Abstract
Osmotin or osmotin-like proteins have been shown to be induced in several plant species in response to various types of biotic and abiotic challenges. The protein is generally believed to be involved in protecting the plant against these stresses. Although some understanding of the possible mechanism underlying the defense function of osmotin against biotic stresses is beginning to emerge, its role in abiotic stress response is far from clear. We have transformed cotton plants with a tobacco-osmotin gene, lacking the sequence encoding its 20 amino acid-long, C-terminal vacuolar-sorting motif, under the control of CaMV 35S promoter. Apoplastic secretion of the recombinant protein was confirmed and the plants were evaluated for their ability to tolerate drought conditions. Under polyethylene glycol-mediated water stress, the osmotin-expressing seedlings showed better growth performance. The transformants showed a slower rate of wilting during drought and faster recovery following the termination of dry conditions in a greenhouse setting. During drought, the leaves from transgenic plants had higher relative water content and proline levels, while showing reduced H2O2 levels, lipid peroxidation and electrolyte leakage. Importantly, following a series of dry periods, the osmotin transformants performed better in terms of most growth and developmental parameters tested. Most relevant, the fiber yield of transgenic plants did not suffer as much as that of their non-transgenic counterparts under drought conditions. The results provide direct support for a protective role of osmotin in cotton plants experiencing water stress and suggest a possible way to achieve tolerance to drought conditions by means of genetic engineering.
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References
Abad LR, D’Urzo MP, Liu D et al (1996) Antifungal activity of tobacco osmotin has specificity and involves plasma membrane permeabilization. Plant Sci 118:11–23. doi:10.1016/0168-9452(96)04420-2
Abraham EM, Huang B, Bonos SA et al (2004) Evaluation of drought resistance for Texas bluegrass, Kentucky bluegrass, and their hybrids. Crop Sci 44:1746–1753
Angeli SD, Altamura MM (2007) Osmotin induces cold protection in olive trees by affecting programmed cell death and cytoskeleton organization. Planta 225:1147–1163. doi:10.1007/s00425-006-0426-6
Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399. doi:10.1146/annurev.arplant.55.031903.141701
Barthakur S, Babu V, Bansal KC (2001) Overexpression of osmotin induces proline accumulation and confers tolerance to osmotic stress in transgenic tobacco. J Plant Biochem Biotechnol 10:31–37
Basal H, Unay A (2006) Water stress in cotton (Gossypium hirsutum L.). Ege Univ Ziraat Fak Derg 43:101–111
Bates LS (1973) Rapid determination of free proline for water-stress studies. Plant Soil 39:205–207. doi:10.1007/BF00018060
Chen C, Dickman M (2005) Proline suppresses apoptosis in the fungal pathogen Colletotrichum trifolii. Proc Natl Acad Sci USA 102:3459–3464. doi:10.1073/pnas.0407960102
Chowdhury SR, Choudhuri MA (1985) Hydrogen peroxide metabolism as an index of water stress tolerance in jute. Physiol Plant 65:503–507. doi:10.1111/j.1399-3054.1985.tb08676.x
Davey MW, Stals E, Panis B et al (2005) High-throughput determination of malondialdehyde in plant tissue. Anal Biochem 347:201–207. doi:10.1016/j.ab.2005.09.041
Dib TA, Monneveux P, Acevedo E et al (1994) Evaluation of proline analysis and chlorophyll fluorescence quenching measurements as drought tolerance indicators in durum wheat (Triticum turgidum L. var. durum). Euphytica 79:65–73. doi:10.1007/BF00023577
Fan L, Zheng S, Wang X (1997) Antisense suppression of phospholipase D retards abscisic acid and ethylene promoted senescence of postharvest Arabidopsis leaves. Plant Cell 9:2183–2196
Garcia-Casado G, Colladaa C, Allona I et al (2000) Characterization of an apoplastic basic thaumatin-like protein from recalcitrant chestnut seeds. Physiol Plant 110:172–180. doi:10.1034/j.1399-3054.2000.110205.x
Grillo S, Leone A, Xu Y et al (1995) Control of osmotin gene expression by ABA and osmotic stress in vegetative tissues of wild-type and ABA-deficient mutants of tomato. Physiol Plant 93:498–504. doi:10.1111/j.1399-3054.1995.tb06849.x
Husaini AM, Abdin MZ (2008) Development of transgenic strawberry (Fragaria x ananassa Dutch.) plants tolerant to salt stress. Plant Sci 174:446–455. doi:10.1016/j.plantsci.2008.01.007
Kaul S, Sharma SS, Mehta IK (2008) Free radical scavenging potential of l-proline: evidence from in vitro assays. Amino Acids 34:315–320. doi:10.1007/s00726-006-0407-x
Kavi Kishor PB, Hong Z, Miao GH et al (1995) Overexpression of Δ1-pyrroline-5-carboxylate synthetase increases proline production and confers osmotolerance in transgenic plants. Plant Physiol 108:1387–1394
Kenton P, Mur LAJ, Atzorn R (1999) (–)-Jasmonic acid accumulation in tobacco hypersensitive response lesions. Mol Plant Microbe Interact 12:74–78. doi:10.1094/MPMI.1999.12.1.74
Khanna-Chopra R, Selote DS (2007) Acclimation to drought stress generates oxidative stress tolerance in drought-resistant than -susceptible wheat cultivar under field conditions. Environ Exp Bot 60:276–283. doi:10.1016/j.envexpbot.2006.11.004
Kononowicz A, Nelson DE, Singh NK et al (1992) Regulation of the osmotin gene promoter. Plant Cell 4:513–524
LaRosa PC, Singh NK, Hasegawa PM et al (1989) Stable NaCl tolerance of tobacco cells is associated with enhanced accumulation of osmotin. Plant Physiol 91:855–861. doi:10.1104/pp.91.3.855
LaRosa PC, Chen Z, Nelson DE et al (1992) Osmotin gene expression is post-transcriptionally regulated. Plant Physiol 100:409–415. doi:10.1104/pp.100.1.409
Liu D, Raghothama KG, Hasegawa PM et al (1994) Osmotin delays development of disease symptoms. Proc Natl Acad Sci USA 91:1888–1892. doi:10.1073/pnas.91.5.1888
Liu D, Rhodes D, D’Urzo MP et al (1996) In vivo and in vitro activity of truncated osmotin that is secreted into the extracellular matrix. Plant Sci 121:123–131. doi:10.1016/S0168-9452(96)04514-1
Luna CM, Pastori GM, Driscoll S et al (2005) Drought controls on H2O2 accumulation, catalase (CAT) activity and CAT gene expression in wheat. J Exp Bot 56:417–423. doi:10.1093/jxb/eri039
McWilliams D (2003) Drought strategies for cotton. Cooperative Extension Service, Circular 582 College of Agriculture and Home Economics. www.cahe.nmsu.edu/pubs/_circulars/CR582.pdf
Miller G, Shulaev V, Mittler R (2008) Reactive oxygen signaling and abiotic stress. Physiol Plant 133:481–489. doi:10.1111/j.1399-3054.2008.01090.x
Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410. doi:10.1016/S1360-1385(02)02312-9
Mittler R, Vanderauwera S, Gollery M et al (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498. doi:10.1016/j.tplants.2004.08.009
Munne-Bosch S, Jubany-Mari T, Alegred L (2001) Drought-induced senescence is characterized by a loss of antioxidant defenses in chloroplasts. Plant Cell Environ 24:1319–1327. doi:10.1046/j.1365-3040.2001.00794.x
Narasimhan ML, Damsz B, Coca MA et al (2001) A plant defense protein induces microbial apoptosis. Mol Cell 8:921–930. doi:10.1016/S1097-2765(01)00365-3
Narasimhan ML, Coca MA, Jin J et al (2005) Osmotin is a homolog of mammalian adiponectin and controls apoptosis in yeast through a homolog of mammalian adiponectin receptor. Mol Cell 17:171–180. doi:10.1016/j.molcel.2004.11.050
Neill SJ, Desikan R, Clarke A et al (2002) Hydrogen peroxide and nitric oxide as signalling molecules in plants. J Exp Bot 53:1237–1247. doi:10.1093/jexbot/53.372.1237
Nishizawa Y, Nishio Z, Nakazono K et al (1999) Enhanced resistance to blast (Magnaporthe grisea) in transgenic Japonica rice by constitutive expression of rice Chitinase. Theor Appl Genet 99:383–390. doi:10.1007/s001220051248
Noori SAS, Sokhansanj A (2008) Wheat plants containing an osmotin gene show enhanced ability to produce roots at high NaCl concentration. Russ J Plant Physiol 55:256–258
Ober ES, Bloa ML, Clark CJA et al (2005) Evaluation of physiological traits as indirect selection criteria for drought tolerance in sugar beet. Field Crops Res 91:231–249. doi:10.1016/j.fcr.2004.07.012
Onishi M, Tachi H, Kojima T (2006) Molecular cloning and characterization of a novel salt-inducible gene encoding an acidic isoform of PR-5 protein in soybean (Glycine max [L.] Merr.). Plant Physiol Biochem 44:574–580. doi:10.1016/j.plaphy.2006.09.009
Ouvrard O, Cellier F, Ferrare K et al (1996) Identification and expression of water stress- and abscisic acid-regulated genes in a drought-tolerant sunflower genotype. Plant Mol Biol 31:819–829. doi:10.1007/BF00019469
Pastori GM, Trippi VS (1992) Oxidative stress induces high rate of glutathione reductase synthesis in a drought-resistant maize strain. Plant Cell Physiol 33:957–961
Quan RD, Shang M, Zhang H et al (2004) Improved chilling tolerance by transformation with betA gene for the enhancement of glycinebetaine synthesis in maize. Plant Sci 166:141–149. doi:10.1016/j.plantsci.2003.08.018
Quisenberry JE, Wendt CW, Berlin JD et al (1985) Potential for using leaf turgidity to select drought tolerance in cotton. Crop Sci 25:294–299
Raghothama KG, Liu D, Nelson DE et al (1993) Analysis of an osmotically regulated pathogenesis-related osmotin gene promoter. Plant Mol Biol 23:1117–1128. doi:10.1007/BF00042346
Rajam MV, Chandola N, Goud PS et al (2007) Thaumatin gene confers resistance to fungal pathogens as well as tolerance to abiotic stresses in transgenic tobacco plants. Biol Plant 51:135–141. doi:10.1007/s10535-007-0026-8
Ramanjulu S, Sudhakar C (2000) Proline metabolism during dehydration in two mulberry genotypes with contrasting drought tolerance. J Plant Physiol 157:81–85
Rathore KS, Sunilkumar G, Campbell LM (2006) Cotton (Gossypium hirsutum L.). In: Wang K (ed) Methods in molecular biology, vol 343: Agrobacterium protocols, vol 1, 2nd edn. Humana Press Inc, Totowa, pp 267–279
Restrepo MA, Freed DD, Carrington JC (1990) Nuclear transport of plant potyviral proteins. Plant Cell 2:987–998
Ringel C, Siebert S, Wienhaus O (2003) Photometric determination of proline in quartz microplates: remarks on specificity. Anal Biochem 313:167–169. doi:10.1016/S0003-2697(02)00565-1
Sairam RK, Deshmukh PS, Saxena DC (1998) Role of antioxidant system in wheat genotypes tolerance to water stress. Biol Plant 41:387–394. doi:10.1023/A:1001898310321
Santen K, Marttila S, Liljeroth E et al (2005) Immunocytochemical localization of the pathogenesis-related PR-1 protein in barley leaves after infection by Bipolaris sorokiniana. Physiol Mol Plant Pathol 66:45–54. doi:10.1016/j.pmpp.2005.04.006
Scarpari LM, Meinhardt LW, Mazzafera P (2005) Biochemical changes during the development of witches’ broom: the most important disease of cocoa in Brazil caused by Crinipellis perniciosa. J Exp Bot 56:865–877. doi:10.1093/jxb/eri079
Schonfeld MA, Johnson RC, Carver BF et al (1988) Water relations in winter wheat as drought resistance indicators. Crop Sci 28:526–553
Shulaev V, Oliver DJ (2006) Virginia bioinformatics metabolic and proteomic markers for oxidative stress. New tools for reactive oxygen species research. Plant Physiol 141:367–372. doi:10.1104/pp.106.077925
Sinclair TR, Ludlow MM (1985) Who taught plants thermodynamics? The unfulfilled potential of plant water potential. Aust J Plant Physiol 12:213–217
Singh NK, Handa AK, Hasegawa PM et al (1985) Proteins associated with adaptation of cultured tobacco cells to NaCl. Plant Physiol 79:126–137. doi:10.1104/pp.79.1.126
Singh NK, Bracker CA, Hasegawa PM et al (1987) Characterization of osmotin. Plant Physiol 85:529–536. doi:10.1104/pp.85.2.529
Singh NK, Nelson DE, Kuhn D et al (1989) Molecular cloning of osmotin and regulation of its expression by ABA and adaptation to low water potential. Plant Physiol 90:1096–1101. doi:10.1104/pp.90.3.1096
Stintzi A, Heitz TS, Kauffmann S et al (1991) Thaumatin-like protein of virus-infected tobacco osmotin. Physiol Mol Plant Pathol 38:137–146. doi:10.1016/S0885-5765(05)80131-6
Sujkowska M, Borucki W, Golinowski W (2007) Localization of expansin-like protein in apoplast of pea (Pisum sativum L.) root nodules during interaction with Rhizobium leguminosarum BV. Viciae 248. Acta Soc Bot Pol 76:17–26
Sunilkumar G, Rathore KS (2001) Transgenic cotton: factors influencing Agrobacterium-mediated transformation and regeneration. Mol Breed 8:37–52. doi:10.1023/A:1011906701925
Sunilkumar G, Campbell LM, Waghela SD, et al. (2009) Expression of anti-K99 scFv in rice tissues and its functional characterization. Trans Res. doi:10.1007/s11248-008-9223-2
Verslues P, Ober E, Sharp R (1998) Root growth and oxygen relations at low water potentials. Impact of oxygen availability in polyethylene glycol solutions. Plant Physiol 116:1403–1412. doi:10.1104/pp.116.4.1403
Verslues P, Agarwal M, Katiyar-Agarwal S et al (2006) Methods and concepts in quantifying resistance to drought, salt and freezing abiotic stresses that affect plant water status. Plant J 45:523–539. doi:10.1111/j.1365-313X.2005.02593.x
Vigers AJ, Wiedemann S, Roberts WK et al (1992) Thaumatin-like pathogenesis-related proteins are antifungal. Plant Sci 83:155–161. doi:10.1016/0168-9452(92)90074-V
Wilkinson JR, Spradling KD, Yoder DW et al (2005) Molecular cloning and analysis of a cotton gene cluster of two genes and pseudo genes for the PR5 protein osmotin. Physiol Mol Plant Pathol 67:68–82. doi:10.1016/j.pmpp.2005.09.006
Woloshuk CP, Meulenhoff SJ, Sela-Buurlage M et al (1991) Pathogen-induced proteins with inhibitory activity toward Phytophthora infestans. Plant Cell 3:619–628
Yamada M, Morishita H, Urano K et al (2005) Effects of free proline accumulation in petunias under drought stress. J Exp Bot 56:1975–1981. doi:10.1093/jxb/eri195
Yun D-J, Zhao Y, Pardo JM et al (1997) Stress proteins on the yeast cell surface determine resistance to osmotin, a plant antifungal protein. Proc Natl Acad Sci USA 94:7082–7087. doi:10.1073/pnas.94.13.7082
Yun DJ, Ibeas JI, Lee H et al (1998) Osmotin, a plant antifungal protein, subverts signal transduction to enhance fungal cell susceptibility. Mol Cell 1:807–817. doi:10.1016/S1097-2765(00)80080-5
Zhou B, Wang J, Guo Z et al (2006) A simple colorimetric method for determination of hydrogen peroxide in plant tissues. Plant Growth Regul 49:113–118. doi:10.1007/s10725-006-9000-2
Zlatev ZS, Lindon FC, Ramalho JC et al (2006) Comparison of resistance to drought of three bean cultivars. Biol Plant 50:389–394. doi:10.1007/s10535-006-0054-9
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This research was supported by funds from Cotton Incorporated and Texas AgriLife Research. We thank Drs. Alois Bell and C. Kenerley for their help with disease resistance assays.
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Parkhi, V., Kumar, V., Sunilkumar, G. et al. Expression of apoplastically secreted tobacco osmotin in cotton confers drought tolerance. Mol Breeding 23, 625–639 (2009). https://doi.org/10.1007/s11032-009-9261-3
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DOI: https://doi.org/10.1007/s11032-009-9261-3