Effects of Water Stress on Carotenoid and Proline Contents in Kale (Brassica oleracea var. acephala) leaves

수분스트레스가 케일 잎의 카로티노이드 및 프롤린 함량에 미치는 영향

  • Lee, Hyo-Joon (Department of Bio-Environmental Chemistry, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Chun, Jin-Hyuk (Department of Bio-Environmental Chemistry, College of Agriculture and Life Sciences, Chungnam National University) ;
  • Kim, Sun-Ju (Department of Bio-Environmental Chemistry, College of Agriculture and Life Sciences, Chungnam National University)
  • 이효준 (충남대학교 농업생명과학대학 생물환경화학과) ;
  • 천진혁 (충남대학교 농업생명과학대학 생물환경화학과) ;
  • 김선주 (충남대학교 농업생명과학대학 생물환경화학과)
  • Received : 2010.05.30
  • Accepted : 2010.06.22
  • Published : 2017.06.30


BACKGROUND : Environmental stress has a major effect on the growth and yields of vegetables, and can significantly affect nutritionally important phytochemicals, causing large economic losses. METHODS AND RESULTS : The present study was aimed at exploring the effects of water stress on the carotenoid and proline contents in kale leaves to understand drought tolerance of kale plants. Kale was randomly divided into two groups at 57 days after sowing (DAS). One of the groups was well-watered (WW) and the other was water stressed (WS). Harvesting of kale leaves was started one day after treatment (58 DAS) and continued for 10 days (~67 DAS). We investigated the status of plant growth (leaf number, length, width, fresh weight) of kale throughout the study. Carotenoid (lutein, ${\alpha}-carotene$, zeaxanthin, ${\beta}-carotene$) and proline contents were analyzed by high-performance liquid chromatography (HPLC). Our results showed that the total carotenoid contents ranged from 926.0 to 1,212.0 mg/kg dry wt. (at 3 and 2 days, respectively) in WW treatment and 887.8 to 1,157.4 mg/kg dry wt. (at 10 and 4 days, respectively) in WS treatment. The ratio of individual carotenoid to the total carotenoid contents of kale leaves was 51.4 for lutein, 4.44 for zeaxanthin, 2.76 for ${\alpha}-carotene$, and 41.4% for ${\beta}-carotene$. Total carotenoid contents showed a significant reduction from 7 days (1,037.2 mg/kg dry wt.) to 10 days (887.8 mg/kg dry wt.) in WS treatment. The lutein content did not show a significant difference in WW between 7 and 10 days after treatment but showed a significant difference in WS treatment. The ${\alpha}-carotene$ content showed no significant difference between the treatments. However, zeaxanthin content was higher during 4-10 days and ${\beta}-carotene$ content was lower during 6-10 days in WS than in WW on each harvest day. In WW, the proline content showed no significant difference, but in WS, the proline content started to increase at 7 days and almost doubled in 10 days. CONCLUSION : The marked increase in zeaxanthin and proline contents in kale leaves indicated that the two phytochemicals are associated with drought tolerance in the plant.


Carotenoids;HPLC analysis;Kale;Proline;Water stress


Supported by : Rural Development Administration


  1. Aktas, L. Y., Turkyilmaz, B., Akca, H., & Parlak, S. (2007). Role of abscisic acid and proline treatment on induction of antioxidant enzyme activities and drought tolerance responses of Laurus nobilis L. seedling. C.U. Fen-Edebiyat Fakultesi Fen Bilimleri Dergisi Cilt 28 Sayi 1.
  2. Balkaya, A., & Yanmaz, R. (2005). Promising kale (Brassica oleracea var. acephala) populations from Black Sea region, Turkey. New Zealand Journal of Crop and Horticultural Science, 33, 1-7.
  3. Bartels, D., & Sunkar, R. (2005). Drought and salt tolerance in plants. Plant Sciences, 24, 23-58.
  4. Bray, E. A. (1997). Plant responses to water deficit. Trends in Plant Science, 2, 48-54.
  5. Chaves, M. M. (1991). Effects of water deficits on carbon assimilation. Journal of Experimental Botany, 42, 1-16.
  6. Delauney, A. J., &Verma, D. P. S. (1993). Proline biosynthesis and osmoregulation in plants. The Plant Journal, 4(2), 215-223.
  7. Demmig-Adams, B., & Adams, W. W. (2002). Antioxidants in photosynthesis and human nutrition. Science, 298, 2149-2153.
  8. Finkelstein, R. R., Gampala, S. S. L, &Rock, C. D. (2002). Abscisic acid signaling in seeds and seedlings. The Plant Cell, 14 (Suppl 1), S15-S45.
  9. Ha, S. H., Jeong, Y. S., Lim, S. H., Kim, J. K., Lee, D. H., Lee, J. Y., & Kim, Y. M. (2012). Carotenoid metabolic engineering in flowering plants. Korean Journal of Science Technology, 30(2), 107-122.
  10. Hare, P. D., Cress, W. A., Staden, J. V. (1998). Dissecting the roles of osmolyte accumulation during stress. Plant, Cell and Environment, 21, 535-553.
  11. Henderson, J. W., Ricker, R. D., Bidlingmeyer, B. A., & Woodward, C. (2000). Rapid, accurate, sensitive, and reproducible HPLC analysis of amino acids. Agilent Technologies Technical Note 5980-1193E.
  12. Heo, J. W., Kim, H. H., Lee, K. J., Yoon, J. B., Lee, J. K., Huh, Y. S., & Lee, K. Y. (2015). Effect of supplementary radiation on growth of greenhouse-grown kales. Korean Journal of Environmental Agriculture, 34(1), 38-45.
  13. Hirschberg, J. (2001). Carotenoid biosynthesis in flowering plants. Current Opinion in Plant Biology, 4, 210-218.
  14. Hong, Y. N. (2009). Introduction of Plant Physiology (4th) (eds. Hopkins, W. G., Huner, N. P. A.), pp. 230, 244, 246-248. World Science Publishing, Seoul, Korea.
  15. Iturbe-Ormaetxe, I., Escuredo, P. R., Arrese-Igor, C., & Becana, M. (1998). Oxidative damage in pea plants exposed to water deficit or paraquat. Plant Physiology, 116, 173-181.
  16. Jaleel, C. A., Manivannan, P., Wahid, A., Farooq, M., Somasundaram, R., & Panneerselvam, R. (2009). Drought stress in plants : A review on morphological characteristics and pigments composition. International Journal of Agriculture and Biology, 11, 100-105.
  17. Jeong, N. R., Chun, J. H., Park, E. J., Lim, Y. H., & Kim, S. J. (2015). Variations of glucosinolates in kale leaves (Brassica oleracea var. acephala) treated with droughtstress in autumn and spring seasons. CNU Journal of Agricultural Science, 42(3), 167-175.
  18. Kang, S. J., & Park, M. (2013). Relationship between relative water content and ascorbate redox enzymes activity in lettuce leaves subjected to soil water stress. Korean Journal of Soil Science and Fertilizer, 46(1), 32-39.
  19. Kim, G. N., & Han, S. H. (2015). Effects on growth, photosynthesis and pigment contents of Liriodendron tulipifera under elevated temperature and drought. Korean Journal of Agricultural and Forest Meteorology, 17(1), 75-84.
  20. Kim, G. N., Han, S. H., Park, G. S. (2014). Differences on growth, photosynthesis and pigment contents of open-pollinated Pinus densiflora families under elevated temperature and drought. Korean Journal of Agricultural and Forest Meteorology, 15(4), 285-296.
  21. Kim, H. K., Chun, J. H., Kim S. J. (2015). Method development and analysis of carotenoid compositions in various tomatoes. Korean Journal of Environmental Agriculture, 34(3), 196-203.
  22. Kishor, P. B. K., Hong, Z., Miao, G. H., Hu, C. A. A., & Verma, D. P. S. (1995). Overexpression of delta-pyrroline-5-carboxylate synthetase increases proline production and confer osmotolerance in transgenic plants. Plant Physiology, 108, 1387-1394.
  23. Kovtun, Y., Chiu, W. L., Tena, G., & Sheen, J. (2000). Functional analysis of oxidative stress-activated mitogen-activated protein kinase cascade in plants. Proceedings of the National Academy of Sciences USA, 97(6), 2940-2945.
  24. Kozlowski, T. T., & Pallardy, S. G. (2002). Acclimation and adaptive responses of woody plants to environmental stresses. The Botanical Review, 68(2), 270-334.[0270:AAAROW]2.0.CO;2
  25. Lefsrud, M., Kopsell, D., Wenzel, A., & Sheehan, J. (2007). Changes in kale (Brassica oleracea L. var. acephala) carotenoid and chlorophyll pigment concentrations during leaf ontogeny. Scientia Horticulturae, 112, 136-141.
  26. Mafakheri, A., Siosemardeh, A., Bahramnejad, B., Struik, P. C., & Sohrabi, Y. (2010). Effect of drought stress on yield, proline and chlorophyll contents in three chickpea cultivars. Australian Journal of Crop Science, 4(8), 580-585.
  27. Mansour, M. F. (1998). Protecion of plasma membrane of onion epidermal cells by glycinebetaine and proline against NaCl stress. Plant Physiology and Biochemistry, 36(10), 767-772.
  28. Oh, C. Y., Han, S. H., Kim, Y. Y., & Lee, J. C. (2005). Changes of drought tolerance and photosynthetic characteristics of Poplulus davidiana dode according to REG concentration. Korean Journal of Agricultural and Forest Meteorology, 7(4), 296-302.
  29. Perng, Z., Lu, Q., & Verma, D. P. S. (1996). Reciprocal regulation of delta 1-pyrroline-5-carboxylate synthetase and proline dehydrogenase genes controls proline levels during and after osmotic stress in plants. Molecular Genetics and Genomics, 253(3), 334-341.
  30. Podse-dek, A. (2007). Natural antioxidants and antioxidant capacity of Brassica vegetables: A review. LWT - Food Science and Technology, 40, 1-11.
  31. Rajendrakumar, C. S., Reddy, B. V., &Reddy, A. R. (1994). Proline-protein interactions: protection of structural and functional integrity of M4 lactate dehydrogenase. Biochemical and biophysical research communications, 201(2), 957-963.
  32. Rudolph, A., Crowe, J. H., &Crowe, L. M. (1986). Effects of three stabilizing agents-proline, betaine, and trehalos-on membrane phospholipids. Archives of Biochemistry and Biophysics, 245(1), 134-143.
  33. Schmidt, S., Zietz, M., Schreiner, M., Rohn, S., & Kroh, L. W., Krumbein, A. (2010). Genotypic and climatic influences on the concentration and composition of flavonoids in kale (Brassica oleracea var. sabellica). Food Chemistry, 119, 1293-1299.
  34. Seki, M., Umezawa, T., Urano, K., & Shinozaki, K. (2007). Regulatory metabolic networks in drought stress responses. Current Opinion in Plant Biology, 10, 296-302.
  35. Seo, M., & Koshiba, T. (2002). Complex regulation of ABA biosynthesis in plants. Trends in Plant Science, 7(1), 41-48.
  36. Verbruggen, N., Hua, X. J., May, M., & Montagu, M. V. (1996). Environmental and developmental signals modulate proline homeostasis: Evidence for a negative transcriptional regulator. Proceedings of the National Academy of Sciences, 93, 8787-8791.
  37. Xiao, X., Xu, X., & Yang, F. (2008). Adaptive responses to progressive drought stress in two Poplulus cathayana populations. Silva Fennica, 42(5), 705-179.
  38. Xiong, L., Wang, R. G., Mao, G,. & Koczan, J. M. (2006). Identification of drought tolerance determinants by genetic analysis of root response to drought stress and abscisic acid. Plant Physiology 142:1065-1074.
  39. Zhu, J. K. (2002). Salt and drought stress signal transduction in plants. Annual Review of Plant Biology, 53, 247-273.