title: Salt tolerance and activity of antioxidative enzymes of transgenic finger millet overexpressing a vacuolar H(+)-pyrophosphatase gene (SbVPPase) from Sorghum bicolor.
abstract: 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.
authors.affiliation: Department of Biochemistry, Sri Venkateswara University, Tirupati 517502, India.
authors.forename: Ediga
authors.initials: E
authors.lastname: Anjaneyulu
authors.affiliation: Department of Genetics, Osmania University, Hyderabad 500007, India.
authors.forename: Palle Surender
authors.initials: PS
authors.lastname: Reddy
authors.affiliation: Department of Genetics, Osmania University, Hyderabad 500007, India.
authors.forename: Merla Srilakshmi
authors.initials: MS
authors.lastname: Sunita
authors.affiliation: Department of Genetics, Osmania University, Hyderabad 500007, India.
authors.forename: Polavarapu B Kavi
authors.initials: PB
authors.lastname: Kishor
authors.affiliation: Department of Biochemistry, Sri Venkateswara University, Tirupati 517502, India. Electronic address: balaji.meriga@gmail.com.
authors.forename: Balaji
authors.initials: B
authors.lastname: Meriga
mesh_terms: D000975:Antioxidants / Q000378:metabolism*; D001483:Base Sequence; D002734:Chlorophyll / Q000378:metabolism; D031748:Eleusine / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant; D018525:Germination; D043564:Inorganic Pyrophosphatase / Q000235:genetics* / Q000378:metabolism; D015227:Lipid Peroxidation; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000502:physiology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011188:Potassium / Q000378:metabolism; D055049:Salt Tolerance; D036226:Seedlings / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000502:physiology; D012639:Seeds / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000502:physiology; D017422:Sequence Analysis, DNA; D012965:Sodium Chloride / Q000378:metabolism / Q000494:pharmacology; D045868:Sorghum / Q000201:enzymology* / Q000235:genetics; D013312:Stress, Physiological; D014617:Vacuoles / Q000201:enzymology
chemical_list: D000975:Antioxidants; D010940:Plant Proteins; D002734:Chlorophyll; D012965:Sodium Chloride; D043564:Inorganic Pyrophosphatase; D011188:Potassium
keywords: Antioxidative enzymes; Finger millet; Na(+) content; NaCl; SbVPPase