Quantitative Gene Expression Analysis of Selected Genes to Screen Drought Tolerance of Selected Hevea Clones

  • I. N. Amarasekara
  • S. P. Withanage
  • I. R. Palihakkara
Keywords: Hevea, stress tolerance, gene expression, Catalase (CAT), Glutathione Peroxidase (GPX), Ascorbate Peroxidase (APX), Real Time PCR

Abstract

Expansion of rubber (Hevea brasiliensis) cultivation to non-traditional areas in Sri Lanka is necessary due to the increase of global demand of natural rubber and limitation of available land in traditional rubber growing areas. To overcome the negative impact of drought stress, which experience in non-traditional rubber growing areas, plants alter their gene expression as an inherent respond strategy. In traditional rubber growing areas, during the period of February to March is considered as wintering period where recorded higher temperature, low rainfall and lower relative humidity (RH%) comparing to the   cropping months of October to December. Therefore, wintering period is more over similar to an environmental condition prevailing in non-traditional areas. Stable clones during wintering period can be expected that having more ability to survive in dry areas too. Therefore, this experiment was conducted to select more stress tolerant clones for non-traditional areas and to evaluate the level of drought responsible genes (Catalase (CAT), ascorbate peroxidase (APX) and Glutathione peroxidase (GPX)) expression of selected Hevea clones under wintering stress compared to the cropping months. Quantitative gene expression during wintering period compared to the wintering period was carried out using Bio-Rad thermal cycler. The Livack method was used to calculate fold difference between control and treatment plants. Then paired t-test was done using minitab 17 software. According to the results, all the genes were significantly up regulated in the clone RRISL Centennial 3 during the wintering period and second highest yield dropped percentage (55%) was recorded in this clone. However, HbAPX and HbCAT gene expressions were significantly upregulated while HbGPX gene expression was significantly downregulated in the clone RRISL 203 and highest yield dropped percentage (56%) during wintering period was recorded in this clone. In contrast the clone RRISL 2006, HbAPX and HbCAT gene expressions were significantly down regulated and upregulation of HbGPX gene expression was not significantly at the P≤0.05 level. This results were different from the expression pattern of other two clones and the clone RRISL 2006 showed lowest yield dropped as around 23% during the wintering period. Leaf falling of this clone also delayed compared to the other clones and might be having more drought tolerant ability than the clones RRISL 203 and RRISL Centennial 3. Therefore, this may be some other genes responsible for the situation and should be studied to confirm the results. According to those results, the clones RRISL 203 and RRISL Centennial 3 can be considered as stress susceptible clones compared to RRISL 2006, which can be considered as stress tolerance clone.

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References

Bandara, P.K.G.S.S. (2018). NNSF Report on Analysis of antioxidant genes expression in tapping panel dryness (TPD) affected rubber trees (Hevea brasiliensis Muell. Arg.) and effect of exogenous application of ascorbic acid on alleviating TPD.

Bartels, D., & Sunkar, R. (2005). Drought and Salt Tolerance in Plants. Critical Reviews in Plant Sciences, 24(1), 23–58. https://doi.org/10.1080/07352680590910410

Chaves, M. M., Maroco, J. P., & Pereira, J. S. (2003). Understanding plant responses to drought—From genes to the whole plant. Functional Plant Biology, 30(3), 239. https://doi.org/10.1071/FP02076

S. D. T. S. Gunasekara, I. R. Palihakkara, & S. P. Withanage. (2020). Study toward the Development of Relationship between Ref Gene Expression and Yield Potential of Hevea Clones. International Journal for Research in Applied Sciences and Biotechnology, 7(5), 98-105. https://doi.org/10.31033/ijrasb.7.5.14

K. L. M. Kalpani, S. P. Withange, & I. R. Palihakkara. (2020). Selection of Superior Genotypes at Early Stage of the Rubber (Hevea brasiliensis) Breeding Cycle. International Journal for Research in Applied Sciences and Biotechnology, 7(4), 146-155. https://doi.org/10.31033/ijrasb.7.4.17

Rubber Research Institute of India, Regional Research Station, Dhenkanal 759 001, India, & B, K. (2017). Assessment of drought tolerance in few clones of natural rubber (Hevea brasiliensis) under dry hot climate of Odisha, India. Journal of Experimental Biology and Agricultural Sciences, 5(1), 106–110. https://doi.org/10.18006/2017.5(1).106.110

Laxa, M., Liebthal, M., Telman, W., Chibani, K., & Dietz, K.-J. (2019). The Role of the Plant Antioxidant System in Drought Tolerance. Antioxidants, 8(4), 94. https://doi.org/10.3390/antiox8040094

Livak, K. J., & Schmittgen, T. D. (2001). Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT Method. Methods, 25(4), 402–408. https://doi.org/10.1006/meth.2001.1262

Luke, L. P., Mohamed Sathik, M. B., Thomas, M., Kuruvilla, L., Sumesh, K. V., & Annamalainathan, K. (2015). Quantitative expression analysis of drought responsive genes in clones of Hevea with varying levels of drought tolerance. Physiology and Molecular Biology of Plants, 21(2), 179–186. https://doi.org/10.1007/s12298-015-0288-0

Nagahawatta, D.P., Attanayake, D.P.S.T.G, Dissanayaka, D.M.S, Withanage, S.P. (2017). Analysis and cloning of the coding region of Rubber Elongation Factor Gene ( ref ) related sequences from Hevea brasiliensis and Hevea spruceana. 21-25.

A. Ranasinghe, I. R. Palihakkara, & A. Nugawela. (2020). Yield Variation during the Period (April to July) 2017 in RRIC 121 & BPM 24 Rubber Clones (Hevea brasiliensis) Grown in WL3 Agro Ecological Zone of Sri Lanka. International Journal for Research in Applied Sciences and Biotechnology, 7(3), 27-34. https://doi.org/10.31033/ijrasb.7.3.5

Shinozaki, K., Yamaguchi-Shinozaki, K., & Seki, M. (2003). Regulatory network of gene expression in the drought and cold stress responses. Current Opinion in Plant Biology, 6(5), 410–417. https://doi.org/10.1016/S1369-5266(03)00092-X

Singh, M., Chandrasekhar, T.R., Nazeer, M.A., Annamalainathan, K. and Mydin, K.K. (2012). Early Performance of some clones of Hevea brasiliensis in the dry subhumid regions of North Konkan. International Rubber Conference, Kovalam, India, pp. 61.

Sri Lanka Export Development Board. (2015). The Natural Rubber Industry of Sri Lanka

Sri Lanka Rubber Secretariat. (2017). Sri Lanka Rubber Industry Development Master Plan 2017-2026.

Somaratne, S., & Dhanapala, A. H. (1996). Potential Impact of Global Climate Change on Forest Distribution in Sri Lanka. In L. Erda, W. C. Bolhofer, S. Huq, S. Lenhart, S. K. Mukherjee, J. B. Smith, & J. Wisniewski (Eds.), Climate Change Vulnerability and Adaptation in Asia and the Pacific (pp. 129–135). Springer Netherlands. https://doi.org/10.1007/978-94-017-1053-4_12

Tardieu, F., & Tuberosa, R. (2010). Dissection and modelling of abiotic stress tolerance in plants. Current Opinion in Plant Biology, 13(2), 206–212. https://doi.org/10.1016/j.pbi.2009.12.012

Verheye, W. (2010). Growth and production of rubber. In W. H. Verheye (Ed.), Land use, land cover and soil sciences. Oxford, UK: UNESCO-EOLSS Publishers. pp. 295–300.

Published
2020-11-03
How to Cite
I. N. Amarasekara, S. P. Withanage, & I. R. Palihakkara. (2020). Quantitative Gene Expression Analysis of Selected Genes to Screen Drought Tolerance of Selected Hevea Clones. International Journal for Research in Applied Sciences and Biotechnology, 7(6), 46-53. https://doi.org/10.31033/ijrasb.7.6.8

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