CC BY
705
Journal of Stress Physiology & Biochemistry, Vol. 8 No. 1 2012, pp. 182-193 ISSN 1997-0838 Original Text Copyright © 2012 by Rahdari, Tavakoli and Hosseini
ORIGINAL ARTICLE
Studying of Salinity Stress Effect on Germination, Proline, Sugar, Protein, Lipid and Chlorophyll Content in Purslane (Portulaca oleracea L.) Leaves
Parvaneh Rahdari1*, Shahrokh Tavakoli2 and Seyed Meysam Hosseini2
1 Department of Biology, Islamic Azad University, Tonekabon Branch, Iran.
2 MSc Plant Physiology, Islamic Azad University, Tonekabon Branch, Iran.
*E mail: Rahdari_parvaneh@yahoo.com
Received December 21, 2011
Approximately 30% of country regions are under influence of salt and one of the main important methods for increasing of hectare performance in salt soils, using of resistant plant against with salt such as medicinal annual plant of Purslane (Portulaca oleraceae L.).Studying of the salt stress effect on germination and some physiological parameters in Purslane was carried out in completely random framework and with three replications and under controlled conditions. In this study, salinity was imposed by sodium chloride (NaCl) and in five levels of framework included of Control (distilled water), 50, 100, 150 and 200 Mmol from NaCl. Studied physiological parameters included of proline, sugar, protein, lipid and chlorophyll in Purslane leaves.In relation with observed germination that increasing of salt level, germination rate was reduced that the least germination rate in 200 Mmol and the highest germination was observed in control which in statistical level 0.01 showed meaningful difference. Also, results showed an increasing of leaf proline and sugar that was synchronize with increasing of sodium chloride, this increasing in statistical level of P<0.01 was meaningful, while blank had the least rate and salt level 200 Mmol was observed. Synchronize with increasing of salinity level, lipid and protein concentration decreasing that was meaningful (P<0.01) which highest of leaf lipid and protein content in control level and least concentration was observed in 200 Mmol of salinity level. Also, salinity stress due to increasing of chlorophyll a and b levels in Purslane leaves which this increasing in leaf chlorophyll b concentrations in 0.01 statistical level was meaningful and the least level chlorophyll a and b in blank and the highest level of chlorophyll a and b by order in 150 and 200 Mmol of NaCl was observed. The results showed that plant for resistance against salinity increasing of proline, sugar and chlorophyll content in leaves are useful for retaining and improving of photosynthesis quality in plants.
Key words: Purslane /Salinity / Carbohydrate /Lipid/Protein
ORIGINAL ARTICLE
Studying of Salinity Stress Effect on Germination, Proline, Sugar, Protein, Lipid and Chlorophyll Content in Purslane (Portulaca oleracea L.) Leaves
Parvaneh Rahdari1*, Shahrokh Tavakoli2 and Seyed Meysam Hosseini2
1 Department of Biology, Islamic Azad University, Tonekabon Branch, Iran.
2 MSc Plant Physiology, Islamic Azad University, Tonekabon Branch, Iran.
*E mail: Rahdari_parvaneh@yahoo.com
Received December 21, 2011
Approximately 30% of country regions are under influence of salt and one of the main important methods for increasing of hectare performance in salt soils, using of resistant plant against with salt such as medicinal annual plant of Purslane (Portulaca oleraceae L.).Studying of the salt stress effect on germination and some physiological parameters in Purslane was carried out in completely random framework and with three replications and under controlled conditions. In this study, salinity was imposed by sodium chloride (NaCl) and in five levels of framework included of Control (distilled water), 50, 100, 150 and 200 Mmol from NaCl. Studied physiological parameters included of proline, sugar, protein, lipid and chlorophyll in Purslane leaves.In relation with observed germination that increasing of salt level, germination rate was reduced that the least germination rate in 200 Mmol and the highest germination was observed in control which in statistical level 0.01 showed meaningful difference. Also, results showed an increasing of leaf proline and sugar that was synchronize with increasing of sodium chloride, this increasing in statistical level of P<0.01 was meaningful, while blank had the least rate and salt level 200 Mmol was observed. Synchronize with increasing of salinity level, lipid and protein concentration decreasing that was meaningful (P<0.01) which highest of leaf lipid and protein content in control level and least concentration was observed in 200 Mmol of salinity level. Also, salinity stress due to increasing of chlorophyll a and b levels in Purslane leaves which this increasing in leaf chlorophyll b concentrations in 0.01 statistical level was meaningful and the least level chlorophyll a and b in blank and the highest level of chlorophyll a and b by order in 150 and 200 Mmol of NaCl was observed. The results showed that plant for resistance against salinity increasing of proline, sugar and chlorophyll content in leaves are useful for retaining and improving of photosynthesis quality in plants.
Key words: Purslane / Salinity / Carbohydrate /Lipid /Protein
One of the most important features of soil and water in agriculture is salinity which it due to presence of more ions (Etesami et al, 2007). Approximately 30% of Iran regions are more than
17 million hectares under influence of salt (Ghassemi et al, 1995), and using of resistant plant against with salinity one of the most effective methods in exploiting and increasing hectares
performance in salt and low-salt soils of arid and semi-arid of the world (Khalesro and Agha alikhani,
2006). Some results mentioned that salinity decrease of sugars store and as a result it can distort the respiration metabolism of embryo growth (Kazerouni, 2005). Salinity with osmosis potential solution reduction can reduce plant growth. It's reason is that alien with water molecules existence in environment, there are ions that preventing of water absorption by plants root and plant face with a water deficit (Tobe et al, 1999). With respect to more than 60 millions hectare from the world regions (25%) of regions in all over the world) face with salinity problem, doing of project in along with decreasing of salt soil damage is necessary (Taffouo et al, 2010). Purslane (Portulaca oleraceae L.) is an annual plant and from Portulacaceae family which it's current name was Purslane are using for reclamation of salt soils. This plant included of much values of Omega-3 fatty acids,hence it is one of the most valuable medical plants in the world.It used as an antiseptic, antisposmodic, diuretic, antipyretic, muscle loose, antioxidant, reinforcing of safety system, blood filtering and for removing of thirsty has medical application (Schuman, 2001).It had a global distribution and taxonomy by eight current plants groups in the world (Liu et al, 2000).It was a main weed for 45 cultivating types in 81 countries of the worlds (Holm et al, 1977). Life cycle of this plant during of 2 to 4 months will complete (Chauhan and Johnson, 2007). Photosynthesis mechanism devoted to C4 and had the ability of converting to photosynthesis mechanism CAM that this factor in great extent is reason of resistance and consistency of Purslane proportional to environmental stresses
(Koch and Kennedy, 1981).With respect to Purslane as a valuable medical plant and consistence with environmental condition and few information in relation with this plant reaction to salinity, performing of this study necessary and important. MATERIALS AND METHODS
For studying of salinity stress effect on germination and physiological factors of Purslane seeds were prepared in Iran Jungle and Pasture Research Institute,Iran. Related studies was carried out in 2010 at Islamic Azad University of Tonekabon under controlled condition. Seeds was put in 5% sodium hypochlorite solution of for 1 minute and then by distilled water washed for antiseptic of it (Falahati, 2006). Seeds was cultivated in petri dish, seedling transported in washed sand pot for controlling of their growth after 3 leaves stage. Hogland solution was added that get solution process performed every one week (table 1). After multi-leaves stage in seedlings, stress for one week was performed then physiological parameters was evaluated. Salinity stress in five levels and by imposing sodium chloride (NaCl); Blank(distilled water), 50, 100,150 and 200 Mmol from NaCl. After passing of stress period, proline was evaluated by Bates method (1973), sugar by Nelson method (1943),protein by Lowery method (1951), chlorophyll by Lichtenthaler method (1987) and for evaluating of lipid by sokseleh set was applied. Experiments in completely random framework and with three replications was performed. Data Analysis,variance and comparison of mean by Dunkan experiment and SPSS software was done and drawing of diagrams was used from Excel software.
Table 1. Contents of nutrient solution used for irrigation of Purslane seedlings
main solution (g/L) Resource Element name
59.025 Ca ( NO3)2.4H 2O Calcium nitrate
25.275 KNO3 Potassium nitrate
61.6 MgSO 4.7 H 2 O Magnesium sulfate
34.025 KH2PO4 Potassium di hydrogen phosphate
0/715 H 3BO3 Boric acid
0/4525 MnCl 2.4 H 2O Manganese chloride
0.055 ZnSO4.5H 2O Zinc sulfate
0.02 CuSO4.5H 2 O Copper sulfate
0.0225 H 2 MoO4. H 2 O Molybdic acid
RESULTS
Results showed that meaningful effect of salinity stress on germination, proline, sugar, leaf lipid, protein and chlorophyll contents in Purslane leaves(P<0.01). In relation with germination percent observed which imposed salinity level occurred meaningful decreasing while the highest observed germination percent in blank and it's least in salinity level 200 Mmol was exist(Figure 1). The free proline level in Purslane leaves with salinity level increasing,had meaningful increasing which the highest rate of proline was observed in 200 Mmol of salinity level and the least proline concentration observed in blank (Figure2).Leaf sugar concentration with increasing of applied NaCl level, increased but didn't see any meaningful difference in leaf carbohydrates concentration in 150 Mmol level, but in 200 Mmol salinity level we can find meaningful was increased which meaningful
difference was recorded with blank level (Figure3).Decreasing of leaf proteins concentrations synchronize with increasing of salinity stress was meaningful, while the highest rate of proteins concentration in blank and the least of proteins rate observed in 200 Mmol level of salinity stress (Figure4). In relation with leaf lipid content observed that with over heading of imposed salinity level,meaningful was decreased occurred in lipid concentrations, while the lowest rate of lipids in 200 Mmol of salinity level and the highest level of that observed in blank (Figure5).But out of expect salinity stress imposing on Purslane due to increasing of chlorophyll a,b in leaves. In relation with chlorophyll a, the highest concentration observed in 150 Mmol of salinity level and the lowest of that was observed in blank and the least chlorophyll b content was observed in blank and highest concentration of that in 200 Mmol of salinity stress were observed ( Figure 6&7).
DISCUSSION
Different studies showed that different levels of salinity stress on germination in a variety of plants which can infer to meaningful decreasing on Vigna radiate L. and Lens culinaris (Kazerouni et al, 2005), Elymus Junceus (Askarian, 2004), forage sorghum and pearl millet (Khalesro et al, 2006). Mainly salinity stress by low osmosis potential and ionic poisonous can reduce germination and for this reason it can influence on germination percent more than drought (Alebrahim et al, 2008).Under salinity stress,plants have mechanisms against with that which accumulation of solution components such as proline one of the primary responses of plant proportion to salinity (Fedina et al, 2002; Yeo,1998). Yazici et al in 2007 was performed of Purslane, he was observed that with increasing of salinity imposed, free proline content in leaves was increased which confirm this result. In other research on barley (Hordeum vulgare L.)was
observed that proline as a reducer component of osmosis pressure in response to increased of salinity (Ueda et al, 2007). Also, increasing of proline content in cotton (Desingh et al, 2007), Paulownia imperialis (Astorga et al, 2010) and wheat (Khan et al, 2009) was synchronised with increasing of salinity level is reported. Increasing of proline level under salinity stress was recognised due to that proline is consistent smolite, macromolecules conserver and remover of active oxygen producing in during of environmental stress(Desingh et al,
2007). Studies showed that proline accumulation in plants was performed by different methods(Yordanov et al, 2003):
1) Stimulating of synthesis from pre-material
2) Reducing of Proline Oxidas activity.
3) Proteins destruction
4) Reducing of proteins structure partnership.
In this research, proteins concentrations with increasing of salinity level showed reduction, of
course in relation with salinity effect on protein synthesis, showed different results which depended on plant species. Terminal content decreasing of solution protein in plants such as Helianthus annuus and Colcus blumei (Yu et al, 1999), Vicica faba and Amaranthus (Doganlar et al, 2010), and also increasing of protein level in plants such as cotton (Jiang et al, 2005), Pancratium maritimum (Khedr et al, 2003) under salinity stress was reported. Vibration in protein solution terminal content, was resourced synthesis of stress proteins (Doganler et al, 2010). Different of physiological studies was offered which under salinity condition of non-structural carbohydrate such as Sucrose and Hexanes accumulated with the rate of this accumulation and concentration increasing in different plant species can be different. This increasing of solution carbohydrates concentration as a result of increasing of starch hydrolyze which it needs to hydrolytic enzymes activity (Bartels et al, 2005). Different studies in plants with salinity stress increasing, sucrose santase enzymes activity increasing and invertase enzymes activity was reduced. Sugar is the main molecule symptoms which influence on different physiological response in regulated genes in photosynthesis, metabolism and defensive Reponses (Crowe et al, 2002). Carbohydrate accumulation in plants tissue under condition of environmental stress due to regulating and osmosis adjustment in current stress (Dhanapackiam & Ilyas, 2010).Increasing of sugar content under condition of salinity stress in such plants as tomato(Amini & Ehsanpour, 2005) and barley (Bagheri & Sadeghipour, 2009) was reported. increasing of sugar under environmental stress was recognized as a result of starch degradation, sugar synthesis by non-photosynthesis pathways, non-converting of these components to other productions and
decreasing of transporting from leaves (Premachandre et al, 1991). Lipids and proteins are major components in membrane which has main role in plant cell resistance in proportional to environmental stress. Environmental stress due to disordering in cohesions of membrane lipids and proteins (Yordanov et al, 2003). Under condition of stress main changes will happen in lipids metabolism (Kesri, 2002). Mono galactosyl di glyceride (MGDG) is main glycol lipid in leaf was effect of intensive stress imposing, was reduced that is express of chloroplast membrane destructions (Yordanov et al, 2003).In other hand, lipid peroxidation was synchronized with increased of the salinity level which had a relation with plants such as Wheat (Hala et al, 2005), Tomato (Neumann, 2001) and Purslane (Yazici et al, 2007) was reported. With changing of leaf chlorophyll content by salinity stress, terminal photosynthesis level, distillation speed and stomatal transport was under control meaningfully (Doganlar et al, 2010).Content reduction of chlorophyll in plants such as Poulownia imperialis (Astorga et al, 2010),Bean (Beinsan et al, 2003) and Carthamus tinctorius (Siddiqi et al, 2009) was reported. The cause of this reduction was the increasing of destructive enzymes called chlorophyllase. Photosynthesis pigments content under condition of salinity will not reduced and some reports due to increasing of leaf chlorophyll level under stress. Increasing of leaf chlorophyll content under salinity stress was reported by Pinheiro et al in 2008 on Ricinus Communis and also by Jamil et al in 2007 on Beta vulgaris L. Mainly the reason increasing of leaf chlorophyll content under salinity stress as a result of the increasing of chloroplast in leaves under stress which occurred for maintaining of plant photosynthesis that from symptoms of plant
resistance in proportion to environmental stress. non-decreasing of chlorophyll content in stress condition expressing of plant resistance against with light damages of chloroplast, while a positive relationship was reported between anti-oxidant enzymes and conserving rate of chlorophyll(Yang et al, 2006). As a result, obtained conclusions symbolic of high resistance of medical plant Purslane are proportional to environmental stresses that can be as a medical plant in low-rain regions and with water limitation and salt regions was respected. ACKNOWLEDGMENTS
This work was supported by Islamic Azad University of Tonekabon Branch of Iran.
REFERENCES
Alebrahim, M.T., Janmohammadi, M., Sharifzade, F.,and Tokasi, S. (2008) Evaluation of salinity and drought stress effects on germination and early growth of Maize inbred lines (Zea Mays L.). EJCP. 1(2): 35-43.
Amini, F., and Ehsanpour, A.A. (2005) Soluble proteins, proline, carbohydrates, and Na+/k+ changes in two tomato (Lycopersicon esculentum Mill.) Cultivars under invitro salt stress. American Journal of Biochemistry and Biotechnology. 1(4): 212-216.
Askarian, M. (2004) The effects of salinity and dryness on germination and seedling establishment in Elymus junceus and Kochia prostrate. Pajouhesh and Sazandegi. J. 64: 7177.
Astorga, G.I., and Melendez, L.A. (2010) Salinity effects on protein content, lipid peroxidation, pigments,and proline in Poulownia imperialis and Paulownia fortune grown in vitro. Electronic Journal of Bitechnology. 13(5): 115.
Bagheri, A., and Sadeghipour, O. (2009) Effects of salt stress on yield, yield components and carbohydrates content in four Hull less Barley
(Hordeum Vulgar L.) cultivars. Journal of Biological sciences. 9(8): 909-912.
Bartels, D., and Ramanjulu, S. (2005) Drought and salt tolerance in plants. Critical Reviews in Plant Sciences. 24: 23-58.
Bates, L.S., Waldren, R.P, and Teare, I.D. (1973) Rapid determination of free proline for water stress studies. Plant and Soil. 39: 205-208.
Beinsan, C., Camen, D., Sumalan, R., and Babau, M. (2003) Study concerning salt stress effect on leaf area dynamics and chlorophyll content in four bean local landraces from Banat area. Faculty of Horticulture. 119: 416-419.
Chauhan, B., and Johnson, D.E.(2007) Germination biology of Portulaca oleracea L. Crop and Environmental Seiences, Division, IRRI.
Crowe, J.H., Carpenter, J.F., Crowe, L.M.,
Anchordouguy, T.J.(2002) Are freezing and dehydration similar stress vectors? A
Comparison of models interaction of stabilizing solutes with biomolecules. Cryobiol, 27 : 219231.
Desingh, R., Kanagaraj, G. (2007) Influence of salinity stress on photosynthesis and
antioxidative systems in two cotton varieties. Gen Appl. Plant Physiol. 33(3-4): 221-234.
Dhanapackiam, S., Ilyas, M. (2010) Effect of salinity on chlorophyll and carbohydrate contents of Sesbania grandiflora seedlings. Indian Journal of Science and Technology. 3(1): 64-66.
Doganlar, Z.B., Demir, K, Basak H., and Gul, I. (2010) Effects of salt stress on pigment and total soluble protein contents of Three different
tomato Cultivars. African Journal of Agricultural Research. 5(15) : 2056-2065.
Etesami, M., and Galeshi, S.(2007) A evaluation reaction of ten genotype of barley in salinity on germination and seedling growth (Hordeum vulgar L.). J. Agric. Sci. Natur. Resour. 15(5): 1-7.
Falahati, A. (2006) Evaluation effect of gamma radiation on increase drought resistance in Rice (oryza Sativa) with invitro. MS.C Thesis. Ferdowsi University of Mashhad. Iran.
Fedina, I.S, Georgieva, K., and Grigorova, I. (2002) Light- Dark changes in proline content of barley leaves under salt stress. Biologia Plantarum. 45(1) : 59-63.
Ghassemi, F., Jackman, A.J., and Nix, H.A. (1995) Salinisation of land and water resources: Human causes extend, management and case studies. UNSW press, Sydney, Australia, and CAB International, Wallingford, UK.
Hala, M., El-Bassiouny, S., and Bekheta, M.A. (2005) Effect of salt stress on relative water content, lipid peroxidation, polyamines, amino acids, and ethylene of two wheat cultivars. International Journal of Agriculture and Biology. 3: 363-368.
Holm, L.G., Plucknett, D.L, Pancho, J.V., and Herberger, J.P. (1977) 'The worlds worst weeds:distribution and biology'. (University of Hawaii Press, Honolulu, Hawaii USA).
Jamil, M., Lee, D.B., Jung, K., Ashraf, M., Lee, S.H., and Rha, E.S.( 2007) Effect of salt stress on germination and early seedling growth of four vegetables species. J. Cent. Eur. Agric. 7 : 273-281.
Jiang, L., Duan, L., Tian, X., Wang, B, and Li, Z. (2005) NaCl salinity stress decreased Bacillus
thuringiensis (Bt) protein content of transgenic Bt cotton (Gossypium Hirsutum L.) seedlings. Environmental and Experimental Botany. 55 (3) : 315-320.
Kazerouni, E., Akramian, M., Tokassi, S., and Eghbali, S. (2005) Physiological effects of salt and drought stresses on germination and seedling growth of lentil (Lens culinaris) and mungbean (Vigna radiate L.). Horticultural. J. 445-448.
Kesri, G., Aid, F., Demandre, C., Kader, J.C, and Mazliak, P. (2002) Drought stress affects chloroplast lipid metabolism in rape (Brassica napus) leaves. Physiol Plant. 115: 221-227.
Khalesro, S., and Aghaalikhani, M. (2006) Effect of salinity and drought stress on seed germination in forage sorghum (Sorghum bicolor L.) and Pearl millet (Pennisetum americanum L.) Horticultural. J. 77 : 153-163.
Khan, M.A., Shirazi, M.U., Alikhan, M., and Ashraf, M. (2009) Role of proline, K/Na ratio and chlorophyll content in salt tolerance of wheat (Triticum aestivum L.). Pak. J. Bot. 41(2) : 633638.
Khedr, A.H., Abbas, A., Wahid, M.A., Quick, W.P., Abogadallah, G.M. (2003) Proline induces:The expression of salt stress responsive proteins and may improve the adaptation of Pancratum maritimum L. to salt stress. J. EXP.Bot. 54 : 2553-2562.
Koch, K.E., and Kennedy, R.A. (1981) Crassulation acid metabolism in the succulent C4 dicot, (Portulaca oleracea L.) under natural environment conditions. Plant Physiol. 69, 757761.
Lichtenthaler, H.K.: Chlorophylls and Carotenoids : Pigments of photosynthetic biomembranes.
Methods Enzymol. 148: 350-382, 1987.
Liu, L., Howe, P., Zhou, Y.F., Xu, Z.Q.,Hocart, C., Zhang, R. ( 2000) Fatty acids and B-Carotene in Australian Purslane (Portulaca oleracea L.) varieties. J. Chromatogr. A. 893, 207-213.
Lowry, O.H., Rosebrough, N.J., Rand, R.J. (1951) Protein measurement with the folinphenol reagent. J. Biol. Chem. 193: 265-275.
Neumann, P.M. (2001) The role of cell wall adjustment in Plant resistance to water deficits. Crop Sci. 35: 1258-1266.
Pinheiro, H.A., Silva, J.V., Endres, L., Dos Santos, B.G. (2008) Leaf gas exchange, chloroplastic pigments and dry matter accumulation in castor bean (Ricinus communis L.) seedlings subjected to salt stress conditions. Crop Prod. 27: 385392.
Premachandre, G.S., Saneoka, H., and Fujta, K. (1991) Osmotic adjustment and stomata response to water deficit in maize. J. Exp. Botany. 43: 1451-1456.
Schuman, M. (2001) Over view of Purslane Edible and Medicinal Herb. NNFA. Today. 15(6) : 12.
Siddiqi, E. H., Ashraf, M., Hussain, M., and Jamil, A. (2009) Assessment of intercultivar Variation for salt tolerance in Safflower (Carthamus tinctorius L.) using gas exchange characteristics as selection criteria. Pak. J.Bot. 41(5) : 22512259.
Taffouo, V.D., Nouck, A.H., Dibong, S.D., and
Amougou, A. (2010) Effects of salinity stress on seedling growth, numeral nutrients, and total chlorophyll of some tomato (Lycopersicum esculentum L.) cultivars. African Journal of Biotechnology. 9 (33): 5366-5372.
Tobe, K., Zhange, L., and Omasa, K. (1999) Effect of NaCl on seed germination of five nonhaplophytic species from a Chinese environment. Seed Sci and Technol. 27 : 851863.
Ueda, A., Yamamoto,Y., and Takabe, T. (2007) Salt stress enhances proline utilization in the apical region of barley roots. Biochemical and Biophysical Researeh. 355 : 61-66.
Yang, X. (2006) Tolerance of photosynthesis hybridization line and its parents grown under field conditions. Plant Sci. 171: 389- 397.
Yazici, I., Turkan, I., Sekmen, A., and Demiral, T. (2007) Salinity tolerance of Purslane (Portulaca oleracea L.) is achieved by enhanced antioxidative system, lower level of lipid peroxidation and proline accumulation. Environmental and Experimental Botany. 1-9.
Yordanov, I., Velikova, V., and Tsonev, T. (2003) Plant responses to drought and stress tolerance. Bulg. J. Plant Physiol. 187-206.
Yu, Q., Rengel, Z.( 1999) Drought and salinity differentially influenced activities of superoxidase dismutase in narrow leafed lupine. Plant Sci. 142 : 1-11.
