Main Article Content
Abstract
This study investigated the physiological and molecular responses of two common bean (Phaseolus vulgaris L.) genotypes, “Balkız” and “Local Genotype”, which differ in their relative heat tolerance, with the “Local Genotype” being more tolerant than “Balkız”, to high-temperature stress during the seedling stage, aiming to identify genotype-specific physiological and biochemical responses and to determine reliable indicators of thermotolerance. Leaf relative water content (RWC), turgor loss, membrane damage, proline levels, total soluble protein (TSP), and the expression of heat shock proteins HSP23 and HSP60 were examined. The “Local Genotype” exhibited greater thermotolerance, with lower membrane damage while the temperature at which 50% injury occurs (LT50) was higher, despite the accumulation of less proline than “Balkız”. Genotype-specific variations detected in protein expression indicate distinct biochemical responses to heat stress. HSP23 showed a rapid, transient increase at moderate temperature (40 °C) in both genotypes, suggesting it has a role in early heat stress response. HSP60 levels were maintained near LT50 temperatures, supporting sustained stress adaptation, but declined at 48 °C, reflecting extreme stress rather than genotype-specific differences. These findings emphasize LT50 and HSP dynamics as key indicators of thermotolerance and provide insights for understanding heat stress responses in common bean seedlings.
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References
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- Arora, R., Pitchay, D.S., Bearce, B.C., 1998: Water stress-induced heat tolerance in geranium leaf tissues: A possible linkage through stress proteins. Physiologia Plantarum 103, 24-34. https://doi.org/10.1034/j.1399-3054.1998.1030104.x
- Arora, R., Wisniewski, M. E., 1994: Cold acclimation in genetically related (sibling) deciduous and evergreen peach (Prunus persica [L.] Batsch) (II. A 60-kilodalton bark protein in cold-acclimated tissues of peach is heat stable and related to the dehydrin family of proteins). Plant Physiology 105(1), 95-101. https://doi.org/10.1104/pp.105.1.95
- Barr, H.D., Weatherley, P.E., 1962: A Re-examination of the Relative turgidity technique for estimating water deficit in leaves, Australian Journal of Biological Sciences 15, 413-428. http://dx.doi.org/10.1071/BI9620413
- Bates, L.S., Waldren, R.P., Teare, I.D., 1973: Rapid determination of free proline for water stress studies. Plant Soil 39, 205-207. https://doi.org/10.1007/BF00018060
- Bita CE, Gerats T., 2013: Plant tolerance to high temperature in a changing environment: scientific fundamentals and production of heat stress-tolerant crops. Frontiers in Plant Science 31, 4:273. https://doi.org/10.3389/fpls.2013.00273
- Blair, M. W., 2013: Mineral biofortification strategies for food staples: the example of common bean. Journal of Agricultural and Food Chemistry 61(35), 8287-8294. https://doi.org/10.1021/jf400774y
- Bradford, M. M., 1976: A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Analytical Biochemistry 72, 248-254.
- Chavez-Arias, C.C, Ligarreto-Moreno, G.A., Restrepo-Díaz, H. 2018: Evaluation of heat stress period duration and the interaction of daytime temperature and cultivar on common bean. Environmental and Experimental Botany 155, 600-608. https://doi.org/10.1016/j.envexpbot.2018.08.012
- da Silva, D.A., Pinto-Maglio, C.A.F., de Oliveira, E.C., dos Reis, R.L.d.M., Carbonell, S.A.M., Chiorato, A.F. 2020: Influence of high temperature on the reproductive biology of dry edible bean (Phaseolus vulgaris L.). Scientia Agricola 77(3), e20180233. https://doi.org/10.1590/1678-992X-2018-0233
- Ergin, S., Gulen, H., Kesici, M., Turhan, E., Ipek, A.N., Köksal, N., 2016: Effects of high temperature stress on enzymatic and nonenzymatic antioxidants and proteins in strawberry plants. Turkish Journal of Agriculture and Forestry 40, 908-917. https://doi.org/10.3906/tar-1606-144
- Haq, S.U., Khan, A., Ali, M. et al., 2019: Knockdown of CaHSP60-6 confers enhanced sensitivity to heat stress in pepper (Capsicum annuum L.). Planta 250, 2127–2145. https://doi.org/10.1007/s00425-019-03290-4
- Harsh, A., Sharma, Y.K., Joshi, U., Rampuria, S., Singh, G., Kumar, S. Sharma, R., 2016: Effect of short-term heat stress on total sugars, proline and some antioxidant enzymes in moth bean (Vigna aconitifolia), Annals of Agricultural Sciences 61(1): 57-64. https://doi.org/10.1016/j.aoas.2016.02.001
- IPCC, Intergovernmental Panel on Climate Change 2023: Climate Change 2023: Synthesis Report. Summary for Policymakers. Contribution of Working Groups I, II and III to the Sixth Assessment Report of the IPCC (Core Writing Team, H. Lee & J. Romero, eds.), Geneva, Switzerland, pp-1-34. https://doi.org/10.59327/IPCC/AR6-9789291691647.001
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- Kumar, S., Kaur, R., Kaur, N., Bhandhari, K., Kaushal, N., Gupta, K., Bains, T.S., Nayyar, H., 2011: Heat-stress induced inhibition in growth and chlorosis in mungbean (Phaseolus aureus Roxb.) is partly mitigated by ascorbic acid application and is related to reduction in oxidative stress. Acta Physiologiae Plantarum 33(6), 2091–2101. https://doi.org/10.1007/s11738-011-0748-2
- Kumar, P., Paul, D., Jhajhriya, S. et al., 2024: Understanding heat-shock proteins’ abundance and pivotal function under multiple abiotic stresses. Journal of Plant Biochemistry and Biotechnology 33, 492-513. https://doi.org/10.1007/s13562-024-00932-x
- Kumari, V. V., Roy, A., Vijayan, R., Banerjee, P., Verma, V. C., Nalia, A., & Hossain, A., 2021: Drought and heat stress in cool-season food legumes in sub-tropical regions: Consequences, adaptation, and mitigation strategies. Plants 10(6), 1038. https://doi.org/10.3390/plants10061038
- McLoughlin F, Kim M, Marshall RS, Vierstra RD, Vierling E., 2019: HSP101 interacts with the proteasome and promotes the clearance of ubiquitylated protein aggregates. Plant Physiology 180(4), 1829-1847. https://doi.org/10.1104/pp.19.00263
- Meena, M., Divyanshu, K., Kumar, S., Swapnil, P., Zehra, A., Shukla, V., Yadav, M., Upadhyay, R.S., 2019: Regulation of L-proline biosynthesis, signal transduction, transport, accumulation and its vital role in plants during variable environmental conditions. Heliyon 5(12), e02952. https://doi.org/10.1016/j.heliyon.2019.e02952
- Mullan, D., Pietragalla, J., 2012: Leaf relative water content. Physiological breeding II: a field guide to wheat phenotyping. CIMMYT, Mexico, 25-27.
- Niu, Y., Xiang, Y., 2018: An overview of biomembrane functions in plant responses to high-temperature stress. Frontiers in Plant Science 9:915. https://doi.org/10.3389/fpls.2018.00915
- Pant, K.K., Naik, J., Barthakur, S., Chandra, V., 2025: High-temperature stress in wheat (Triticum aestivum L.): unfolding the impacts, tolerance and methods to mitigate the detrimental effects. Cereal Research Communications. https://doi.org/10.1007/s42976-025-00634-7
- Rani, B., Kumari, N., Jain, V., Dhawan, K., Avtar, R., 2016: Heat stress induced changes in protein profile of Indian mustard (Brassica juncea L.). Journal of Oilseed Brassica 1(1), 302-305. https://epubs.icar.org.in/index.php/JOB/article/view/159031
- Raza, A., Charagh, S., Abbas, S., Hassan, M.U., Saeed, F., Haider, S., Sharif, R., Anand, A., Corpas, F.J., Jin, W., Varshney, R.K., 2023: Assessment of proline function in higher plants under extreme temperatures. Plant Biology 25(3), 379-395. https://doi.org/10.1111/plb.13510
- Roy, S., Mishra, M., Dhankher, O.P., Singla-Pareek, S.L., Pareek, A., 2019: Molecular Chaperones: Key Players of Abiotic Stress Response in Plants. In: Rajpal, V., Sehgal, D., Kumar, A., Raina, S. (eds) Genetic Enhancement of Crops for Tolerance to Abiotic Stress: Mechanisms and Approaches, Vol. I. Sustainable Development and Biodiversity, vol 20. Springer, Cham. https://doi.org/10.1007/978-3-319-91956-0_6
- Scharf, K. D., Berberich, T., Ebersberger, I., Nover, L., 2012: The plant heat stress transcription factor (Hsf) family: structure, function and evolution. Biochimica et Biophysica Acta (BBA)-Gene Regulatory Mechanisms 1819(2), 104-119. https://doi.org/10.1016/j.bbagrm.2011.10.002
- Shen, S., Jing, Y., Kuang, T., 2003: Proteomics approach to identify wound-response related proteins from rice leaf sheath, Proteomics 3 (4), 527-535. https://doi.org/10.1002/pmic.200390066
- Soltani, A., Weraduwage, S.M., Sharkey, T.D., Lowry, D.B., 2019: Elevated temperatures cause loss of seed set in common bean (Phaseolus vulgaris L.) potentially through the disruption of source-sink relationships. BMC Genomics 20(1), 312. https://doi.org/ 10.1186/s12864-019-5669-2
- Soltys-Kalina D, Plich J, Strzelczyk-Żyta D, Śliwka J, Marczewski W., 2016: The effect of drought stress on the leaf relative water content and tuber yield of a half-sib family of 'Katahdin'-derived potato cultivars. Breed Sci. 66(2), 328-31. https://doi.org/10.1270/jsbbs.66.328
- Spormann, S., Schneider, T., Kreuzwieser, J., 2023: Accumulation of proline in plants under contaminated soils—Are we on the same page? Antioxidants 12(3), 666. https://doi.org/10.3390/antiox12030666
- Tiwari, B., Kalim, S., Bangar, P., Kumari, R., Kumar, S., Gaikwad, A., Bhat, K.V., 2018: Physiological, biochemical, and molecular responses of thermotolerance in moth bean (Vigna aconitifolia (Jacq.) Marechal). Turkish Journal of Agriculture and Forestry 42 (3), 176-184. https://doi.org/10.3906/tar-1709-1
- Tokyol, A., Turhan, E., 2019: Heat stress tolerance of some green bean (Phaseolus vulgaris L.) genotypes. Scientific Papers. Series A. Agronomy, Vol. LXII, 1, 2019, 472-479.
- Vargas, Y., Mayor-Duran, V. M., Buendia, H. F., Ruiz-Guzman, H., Raatz, B., 2021: Physiological and genetic characterization of heat stress effects in a common bean RIL population. Plos one 16(4), e0249859. https://doi.org/10.1371/journal.pone.0249859
- Wahid, A., Close, T. J., 2007: Expression of dehydrins under heat stress and their relationship with water relations of sugarcane leaves. Biologia Plantarum 51(1), 104–109.
- Xu, Y., Zhan, C., Huang, B., 2011: Heat shock proteins in association with heat tolerance in grasses. International Journal of Proteomics, 1-11. https://doi.org/10.1155/2011/529648
- Zhao, C., Liu, B., Piao, S., Wang, X., Lobell, D.B., Huang, Y., … Asseng, S., 2017: Temperature increase reduces global yields of major crops in four independent estimates. Proceedings of the National Academy of Sciences 114, 35, 9326-9331. https://doi.org/10.1073/pnas.1701762114
- Zhao, J., Lu, Z., Wang, L., Jin, B., 2021: Plant responses to heat stress: physiology, transcription, noncoding RNAs, and epigenetics. International Journal of Molecular Sciences 22(1), 117. https://doi.org/10.3390/ijms22010117
References
Ali, M., Ayyub, C. M., Amjad, M., Ahmad, R., 2019: Evaluation of thermo-tolerance potential in cucumber genotypes under heat stress. Pakistan Journal of Agricultural Sciences 56(1), 53-61.
Arora, R., Pitchay, D.S., Bearce, B.C., 1998: Water stress-induced heat tolerance in geranium leaf tissues: A possible linkage through stress proteins. Physiologia Plantarum 103, 24-34. https://doi.org/10.1034/j.1399-3054.1998.1030104.x
Arora, R., Wisniewski, M. E., 1994: Cold acclimation in genetically related (sibling) deciduous and evergreen peach (Prunus persica [L.] Batsch) (II. A 60-kilodalton bark protein in cold-acclimated tissues of peach is heat stable and related to the dehydrin family of proteins). Plant Physiology 105(1), 95-101. https://doi.org/10.1104/pp.105.1.95
Barr, H.D., Weatherley, P.E., 1962: A Re-examination of the Relative turgidity technique for estimating water deficit in leaves, Australian Journal of Biological Sciences 15, 413-428. http://dx.doi.org/10.1071/BI9620413
Bates, L.S., Waldren, R.P., Teare, I.D., 1973: Rapid determination of free proline for water stress studies. Plant Soil 39, 205-207. https://doi.org/10.1007/BF00018060
Bita CE, Gerats T., 2013: Plant tolerance to high temperature in a changing environment: scientific fundamentals and production of heat stress-tolerant crops. Frontiers in Plant Science 31, 4:273. https://doi.org/10.3389/fpls.2013.00273
Blair, M. W., 2013: Mineral biofortification strategies for food staples: the example of common bean. Journal of Agricultural and Food Chemistry 61(35), 8287-8294. https://doi.org/10.1021/jf400774y
Bradford, M. M., 1976: A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Analytical Biochemistry 72, 248-254.
Chavez-Arias, C.C, Ligarreto-Moreno, G.A., Restrepo-Díaz, H. 2018: Evaluation of heat stress period duration and the interaction of daytime temperature and cultivar on common bean. Environmental and Experimental Botany 155, 600-608. https://doi.org/10.1016/j.envexpbot.2018.08.012
da Silva, D.A., Pinto-Maglio, C.A.F., de Oliveira, E.C., dos Reis, R.L.d.M., Carbonell, S.A.M., Chiorato, A.F. 2020: Influence of high temperature on the reproductive biology of dry edible bean (Phaseolus vulgaris L.). Scientia Agricola 77(3), e20180233. https://doi.org/10.1590/1678-992X-2018-0233
Ergin, S., Gulen, H., Kesici, M., Turhan, E., Ipek, A.N., Köksal, N., 2016: Effects of high temperature stress on enzymatic and nonenzymatic antioxidants and proteins in strawberry plants. Turkish Journal of Agriculture and Forestry 40, 908-917. https://doi.org/10.3906/tar-1606-144
Haq, S.U., Khan, A., Ali, M. et al., 2019: Knockdown of CaHSP60-6 confers enhanced sensitivity to heat stress in pepper (Capsicum annuum L.). Planta 250, 2127–2145. https://doi.org/10.1007/s00425-019-03290-4
Harsh, A., Sharma, Y.K., Joshi, U., Rampuria, S., Singh, G., Kumar, S. Sharma, R., 2016: Effect of short-term heat stress on total sugars, proline and some antioxidant enzymes in moth bean (Vigna aconitifolia), Annals of Agricultural Sciences 61(1): 57-64. https://doi.org/10.1016/j.aoas.2016.02.001
IPCC, Intergovernmental Panel on Climate Change 2023: Climate Change 2023: Synthesis Report. Summary for Policymakers. Contribution of Working Groups I, II and III to the Sixth Assessment Report of the IPCC (Core Writing Team, H. Lee & J. Romero, eds.), Geneva, Switzerland, pp-1-34. https://doi.org/10.59327/IPCC/AR6-9789291691647.001
Jha, U.C.,Priya, M., Naik, Y.D., Nayyar, H., Thudi, M., Punnuri, S.M., Siddique, K. H.M. Prasad, P.V.V., 2024: Major abiotic stresses on quality parameters in grain legumes: Impacts and various strategies for improving quality traits, Environmental and Experimental Botany, 228, 105978. https://doi.org/10.1016/j.envexpbot.2024.105978
Kavi Kishor, P. B., Kumari, P. H., Polavarapu, B., 2022: Intriguing role of proline in redox potential conferring heat stress tolerance in plants. Frontiers in Plant Science 13, 867531. https://doi.org/10.3389/fpls.2022.867531
Kosová, K., Vítámvás, P., Prášil, I.T., Renaut, J 2011: Plant proteome changes under abiotic stress — Contribution of proteomics studies to understanding plant stress response. Journal of Proteomics, 74 (8), 1301-1322. https://doi.org/10.1016/j.jprot.2011.02.006
Kumar, S., Kaur, R., Kaur, N., Bhandhari, K., Kaushal, N., Gupta, K., Bains, T.S., Nayyar, H., 2011: Heat-stress induced inhibition in growth and chlorosis in mungbean (Phaseolus aureus Roxb.) is partly mitigated by ascorbic acid application and is related to reduction in oxidative stress. Acta Physiologiae Plantarum 33(6), 2091–2101. https://doi.org/10.1007/s11738-011-0748-2
Kumar, P., Paul, D., Jhajhriya, S. et al., 2024: Understanding heat-shock proteins’ abundance and pivotal function under multiple abiotic stresses. Journal of Plant Biochemistry and Biotechnology 33, 492-513. https://doi.org/10.1007/s13562-024-00932-x
Kumari, V. V., Roy, A., Vijayan, R., Banerjee, P., Verma, V. C., Nalia, A., & Hossain, A., 2021: Drought and heat stress in cool-season food legumes in sub-tropical regions: Consequences, adaptation, and mitigation strategies. Plants 10(6), 1038. https://doi.org/10.3390/plants10061038
McLoughlin F, Kim M, Marshall RS, Vierstra RD, Vierling E., 2019: HSP101 interacts with the proteasome and promotes the clearance of ubiquitylated protein aggregates. Plant Physiology 180(4), 1829-1847. https://doi.org/10.1104/pp.19.00263
Meena, M., Divyanshu, K., Kumar, S., Swapnil, P., Zehra, A., Shukla, V., Yadav, M., Upadhyay, R.S., 2019: Regulation of L-proline biosynthesis, signal transduction, transport, accumulation and its vital role in plants during variable environmental conditions. Heliyon 5(12), e02952. https://doi.org/10.1016/j.heliyon.2019.e02952
Mullan, D., Pietragalla, J., 2012: Leaf relative water content. Physiological breeding II: a field guide to wheat phenotyping. CIMMYT, Mexico, 25-27.
Niu, Y., Xiang, Y., 2018: An overview of biomembrane functions in plant responses to high-temperature stress. Frontiers in Plant Science 9:915. https://doi.org/10.3389/fpls.2018.00915
Pant, K.K., Naik, J., Barthakur, S., Chandra, V., 2025: High-temperature stress in wheat (Triticum aestivum L.): unfolding the impacts, tolerance and methods to mitigate the detrimental effects. Cereal Research Communications. https://doi.org/10.1007/s42976-025-00634-7
Rani, B., Kumari, N., Jain, V., Dhawan, K., Avtar, R., 2016: Heat stress induced changes in protein profile of Indian mustard (Brassica juncea L.). Journal of Oilseed Brassica 1(1), 302-305. https://epubs.icar.org.in/index.php/JOB/article/view/159031
Raza, A., Charagh, S., Abbas, S., Hassan, M.U., Saeed, F., Haider, S., Sharif, R., Anand, A., Corpas, F.J., Jin, W., Varshney, R.K., 2023: Assessment of proline function in higher plants under extreme temperatures. Plant Biology 25(3), 379-395. https://doi.org/10.1111/plb.13510
Roy, S., Mishra, M., Dhankher, O.P., Singla-Pareek, S.L., Pareek, A., 2019: Molecular Chaperones: Key Players of Abiotic Stress Response in Plants. In: Rajpal, V., Sehgal, D., Kumar, A., Raina, S. (eds) Genetic Enhancement of Crops for Tolerance to Abiotic Stress: Mechanisms and Approaches, Vol. I. Sustainable Development and Biodiversity, vol 20. Springer, Cham. https://doi.org/10.1007/978-3-319-91956-0_6
Scharf, K. D., Berberich, T., Ebersberger, I., Nover, L., 2012: The plant heat stress transcription factor (Hsf) family: structure, function and evolution. Biochimica et Biophysica Acta (BBA)-Gene Regulatory Mechanisms 1819(2), 104-119. https://doi.org/10.1016/j.bbagrm.2011.10.002
Shen, S., Jing, Y., Kuang, T., 2003: Proteomics approach to identify wound-response related proteins from rice leaf sheath, Proteomics 3 (4), 527-535. https://doi.org/10.1002/pmic.200390066
Soltani, A., Weraduwage, S.M., Sharkey, T.D., Lowry, D.B., 2019: Elevated temperatures cause loss of seed set in common bean (Phaseolus vulgaris L.) potentially through the disruption of source-sink relationships. BMC Genomics 20(1), 312. https://doi.org/ 10.1186/s12864-019-5669-2
Soltys-Kalina D, Plich J, Strzelczyk-Żyta D, Śliwka J, Marczewski W., 2016: The effect of drought stress on the leaf relative water content and tuber yield of a half-sib family of 'Katahdin'-derived potato cultivars. Breed Sci. 66(2), 328-31. https://doi.org/10.1270/jsbbs.66.328
Spormann, S., Schneider, T., Kreuzwieser, J., 2023: Accumulation of proline in plants under contaminated soils—Are we on the same page? Antioxidants 12(3), 666. https://doi.org/10.3390/antiox12030666
Tiwari, B., Kalim, S., Bangar, P., Kumari, R., Kumar, S., Gaikwad, A., Bhat, K.V., 2018: Physiological, biochemical, and molecular responses of thermotolerance in moth bean (Vigna aconitifolia (Jacq.) Marechal). Turkish Journal of Agriculture and Forestry 42 (3), 176-184. https://doi.org/10.3906/tar-1709-1
Tokyol, A., Turhan, E., 2019: Heat stress tolerance of some green bean (Phaseolus vulgaris L.) genotypes. Scientific Papers. Series A. Agronomy, Vol. LXII, 1, 2019, 472-479.
Vargas, Y., Mayor-Duran, V. M., Buendia, H. F., Ruiz-Guzman, H., Raatz, B., 2021: Physiological and genetic characterization of heat stress effects in a common bean RIL population. Plos one 16(4), e0249859. https://doi.org/10.1371/journal.pone.0249859
Wahid, A., Close, T. J., 2007: Expression of dehydrins under heat stress and their relationship with water relations of sugarcane leaves. Biologia Plantarum 51(1), 104–109.
Xu, Y., Zhan, C., Huang, B., 2011: Heat shock proteins in association with heat tolerance in grasses. International Journal of Proteomics, 1-11. https://doi.org/10.1155/2011/529648
Zhao, C., Liu, B., Piao, S., Wang, X., Lobell, D.B., Huang, Y., … Asseng, S., 2017: Temperature increase reduces global yields of major crops in four independent estimates. Proceedings of the National Academy of Sciences 114, 35, 9326-9331. https://doi.org/10.1073/pnas.1701762114
Zhao, J., Lu, Z., Wang, L., Jin, B., 2021: Plant responses to heat stress: physiology, transcription, noncoding RNAs, and epigenetics. International Journal of Molecular Sciences 22(1), 117. https://doi.org/10.3390/ijms22010117
