Abstract
Salicornia brachiata Roxb., an extreme halophyte, is a naturally adapted higher plant model for additional gene resources to engineer salt tolerance in plants. Ascorbate peroxidase (APX) plays a key role in protecting plants against oxidative stress and thus confers abiotic stress tolerance. A full-length SbpAPX cDNA, encoding peroxisomal ascorbate peroxidase, was cloned from S. brachiata. The open reading frame encodes for a polypeptide of 287 amino acid residues (31.3-kDa protein). The deduced amino acid sequence of the SbpAPX gene showed characteristic peroxisomal targeting sequences (RKRAI) and a C-terminal hydrophobic region of 39 amino acid residues containing a transmembrane domain (TMD) of 23 amino acid residues. Northern blot analysis showed elevated SbpAPX transcript in response to salt, cold, abscisic acid and salicylic acid stress treatments. The SbpAPX gene was transformed to tobacco for their functional validation under stresses. Transgenic plants over-expressing SbpAPX gene showed enhanced salt and drought stress tolerance compared to wild-type plants. Transgenic plants showed enhanced vegetative growth and germination rate both under normal and stressed conditions. Present study revealed that the SbpAPX gene is a potential candidate, which not only confers abiotic stress tolerance to plants but also seems to be involved in plant growth.
Similar content being viewed by others
References
Alscher RG, Donahue JL, Cramer CL (1997) Reactive oxygen species and antioxidants: relationships in green cells. Physiol Plantarum 100:224–233
Asada K (1992) Ascorbate peroxidase—a hydrogen peroxide scavenging enzyme in plants. Physiol Plantarum 85:235–241
Asada K (1994) Production and action of active oxygen species in photosynthetic tissues. In: Foyer CH, Mullineaux PM (eds) Causes of photooxidative stress and amelioration of defense systems in plants. CRC Press, Boca Raton, FL, pp 77–104
Asada K (1997) The role of ascorbate peroxidase and monodehydroascorbate reductase in H2O2 scavenging in plants. In: Scandalios JG (ed) Oxidative stress and the molecular biology of antioxidant defenses. Cold Spring Harbor Laboratory Press, New York, USA, pp 715–135
Ashraf M (2009) Biotechnological approach of improving plant salt tolerance using antioxidants as markers. Biotechnol Adv 27:84–93
Barba-Espin G, Diaz-Vivancos P, Job D, Belghazi M, Job C, Hernández JA (2011) Understanding the role of H2O2 during pea seed germination: a combined proteomic and hormone profiling approach. Plant Cell Environ 34:1907–1919
Bohnert HJ, Sheveleva E (1998) Plant stress adaptations—making metabolism move. Curr Opin Plant Biol 3:267–274
Bonifacio A, Martins MO, Ribeiro CW, Fontenele AV, Carvalho FE, Margis–Pinheiro M, Silveira JA (2011) Role of peroxidases in the compensation of cytosolic ascorbate peroxidase knockdown in rice plants under abiotic stress. Plant Cell Environ 34:1705–1722
Bradford MM (1976) Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Bressan R, Bohnert H, Zhu JK (2009) Abiotic stress tolerance: from gene discovery in model organisms to crop improvement. Mol Plant 2:1–2
Bunkelmann JR, Trelease RN (1996) Ascorbate peroxidase: a prominent membrane protein in oilseed glyoxysomes. Plant Physiol 110:589–598
Chaturvedi AK, Mishra A, Tiwari V, Jha B (2012) Cloning and transcript analysis of type 2 metallothionein gene (SbMT-2) from extreme halophyte Salicornia brachiata and its heterologous expression in E. coli. Gene 499:280–287
Chen Y, Wang H, Wang X, Cao A, Chen P (2006) Cloning and expression of peroxisomal ascorbate peroxidase gene from wheat. Mol Biol Rep 33:207–213
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 161:156–159
Corpas FJ, Gómez M, Hernández JA, delRío LA (1993) Metabolism of activated oxygen in peroxisomes from two Pisum sativum L. cultivars with different sensitivity to sodium chloride. J Plant Physiol 141:160–165
Dellaporta SL, Wood J, James BH (1983) A plant DNA minipreparation (version II). Plant Mol Biol Rep 1:19–21
delRío LA, Pastori GM, Palma JM, Sandalio LM, Sevilla F, Corpas FJ, Jiménez A, López-Huertas E, Hernández JA (1998) The activated oxygen role of peroxisomes in senescence. Plant Physiol 116:1195–1200
Díaz-Vivancos P, Rubio M, Mesonero V, Periago PM, Ros Barceló A, Martínez-Gómez P, Hernández JA (2006) The apoplastic antioxidant system in Prunus: response to plum pox virus. J Exp Bot 57:3813–3824
Flowers TJ (2004) Improving crop salt tolerance. J Exp Bot 55:307–319
Foyer CH, Lelandais M, Kunert KJ (1994) Photooxidative stress in plants. Physiol Plantarum 92:696–717
Gest N, Gautier H, Stevens R (2013) Ascorbate as seen through plant evolution: the rise of a successful molecule? J Exp Bot 64:33–53
Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930
Hajiboland R (2012) Effect of micronutrient deficiencies on plants stress responses. In: Ahmad P, Prasad MNV (eds) Abiotic stress responses in plants. Springer, New York, pp 283–329
Hasanuzzaman M, Hossain MA, daSilva JAT, Fujita M (2012) Plant response and tolerance to abiotic oxidative stress: antioxidant defense is a key factor. In: Venkateswarlu B, Shanker AK, Shanker C, Maheswari M (eds) Crop stress and its management: perspectives and strategies. Springer, Netherlands, pp 261–315
Hernández JA, Corpas FJ, Gómez M, delRío LA, Sevilla F (1993) Salt-induced oxidative stress mediated by activated oxygen species in pea leaf mitochondria. Physiol Plantarum 89:103–110
Hernández JA, Olmos E, Corpas FJ, Sevilla F, delRío LA (1995) Salt-induced oxidative stress in chloroplast of pea plants. Plant Science 105:151–167
Holsters M, DeWaele D, Depicker A, Messens E, Montagu MV, Schell J (1978) Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet 163:181–187
Horsch RB, Fry JE, Hoffman NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring gene into plants. Science 227:1229–1232
Hu L, Li H, Pang H, Fu J (2012) Responses of antioxidant gene, protein and enzymes to salinity stress in two genotypes of perennial ryegrass (Lolium perenne) differing in salt tolerance. J Plant Physiol 169:146–156
Ishikawa T, Sakai K, Takeda T, Yoshimura K, Shigeoka S (1996) cDNA encoding spinach stromal and thylakoid-bound ascorbate peroxidase, differing in the presence or absence of their 3′-coding regions. FEBS Lett 384:289–293
Jha B, Sharma A, Mishra A (2011) Expression of SbGSTU (tau class glutathione S-transferase) gene isolated from Salicornia brachiata in tobacco for salt tolerance. Mol Biol Rep 38:4823–4832
Jha B, Singh N, Mishra A (2012) Proteome profiling of seed storage proteins reveals the nutritional potential of Salicornia brachiata Roxb., an extreme halophyte. J Agric Food Chem 60:4320–4326
Jha B, Mishra A, Jha A, Joshi M (2013) Developing transgenic Jatropha using the SbNHX1 gene from an extreme halophyte for cultivation in saline wasteland. PlosOne 8:e71136
Jiménez A, Hernandez JA, delRío LA, Sevilla F (1997) Evidence for the presence of the ascorbate-glutathione cycle in mitochondria and peroxisomes of pea leaves. Plant Physiol 114:275–284
Jiménez A, Hernandez JA, Pastori G, delRío LA, Sevilla F (1998) Role of the ascorbate-glutathione cycle of mitochondria and peroxisomes in the senescence of pea leaves. Plant Physiol 118:1327–1335
Joshi M, Mishra A, Jha B (2011) Efficient genetic transformation of Jatropha curcas L. by microprojectile bombardment using embryo axes. Ind Crop Prod 33:67–77
Joshi M, Mishra A, Jha B (2012) NaCl plays a key role for in vitro micropropagation of Salicornia brachiata, an extreme halophyte. Ind Crops Prod 35:313–316
Karpinska B, Wisgle G, Karpinski S (2001) Antagonistic effects of hydrogen peroxide and glutathione on acclimation to excess excitation energy in Arabidopsis. IUBMB Life 50:21–26
Kavitha K, Venkataraman G, Parida A (2008) An oxidative and salinity stress induced peroxisomal ascorbate peroxidase from Avicennia marina: molecular and functional characterization. Plant Physiol Biochem 46:794–804
Kosová K, Vítámvás P, Urban MO, Prášil IT (2013) Plant proteome responses to salinity stress—comparison of glycophytes and halophytes. Funct Plant Biol 40:775–786
Laureau C, De Paepe R, Latouche G, Moreno–Chacón M, Finazzi G, Kuntz M, Cornic G, Streb P (2013) Plastid terminal oxidase (PTOX) has the potential to act as a safety valve for excess excitation energy in the alpine plant species Ranunculus glacialis L. Plant Cell Environ 36:1296–1310
Li YJ, Hai RL, Du XH, Jiang XN, Lu H (2009) Over-expression of a Populus peroxisomal ascorbate peroxidase (PpAPX) gene in tobacco plants enhances stress tolerance. Plant Breed 128:404–410
Lin KH, Pu SF (2010) Tissue- and genotype-specific ascorbate peroxidase expression in sweet potato in response to salt stress. Biol Plantarum 54:664–670
Lisenbee CS, Heinze M, Trelease RN (2003) Peroxisomal ascorbate peroxidase resides within a subdomain of rough endoplasmic reticulum in wild-type Arabidopsis cells. Plant Physiol 132:870–882
López-Huertas E, Charlton WL, Johnson B, Graham IA, Baker A (2000) Stress induces peroxisome biogenesis genes. EMBO J 19:6770–6777
Mishra A, Jha B (2011) Antioxidant response of the microalga Dunaliella salina under salt stress. Bot Mar 54:195–199
Mishra A, Joshi M, Jha B (2013) Oligosaccharide mass profiling of nutritionally important Salicornia brachiata, an extreme halophyte. Carbohydr Polym 92:1942–1945
Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410
Mullen RT, Lisenbee CS, Miernyk JA, Trelease RN (1999) Peroxisomal membrane ascorbate peroxidase is sorted to a membranous network that resembles a subdomain of the endoplasmic reticulum. The Plant Cell 11:2167–2185
Mullen RT, Trelease RN (2000) The sorting signals for peroxisomal membrane-bound ascorbate peroxidase are within its C-terminal tail. J Biolo Chem 275:16337–16344
Munné-Bosch S, Queval G, Foyer CH (2013) The impact of global change factors on redox signaling underpinning stress tolerance. Plant Physiol 161:5–19
Noctor G, Foyer CH (1998) Ascorbate and glutathione: keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49:249–279
Pandey S, Mishra A, Patel MK, Jha B (2013) An efficient method for Agrobacterium-mediated genetic transformation and plant regeneration in cumin (Cuminum cyminum L.). Appl Biochem Biotechnol 171:1–9
Ray PD, Huang BW, Tsuji Y (2012) Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cell Signal 24:981–990
Ros Barceló A, Gómez-Ros LV, Ferrer MA, Hernández JA (2006) The apoplastic antioxidant enzymatic system in the wood-forming tissues of trees. Trees-Struct Funct 20:145–156
Sarowar S, Kim EN, Kim YJ, Ok SH, Kim KD, Hwang BK, Shin JS (2005) Overexpression of a pepper ascorbate peroxidase-like 1 gene in tobacco plants enhances tolerance to oxidative stress and pathogens. Plant Sci 169:55–63
Scandalios JG (1993) Oxygen stress and superoxide dismutase. Plant Physiol 101:7–12
Sharma P, Jha AB, Dubey RS, Pessarakli M (2012) Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions. J Bot Article ID:217037
Shi WM, Muramoto Y, Ueda A, Takabe T (2001) Cloning of peroxisomal ascorbate peroxidase gene from barley and enhanced thermotolerance by overexpressing in Arabidopsis thaliana. Gene 273:23–27
Singh N, Mishra A, Joshi M, Jha B (2010) Microprojectile bombardment mediated genetic transformation of embryo axes and plant regeneration in cumin (Cuminum cyminum L.). Plant Cell Tiss Organ Cult 103:1–6
Suzuki N, Koussevitzky S, Mittler R, Miller G (2012) ROS and redox signalling in the response of plants to abiotic stress. Plant Cell Environ 35:259–270
Tan M, Lu J, Zhang A, Hu B, Zhu X, Li W (2011) The distribution and cooperation of antioxidant (iso) enzymes and antioxidants in different subcellular compartments in maize leaves during water stress. J Plant Growth Regul 30:255–271
Tewari RK, Hadacek F, Sassmann S, Lang I (2013) Iron deprivation-induced reactive oxygen species generation leads to non-autolytic PCD in Brassica napus leaves. Environ Exp Bot 91:74–83
Wang J, Zeng Q, Zhu J, Liu G, Tang H (2013) Dissimilarity of ascorbate–glutathione (AsA–GSH) cycle mechanism in two rice (Oryza sativa L.) cultivars under experimental free-air ozone exposure. Agric Ecosyst Environ 165:39–49
Wang J, Zhang H, Allen RD (1999) Overexpression of an Arabidopsis peroximal ascorbate gene increases protection against oxidative stress. Plant Cell Physiol 40:725–732
Wei ZF, Luo M, Zhao CJ, Li CY, Gu CB, Wang W, Zu YG, Efferth T, Fu YJ (2013) UV-induced changes of active components and antioxidant activity in postharvest pigeon pea [Cajanus cajan (L.) Millsp.] leaves. J Agric Food Chem 61:1165–1171
Xiong L, Zhu JK (2002) Molecular and genetic aspects of plant responses to osmotic stress. Plant Cell Environ 25:131–139
Yamaguchi K, Mori H, Nishimura M (1995) A novel isoenzyme of ascorbate peroxidase localized on glyoxysomal and leaf peroxisomal membranes in pumpkin. Plant Cell Physiol 36:1157–1162
Yan N, Xu XF, Wang ZD, Huang JZ, Guo DP (2013) Interactive effects of temperature and light intensity on photosynthesis and antioxidant enzyme activity in Zizania latifolia Turcz. plants. Photosynthetica 51:127–138
Zhang H, Wang J, Nickel U, Allen RD, Goodman HM (1997) Cloning and expression of an Arabidopsis gene encoding a putative peroxisomal ascorbate peroxidase. Plant Mol Biol 34:967–971
Zhang Z, Zhang Q, Wu J, Zheng X, Zheng S, Sun X, Qiu Q, Lu T (2013) Gene knockout study reveals that cytosolic ascorbate peroxidase 2 (OsAPX2) plays a critical role in growth and reproduction in rice under drought, salt and cold stresses. PloS one 8:e57472
Acknowledgments
The financial support received from CSIR, New Delhi (BSC0107–PlaGen) for carrying out this study is thankfully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
ESM 1
(DOC 4864 kb)
Rights and permissions
About this article
Cite this article
Singh, N., Mishra, A. & Jha, B. Over-expression of the Peroxisomal Ascorbate Peroxidase (SbpAPX) Gene Cloned from Halophyte Salicornia brachiata Confers Salt and Drought Stress Tolerance in Transgenic Tobacco. Mar Biotechnol 16, 321–332 (2014). https://doi.org/10.1007/s10126-013-9548-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10126-013-9548-6