Expression analysis and physiological response of sunTIP7 aquaporin gene to different water regimes in sunflower

Document Type : Research Paper


1 Department of Plant Production and Genetics, Faculty of Agriculture, University of Zanjan, Zanjan, Iran.

2 Instituto de Recursos Naturales y Agrobiologia (IRNAS), Consejo Superior de Investigaciones Científicas (CSIC), Seville, Spain.


The response of a plant to drought stress is positively related to physiological traits and gene expression. Various recent studies suggest that membrane channel proteins, named aquaporin (AQPs), by affecting stomatal conductance behavior could be involved in plant responses to stress conditions. The sunTIP7 gene is a member of AQPs protein that is included in different environmental stress such as drought stress. In this study in order to investigate the sunTIP7 gene expression and its relation with relative water content (RWC), stomatal conductance, shoot fresh and dry weight, root area, chlorophyll index (SPAD) and electrolyte leakage (EL), six selected sunflower (Helianthus annuus L.) inbred lines were planted in a greenhouse under normal irrigation and water deficit conditions. The water deficit treatments were applied during 4th leaflet to flowering stage. Drought conditions reduced crop growth and physiological traits. The highest stomatal conductance was found in the C138 line under both conditions. Sunflower lines had different fold change expression of sunTIP7 gene under drought stress. The expression of the sunTIP7 genewas the lowest in C138, and this downregulation may explain its highest stomatal conductance. However, there was not any clear relationship between physiological traits and expression of sunTIP7 gene in all six sunflower inbred lines. These results suggest that drought tolerance in sunflower is a complex trait and there is no simple molecular explanation for drought tolerance in sunflower lines.


Article Title [فارسی]

تجزیه بیان و پاسخ فیزیولوژیک ژن آکواپورین sunTIP7 تحت رژیم های مختلف آبیاری در آفتابگردان

Abstract [فارسی]

پاسخ گیاه به تنش خشکی با صفات فیزیولوژیکی و بیان ژن رابطه مثبتی دارد. مطالعات متعدد اخیر نشان می­دهد که پروتئین­های کانال غشایی به نام  Aquaporin (AQPs) با تأثیر بر رفتار هدایت روزنه می­ توانند در پاسخ گیاهان به شرایط تنش نقش داشته باشند. ژنsunTIP7  عضو خانواده AQPs  می ­باشد  که در تنش­ های مختلف محیطی از جمله تنش خشکی درگیر می ­باشد. در این مطالعه به منظور بررسی بیان ژن sunTIP7 و ارتباط آن با صفات محتوای آب نسبی (RWC)، هدایت روزنه ­ا­ی، وزن تر و خشک بخش هوایی گیاه، سطح ریشه، شاخص کلروفیل (SPAD) و نشت الکترولیتی در شرایط تنش خشکی، شش لاین اینبرد انتخابی آفتابگردان (Helianthus annuus L.) در شرایط آبیاری نرمال و کم ­آبیاری در گلخانه کشت شدند. تیمار کم ­آبیاری در مرحله چهار برگی تا زمان گلدهی اعمال گردید و موجب کاهش صفات فیزیولوژیک و رشدی گیاه شد. بیشترین میزان هدایت روزنه­ ا­ی در هر دو شرایط در لاین C138 مشاهده شد. مقدار بیان ژن sunTIP7  در لاین­ های آفتابگردان در شرایط تنش متفاوت بود.  کمترین میزان بیان این ژن در لاین C138 مشاهده شد که این کاهش بیان احتمالا با هدایت روزنه­­ ای بالا در این لاین مرتبط است. با این حال رابطه مشخصی بین بیان صفات فیزیولوژیک و بیان ژن sunTIP7 در لاین­ های مورد بررسی مشاهده نشد. این نتایج نشان می­ دهد که تحمل به خشکی در آفتابگردان یک صفت پیچیده بوده و یک راهکار مولکولی ساده قادر به توضیح تحمل خشکی در لاین­ های اینبرد آفتابگردان نیست.

Keywords [فارسی]

  • آفتابگردان
  • آکواپورین
  • تنش خشکی
  • هدایت روزنه ای
  • sunTIP7
Aharon R, Shahak Y, Wininger S, Bendov R, Kapulnik Y and Galili G, 2003. Overexpression of a plasma membrane aquaporin in transgenic tobacco improves plant vigor under favorable growth conditions but not under drought or salt stress. The Plant Cell 15(2): 439-447.
Alberio C, Izquierdo NG, Galella T, Zuil S, Reid R, Zambelli A and Aguirrezábal LAN, 2016. A new sunflower high oleic mutation confers stable oil grain fatty acid composition across environments. European Journal of Agronomy 73: 25-33.
Aguado A, Capote N, Romero F, Dodd IC and Colmenero-Flores JM, 2014. Physiological and gene expression responses of sunflower (Helianthus annuus L.) plants differ according to irrigation placement. Plant Science 227: 37-44.
Alexandersson E, Fraysse L, Sjövall-Larsen S, Gustavsson S, Fellert M, Karlsson M, Johanson U and Kjellbom P, 2005. Whole gene family expression and drought stress regulation of aquaporins. Plant Molecular Biology 59: 469-484.
Bajji M, Kinet J-M and Lutts S, 2002. The use of the electrolyte leakage method for assessing cell membrane stability as a water stress tolerance test in durum wheat. Plant Growth Regulation 36: 61-70.
Barrieu F, Marty-Mazars D, Thomas D, Chaumont F, Charbonnier M and Marty F, 1999. Desiccation and osmotic stress increase the abundance of mRNA of the tonoplast aquaporin BobTIP26-1 in cauliflower cells. Planta 209(1): 77-86.
Blanchet R, Marty JR, Merrien A and Puech J, 1981. Main factors limiting sunflower yield in dry areas. In: Bunting ES (eds) Production and Utilization of Protein in Oilseed Crops. Pp. 205-226. World Crops: Production, Utilization, and Description, vol 5. Springer, Netherlands.
Blum A and Ebercon A, 1981. Cell membrane stability as a measure of drought and heat tolerance in wheat. Crop Science 21: 43-47.
Blum A, Gozlan G and Mayer J, 1981. The manifestation of dehydration avoidance in wheat breeding germplasm. Crop Science 21: 495-499.
Cui X-H, Hao F-S, Chen H, Chen J and Wang X-C, 2008. Expression of the Vicia faba VfPIP1 gene in Arabidopsis thaliana plants improves their drought resistance. Journal of Plant Research 121(2): 207-214.
Danielson JÅ and Johanson U, 2008. Unexpected complexity of the aquaporin gene family in the moss Physcomitrella patens. BMC Plant Biology 8: 45,
Forrest KL and Bhave M, 2008. The PIP and TIP aquaporins in wheat form a large and diverse family with unique gene structures and functionally important features. Functional and Integrative Genomics 8: 115-133.
Fraysse LC, Wells B, McCann MC and Kjellbom P, 2005. Specific plasma membrane aquaporins of the PIP1 subfamily are expressed in sieve elements and guard cells. Biology of the Cell 97(7): 519-434.
Galmés J, Pou A, Alsina MM, Tomas M, Medrano H and Flexas J, 2007. Aquaporin expression in response to different water stress intensities and recovery in Richter-110 (Vitis sp.): relationship with ecophysiological status. Planta 226: 671-681.
Ghaffari M, Toorchi M, Valizadeh M and Shakiba MR, 2012. Morpho-physiological screening of sunflower inbred lines under drought stress condition. Turkish Journal of Field Crops 17(2): 185-190.
Ghobadi M, Taherabadi S, Ghobadi M-E, Mohammadi G-R and Jalali-Honarmand S, 2013. Antioxidant capacity, photosynthetic characteristics and water relations of sunflower (Helianthus annuus L.) cultivars in response to drought stress. Industrial Crops and Products 50: 29-38.
Gustavsson S, Lebrun A-S, Nordén K, Chaumont F and Johanson U, 2005. A novel plant major intrinsic protein in Physcomitrella patens most similar to bacterial glycerol channels. Plant Physiology 139: 287-295.
Hachez C, Heinen RB, Draye X and Chaumont F, 2008. The expression pattern of plasma membrane aquaporins in maize leaf highlights their role in hydraulic regulation. Plant Molecular Biology 68: 337,
Huang R-F, Zhu M-J, Kang Y, Chen J and Wang X-C, 2002. Identification of plasma membrane aquaporin in guard cells of Vicia faba and its role in stomatal movement. Acta Botanica Sinica 44(1): 42-48.Top of Form
Johanson U, Karlsson M, Johansson I, Gustavsson S, Sjövall S, Fraysse L, Weig AR and Kjellbom P, 2001. The complete set of genes encoding major intrinsic proteins in Arabidopsis provides a framework for a new nomenclature for major intrinsic proteins in plants. Plant Physiology 126: 1358-1369.
Johansson I, Karlsson M, Johanson U, Larsson C and Kjellbom P, 2000. The role of aquaporins in cellular and whole plant water balance. Biochimica et Biophysica Acta (BBA)-Biomembranes 1465(1–2): 324-342.
Kirch H-H, Vera-Estrella R, Golldack D, Quigley F, Michalowski CB, Barkla BJ and Bohnert HJ, 2000. Expression of water channel proteins in Mesembryanthemum crystallinum. Plant Physiology 123: 111-124.
Leonhardt N, Kwak JM, Robert N, Waner D, Leonhardt G and Schroeder JI, 2004. Microarray expression analyses of Arabidopsis guard cells and isolation of a recessive abscisic acid hypersensitive protein phosphatase 2C mutant. The Plant Cell 16: 596-615.
Lian H-L, Yu X, Ye Q, Ding X-S, Kitagawa Y, Kwak SS, Su WA, Tang ZC and Ding XS, 2004. The role of aquaporin RWC3 in drought avoidance in rice. Plant and Cell Physiology 45(4): 481-489.
Lim CW, Kim J-H, Baek W, Kim BS and Lee SC, 2012. Functional roles of the protein phosphatase 2C, AtAIP1, in abscisic acid signaling and sugar tolerance in Arabidopsis. Plant Science 187: 83-88.
Liu Q, Umeda M and Uchimiya H, 1994. Isolation and expression analysis of two rice genes encoding the major intrinsic protein. Plant Molecular Biology 26: 2003-2007.
Livak KJ and Schmittgen TD, 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25(4): 402-408.
Lutts S, Kinet JM and Bouharmont J, 1996. NaCl-induced senescence in leaves of rice (Oryza sativa L.) cultivars differing in salinity resistance. Annals of Botany 78: 389-398.
Mardani R, Yousefi AR and Fotovat R, 2012. Using image analysis to study the response of wild barley to different concentrations of wheat aqueous extract. International Journal of Agriculture: Research and Review 2(6): 774-782.
Martre P, Morillon R, Barrieu F, North GB, Nobel PS and Chrispeels MJ, 2002. Plasma membrane aquaporins play a significant role during recovery from water deficit. Plant Physiology 130: 2101-2110.
Moshelion M, Halperin O, Wallach R, Oren R and Way DA, 2015. Role of aquaporins in determining transpiration and photosynthesis in water‐stressed plants: crop water‐use efficiency, growth and yield. Plant, Cell and Environment 38(9): 1785-1793.
Oliviusson P, Salaj J and Hakman I, 2001. Expression pattern of transcripts encoding water channel-like proteins in Norway spruce (Picea abies). Plant Molecular Biology 46(3): 289-299.
Pekcan V, Evci G, Yilmaz MI, Balkan Nalcaiyi AS, Erdal SC, Cicek N, Arslan O, Ekmekci Y and Kaya Y, 2016. Effects of drought stress on sunflower stems and roots. Pp. 53-59. International Conference on Chemical, Agricultural and Life Sciences (CALS-16), Feb. 4-5, Bali , Indonesia.
Pfaffl MW, Horgan GW and Dempfle L, 2002. Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Research 30(9): e36. doi: 10.1093/nar/30.9.e36.
Poormohammad Kiani S, Grieu P, Maury P, Hewezi T, Gentzbittel L and Sarrafi A, 2007. Genetic variability for physiological traits under drought conditions and differential expression of water stress-associated genes in sunflower (Helianthus annuus L.). Theoretical and Applied Genetics 114: 193-207.
Rauf S. 2008. Breeding sunflower (Helianthus annuus L.) for drought tolerance. Communications in Biometry and Crop Science 3(1): 29-44.
Sarda X, Tousch D, Ferrare K, Cellier F, Alcon C, Dupuis JM, Casse F and Lamaze T, 1999. Characterization of closely related delta-TIP genes encoding aquaporins which are differentially expressed in sunflower roots upon water deprivation through exposure to air. Plant Molecular Biology 40(1): 179-191.
Sarda X, Tousch D, Ferrare K, Legrand E, Dupuis JM, Casse-Delbart F and Lamaze T, 1997. Two TIPlike genes encoding aquaporins are expressed in sunflower guard cells. The Plant Journal 12(5): 1103-1111.
Skoric D, 2009. Sunflower breeding for resistance to abiotic stresses. Helia 32(50): 1-15.
Uehlein N, Otto B, Hanson DT, Fischer M, McDowell N and Kaldenhoff R, 2008. Function of Nicotiana tabacum aquaporins as chloroplast gas pores challenges the concept of membrane CO2 permeability. The Plant Cell 20: 648-657.
Venkatesh J, Yu J-W, Gaston D and Park SW, 2015. Molecular evolution and functional divergence of x-intrinsic protein genes in plants. Molecular Genetics and Genomic 290(2): 443-460.
Wasson AP, Richards RA, Chatrath R, Misra SC, Prasad SV, Rebetzke GJ, Kirkegaard JA,
Christopher J and Watt M, 2012. Traits and selection strategies to improve root systems and water uptake in water-limited wheat crops. Journal of Experimental Botany 63: 3485-3498.
Wei W, Alexandersson E, Golldack D, Miller AJ, Kjellbom PO and Fricke W, 2007. HvPIP1;6, a barley (Hordeum vulgare L.) plasma membrane water channel particularly expressed in growing compared with non-growing leaf tissues. Plant and Cell Physiology 48(8): 1132-1147.
Yamada S, Katsuhara M, Kelly WB, Michalowski CB and Bohnert HJ, 1995. A family of transcripts encoding water channel proteins: tissue-specific expression in the common ice plant. The Plant Cell 7(8): 1129-1142.
Yamaguchi-Shinozaki K, Koizumi M, Urao S and Shinozaki K, 1992. Molecular cloning and characterization of 9 cDNAs for genes that are responsive to desiccation in Arabidopsis thaliana: sequence analysis of one cDNA clone that encodes a putative transmembrane channel protein. Plant and Cell Physiology 33(3): 217-224.
Zaharieva M, Gaulin E, Havaux M, Acevedo E and Monneveux P, 2001. Drought and heat responses in the wild wheat relative Aegilops geniculata Roth: potential interest for wheat improvement. Crop Science 41: 1321-1329.
Zardoya R and Villalba S, 2001. A phylogenetic framework for the aquaporin family in eukaryotes. Journal of Molecular Evolution 52(5): 391-404.