Isolation of gene conferring salt tolerance from halophilic bacteria of Lunsu, Himachal Pradesh, India
Tóm tắt
Halophiles offer an attractive source of genes conferring salt tolerance. Halobacillus trueperi SS1 strain of Lunsu, Himachal Pradesh, India, a strict halophile, was exploited to isolate and clone the genes for salt tolerance. The genomic library of BamH1 digest of H. trueperi SS1 was constructed in pUC19, and recombinants were screened for salt tolerance on an LB medium containing ampicillin (100 μg/ml) and NaCl (0 to 1.5 M). One recombinant clone named as salt-tolerant clone (STC) conferred salt tolerance to host Escherichia coli/DH5α, which showed growth in the LB medium supplemented with ampicillin and 1.2 M NaCl. Restriction digestion and PCR analysis revealed the presence of an insert of approximately 2000 bp in the STC. DNA sequencing of the 2-kb insert on both strands yielded a sequence of 2301 nucleotides. Protein BLAST analysis of 2301-bp sequence of H. trueperi SS1 present in STC showed 97% identity to multidrug transport ATP binding/permease protein of Halobacillus karajensis. The insert contained in STC was subcloned into pGEX4T2 vector, and the recombinant clone STC/pGEX4T2 conferred salt tolerance to the bacterial host E. coli. The present study led to the isolation of salt tolerance gene encoding a putative multidrug transport ATP binding/permease protein from H. trueperi SS1. The salt tolerance gene can be subcloned for transferring salt tolerance traits into agricultural crop plants for cultivation in saline and coastal lands.
Tài liệu tham khảo
Flowers TJ, Yeo AR (1995) Breeding for salinity resistance in crop plants—where next? Aust J Plant Physiol 22:875–884
Flowers TJ (2004) Improving crop salt tolerance. J Exp Bot 55:307–319
Zuleta LFG, Italiani VCS, Marques MV (2003) Isolation and characterization of NaCl-sensitive mutants of Caulobacter crescentus. Appl Environ Microbiol 69:3029–3035
Naughton LM, Blumerman SL, Carlberg M, Boyd EF (2009) Osmoadaptation among Vibrio species and unique genomic features and physiological responses of Vibrio parahaemolyticus. Appl Environ Microbiol 75:2802–2810
Kapley A, Purohit HJ, Chhatre S, Shanker R, Chakrabarti T, Khanna P (1999) Osmotolerance and hydrocarbon degradation by a genetically engineered microbial consortium. Bioresour Technol 67:241–245
Klahn S, Marquardt DM, Rollwitz I, Hagemann M (2009) Expression of the ggpPS gene for glucosylglycerol biosynthesis from Azotobacter vinelandii improves the salt tolerance of Arabidopsis thaliana. J Exp Bot 60:1679–1689
Kapardar RK, Ranjan R, Grover A, Puri M, Sharma R (2010) Identification and characterization of genes conferring salt tolerance to Escherichia coli from pond water metagenome. Bioresour Technol 101:3917–3924
Xu D, Duan X, Wang B, Hong B, Ho HD, Wu R (1996) Expression of a late embryogenesis abundant protein gene, HVA1, from barley confers tolerance to water deficit and salt stress in transgenic rice. Plant Physiol 110:249–257
Kishor PB, Hong Z, Miao G, Hu CA, Verma DPS (1995) Overexpression of D-pirroline-5-carboxylate synthetase increases proline production and confers osmotolerance in transgenic plants. Plant Physiol 108:1387–1394
Kasuga M, Liu Q, Miuras S, Yamaguchi-Shinozaki K, Shinozaki K (1999) Improving plant drought, salt and freezing tolerance by gene transfer of a single stress-inducible transcription factor. Nat Biotechnol 17:287–291
Apse MP, Aharon GS, Snedden WA, Blumwald E (1999) Salt tolerance conferred by overexpression of a vacuolar Na+ / H+ antiport in Arabidopsis. Sci 285:1256–1258
Gisbert C, Ana M, Rus AM, Carmen Bolarín MC, López-Coronado JM, Arrillaga I, Manuel Caro M, Serrano R, Moreno V (2000) The yeast HAL1 gene improves salt tolerance of transgenic tomato. Plant Physiol 123(1):393–402
Empadinhas N, da Costa MS (2008) Osmoadaptation mechanisms in prokaryotes: distribution of compatible solutes. Int Microbiol 11:151–161
Cen H, Ye W, Liu Y, Li D, Wang K, Zhang W (2016) Overexpression of a chimeric gene, OsDST-SRDX, improved salt tolerance of perennial ryegrass. Sci Rep 6:27320. https://doi.org/10.1038/srep27320
Dou M, Fan S, Yang S, Huang R, Yu H, Feng X (2017) Overexpression of AmRosea1 gene confers drought and salt tolerance in rice. Int J Mol Sci 18(1):2. https://doi.org/10.3390/ijms18010002
Gupta S, Sharma P, Dev K, Srivastava M, Sourirajan A (2015) A diverse group of halophilic bacteria exist in Lunsu, a natural salt water body of Himachal Pradesh, India. Springer Plus 4:274. https://doi.org/10.1186/s40064-015-1028-1
Gupta S, Sharma P, Dev K, Sourirajan A (2016) Halophilic bacteria of Lunsu produce an array of industrially important enzymes with salt-tolerant activity. Biochem Res Int 9237418:10. https://doi.org/10.1155/2016/9237418
Sambrook J, Fritsch EF, Maniatis T (2009) Molecular cloning: a laboratory manual by Joe Sambrook Third Edition. Cold Spring Harbor Laboratory Press
Ahmed V, Manoj K, Verma MK, Gupta S, Mandhan V, Chauhan N (2018) Metagenomic profiling of soil microbes to mine salt stress tolerance genes. Front Microbiol. https://doi.org/10.3389/fmicb.2018.00159
Yamada A, Tsutsumi K, Tanimoto S, Ozeki Y (2003) Plant RelA/SpoT homolog 375 confers salt tolerance in Escherichia coli and Saccharomyces cerevisiae. Plant Cell 44(1):3–9
Young J, Holland IB (1999) ABC transporters: bacterial exporters-revisited five years on. Biochim Biophys Acta Biomembr 1461:177–200
Higgins CF (1992) ABC Transporters: from microorganisms to man. Annu Rev Cell Dev Biol 8:67–113
Culligan EP, Sleator RD, Julian R, Marchesi JR, Hill C (2012) Functional metagenomics reveals novel salt tolerance loci from the human gut microbiome. ISME J 6:1916–1925
Deshnium P, Los DA, Hayashi H, Mustardy L, Murata N (1995) Transformation of Synechococcus with a gene for choline oxidase enhances tolerance to salt stress. Plant Mol Biol 29:897–907
Gupta S, Sharma P, Dev K, Baumler DJ, Sourirajan A (2019) Draft genome sequence of Halobacillus trueperi SS1 isolated from Lunsu, a salt water body of North West Himalayas. Microbiol 8(10):e01710–e01718. https://doi.org/10.1128/MRA.01710-18
Vaidya S, Dev K, Sourirajan A (2018) Distinct osmoadaptation strategies in the strict halophilic and halotolerant bacteria isolated from Lunsu salt water body of North West Himalayas. Curr Microbiol 75(7):888–895. https://doi.org/10.1007/s00284-018-1462-8