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Low and High Temperature Induced Sugar Accumulation in Pui Vegetable (basella Alba)

Md. M. Islam
Department of Biochemistry and Molecular Biology, Laboratory of Protein and Enzyme Research University of Rajshahi, Rajshahi-6205, Bangladesh

Md. S. Haque
Department of Biochemistry and Molecular Biology, Laboratory of Protein and Enzyme Research University of Rajshahi, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

The formation of sugars in plants is an essential aspect of metabolism and is altered by different ways although the mechanism is not clarified. Here, we used pot experiment for cultivation of vegetable Basella alba and examined the effects of low and high temperature on sugar, reducing sugar and non reducing sugar level in leaf. Cold acclimation (8^oC) causes the accumulation of sugar for 24, 48 and 72 h however, the higher accumulation was observed after 72 h of the treatment. Reducing and non reducing sugars were also increased time dependently in response to cold. Whenever the plants were exposed to high temperature (45^oC), the similar stimulatory and more pronounced effects on sugar accumulation were obtained for 24, 48 and 72 h periods and the accumulation of sugars in leaf were found time dependently up to 72 h compared to the respective controls. The reducing and non reducing sugars in leaf were accumulated similarly in response to high temperature. The increased sugar accumulation might be due to the oxidative stress caused by such extreme alteration of temperature and may play the critical role in survival of this species of plant in such adverse environment.

Keywords: Adaptive response, Cold acclimation, High temperature, Metabolic effects, Sugar accumulation

Location: Department of Biochemistry and Molecular Biology, Laboratory of Protein and Enzyme Research University of Rajshahi, Rajshahi

Start Date:

End Date:

Program Area: Risk Management in Agriculture

Commodity: Vegetables

Objectives:

To find the role of cold acclimation and exposure of high temperature on the regulation of metabolic activities regarding the accumulation of sugars in Pui vegetables.

Methodology:

Plant Materials and Low Temperature Treatment - For this experiment, two plastic pots were used; each pot size was 70 cm in diameter and 24 cm in height. An adequate amount of soil was taken in each plastic pot and the plastic pots were seeded with Basella alba. For the germination of seeds, the following points were carried out: i) the strong seeds were selected; the seeds were added to normal water and the floating seeds were discarded; ii) the seeds were kept in normal water with temperature below 37^oC for overnight; iii) the seeds which were swollen by water absorption, were expected to be effective for germination; iv) the seeds were seeded in the pots prepared with soil and the efficiency of seed germination was 65%- 75%. After 30 days of germination, the two different pots were described as control and low temperature induced plant. Control pot was used for 24, 48 and 72 h treatments in the room temperature without cold acclimation. The second pot was used for 24, 48 and 72 h duration in the temperature controlled cooling chamber and given cold exposure (8^oC) with full aeration. After the treatments, leaves were collected consecutively from each pot for 24, 48 and 72 h duration and kept in -80^oC. Plant Materials and High Temperature Treatment - In separate experiments, another two plastic pots were prepared with soil and seeded similarly with Basella alba. After 20 days of germination, the two different pots were described as control and high temperature induced plant. Control pot was used for 24, 48 and 72 h treatments in room temperature, however, the temperature was maintained 30oC by using air cooling system (AC) already fixed in the room and without giving any high temperature. The second pot was used similarly for 24, 48 and 72 h duration in the plastic chamber and was exposed to 45oC with full aeration along with sufficient water. To maintain this temperature, electric bulbs (2×200 W) were connected to the chamber. After the treatments, the leaves were collected consecutively from each pot for 24, 48 and 72 h duration and kept in -80^oC. Assay of Sugar Metabolites in Leaf - Approximately, 3~4 g of cold exposed and their respective control leaves after 24, 48 and 72 h periods were used for determination of sugar, reducing sugar and non reducing sugar while approximately 2~4 g of high temperature induced leaves and their respective controls were used similarly. The sugar and reducing sugar content in leaf was determined colorimetrically. Briefly, leaves were plunged into boiling ethyl alcohol and allowed to boil for 10 minutes (10 mL of ethanol was used for each gram of leaf). The extract was cooled, crushed in a mortar with pestle and was filtered through two layers of muslin cloth and re-extracted the crude leaf for three minutes in hot 80% ethanol, using 2 to 3 mL of ethanol for each gram of leaf sample. The second extraction ensured complete removal of alcohol soluble substances. The extract was cooled again and passed through muslin cloth. Both the extracts were filtered through Whatmann No. 41 filter paper. The volume of the extract was evaporated to about ¼ of the volume over a stream bath and cooled. The reduced volume of the extract was transferred to a 100 mL volumetric flask and made up to the mark with distilled water. Aliquot of leaf extract (1 mL) was taken into a test tube, 4 mL of anthrone reagent (0.2% in concentrated H2SO4) was added and mixed well. The tubes were heated in a water bath and cooled. To prevent water loss, glass marbles were placed on top of each tube. A blank was prepared by taking 1 mL of water and 4 mL of anthrone reagent in a tube and treated similarly. The absorbance of the blue-green solution was measured at 680 nm in a colorimeter. The amount of free sugar was calculated from the standard glucose concentration (10 mg dL^-1) and was expressed as mg 100 g^-1 of leaf. For determination of reducing sugar content in leaf, aliquot of the extract (10 mL) was pipetted into test tubes and 3 mL of DNS (dinitrosalicylic acid) reagent (1g of DNS, 200 mg of crystalline phenol and 50 mg of sodium sulphate were placed in a beaker and mixed with 100 mL of 1% NaOH solution) were added to each of the tubes and mixed well. The tubes were heated for 5 minutes in a boiling water bath to develop the color and 1 mL of 40% Rochelle salt was added when the contents of the tubes were still worm and cooled with tap water. A blank was prepared by taking 10 mL of water and 3 mL of DNS reagent in a tube and treated similarly. The absorbance of the solutions was measured at 575 nm in a colorimeter. The amount of reducing sugar was calculated from the standard glucose concentration (10 mg dL^-1). Non reducing sugar content in leaf was determined from the difference of total sugar and reducing sugar content in the different treated plants and their respective controls. Statistical Analysis - Results of the experiments were expressed as mean and standard error of different groups. The differences between the mean values were evaluated by ANOVA followed by paired t-test using SPSS software.

Source of Information: Thai Journal of Agricultural Science 2013, 46(2): 85-94

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Temperature variations particularly low and high temperature, have been applied to the plant Basella alba to find the sugar accumulation during this critical environment. In fact, in such an adverse situation, plants survive, however accumulation of sugar was observed in leaves of this species of plant. Sugar, reducing sugar and non reducing sugars might be the major energy source during these critical circumstances. To make an adaptive response to the environment, these accumulated sugars play a critical role. Although several factors might be involved in this respect, however, these two environmental factors are found to be predominantly involved. Our investigations will give the concept to find the new strategy for survival of the plant species in the critical situation like cold and drought.

Knowledge Management: Journal