Abstract
The vacuolar Na+/H+ antiporter is known to alleviate saline stress by sequestering Na+ in both wild-type Arabidopsis and rice and when over-expressed in many transgenic plants. Here we report on the effect of the NHX1 transgene on the salt tolerance properties it confers to a rice landrace and a commercial cultivar suitable for the dry winter season, but which suffers loss due to seasonal stresses, particularly in the coastal areas. The Nipponbare Na+/H+ antiporter 1.9 kb cDNA was cloned into pCAMBIA1305.1 under the control of the CaMV35S promoter and transformed into tissue-culture-responsive rice landrace Binnatoa (BA). The best-expressing transgenic line at T2 was found to be significantly tolerant at the seedling stage and was advanced to T3. The transgene was then transferred to the tissue-culture recalcitrant farmer-popular commercial rice genotype, BRRIdhan 28 (BR28) by crossing. The data generated both from semi-quantitative RT-PCR and western blot hybridization revealed that the transgene showed similar expression in the crossbred BR28 plants and BA transgenic line. Comparative stress tolerance tests, however, revealed that the BR28 crossbred lines were significantly less tolerant than its transgenic parent BA at both seedling and reproductive stages. A single successful transgenic event may therefore not show the same performance in the recipient genetic background, if introgressed by crossing.
Similar content being viewed by others
References
Aldemita RR, Hodges TK (1996) Agrobacterium tumefaciens-mediated transformation of japonica and indica rice varieties. Planta 199:612–617
Amin M, Elias S, Hossain A, Ferdousi A, Rahman MS, Tuteja N, Seraj Z (2012) Over-expression of a DEAD-box helicase, PDH45, confers both seedling and reproductive stage salinity tolerance to rice (Oryza sativa L.). Mol Breed 30:345–354. doi:10.1007/s11032-011-9625-3
Apse MP, Aharon GS, Snedden WA, Blumwald E (1999) Salt tolerance conferred by overexpression of a vacuolar Na+/H+ antiport in Arabidopsis. Science 285:1256–1258
Apse MP, Sottosanto JB, Blumwald E (2003) Vacuolar cation/H+ exchange, ion homeostasis, and leaf development are altered in a T-DNA insertional mutant of AtNHX1, the Arabidopsis vacuolar Na+/H+ antiporter. Plant J 36:229–239
Barragán V et al (2012) Ion exchangers NHX1 and NHX2 mediate active potassium uptake into vacuoles to regulate cell turgor and stomatal function in Arabidopsis. Plant Cell (Online) 24:1127–1142
Bassil E et al (2011) The Arabidopsis Na+/H+ antiporters NHX1 and NHX2 control vacuolar pH and K+ homeostasis to regulate growth, flower development, and reproduction. Plant Cell (Online) 23:3482–3497
Blumwald E, Poole RJ (1987) Salt tolerance in suspension cultures of sugar beet induction of Na+/H+ antiport activity at the tonoplast by growth in salt. Plant Physiol 83:884–887
Chen H, An R, Tang J-H, Cui X-H, Hao F-S, Chen J, Wang X-C (2007) Over-expression of a vacuolar Na+/H+ antiporter gene improves salt tolerance in an upland rice. Mol Breed 19:215–225
Cheng M, Lowe BA, Spencer TM, Ye X, Armstrong CL (2004) Factors influencing Agrobacterium-mediated transformation of monocotyledonous species. In Vitro Cell Dev Biol-Plant 40:31–45
Deinlein U, Stephan AB, Horie T, Luo W, Xu G, Schroeder JI (2014) Plant Salt-Toler Mech Trends Plant Sci 19:371–379
Doyle J (1991) DNA Protocols for Plants. In: Hewitt G, Johnston AB, Young JP (eds) Molecular techniques in taxonomy, vol 57. NATO ASI series. Springer Berlin Heidelberg, pp 283–293. doi:10.1007/978-3-642-83962-7_18
Fukuda A, Nakamura A, Tanaka Y (1999) Molecular cloning and expression of the Na+/H+ exchanger gene in Oryza sativa. Biochim Biophys Acta (BBA)-Gene Struct Expr 1446:149–155
Fukuda A, Nakamura A, Tagiri A, Tanaka H, Miyao A, Hirochika H, Tanaka Y (2004) Function, intracellular localization and the importance in salt tolerance of a vacuolar Na+/H+ antiporter from rice. Plant Cell Physiol 45:146–159
Gaxiola RA, Rao R, Sherman A, Grisafi P, Alper SL, Fink GR (1999) The Arabidopsis thaliana proton transporters, AtNhx1 and Avp1, can function in cation detoxification in yeast. Proc Natl Acad Sci 96:1480–1485
Gelvin SB (2003) Agrobacterium-mediated plant transformation: the biology behind the “gene-jockeying” tool. Microbiol Mol Biol Rev 67:16–37
Godfray HCJ et al (2010) Food security: the challenge of feeding 9 billion people. Science 327:812–818
Hossain M, Jaim W, Alam MS, Rahman AM (2013) Rice biodiversity in Bangladesh
Khanna HK, Raina SK (1999) Agrobacterium-mediated transformation of indica rice cultivars using binary and superbinary vectors. Funct Plant Biol 26:311–324
Kyozuka J, Otoo E, Shimamoto K (1988) Plant regeneration from protoplasts of indica rice: genotypic differences in culture response. Theor Appl Genet 76:887–890
Leidi EO et al (2010) The AtNHX1 exchanger mediates potassium compartmentation in vacuoles of transgenic tomato. Plant J 61:495–506
Li J-Y, He X-W XuL, Zhou J, Wu P, Shou H-X, Zhang F-C (2008) Molecular and functional comparisons of the vacuolar Na+/H+ exchangers originated from glycophytic and halophytic species. J Zhejiang Univ Sci B 9:132–140
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Maathuis FJ (2014) Sodium in plants: perception, signalling, and regulation of sodium fluxes. J Exp Bot 65:849–858
Maggio A, Zhu J-K, Hasegawa PM, Bressan RA (2006) Osmogenetics: aristotle to arabidopsis. Plant Cell (Online) 18:1542–1557
Moradi F, Ismail AM (2007) Responses of photosynthesis, chlorophyll fluorescence and ROS-scavenging systems to salt stress during seedling and reproductive stages in rice. Ann Bot 99:1161–1173
Munns R, Tester M (2008) Mechanisms of salinity tolerance. Ann Rev Plant Biol 59:651–681
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497
Nishi T, Yamada Y, Takahashi E (1973) The role of auxins in differentiation of rice tissues culture in vitro. J Plant Res 86:183–188
Ogawa T, Vernon LP, Mollenhauer HH (1969) Properties and structure of fractions prepared from Anabaena variabilis by the action of Triton X-100. Biochim Biophys Acta (BBA)-Bioenerget 172:216–229
Pardo JM, Cubero B, Leidi EO, Quintero FJ (2006) Alkali cation exchangers: roles in cellular homeostasis and stress tolerance. J Exp Bot 57:1181–1199
Rasul N, Ali K, Islam R, Seraj Z (1997) Transformation of an Indica Rice cultivar Binnatoa with Agrobacterium tumefaciens. Plant Tissue Cult 7:71–80
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: a laboratory manual, vol 1, 2nd edn. Cold Spring Harbor Laboratory Press
Seraj ZI, Islam Z, Faruque MO, Devi T, Ahmed S (1997) Identification of the regeneration potential of embryo derived calluses from various indica rice varieties. Plant Cell Tissue Organ Cult 48:9–13
Shoeb F, Yadav J, Bajaj S, Rajam M (2001) Polyamines as biomarkers for plant regeneration capacity: improvement of regeneration by modulation of polyamine metabolism in different genotypes of indica rice. Plant Sci 160:1229–1235
Wu Y-Y, Chen Q-J, Chen M, Chen J, Wang X-C (2005) Salt-tolerant transgenic perennial ryegrass (Lolium perenne L.) obtained by Agrobacterium tumefaciens-mediated transformation of the vacuolar Na+/H+ antiporter gene. Plant Sci 169:65–73
Xue Z-Y, Zhi D-Y, Xue G-P, Zhang H, Zhao Y-X, Xia G-M (2004) Enhanced salt tolerance of transgenic wheat (Tritivum aestivum L.) expressing a vacuolar Na+/H+ antiporter gene with improved grain yields in saline soils in the field and a reduced level of leaf Na+. Plant Sci 167:849–859
Yamaguchi T, Hamamoto S, Uozumi N (2013) Sodium transport system in plant cells Frontiers in plant science 4
Yokoi S, Quintero FJ, Cubero B, Ruiz MT, Bressan RA, Hasegawa PM, Pardo JM (2002) Differential expression and function of Arabidopsis thaliana NHX Na+/H+ antiporters in the salt stress response. Plant J 30:529–539
Yoshida S, Forno D, Cock J, Gomez K (1976) Routine procedure for growing rice plants in culture solution. Laboratory manual for physiological studies of rice, pp 61–66
Zhang H-X, Blumwald E (2001) Transgenic salt-tolerant tomato plants accumulate salt in foliage but not in fruit. Nat Biotechnol 19:765–768
Zhang H-X, Hodson JN, Williams JP, Blumwald E (2001) Engineering salt-tolerant Brassica plants: characterization of yield and seed oil quality in transgenic plants with increased vacuolar sodium accumulation. Proc Natl Acad Sci 98:12832–12836
Acknowledgments
Funds for this work including fellowships, consumables and equipments were received from the Bangladesh Chapter of USDA under the 416 (B) grant aid. Thanks to Md. Sazzadur Rahman, Senior Scientific Officer, for arranging for the Na and K ion content determination at BRRI. Thanks to Md. Shamim Hossain for the crossing work and taking care of the rice plants.
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by T. Moriguchi.
S. Biswas and S. Razzaque contributed equally.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Biswas, S., Razzaque, S., Elias, S.M. et al. Effect of the vacuolar Na+/H+ antiporter transgene in a rice landrace and a commercial rice cultivar after its insertion by crossing. Acta Physiol Plant 37, 1730 (2015). https://doi.org/10.1007/s11738-014-1730-6
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11738-014-1730-6