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Effect of Salinity Stress on Glutathione –s-transferases (gsts) of Maize

R. N. Remme
MS student,
Department of Plant Breeding and Genetics, BSMRAU, Gazipur

N. A. Ivy
Assoc. Professor
Department of Plant Breeding and Genetics, BSMRAU, Gazipur

MAK Mian
Professor
Department of Plant Breeding and Genetics, BSMRAU, Gazipur

Dr. Md. Motiar Rohman
Senior Scientific Officer
Plant Breeding Division, BARI, Gazipur

Abstract/ Executive Summary:

The present study was undertaken to explore the responses of maize GST to salinity. GST activities were found to induce with the increasing of salinity levels and duration of stress. At 3 day, 5 day and 7 day stress most of all the salinity concentration (4 dS/m, 8 dS/m and 12 dS/m) increased the GST activities over control. However 16 dS/m salinity decreased the activity over control. In foliar application of proline GST activities were increased gradually over control and NaCl treatment at 3 day, 5 day and 7 day stress in all the salinity level. In betaine application at 4 dS/m, 8 dS/m, 12 dS/m and 16 dS/m salinity also increased GST activity were gradually over control at 3 day, 5 day and 7 day stress. It also increases the GST activity during the stress period over NaCl stress in all the salinity levels, except 12 dS/m and 16 dS/m salinity at 3 day stress. When both proline and betaine applied together in stressed seedlings at 4 dS/m, 8 dS/m, 12 dS/m and 16 dS/m salinity increased the GST activity at 3 day, 5 day and 7 day stress over control. Though the combination of 15 mM proline and betaine decreased the activities at 3 day and 7 day stress at 4 dS/m, 8 dS/m and 12 dS/m at 3 day stress GST activity increased at other salinity stress in different stress period. To examine the detoxification role of maize GST, concentration of MDA (peroxident product of poly unsaturated fatty acid) in the stressed maize seedlings was studied. In all the salinity levels, MDA contents increased over control. Supplementation of proline and betaine reduced the concentrations of MDA over salinity stresses. The simultaneous increased GST activities and lower MDA concentration suggested the detoxification role of maize GST on cytotoxic MDA.

Keywords: Glutathione –S-transferases, Maize, NaCl, Melondialdehide

Location: Molecular Breeding Lab, Plant Breeding Division, BARI

Start Date: 00-04-2011

End Date: 00-08-2012

Program Area: Risk Management in Agriculture

Commodity: Maize

Objectives:

To explore the responses of maize GST to salinity. GST activities were found to induce with the increasing of salinity levels and duration of stress.

Methodology:

Experimental site: This study was carried out in the Molecular Breeding Laboratory of Bangladesh Agricultural Research Institute (BARI), Joydebpur, Gazipur during the period from April, 2011 to August, 2012.

Plant materials: Seedlings of maize (BARI Hybrid Maize 7) were used as plant material.

Chemicals: All of the chemicals except Hydroxylapaptite, NaCl (Merck, Germany) were used manufactured from Sigma-Aldrich, USA.

Stress treatments: Maize seedlings were grown in plastic pots. Five days old seedlings were subjected for stress implementation. Salinity levels (4, 8 and 12 dS/m) were maintained by adding NaCl and water. A control treatment (with adding NaCl) was also growing under same condition. In NaCl stressed seedlings 15 mM proline and 15 mM betaine were applied as foliar spray twice daily. Digital EC meter was used to measured level of salinity, and the level was expressed at field capacity.

Preparation of soluble protein:To assay the activity level of GST in maize seedlings five gram of sample was homogenized in an equal volume of 50 mM potassium phosphate buffer (pH 7.0) containing 100 mM KCl, 1% (w/v) ascorbate and 10 % (w/v) glycerol with mortar and pestle. The homogenates were centrifuged at 11,500 x g for 10 min and the supernatant was used as a soluble protein solutions.

Measurement of lipid peroxidation:The level of lipid peroxidation was measured by estimating MDA, a decomposition product of the peroxidized polyunsaturated fatty acid component of the membrane lipid, using thiobarbituric acid (TBA) as the reactive material following the method of Heath and Packer (1968) with slight modifications. The seedlings (2.5 g) were homogenized in 15 ml 5% (w/v) trichloroacetic acid (TCA) and the homogenate was centrifuged at 11,500 rpm for 10 min. One ml supernatant was mixed with 4 ml of TBA reagent (0.5% of TBA in 20% TCA). The reaction mixture was heated at 95°C for 30 min in a water bath and then quickly cooled in an ice bath and centrifuged at 11,500g for 15 min. The absorbance of the colored supernatant was measured at 532 nm and was corrected for non-specific absorbance at 600 nm. The concentration of MDA was calculated by using the extinction coefficient of 155 mM-1 cm-1 and expressed as nmol of MDA g-1 fresh weight.

Source of Information: IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS) e-ISSN: 2319-2380, p-ISSN: 2319-2372. Volume 4, Issue 4 (Jul. - Aug. 2013), PP 42-52 www.iosrjournals.org

Article Link (Online Journal):

Funding Source:

1. Government

Budget: One lac taka only

Total Budget (Tk.) : One lac taka only

Achievement/Findings:

Maize GST was found to be regulated with organic hydroperoxide.

Knowledge Management: Journal