Construction of cDNA library from Prunus campanulata leaves and preliminary expressed sequence tag (EST) analysis during cold stress
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Ying-Hui Zhang
Abstract
The molecular mechanisms underlying cold-resistance in Prunus campanulata Maxim. (P. campanulata) are not fully understood. This study aimed to establish a full-length library and analyze expressed sequence tags (ESTs) to provide tools to investigate the mechanisms of P. campanulata growth at low temperatures. Based on the switching mechanism at 5’end of RNA transcript technology, a full-1ength cDNA library was generated from young leaves of P. campanulata after 72 h treatment at 1◦C, and a preliminary EST analysis was carried out. Quantitative reverse transcription polymerase chain reaction was used to assess the expression of selected cold-related genes. The cDNA library titer was 1.2 × 106 cfu/mL−1, with a recombinant rate of 96%. The average size of inserted cDNA fragments was 1.3 Kb. EST data revealed the existence of 834 clones representing a total of 667 unigenes, including 574 singletons and 93 contigs. Blast analysis identified 475 unigenes with known and putative functions. Based on similarity search and GO annotation, 84 unigenes were associated with “response to stimuli”, suggesting that cold stress induced significant alterations in gene expression in P. campanulata cultivated at 1◦C for 72 h. Interestingly, DRP, MYB, HSP, GPX and GA20-ox gene expression was significantly up-regulated in plants cultivated at low temperature, while transcript levels of TIL, CDPK were decreased. P. campanulata cultivating at low temperature express genes associated with “response to stimuli”, and in particular DRP, MYB, HSP, GPX and GA20-ox gene are up-regulated while TIL, CDPK are downregulated in response to low temperature-stress
References
Ainsworth C., Tarvis M. & Clark J. 1993. Isolation and analysis of a cDNA clone encoding the small subunit of ADP-glucose pyrophosphorylase from wheat. Plant Mol. Biol. 23: 23-33.10.1007/BF00021416Suche in Google Scholar
Chen B.-J.,Wang Y., Hu Y.-L., Wu Q. & Lin Z.-P. 2005. Cloning and characterization of a drought-inducible MYBgene from- Boea crassifolia. Plant Sci. 168: 493-500.10.1016/j.plantsci.2004.09.013Suche in Google Scholar
Chen Z., Lu Y.L. & Wu W.X. 2011. Research advance,prospect and breeding strategy of Cerasus campanulata Maxim. J. of Nanjing Forestry University (Natural Sciences Edition), 36: 1-5.Suche in Google Scholar
Chen Z.H., Xiao M. & Chen X. 2008. Change in flowering dates of japanese cherry blossoms (Prunus campanulata Maxim.) in Wuhan University campus and its relationship with variability of winter temperature. Acta Ecol. Sinica 28: 5209-5217.10.1016/S1872-2032(09)60006-8Suche in Google Scholar
Cheong E.J. 2012. Biotechnological approaches for improvement and conservation of Ispecies. Plant Biotech. Rep. 6: 17-28.10.1007/s11816-011-0195-ySuche in Google Scholar
Da Silva C., Campos T., Scaloppi E., Maluf M., Gon¸calves P. & Souza A. 2011. Construction and analysis of a leaf cDNA library from cold stressed rubber tree clones. BMC Proc. 5: 24.10.1186/1753-6561-5-S7-P24Suche in Google Scholar
De los Reyes B., Morsy M., Gibbons J., Varma T., Antoine W., McGrath J., Halgren R. & Redus M. 2003. A snapshot of the low temperature stress transcriptome of developing rice seedlings (Oryza sativa L.) via ESTs from subtracted cDNA library. Theor. Appl. Genet. 107: 1071-1082. 10.1007/s00122-003-1344-7Suche in Google Scholar
Editorial Board for Flora (the Chinese Academy of Sciences) 1986. Flora of China. Science Press, Beijing, China.Suche in Google Scholar
Frenette Charron J.B., Breton G., Badawi M. & Sarhan F. 2002. Molecular and structural analyses of a novel temperature stress-induced lipocalin from wheat and Arabidopsis. FEBS Lett. 517: 129-132.10.1016/S0014-5793(02)02606-6Suche in Google Scholar
Futamura N., Totoki Y., Toyoda A., Igasaki T., Nanjo T., Seki M., Sakaki Y., Mari A., Shinozaki K. & Shinohara K. 2008. Characterization of expressed sequence tags from a full-length enriched cDNA library of Cryptomeria japonica male strobili. BMC Genomics 9: 383.10.1186/1471-2164-9-383Suche in Google Scholar PubMed PubMed Central
Imanishi J., Nakayama A., Suzuki Y., Imanishi A., Ueda N., Morimoto Y. & Yoneda M. 2010. Nondestructive determination of leaf chlorophyll content in two flowering cherries using reflectance and absorptance spectra. Landsc. Ecol. Eng. 6: 219-234.10.1007/s11355-009-0101-8Suche in Google Scholar
Jung H.A., Kim A.R., Chung H.Y. & Choi J.S. 2002. In vitro antioxidant activity of some selected Prunus species in Korea. Arch. Pharm. Res. 25: 865-872.10.1007/BF02977006Suche in Google Scholar PubMed
Kang S.-G., Jeong H.K. & Suh H.S. 2004. Characterization of a new member of the glutathione peroxidase gene family in Oryza sativa. Mol. Cells 17: 23-28.Suche in Google Scholar
Kruse E., Mock H.P. & Grimm B. 1997. Isolation and characterisation of tobacco (Nicotiana tabacum) cDNA clones encoding proteins involved in magnesium chelation into protoporphyrin IX. Plant Mol. Biol. 35: 1053-1056.Suche in Google Scholar
Li H.C., Guo X.L., Wang D.M. & Li G.L. 2010. Responses of HSP70 gene expression to temperature stresses in maize (Zea mays L.). Agric. Univ. of Hebei. 33: 12-15.Suche in Google Scholar
Liu M., Shi J. & Lu C. 2013. Identification of stress-responsive genes in Ammopiptanthus mongolicus using ESTs generated from cold-and drought-stressed seedlings. BMC Plant. Biol. 13: 88.10.1186/1471-2229-13-88Suche in Google Scholar
Lu Y.L., Chen Z. & Shi J.S. 2006. Research advance,prospect and breeding strategy of Cerasus campanulata Maxim. J. of Nanjing Forestry University Natural (Sciences Edition). 30: 115-119.Suche in Google Scholar
Phukon M., Namdev R., Deka D., Modi M.K. & Sen P. 2012. Construction of cDNA library and preliminary analysis of expressed sequence tags from tea plant [Camellia sinensis (L.) O. Kuntze]. Gene 506: 202-206.10.1016/j.gene.2012.06.055Suche in Google Scholar
Short J.M. & Sorge J.A. 1992. In vivo excision properties of bacteriophage lambda ZAP expression vectors. Methods Enzymol. 216: 495-508.10.1016/0076-6879(92)16045-LSuche in Google Scholar
Singh R.K., Singh S., Pandey P., Anandhan S., Goyary D., Pande V. & Ahmed Z. 2013. Construction of cold induced subtracted cDNA library from Cicer microphyllum and transcript characterization of identified novel wound induced gene. Protoplasma 250: 459-469.10.1007/s00709-012-0429-zSuche in Google Scholar PubMed
Tong Z., Qu S., Zhang J., Wang F., Tao J., Gao Z. & Zhang Z. 2012. A modified protocol for RNA extraction from different peach tissues suitable for gene isolation and real-time PCR analysis. Mol. Biotechnol. 50: 229-236.10.1007/s12033-011-9433-3Suche in Google Scholar PubMed
Tsai T.M., Chen Y.R., Kao T.W., Tsay W.S., Wu C.P., Huang D.D., Chen W.H., Chang C.C. & Huang H.J. 2007. PaCDPK1, a gene encoding calcium-dependent protein kinase from orchid, Phalaenopsis amabilis, is induced by cold, wounding, and pathogen challenge. Plant Cell. Rep. 26: 1899-1908.10.1007/s00299-007-0389-5Suche in Google Scholar PubMed
Wang H., Zou Z., Wang S. & Gong M. 2013. Global analysis of transcriptome responses and gene expression profiles to cold stress of Jatropha curcas L. PLoS One. 8: e82817.10.1371/journal.pone.0082817Suche in Google Scholar PubMed PubMed Central
Wang K., Wang Y., Bao Y.L., Meng X.Y., Wu G., Yi J.W. & Li Y.X. 2006. Construction and sequence analysis of a fulllength cDNA library of panax ginseng leaf tissues. J. Mol. Sci. 22: 58-62.Suche in Google Scholar
Wei H., Dhanaraj A.L., Rowland L.J., Fu Y., Krebs S.L. & Arora R. 2005. Comparative analysis of expressed sequence tags from cold-acclimated and non-acclimated leaves of Rhododendron catawbiense Michx. Planta 221: 406-416.10.1007/s00425-004-1440-1Suche in Google Scholar PubMed
Wilkie S.E., Roper J.M., Smith A.G. & Warren M.J. 1995. Isolation, characterisation and expression of a cDNA clone encoding plastid aspartate aminotransferase from Arabidopsis thaliana. Plant Mol. Biol. 27: 1227-1233.Suche in Google Scholar
Xu Y. 2009. A simplified method for quick screening positive clones and foreign DNA orientation by grouping primers PCR. J. of Chongqing University of Science and Technology (Natural Sciences Edition). 11: 52-54.Suche in Google Scholar
Zhang Y F.J. & Gu R. 2009. Isolation and analysis of cold stress inducible genes in Zea mays by suppression subtractive hybridization and cDNA microarray. Plant Mol. Biol. Report. 27: 38-49.10.1007/s11105-008-0055-2Suche in Google Scholar
Zhang Y., Rong J., Chen L., Li S., He T., Chen L. & Zheng Y. 2013. Response of chlorophyll fluorescence parameters to different temperature stresses in Prunus campanulata Maxim. and Prunus yedoensis Matsum. seedlings. J. Food, Agricult. Environ. 11: 2797-2802. Suche in Google Scholar
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Artikel in diesem Heft
- Down into the Earth: microbial diversity of the deepest cave of the world
- Factors influencing synergistic antimicrobial activity of thymol and nisin against Shigella spp. in sugarcane juice
- Effects of low-temperature hardening on the biochemical response of winter oilseed rape seedlings inoculated with the spores of Leptosphaeria maculans
- Mitochondrial structures during seed germination and early seedling development in Arabidopsis thaliana
- Transcriptome analysis for identification of indigo biosynthesis pathway genes in Polygonum tinctorium
- Different components of plant diversity suggest the protection of a large area for the conservation of a riparian ecosystem
- Anatomical adaptations of the desert species Stipa lagascae against drought stress
- Effects of ammonium ion on cell growth and biosynthesis of shikonin derivatives in callus tissues of Arnebia euchroma
- cDNA cloning, heterologous expression and characterization of a cell wall invertase from copper tolerant population of Elsholtzia haichowensis
- Construction of cDNA library from Prunus campanulata leaves and preliminary expressed sequence tag (EST) analysis during cold stress
- Phylogenetic utility of the geometric model of the body form in leeches (Clitellata: Hirudinida)
- Substrate choice by the alien snail Ferrissia fragilis (Gastropoda: Planorbidae) in an industrial area: A case study in a forest pond (Southern Poland)
- History of two critically endangered grassland snails (Pulmonata: Helicellinae) in the Czech Republic with first molecular data on extinct populations
- Monteustium marezensis gen. n., sp. n. and the first record of Italustiun eframi (Acari: Prostigmata: Erythraeidae: Balaustiinae) from Montenegro
- Lithobius (Ezembius) laevidentata sp. n., a new species (Chilopoda: Lithobiomorpha: Lithobiidae) from the Northwest region of China
- Aphids in jeopardy? Aphid communities on xerothermic habitats
- Habitat and weather requirements of diurnal raptors wintering in river valleys
