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Journal of Integrative Agriculture  2017, Vol. 16 Issue (10): 2197-2205    DOI: 10.1016/S2095-3119(16)61515-0
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Drought-induced responses of organic osmolytes and proline metabolism during pre-flowering stage in leaves of peanut (Arachis hypogaea L.)
ZHANG Ming, WANG Li-feng, ZHANG Kun, LIU Feng-zhen, WAN Yong-shan
Key Laboratory of Crop Ecophysiology and Farming System, Ministry of Agriculture/College of Agronomy, Shandong Agricultural University,Tai’an 271018, P.R.China
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Abstract  Peanut (Arachis hypogaea L.), an improtant oil crop, usually encounters drought stress in the process of growth and development, especially at pre-flowering stage.  In order to gain insight into the drought tolerance potentials based on osmolyte accumulation and metabolism of proline aspects of peanut, pot experiments were conducted with a split-plot design in Tai’an, Shangdong Province, China in 2013 and 2014.  Pre-flowering drought (PFD) stress and optinum irrigation (control, CK) were served as the main plots and the two peanut cultivars Shanhua 11 and Hua 17 served as sub-plots.  Shanhua 11 was drought-tolerant cultivar and Hua 17 was drought-sensitive.  The content of soluble sugars, soluble protein, free proline and other free amino acids, the activities of enzymes involved in proline metabolism, and malondialdehyde (MDA) content and ion leakage were all investigated in the two cultivars at pre-flowering stage.  Results showed that PFD stress significantly increased the levels of soluble protein, free proline and free amino acid, and increased Δ1-pyrroline-5-carboxylate synthetase (P-5-CS, EC activity in the leaves of drought-tolerant and drought-sensitive cultivars.  The activity of proline dehydrogenase (proDH) (EC decreased under PFD stress in both cultivars.  The leaves of the tolerant cultivar maintained higher increments of osmolyte levels, lower increments of MDA content and ion leakage, as well as a higher increased proportion of P-5-CS activity and higher inhibited proportion of proDH activity under water stress compared with the drought-sensitive cultivar.  The study suggests that proline accumulation in peanut leaves under PFD can be explained by the higher enhanced activities of P-5-CS and higher inhibition of proDH.  The results will provide useful information for genetic improvement of peanut under drought tolerance.
Keywords:  drought stress        peanut (Arachis hypogaea L.)        Δ1-pyrroline-5-carboxylate synthetase (P-5-CS)        δ-ornithine transaminase (OAT)        proline dehydrogenase (proDH)  
Received: 22 September 2016   Accepted:

We acknowledge financial support from the National Natural Science Foundation of China (31201167), the earmarked foud for the China Agriculture Research System (CARS-14), and Taishan Scholar Seed Industry Projects in Shandong Province, China (Shandong [2014] 126).

Corresponding Authors:  Correspondence WAN Yong-shan, Tel/Fax: +86-538-8241540, E-mail:    
About author:  ZHANG Ming, E-mail:;

Cite this article: 

ZHANG Ming, WANG Li-feng, ZHANG Kun, LIU Feng-zhen, WAN Yong-shan. 2017. Drought-induced responses of organic osmolytes and proline metabolism during pre-flowering stage in leaves of peanut (Arachis hypogaea L.). Journal of Integrative Agriculture, 16(10): 2197-2205.

Ahmed I M, Dai H, Zheng W, Cao F, Zhang G, Sun D, Wu F. 2013. Genotypic differences in physiological characteristics in the tolerance to drought and salinity combined stress between Tibetan wild and cultivated barley. Plant Physiology and Biochemistry, 63, 49–60.

Ashraf M, Harris P J C. 2004. Potential biochemical indicators of salinity tolerance in plants. Plant Science, 166, 3–16.

Bates L S, Waldren R P, Teare I D. 1973. Rapid determination of free proline for water-stress studies. Plant and Soil, 39, 205–207.

Bradford M M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254.

Chaitanya K V, Rasineni G K, Reddy A R. 2009. Biochemical responses to drought stress in mulberry (Morus alba L.): evaluation of proline, glycine betaine and abscisic acid accumulation in five cultivars. Acta Physiologiae Plantarum, 31, 437–443.

Charest C, Phan C T. 1990. Cold acclimation of wheat (Triticum aestivum): Properties of enzymes involved in proline metabolism. Physiologia Plantarum, 80, 159–168.

Clifford S C, Arndt S K, Corlett J E, Joshi S, Sankhla N, Popp M, Jones H G. 1998. The role of solute accumulation, osmotic adjustment and changes in cell wall elasticity in drought tolerance in Ziziphus Mauritiana (Lamk.). Journal of Experimental Botany, 49, 967–977.

Efeo?lu B, Ekmekci Y, Cicek N. 2009. Physiological responses of three maize cultivars to drought stress and recovery. South African Journal of Botany, 75, 34–42.

Gunes A, Inal A, Adak MS, Bagci E G, Cicek N, Eraslan F. 2008. Effect of drought stress implemented at pre- or post-anthesis stage on some physiological parameters as screening criteria in chickpea cultivars. Russian Journal of Plant Physiology, 55, 59–67.

Hernández J A, Almansa M S. 2002. Short-term effects of salt stress on antioxidant systems and leaf water relations of pea leaves. Physiologia Plantarum, 115, 251–257.

Hessini K, Martínez J P, Gandour M, Albouchi A, Soltani A, Abdelly C. 2009. Effect of water stress on growth, osmotic adjustment, cell wall elasticity and water-use efficiency in Spartina alterniflora. Environmental and Experimental Botany, 67, 312–319.

Hoekstra F A, Golovina E A, Buitink J. 2001. Mechanisms of plant desiccation tolerance. Trends in Plant Science, 6, 431–438.

Iqbal N, Umar S, Khan N A, Khan M I R. 2014. A new perspective of phytohormones in salinity tolerance: regulation of proline metabolism. Environmental and Experimental Botany, 100, 34–42.

Jiang Y W, Huang B R. 2002. Protein alterations in tall fescue in response to drought stress and abscisic acid. Crop Science, 42, 202–207.

Kandpal R P, Vaidyanathan C S, Kumar M U, Sastry K S K, Rao N A. 1981. Alterations in the activities of the enzymes of proline metabolism in Ragi (Eleusine coracana) leaves during water stress. Journal of Biosciences, 3, 361–370.

Khan M I R, Iqbal N, Masood A, Khan N A. 2012. Variation in salt tolerance of wheat cultivars: role of glycinebetaine and ethylene. Pedosphere, 22, 746–754.

Klíma M, Vítámvás P, Zelenková S, Vyvadilová M, Prášil I T. 2012. Dehydrin and proline content in Brassica napus and B. carinata under cold stress at two irradiances. Biologia Plantarum, 56, 157–161.

Kocheva K, Lambrev P, Georgiev G, Goltsev V, Karabaliev M. 2004. Evaluation of chlorophyll fluorescence and membrane injury in the leaves of barley cultivars under osmotic stress. Bioelectrochemistry, 63, 121–124.

Kumar S G, Reddy A M, Sudhakar C. 2003. NaCl effects on proline metabolism in two high yielding genotypes of mulberry (Morus alba L.) with contrasting salt tolerance. Plant Science, 165, 1245–1251.

Liu C C, Liu Y G, Guo K, Fan D Y, Li G Q, Zheng Y R, Yu L F, Yang R. 2011. Effect of drought on pigments, osmotic adjustment and antioxidant enzymes in six woody plant species in karst habitats of southwestern China. Environmental and Experimental Botany, 71, 174–183.

Mardeh A S, Ahmadi A, Poustini K, Mohammadi V. 2006. Evaluation of drought resistance indices under various environmental conditions. Field Crops Research, 98, 222-229.

McCue K F, Hanson A D. 1990. Drought and salt tolerance: towards understanding and application. Trends in Biotechnology, 8, 358–362.

Misra N, Gupta A K. 2005. Effect of salt stress on proline metabolism in two high yielding genotypes of green gram. Plant Science, 169, 331–339.

Moore S, Stein W H. 1954. A modified ninhydrin reagent for the photometric determination of amino acids and related compounds. Journal of Biological Chemistry, 211, 907–913.

Morales C G, Pino M T, Pozo A D. 2013. Phenological and physiological responses to drought stress and subsequent rehydration cycles in two raspberry cultivars. Scientia Horticulturae, 162, 234–241.

Mundree S G, Baker B, Mowla S, Peters S, Marais S, Willigen C V, Govender K, Maredza A, Muyanga S, Farrant J M. 2002. Physiological and molecular insights into drought tolerance. African Journal of Biotechnology, 1, 28–38.

Nayyar H. 2003. Accumulation of osmolytes and osmotic adjustment in water-stressed wheat (Triticum aestivum) and maize (Zea mays) as affected by calcium and its antagonists. Environmental and Experimental Botany, 50, 253–264.

Nazar R, Iqbal N, Syeed S, Khan N A. 2011. Salicylic acid alleviates decreases in photosynthesis under salt stress by enhancing nitrogen and sulfur assimilation and antioxidant metabolism differentially in two mungbean cultivars. Journal of Plant Physiology, 168, 807–815.

Padmavathi T A V, Rao D M. 2013. Differential accumulation of osmolytes in 4 cultivars of peanut (Arachis hypogaea L.) under drought stress. Journal of Crop Science and Biotechnology, 16, 151–159.

Parida A K, Dagaonkar V S, Phalak M S, Aurangabadkar L P. 2008. Differential responses of the enzymes involved in proline biosynthesis and degradation in drought tolerant and sensitive cotton genotypes during drought stress and recovery. Acta Physiologiae Plantarum, 30, 619–627.

Parida A K, Dagaonkar V S, Phalak M S, Umalkar G V, Aurangabadkar L P. 2007. Alterations in photosynthetic pigments, protein and osmotic components in cotton genotypes subjected to short-term drought stress followed by recovery. Plant Biotechnology Reports, 1, 37–48.

Parida A K, Jha B. 2013a. Inductive responses of some organic metabolites for osmotic homeostasis in peanut (Arachis hypogaea L.) seedlings during salt stress. Acta Physiologiae Plantarum, 35, 2821–2832.

Parida A K, Jha B. 2013b. Physiological and biochemical responses reveal the drought tolerance efficacy of the halophyte Salicornia brachiata. Journal of Plant Growth Regulation, 32, 342–352.

Patakas A, Nikolaou N, Zioziou E, Radogloub K, Noitsakisc B. 2002. The role of organic solute and ion accumulation in osmotic adjustment in drought-stressed grapevines. Plant Science, 163, 361–367.

Pimratch S, Jogloy S, Vorasoot N, Toomsan B, Patanothai A, Holbrook C C. 2008. Relationship between biomass production and nitrogen fixation under drought-stress conditions in peanut genotypes with different levels of drought resistance. Journal of Agronomy and Crop Science, 194, 15–25.

Rai V K. 2002. Role of amino acids in plant responses to stresses. Biologia Plantarum, 45, 481–487.

Ramanjulu S, Bartels D. 2002. Drought and desiccation induced modulation of gene expression in plants. Plant, Cell & Environment, 25, 141–151.

Ramanjulu S, Sudhakar C. 2000. Proline metabolism during dehydration in two mulberry genotypes with contrasting drought tolerance. Journal of Plant Physiology, 157, 81–85.

Reddy A R, Chaitanya K V, Jutur P P, Sumithra K. 2004. Differential antioxidative responses to water stress among five mulberry (Morus alba L.) cultivars. Environmental and Experimental Botany, 52, 33–42.

Rosas-Anderson P, Shekoofa A, Sinclair T R, Balota M, Isleib T G, Tallury S, Rufty T. 2014. Genetic variation in peanut leaf maintenance and transpiration recovery from severe soil drying. Field Crops Research, 158, 65–72.

Roosens N H, Thu T T, Iskandar H M, Iskandar H M, Jacobs M. 1998. Isolation of the ornithine-δ-aminotransferase cDNA and effect of salt stress on its expression in Arabidopsis thaliana. Plant Physiology, 117, 263–271.

Sakuraba H, Takamatsu Y, Satomura T, Kawakami R, Ohshima T. 2001. Purification, characterization, and application of a novel dye-linked L-proline dehydrogenase from a hyperthermophilic archaeon, Thermococcus profundus.Applied and Environmental Microbiology, 67, 1470–1475.

Sánchez F J, Manzanares M, Andres E F D, Tenorio J L, Ayerbe L. 1998. Turgor maintenance, osmotic adjustment and soluble sugar and proline accumulation in 49 pea cultivars in response to water stress. Field Crops Research, 59, 225–235.

Serraj R, Sinclair T R. 2002. Osmolyte accumulation: can it really help increase crop yield under drought conditions? Plant, Cell & Environment, 25, 333–341.

Sheen J. 1999. C4 gene expression. Annual Review of Plant Biology, 50, 187–217.

Sperdouli I, Moustakas M. 2012. Interaction of proline, sugars, and anthocyanins during photosynthetic acclimation of Arabidopsis thaliana to drought stress. Journal of Plant Physiology, 169, 577–585.

Sumithra K, Jutur P P, Carmel B D, Reddy A R. 2006. Salinity-induced changes in two cultivars of Vigna radiata: responses of antioxidative and proline metabolism. Plant Growth Regulation, 50, 11–22.

Toscano S, Scuderi D, Giuffrida F, Romano D. 2014. Responses of Mediterranean ornamental shrubs to drought stress and recovery. Scientia Horticulturae, 178, 145–153.

Valentovi? P, Luxová M, Kolarovi? L, O.Gašparíková O. 2006. Effect of osmotic stress on compatible solutes content, membrane stability and water relations in two maize cultivars. Plant Soil and Environment, 52, 186–191.

Vendruscolo E C G, Schuster I, Pileggi M, Scapim C A, Molinari H B C, Marur C J, Vieira L G E. 2007. Stress-induced synthesis of proline confers tolerance to water deficit in transgenic wheat. Journal of Plant Physiology, 164, 1367–1376.

Verslues P E, Sharp R E. 1999. Proline accumulation in maize (Zea mays L.) primary roots at low water potentials. II. Metabolic source of increased proline deposition in the elongation zone. Plant Physiology, 119, 1349–1360.

Wang H Z, Ma J, Li X Y, Li Y, Zhang R P, Wang R Q. 2007. Relationship between some physiological and biochemical characteristics and drought tolerance at rice flowering stage. Scientia Agricultura Sinica, 40, 188–193. (in Chinese)

Xu Z Z, Zhou G S, Wang Y L, Han G X, Li Y J. 2008. Changes in chlorophyll fluorescence in maize plants with imposed rapid dehydration at different leaf ages. Journal of Plant Growth Regulation, 27, 83–92.

Xue X N, Liu A H, Hua X J. 2009. Proline accumulation and transcriptional regulation of proline biothesynthesis and degradation in Brassica napus. Biochemistry and Molecular Biology Reports, 42, 28–34.

Yemm E W, Willis A J. 1954. The estimation of carbohydrates in plant extracts by anthrone. Biochemical Journal, 57, 508.

Yoshiba Y, Kiyosue T, Nakashima K, Yamaguchi-Shinozaki K, Shinozaki K. 1997. Regulation of levels of proline as an osmolyte in plants under water stress. Plant and Cell Physiology, 38, 1095–1102.

Zhang C S, Lu Q, Verma D P S. 1995. Removal of feedback inhibition of Δ1-pyrroline-5-carboxylate synthetase, a bifunctional enzyme catalyzing the first two steps of proline biosynthesis in plants. Journal of Biological Chemistry, 270, 20491–20496.

Zhang H M, Zhang L S, Liu L, Zhu W N, Yang W B. 2013. Changes of dehydrin profiles induced by drought in winter wheat at different developmental stages. Biologia Plantarum, 57, 797–800.

Zhou Y H, Lam H M, Zhang J H. 2007. Inhibition of photosynthesis and energy dissipation induced by water and high light stresses in rice. Journal of Experimental Botany, 58, 1207–1217.
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