Molecular characterization and functional analysis of a cation-chloride cotransporter gene from trifoliate orange (Poncirus trifoliata L.)
Tóm tắt
A cation-chloride cotransporter gene PtrCCC from trifoliate orange was identified, its ectopic expression confers salt tolerance in transgenic tobacco. Salinity is one of the most serious environmental stresses that limit crop growth. Cation-chloride cotransporters (CCCs) are membrane proteins that are involved in plant development and salt tolerance. Here we provide an insight into the molecular characterization and functional analysis of CCC-encoding gene (PtrCCC) from trifoliate orange (Poncirus trifoliata L.). The open-reading frame of PtrCCC is 2943 bp in length, which encodes a protein of 980 amino acids. Phylogenetic analysis revealed that PtrCCC is closely related to CCCs from its allied genera as well as woody perennials. Quantitative real-time PCR (qRT-PCR) analysis demonstrated that PtrCCC was expressed in all tested tissues, with higher expression level in root tip and leaf tip. Its expression was up-regulated in the leaves of trifoliate orange under the presence of KCl conditions. Ectopic expression of PtrCCC gene in tobacco exhibited an improvement in salt tolerance. The transgenic plants had higher dry biomass in the roots and shoots than that in wild type plants. Moreover, under KCl and NaCl + KCl stress conditions, the accumulation of shoot Cl− was remarkably decreased in the transgenic lines as compared with the wild type plants. Collectively, these results suggest that PtrCCC plays an important role in salt tolerance, which is partially attributed to its role in ions homeostasis.
Tài liệu tham khảo
Aloni R, Schwalm K, Langhans M, Ullrich CI (2003) Gradual shifts in sites of free-auxin production during leaf-primordium development and their role in vascular differentiation and leaf morphogenesis in Arabidopsis. Planta 216:841–853
Brumós J, Colmenero-Flores JM, Conesa A, Izquierdo P, Sánchez G, Iglesias DJ, López-Climent MF, Gómez-Cadenas A, Talón M (2009) Membrane transporters and carbon metabolism implicated in chloride homeostasis differentiate salt stress responses in tolerant and sensitive citrus rootstocks. Funct Integr Genomics 9:293–309
Brumós J, Talon M, Bouhlal R, Colmenero-Flores JM (2010) Cl– homeostasis in includer and excluder citrus rootstocks: transport mechanisms and identification of candidate genes. Plant Cell Environ 33:2012–2027
Chen ZC, Yamaji N, Kashino-Fujii M, Ma JF (2016) A cation-chloride cotransporter gene is required for cell elongation and osmoregulation in rice. Physiol Plant 171:494–507
Colmenero-Flores JM, Martinez GG, Vazquez N, Iglesias DJ, Brumos J, Talon M (2007) Identification and functional characterization of cation-chloride cotransporters in plants. Plant J 50:278–292
Conde A, Chaves MM, Gerós H (2011) Membrane transport, sensing and signaling in plant adaptation to environmental stress. Plant Cell Physiol 52:1583–1602
Gamba G (2005) Molecular physiology and pathophysiology of electroneutral cation-chloride cotransporters. Physiol Rev 85:423–493
Haas M (1994) The Na–K–Cl cotransporters. Am J Physiol 267:869–885
Harling H, Czaja I, Schell J, Walden R (1997) Plant cation-chloride co-transporter promoting auxin-independent tobacco protoplast division. Embo J 16:5855–5866
Hebert SC, Mount DB, Gamba G (2004) Molecular physiology of cation-coupled Cl– cotransport: the SLC12 family. Pflugers Arch Eur J Physiol 447:580–593
Henderson SW, Wege S, Qiu J, Blackmore DH, Walker AR, Tyerman SD, Walker RR, Gilliham M (2015) Grapevine and Arabidopsis cation-chloride cotransporters localize to the Golgi and trans-Golgi network and indirectly influence long-distance ion transport and plant salt tolerance. Plant Physiol 169:2215–2229
Kahle KT, Staley KJ, Nahed BV, Gamba G, Hebert SC, Lifton RP, Mount DB (2008) Roles of the cation-chloride cotransporters in neurological disease. Nat Clin Pract Neuro 4:490–503
Kong XQ, Gao XH, Sun W, An J, Zhao YX, Zhang H (2011) Cloning and functional characterization of a cation-chloride cotransporter gene OSCCC1. Plant Mol Biol 75:567–578
Lindinger MI, Leung M, Trajcevski KE, Hawke TJ (2011) Volume regulation in mammalian skeletal muscle: the role of sodium-potassium-chloride cotransporters during exposure to hypertonic solutions. J Physiol 589:2887–2899
Maas EV (1993) Salinity and citriculture. Tree Physiol 12:195–216
Moes AD, van der Lubbe N, Zietse R, Loffing J, Hoorn EJ (2014) The sodium chloride cotransporter SLC12A3: new roles in sodium, potassium, and blood pressure regulation. Pflugers Arch 466:107–118
Moya JL, Primo-Millo E, Talon M (1999) Morphological factors determining salt tolerance in citrus seedlings: the shoot to root ratio modulates passive root uptake of chloride ions and their accumulation in leaves. Plant Cell Environ 22:1425–1433
Moya JL, Gomez-Cadenas A, Primo-Millo E, Talon M (2003) Chloride absorption in salt-sensitive Carrizo citrange and salt-tolerant Cleopatra mandarin citrus rootstocks is linked to water use. J Exp Bot 54:825–833
Munns R (2005) Genes and salt tolerance: bringing them together. New Phytol 167:645–663
Munns R, Tester M (2008) Mechanisms of salinity tolerance. Annu Rev Plant Biol 59:651–681
Parida AK, Jha B (2010) Salt tolerance mechanisms in mangroves: a review. Trees 24:199–217
Sairam RK, Tyagi A (2004) Physiology and molecular biology of salinity stress tolerance in plants. Curr Sci 86:407–421
Shalhevet J, Yaron D, Horowitz U (2015) Salinity and citrus yield-an analysis of results from a salinity survey. J Hortic Sci 49:15–27
Storey R, Walker RR (1998) Citrus and salinity. Sci Hort 78:39–81
Syvertsen JP, Melgar JC, Garcia-Sanchez F (2010) Salinity tolerance and leaf water use efficiency in citrus. J Am Soc Hortic Sci 135:33–39
Teakle NL, Tyerman SD (2010) Mechanisms of Cl– transport contributing to salt tolerance. Plant Cell Environ 33:566–589
Toyoda H, Yamada J, Ueno S, Okabe A, Kato H, Sato K, Hashimoto K, Fukuda A (2005) Differential functional expression of cation-Cl– cotransporter mRNAs (KCC1, KCC2, and NKCC1) in rat trigeminal nervous system. Mol Brain Res 133:12–18
Wei QJ, Liu YZ, Zhou GF, Li QH, Yang CQ, Peng SA (2013) Overexpression of CsCLCc, a chloride channel gene from Poncirus trifoliata, enhances salt tolerance in Arabidopsis. Plant Mol Biol Rep 31:1548–1557
White PJ, Broadley MR (2001) Chloride in soils and its uptake and movement within the plant: a review. Ann Bot-London 88:967–988
Xu Q, Chen LL, Ruan X, Chen D, Zhu A, Chen C, Bertrand D, Jiao WB, Hao BH, Lyon MP, Chen J, Gao S, Xing F, Lan H, Chang JW, Ge X, Lei Y, Hu Q, Miao Y, Wang L, Xiao S, Biswas MK, Zeng W, Guo F, Cao H, Yang X, Xu XW, Cheng YJ, Xu J, Liu JH, Luo OJ, Tang Z, Guo WW, Kuang H, Zhang HY, Roose ML, Nagarajan N, Deng XX, Ruan Y (2012) The draft genome of sweet orange (Citrus sinensis). Nat Genet 45:59–66