Abstract
Dunaliella salina (Dunal), a unicellular green alga which lacks a rigid polysaccharide cell wall, can thrive in hypersaline environments. To better understand its salt tolerance mechanism, we first constructed a high-quality cDNA library with 55 % novel genes for D. salina and then screened this library, isolating 37 unique salt tolerance genes. Eighteen of these newly isolated genes were annotated using the public databases, including a number of conventional salt tolerance genes, such as spermdine synthetase, protein disulfide isomerase, cyclophilin CYP1, F-box protein SKIP5, among others. The other 19 isolated genes were novel salt tolerance genes. Further annotation of these data may help us elucidate the salt tolerance mechanism of Dunaliella salina.
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Alca’zar R, Marco F, Cuevas JC, Patron M, Ferrando A, Canavate P, Tiburcio AF, Altabella T (2006) Involvement of polyamines in plant response to abiotic stress. Biotechnol Lett 28:1867–1876. doi:10.1007/s10529-006-9179-3
Alkayala F, Albionb RL, Tillettb RL, Hathwaikb LT, Lemosb MS, Cushmanb JC (2010) Expressed sequence tag (EST) profiling in hyper saline shocked Dunaliella salina reveals high expression of protein synthetic apparatus components. Plant Sci 179:437–449. doi:10.1016/j.plantsci.2010.07.001
Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402. doi:10.1093/nar/25.17.3389
Altschul SF, Wootton JC, Gertz EM, Agarwala R, Morgulis A, Schäffer AA, Yu YK (2005) Protein database searches using compositionally adjusted substitution matrices. FEBS J 272:5101–5109. doi:10.1111/j.1742-4658.2005.04945.x
Amotz BA, Avron M (1973) The role of glycerol in the osmotic regulation of the halophilic alga Dunaliella parva. Plant Physiol 51:875–878. doi:10.1104/pp. 51.5.875
Aron MB, Bryant SH (2004) CD-Search: protein domain annotations on the fly. Nucleic Acids Res 32:W327–W331. doi:10.1093/nar/gkh454
Aron MB, Anderson JB, Chitsaz F, Derbyshire MK, Carol DS, Fong JH, Geer LY, Geer RC, Gonzales NR, Gwadz M, He S, Hurwitz DI, Jackson JD, Ke Z, Lanczycki CJ, Liebert CA, Liu C, Lu F, Lu S, Marchler GH, Mullokandov M, Song JS, Tasneem A, Thanki N, Yamashita RA, Zhang D, Zhang N, Bryant SH (2009) CDD: specific functional annotation with the conserved domain database. Nucleic Acids Res 37:D205–D210. doi:10.1093/nar/gkn845
Aron MB, Lu S, Anderson JB, Chitsaz F, Derbyshire MK, Craol DS, Fong JH, Geer LY, Geer RC, Gonzales NR, Gwadz M, Hurwitz DI, Jackson JD, Ke Z, Lanczycki CJ, Lu F, Marchler GH, Mullokandov M, Omelchenko MV, Robertson CL, Song JS, Thanki N, Yamashita RA, Zhang D, Zhang N, Zheng C, Bryant SH (2011) CDD: a conserved domain database for the functional annotation of proteins. Nucleic Acids Res 39:D225–D233. doi:10.1093/nar/gkq1189
Ashburner M, Ball CA, Blake JA, Botstein D, Butler H, Cherry JM, Davis AP, Dolinski K, Dwight SS, Eppig JT, Harris MA, Hill DP, Issel-Tarver L, Kasarskis A, Lewis S, Matese JC, Richardson JE, Ringwald M, Rubin GM, Sherlock G (2000) Gene ontology: tool for the unification of biology. Nat Genet 25:25–29
Azachi M, Sadka A, Fisher M, Goldshlag P, Gokhman I, Zamir A (2002) Salt induction of fatty acid elongase and membrane lipid modifications in the extreme halotolerant alga Dunaliella salina. Plant Physiol 129:1320–1329. doi:10.1104/pp. 001909
Baird TD, Delorenzo ME (2009) Descriptive and mechanistic toxicity of conazole fungicides using the model test alga Dunaliella tertiolecta (Chlorophyceae). Environ Toxicol 25:213–220. doi:10.1002/tox.20493
Blair MW, Fernandez AC, Ishitani M, Moreta D, Seki M, Ayling S, Shinozaki K (2011) Construction and EST sequencing of full-length, drought stress cDNA libraries for common beans (Phaseolus vulgaris L.). BMC Plant Biol 11:171–185. doi:10.1186/1471-2229-11-171
Chen AP, Wang GL, Qu ZL, Lu CH, Liu N, Wang F, Xia GX (2007) Ectopic expression of ThCYP1, a stress-responsive cyclophilin gene from Thellungiella halophila, confers salt tolerance in fission yeast and tobacco cells. Plant Cell Rep 26:237–245. doi:10.1007/s00299-006-0238-y
Chen H, Lao YM, Jiang JG (2011) Effects of salinities on the gene expression of a (NAD+)-dependent glycerol-3-phosphate dehydrogenase in Dunaliella salina. Sci Total Environ 409:1291–1297
Chou IT, Gasser CS (1997) Characterization of the cyclophilin gene family of Arabidopsis thaliana and phylogenetic analysis of known cyclophilin proteins. Plant Mol Biol 35:873–887. doi:10.1023/A:1005930024796
Di Matteo A, Scandurra FM, Testa F, Forte E, Sarti P, Brunori M, Giuffre A (2008) The O2-scavenging flavodiiron protein in the human parasite Giardia intestinalis. J Biol Chem 283:4061–4069. doi:10.1074/jbc.M705605200
Dreher K, Callis J (2007) Ubiquitin, hormones and biotic stress in plants. Ann Bot 99:787–822. doi:10.1093/aob/mcl255
Farras R, Ferrando A, Jasik J, Kleinow T, Okresz L, Tiburcio A, Salchert K, del Pozo C, Schell J, Koncz C (2001) SKP1-SnRK protein kinase interactions mediate proteasomal binding of a plant SCF ubiquitin ligase. EMBO J 20:2742–2756. doi:10.1093/emboj/20.11.2742
Frand AR, Kaiser CA (1998) The ERO1 gene of yeast is required for oxidation of protein dithiols in the endoplasmic reticulum. Mol Cell 1:161–170. doi:10.1016/S1097-2765(00)80017-9
Freeman M (2001) Pin-pointing MAPK signaling. Nat Cell Biol 3:E136–E137. doi:10.1038/35078599
Garcıá F, Freile-Pelegrín Y, Robledo D (2007) Physiological characterization of Dunaliella sp. (Chlorophyta, Volvocales) from Yucatan, Mexico. Bioresource Technol 98:1359–1365. doi:10.1016/j.biortech.2006.05.051
Gouveia L, Oliveira AC (2009) Microalgae as a raw material for biofuels production. J Ind Microbiol Biotechnol 36:269–274. doi:10.1007/s10295-008-0495-6
Goyal A (2007a) Osmoregulation in Dunaliella, Part I: Effects of osmotic stress on photosynthesis, dark respiration and glycerol metabolism in Dunaliella tertiolecta and its salt-sensitive mutant (HL 25/8). Plant Physiol Biochem 45:696–704. doi:10.1016/j.plaphy.2007.05.008
Goyal A (2007b) Osmoregulation in Dunaliella. Part II: photosynthesis and starch contribute carbon for glycerol synthesis during a salt stress in Dunaliella tertiolecta. Plant Physiol Biochem 45:705–710. doi:10.1016/j.plaphy.2007.05.008
Groppa MD, Benavides MP (2008) Polyamines and abiotic stress: recent advances. Amino Acids 34:35–45. doi:10.1007/s00726-007-0501-8
Huang X, Madan A (1999) CAP3: A DNA sequence assembly program. Genome Res 9:868–877. doi:10.1101/gr.9.9.868
Hunter T (1998) Prolyl Isomerases and nuclear function. Cell 92:141–143. doi:10.1016/S0092-8674(00)80906-X
Jahnke LS (1999) Massive carotenoid accumulation in Dunaliella bardawil induced by ultraviolet-A radiation. J Photochem Photobiol B Biol 48:68–74. doi:10.1016/S1011-1344(99)00012-3
Jimenez C, Berl T, Rivard CJ, Edelstein CL, Capasso JM (2004) Phosphorylation of MAP kinase-like proteins mediate the response of the halotolerant alga Dunaliella viridis to hypertonic shock. Biochim Biophys Acta 1644:61–69. doi:10.1016/j.bbamcr.2003.10.009
Kasukabe Y, He L, Nada K, Misawa S, Ihara I, Tachibana S (2004) Overexpression of spermidine synthase enhances tolerance to multiple environmental stresses and up-regulates the expression of various stress-regulated genes in transgenic Arabidopsis thaliana. Plant Cell Physiol 45:712–722. doi:10.1093/pcp/pch083
Lamantia ML, Lennarz WJ (1993) The essential function of yeast protein disulfide isomerase does not reside in its isomerase activity. Cell 74:899–908. doi:10.1016/0092-8674(93)90469-7
Lee Y, Tsai J, Sunkara S, Karamycheva S, Pertea G, Sultana R, Antonescu V, Chan A, Cheung F, Quackenbush J (2005) The TIGR Gene Indices: clustering and assembling EST and known genes and integration with eukaryotic genomes. Nucleic Acids Res 33:D71–D74. doi:10.1093/nar/gki064
Li C, Jiang L, Shao Y, Wang W (2005) Biostatistics. In: ANOVA, 3rd ern. Sciencep, Beijing, pp 91–104
Liska AJ, Shevchenko A, Pick U, Katz A (2004) Enhanced photosynthesis and redox energy production contribute to salinity tolerance in Dunaliella as revealed by homology-based proteomics. Plant Physiol 136:2806–2817. doi:10.1104/pp.104.039438
Lu KP, Liou YC, Zhou XZ (2002) Pinning down proline-directed phosphorylation signaling. Trends Cell Biol 12:164–172. doi:10.1016/S0962-8924(02)02253-5
Martínez IM, Chrispeels MJ (2003) Genomic analysis of the unfolded protein response in Arabidopsis shows its connection to important cellular processes. Plant Cell 15:561–566. doi:10.1105/tpc.007609
Miyasaka H, Kanaboshi H, Ikeda K (2000) Isolation of several anti-stress genes from the halotolerant green alga Chlamydomonas by simple functional expression screening with Escherichia coli. World J Microbiol Biotechnol 16:23–29. doi:10.1023/A:1008982332139
Mogedas B, Casal C, Forján E, Vílchez C (2009) β-Carotene production enhancement by UV-A radiation in Dunaliella bardawil cultivated in laboratory reactors. J Biosci Bioeng 108:47–51. doi:10.1016/j.jbiosc.2009.02.022
Oren A (2005) A hundred years of Dunaliella research: 1905–2005. Saline Systems 1:2–15. doi:10.1186/1746-1448-1-2
Pang Q, Chen S, Dal S, Chen Y, Wang Y, Yan X (2010) Comparative proteomics of salt tolerance in Arabidopsis thaliana and Thellungiella halophila. J Proteome Res 9:2584–2589. doi:10.1021/pr100034f
Romano PG, Horton P, Gray JE (2004) The Arabidopsis cyclophilin gene family. Plant Physiol 134:1268–1272. doi:10.1104/pp. 103.022160
Ruan SL, Ma HS, Wang SH, Fu YP, Xin Y, Liu WZ, Wang F, Tong JX, Wang SZ, Chen HZ (2011) Proteomic identification of OsCYP2, a rice cyclophilin that confers salt tolerance in rice (Oryza sativa L.) seedlings when overexpressed. BMC Plant Biol 11:1–34. doi:10.1186/1471-2229-11-34
Sahia C, Singha A, Blumwaldb E, Grovera A (2006) Beyond osmolytes and transporters: novel plant salt-stress tolerance-related genes from transcriptional profiling data. Physiol Plant 127:1867–1875. doi:10.1111/j.1399-3054.2005.00610.x
Sekhar K, Priyanka B, Reddy VD, Rao KV (2010) Isolation and characterization of a pigeonpea cyclophilin (CcCYP) gene, and its over-expression in Arabidopsis confers multiple abiotic stress tolerance. Plant Cell Environ 33:1324–1338. doi:10.1111/j.1365-3040.2010.02151.x
Takagi M, Karseno YT (2006) Effect of salt concentration on intracellular accumulation of lipids and triacylglyceride in marine microalgae Dunaliella cells. J Biosci Bioeng 101:223–226. doi:10.1263/jbb.101.223
Tanaka S, Ikeda K, Ono MHM (2002) Isolation of several anti-stress genes from a mangrove plant Avicennia marina. World J Microbiol Biotechnol 18:801–804. doi:10.1023/A:1020485227270
Wen XP, Pang XM, Matsuda N, Kita M, Inoue H, Hao Y, Honda C, Moriguchi T (2008) Over-expression of the apple spermidine synthase gene in pear confers multiple abiotic stress tolerance by altering polyamine titers. Transgenic Res 17:251–260. doi:10.1007/s11248-007-9098-7
Ye ZW, Jiang JG, Wu GH (2008) Biosynthesis and regulation of carotenoids in Dunaliella: progresses and prospects. Biotechnol Adv 26:352–360. doi:10.1016/j.biotechadv
Zhang Z, Schwartz S, Wagner L, Miller W (2000) A greedy algorithm for aligning DNA sequences. J Comput Biol 7:203–214. doi:10.1089/10665270050081478
Zhang XX, Liu SK, Takano T (2008) Two cysteine proteinase inhibitors from Arabidopsis thaliana, AtCYSa and AtCYSb, increasing the salt, drought, oxidation and cold tolerance. Plant Mol Biol 68:131–143. doi:10.1007/s11103-008-9357-x
Zhao R, Cao Y, Xu H, Lv LF, Qiao DR, Cao Y (2011) Analysis of espressed sequence tags from the green alga Dunaliella salina (Chalrophyta). J Phycol 47:1454–1460. doi:10.1111/j.1529-8817.2011.01071.x
Zhu J (2001) Plant salt tolerance. Trends Plant Sci 6:66–71. doi:10.1016/S1360-1385(00)01838-0
Acknowledgments
We thank Professor Guangxiao Yang for his carefully reading of the manuscript. This research was supported in part by the 863 Grant of Ministry of Science and Technology (P. R. China, No. 2007AA09Z449), grants from NSFC (P. R. China, No. 30971608; No. 81272210), NSF of the Hubei province (P. R. China, 2009CDB074), the Specific Key Project of Novel Medicine Discovery (P. R. China, 2009ZX09301-014) and International Collaboration Programs of Wuhan Science and Technology Bureau (P. R. China, No. 201070934334).
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Liu, J., Zhang, D. & Hong, L. Isolation, characterization and functional annotation of the salt tolerance genes through screening the high-quality cDNA library of the halophytic green alga Dunaliella salina (Chlorophyta). Ann Microbiol 65, 1293–1302 (2015). https://doi.org/10.1007/s13213-014-0967-z
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DOI: https://doi.org/10.1007/s13213-014-0967-z