Study of total protein content, soluble sugar, proline content and P5CS gene expression in leaves of three wheat cultivars under drought stress

Document Type : Research Paper

Authors

1 . M.Sc. Agricultural Biotechnology Faculty of Agricultural Sciences, University of Mohaghegh Ardabili, Ardabil, Iran

2 Department of Agronomy & Plant Breeding, Faculty of Agriculture & Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran.

3 PhD. Student of genetic molecular, Faculty of Agriculture & Natural Resources, University of Mohaghegh Ardabili, Ardabil, Iran.

Abstract

Objective
The present study was designed to identify the physiological and molecular responses of drought stress induced metabolism in plants. Since the studied plants include resistant and susceptible wheat crop, it is important to know the drought tolerance mechanisms in these cultivars in order to improve the resistance and tolerance of drought stress to the climate in Iran.
 
Materials and Methods
For this purpose, a factorial experiment based on completely randomized design with three replications was conducted in Mohaghegh Ardebil University in 2018-2019. The main factors included drought stress (35, 60 and 85% of field capacity (control)) and sub factor included three wheat cultivars (Pishgam, Pishtaz and Baharan). Drought stress in the three-leaf stage was applied for 10 days and then the seedlings were sampled to check the total protein content of the solution, the sugar content, proline content and the expression of proline-5-D gene expression -5-carboxyxyl synthase (P5CS) of the leaf tissue sample.
 
Results 
The results showed that with increasing severity of drought stress, the amount of proline, soluble sugar and total protein increased. The highest and lowest levels of total protein, respectively, belonged to the leading cultivar under conditions of strict control and stress. The highest amount of soluble sugar (75.76 and 91.66 mg / g, respectively, wet weight of leaves) in spring wheat and severe drought stress and the lowest (57.59 and 48.7 mg / g, respectively). Examination of P5SC gene expression showed that the number of transcripts of this gene increases under stress and this shows that this gene plays an important role in responding to stress in plants. Baharan showed higher drought resistance than other cultivars by increasing P5CS gene expression and significant proline production and accumulation under drought stress.
 
Conclusions
In general, it is inferred that drought-induced expression key gene involved in the biosynthesis of proline (P5CS) cause increased levels of proline leaves and also with the accumulation of soluble sugar probably causes more stress tolerance in cultivars.

Keywords


 
احسنی محمدرضا؛ محمدآبادی محمدرضا؛ اسدی فوزی مسعود و همکاران (1398) بیان ژن لپتین در بافت چربی زیرپوستی گاوهای هلشتاین با استفاده از Real Time PCR‎. مجله بیوتکنولوژی کشاورزی 11،150-135.
توحیدی نژاد فاطمه، محمدآبادی محمدرضا، اسمعیلی‌زاده کشکوئیه علی، نجمی نوری عذرا (1393) مقایسه سطوح مختلف بیان ژنRheb  در بافت‌های مختلف بز کرکی راینی. مجله بیوتکنولوژی کشاورزی 6، 50-35. 
جعفری دره در امیر حسین، محمدآبادی محمدرضا، اسمعیلی زاده کشکوئیه علی، ریاحی مدوار علی (1395) بررسی بیان ژن CIB4  در بافت‌های مختلف گوسفند کرمانی با استفاده از Real Time qPCR. مجله پژوهش در نشخوارکنندگان 4، 132-119.
سادات اسیلان کمال (1395) تاثیر تنش کمبود آب بر محتوای قندهای محلول، پرولین، سبزینه برگ و پروتئین دانه در برخی دورگ‌های آفتابگردان (Helianthus annuus L.). علوم گیاهان زراعی ایران. 47، 184-175.
فتحی امیرخیز کیوان؛ امینی دهقنی مجید؛ مدرس ثانوی سید علی محمد؛ حشمتی سیاوش (1390) اثر کاربرد خاکی و برگی عنصر آهن (Fe) بر برخی خصوصیات بیوشیمیایی گلرنگ (Carthamus tinctorius L.)، تحت دو رژیم رطوبتی. مجله علوم گیاهان زراعی ایران. 42، 509-518.
محمدآبادی محمدرضا (1398) بیان ژن کالپاستاتین در بز کرکی راینی با استفاده از Real Time PCR. مجله بیوتکنولوژی کشاورزی 11،234-219.
محمدآبادی محمدرضا؛ کرد محبوبه؛ نظری محمود (1397) مطالعه بیان ژن لپتین در بافت‌های مختلف گوسفند کرمانی با استفاده از Real Time PCR‎. مجله بیوتکنولوژی کشاورزی 10،122-111.
هادی هاشم؛ سیدشریفی رئوف؛ نامور علی. (1395) محافظ‌های گیاهی و تنش‌های غیر زیستی. انتشارات دانشگاه ارومیه، ارومیه.
References
Ashraf M, Foolad MR (2007) Roles of glycinebetaine and proline in improving plant abiotic stress resistance. Env Exp Bot 59, 206-216.
Ahsani MR, Mohammadabadi MR, Asadi Fozi M, et al. (2019) Leptin gene expression in subcutaneous adipose tissue of Holstein dairy cattle using Real Time PCR. Agric Biotechnol J 11, 135-150 (In Persian).
Aminafshar M, Bahrampour V, Baghizadeh A, et al. (2014) CD44 gene expression in mature, immature oocytes and fetal kermani, baluchi sheep and rayeni, tali goats. J cell Anim Biol 8, 156-160.
Bates LS, Waldren RP, Teare ID (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39, 205-207.
Bradford MM (1976). A rapid and sensitive for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72, 248-254.
Bajii M, Lutts S, Kinet J. (2011) Water deficit effect on solute contribution toosmotic adjustment as a function of leaf ageing in three wheat cultivars performingdifferently in arid conditions. Plant Sci 160, 669-681.
Chen J, Zhang X, Jing R et al. (2010). Cloning and genetic diversity analysis of a new P5CS gene from common bean (Phaseolus vulgaris L.). Theory Appl Gen 120, 1393-1404.
Donaldson E, Schillinger WE, Dofing SM (2001) Strwa production and grain yield relationships in winter wheat. Crop Sci 41, 100-106.
Emam Y (2011) Cereal Production. (4th Eds). Shiraz University Press, Shiraz. 190 p. (In Persian)
Fathi Amirkhiz K, Amini Dehaghi M, Modarres Sanavi SAM, Heshmati S (2011) The Effects of Soil and Foliar Application of Fe on some Biochemical Characteristics of Safflower (Carthamus tinctorius L.) under Two Irrigation Regimes. Iran J Field Crop Sci 42, 509-518. (In Persian)
Farooq M, Wahid A, Kobayashi N et al. (2009) Plant drought stress: effects, mechanisms and management. Agr Sust Dev 29, 185-212.
Fujita T, Maggio A, Rios MG et al. (2003). Identification of regions of the tomato - glutamyl kinase that are involved in allosteric regulation by proline. J Biol Chem Pp, 278. 
Gomes FP, Oliva MA, Mielke MS et al. (2010) Osmotic adjustment, proline accumulation and membrane stability in leaves of Cocos nuciera submitted to drought stress. Sci Hort 126, 379-384.
Hur J, Jung KH, Lee CH, An G (2004) Stress- inducible OsP5CS2 gene is essential for salt and cold tolerance in rice. Plant Sci 167, 417-426.
Hanson H, Bolaugh NE, Anderson RG (1982). Wheat in the Third World. West view press Inc. Boulder, Colorado, USA. P.13
Hadi H, Seyed Sharifi R, Namvar AS (2016) Plant protection and abiotic stresses. Urmia University Press, Urmia (In Persian)
Hong Z, Lakkineni K, Zhang Z, Verma DPS (2005) Removal of feedback inhibition of Δ1-pyrroline-5-carboxylate synthetase results in increased proline accumulation and protection of plants from osmotic stress. Plant Physiol 122, 1129-1136.
Irigoyen JJ, Emerich DW, Sanchez-Diaz M (1992) Water stress inducedchanges in concentrations of proline and total soluble sugars in nodulated alfalfa (Medicago sativa L.) plants. Plant Physiol 84, 55-60.
Jafari Darehdor AH, Mohammadabadi MR, Esmailizadeh AK, Riahi Madvar A (2016) Investigating expression of CIB4 gene in different tissues of Kermani Sheep using Real Time qPCR. J Rumin Res 4, 119-132 (In Persian).
KaviKishor PB, Sangam S, Amrutha RN et al. (2005) Regulation of proline biosynthesis, degradation, uptake and transport in higher plants: Its implications in plant growth and stress tolerance. Curr Sci 88, 424-438.
Kavar T, Maras M, Kidric M et al. (2008) Identification of genes involved in the response of leaves of Phaseolus vulgaris to drought stress. Mol Bree 21, 159-172.
Movludi A, Ebadi A, Jahanbakhsh S et al. (2014) The effect of water deficit and nitrogen on the antioxidant enzymes activity and quantum yield of barley (Hordeum vulgare L.). Notulae Botanicae Horti Agrobotanici Cluj-Napoca 42, 398-404.
Mohammadabadi MR (2019) Expression of calpastatin gene in Raini Cashmere goat using Real Time PCR. Agric Biotechnol J 11, 219-235 (In Persian).
Mohammadabadi MR, Jafari AHD, Bordbar F (2017) Molecular analysis of CIB4 gene and protein in Kermani sheep. Brazil J Medic Biol Res 5, e6177.
Mohammadabadi MR, Kord M, Nazari M (2018) Studying expression of leptin gene in different tissues of Kermani Sheep using Real Time PCR. Agric Biotechnol J 10 ,111-122 (In Persian).
Mohammadabadi MR, Tohidinejad F (2017) Charachteristics determination of Rheb gene and protein in Raini Cashmere goat. Iran J Appl Anim Sci 7, 289-295.
Munns R, Tester M (2008) Mechanisms of Salinity Tolerance. Plant Biol 59, 651-681.
Nanjo T, Masatomo K, Yoshiba Y et al. (1999) Biological function of proline in morphogenesis and osmotolerance revealed in antisense transgenic Arabidopsis thaliana. Plant J 18, 185.
Pirasteh-Anosheh H, Emam Y (2012) The role of plant growth regulators in enhancing crop yield under saline conditions: from theory to practice. Iran J Crop Sci 21, 188-209. (In Persian).
Patger M, Bragato C, Brix H (2005) Tolerance and physiological responses of phragmites australis to water deficit. Aquatic Bot 81, 285-299.
Porcel R, Azcón R, Ruiz-Lozano JM (2004) Evaluation of the role of genes encoding for Δ1-pyrroline-5-carboxylate synthetase (P5CS) during drought stress in arbuscular mycorrhizal Glycine max and Lactuca sativa plants. Physiol Mol Plant Pathology 65, 211-221. ‏
Qin Y, Wang M, Tian Y et al. (2012) Over-expression of TaMYB33 encoding a novel wheat MYB transcription factor increases salt and drought tolerance in Arabidopsis. Mol Biol Rep 39, 7183-7192. ‏
Ruijter JM, Ramakers C, Hoogaars WMH, Karlen Y et al. (2009) Amplification efficiency: Linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Res 37, e45. 
Ranjan R, Bohra SP, Jeet AM (2001) Plant Senescence. Jodhpur, agrobios. Pp, 18-42.
Secenji M, Lendvai A, Hajósné Z et al. (2005) Experimental system for studying long-term drought stress adaptation of wheat cultivars. Acta Biol Szegediensis 49, 51-52.
Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protocols, 3, 1101-1108.
Shinozaki K, Yamagushi-Shinozaki M, Seki M (2003) Regulatory network of gene expression in the drought and cold stress responses. Curr Opinion Plant Biol 6, 410-417.
Shi H, Lee BH, Wu SJ, Zhu JK (2003) Overexpression of a plasma membrane Na+/H+ antiporter gene improves salt tolerance in Arabidopsis thaliana. Nat biotech, 21(1), 81. ‏
Silva-Ortega CO, Ochoa-Alfaro AE, Reyes-Agüero JA (2008). Salt stress increases the expression of p5cs gene and induces proline accumulation in cactus pear. Plant Physiol Biochem 46, 82-92.
Silveira JAG, Arau´ jo SAM, Lima JPMS, vie´gas RA (2010) Roots and leaves display contrasting osmotic adjustment mechanisms in response to NaCl-salinity in Atriplexnummularia. Env Exp Bot 66,1-8.
Sudhakar S, Li Y, Katz MS, Elango N (2001) Translational regulation is a control point in RUNX2/Cbfa1 gene expression. Biochem Biophys Res Commun 289, 616-22.
Szabados L, Savoure A (2010) Proline: a multifunctional amino acid. Trends Plant Scencei 15.
Sadat Asilan K (2016) Effect of water deficit stress on soluble sugars, proline, protein and chlorophyll content in Sunflower (Helianthus annuus L.) hybrids. Iran J Filed Crop Sci 47, 184-175. (In Persian)
Seki M, Narusaka M, Ishida J et al. (2005) Monitoring the expression proles of 7000 Arabidopsis genes under drought, cold and high-salinity stresses using a full-length cDNA microarray. Plant J 31, 279-292.
Tang Y, Liu M, Gao S et al. (2012) Molecular characterization of novel TaNAC genes in wheat and overexpression of TaNAC2a confers drought tolerance in tobacco. Physiol plantarum 144, 210-224.
Turkan I (2011) Plant responses to drought and salinity stress, Development in a post-Genomic era. Adv Bot Res, 593p.
Tohidi nezhad F, Mohammadabadi MR, Esmailizadeh AK, Najmi Noori A (2015) Comparison of different levels of Rheb gene expression in different tissues of Raini Cashmir goat. Agric Biotechnol J 6, 35-50 (In Persian).
Xiong L, Schumaker KS, Zhu JK (2002) Cell signaling during cold, drought, and salt stress. plant cell 14, 165-183. ‏
Zhang H, Mao X, Jing R et al. (2010) Characterization of a common wheat (Triticum aestivum L.) TaSnRK2. 7 gene involved in abiotic stress responses. J Exp Bot 62, 975-988.