyll contents as well as the enzyme activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and glutathione reductase (GR) increased by 25–100% in transgenics, while malondialdehyde (MDA) showed a 2–4-fold decrease. The increased activities of antioxidant enzymes and the reduction in the MDA content suggest efficient scavenging of reactive oxygen species (ROS) in transgenics and, as a consequence, probably alleviation of salt stress. Also, the leaf tissues of the transgenics accumulated 1.5–2.5-fold higher Na+ and 0.4–0.8-fold higher K+ levels. Together, these results clearly demonstrate that overexpression of SbVPPase in transgenic finger millet enhances the plant's performance under salt stress.
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pubElsevier GmbH
pmid24877670
doi10.1016/j.jplph.2014.02.001