Abstract
A cDNA library was constructed from the newly developed young leaves of Aralia elata Seem, and a total of 2,755 cDNAs were partially sequenced. Sequences with high quality greater than 150 bp after trimming vector and ambiguous sequences resulted in 2,689 ESTs. These ESTs were clustered into 2,010 unigenes consisting of 376 contigs with two or more ESTs and 1,634 singletons. A homology search with BLASTX against the NCBI database identified 1,256 (63%) unigenes homologous to proteins of known or putative function, 350 (17%) genes assumed as proteins with unknown function, and 404 (20%) unigenes with no significant match to any protein sequence, including sequences with matches greater than the E value of −105 in the NCBI database. The expression level of each gene was analyzed based on the number of cDNA clones in each contig composed of at least 10 ESTs. The most abundant gene identified was acid phosphatase. The genes related to chlorophyll a-b binding protein, elongation factor 1-alpha, ribulose 1,5-bisphosphate carboxylase small subunit and cyclophilin were also plentiful. The 1,259 unigenes matched with proteins of known or putative function were classified into a functional category using the MIPS BLAST program and 616 unigenes were assigned to putative biochemical functions. Furthermore, five, sixteen and twelve unigenes in our EST set were assumed as proteins involved in squalene synthesis, cytochrome P450 and glycosyltransferase, respectively. These genes should be further explored for their involvement in the saponin biosynthesis pathway in A. elata Seem.
Similar content being viewed by others
Literature Cited
Brenner, E.D., M.S. Katari, D.W. Stevenson, S.A. Rudd, A.W. Douglas, W.N. Moss, R.W. Twigg, S.J. Runko, G.M. Stellari, W.R. McCombie, and G.M. Coruzzi. 2005. EST analysis in Ginkgo biloba: An assessment of conserved developmental regulators and gymnosperm specific genes. BMC Genomics 143:1–10.
Fung, M.C., M.T. Lau, and X.G. Chen. 2002. Expressed sequence tag (EST) analysis of a Schistosoma japonicum cercariae cDNA library. Acta Trop. 82:215–24.
Huber, D.P., R.N. Philippe, K.A. Godard, R.N. Sturrock, and J. Bohlmann. 2005. Characterization of four terpene synthase cDNAs from methyl jasmonate-induced Douglas-fir, Pseudotsuga menziesii. Phytochemistry 66:1427–1439.
Jantasuriyarat, C., M. Gowda, K. Haller, J. Hatfield, G. Stahlberg, E. Lu, B. Zhou, H. Li, H. Kim, Y. Yu, R.A. Dean, R.A. Wing, C. Soderlund, and G.L. Wang. 2005. Large-scale identification of expressed sequence tags involved in rice and rice blast fungus interaction. Plant Physiol. 138:105–115.
Jenner, H., B. Townsend, and A. Osbourn. 2005. Unravelling triterpene glycoside synthesis in plants: Phytochemistry and functional genomics join forces. Planta 220:503–506.
Jouannic, S., X. Argout, F. Lechauve, C. Fizames, A. Borgel, F. Morcillo, F. Aberlenc-Bertossi, Y. Duval, and J. Tregear. 2005. Analysis of expressed sequence tags from oil palm (Elaeis guineensis). FEBS Lett. 579:2709–2714.
Kim, J.S., S.H. Shim, S. Chae, S.J. Han, S.S. Kang, K.H. Son, H.W. Chang, H.P. Kim, and K. Bae. 2005. Saponins and other constituents from the leaves of Aralia elata. Chem. Pharm. Bull. 53:696–700.
Kohara, A., C. Nakajima, K. Hashimoto, T. Ikenaga, H. Tanaka, Y. Shoyama, S. Yoshida, and T. Muranaka. 2005. A novel glucosyltransferase involved in steroid saponin biosynthesis in Solanum aculeatissimum. Plant Mol. Biol. 57:225–239.
Kushiro, T., M. Shibuya, and Y. Ebizuka. 1998. Beta-amyrin synthase cloning of oxidosqualene cyclase that catalyzes theformation of the most popular triterpene among higher plants. Eur. J. Biochem. 256:238–244.
Liu, P.G. and Q. Yang. 2005. Identification of genes with a biocontrol function in Trichoderma harzianum mycelium using the expressed sequence tag approach. Res. Microbiol. 156:416–423.
Ma, Z.Q., S.J. Song, W. Li, and S.X. Xu. 2005. Two new saponins from the bud of Aralia elata (Miq.) Seem. J. Asian Nat. Prod. Res. 7:817–821.
Mou, C.Y., S.C. Zhang, J.H. Lin, W.L. Yang, W.Y. Wu, J.W. Wei, X.K. Wu, J.C. Du, Z.Y. Fu, L.T. Ye, Y. Lu, X.J. **e, Y.L. Wang, and A.L. Xu. 2002. EST analysis of mRNAs expressed in neurula of Chinese amphioxus. Biochem. Bioph. Res. Co. 299:74–84.
Murata, J., D. Bienzle, J.E. Brandle, C.W. Sensen, and V. De Luca. 2006. Expressed sequence tags from Madagascar periwinkle (Catharanthus roseus). FEBS Lett. 580:4501–4507.
Osbourn, A.E. 2003. Saponins in cereals. Phytochemistry 62:1–4.
Palazon, J., E. Moyano, M. Bonfill, L.T. Osuna, R.M. Cusido, and M.T. Pinol. 2006. Effect of organogenesis on steroidal saponin biosynthesis in calli cultures of Ruscus aculeatus. Fitoterapia 77: 216–220.
Park, J.S., J.B. Kim, B.S. Hahn, K.H. Kim, S.H. Ha, J.B. Kim, and Y.H. Kim. 2004. EST analysis of genes involved in secondary metabolism in Camellia sinensis (tea), using suppression subtractive hybridization. Plant Sci. 116:953–961.
Qi, X., S. Bakht, M. Leggett, C. Maxwell, R. Melton, and A. Osbourn. 2004. A gene cluster for secondary metabolism in oat: Implications for the evolution of metabolic diversity in plants. P. Natl. Acad. Sci. USA. 101:8233–8238.
Rudd, S. 2003. Expressed sequence tags: Alternative or complement to whole genome sequences? Trends Plant Sci. 8:321–329.
Seo, J.W., J.H. Jeong, C.G. Shin, S.C. Lo, S.S. Han, K.W. Yu, E. Harada, J.Y. Han, and Y.E. Choi. 2005. Overexpression of squalene synthase in Eleutherococcus senticosus increasesphytosterol and triterpene accumulation. Phytochemistry 66:869–877.
Shelton, D., D. Leach, P. Baverstock, and R. Henry. 2002. Isolation of genes involved in secondary metabolism from Melaleuca alternifolia (Cheel) using expressed sequence tags (ESTs). Plant Sci. 162:9–15.
Shelton, D., D. Zabaras, S. Chohan, S.G. Wyllie, P. Baverstock, D. Leach, and R. Henry. 2004. Isolation and partial characterization of a putative monoterpene synthase from Melaleuca alternifolia. Plant Physiol. Bioch. 42:875–882.
Song, S.J., N. Nakamura, C.M. Ma, M. Hattori, and S.X. Xu. 2001. Five saponins from the root bark of Aralia elata. Phytochemistry 56:491–497.
Sterky, F., R.R. Bhalerao, P. Unneberg, B. Segerman, P. Nilsson, A.M. Brunner, L. Charbonnel-Campaa, J.J. Lindvall, K. Tandre, S.H. Strauss, B. Sundberg, P. Gustafsson, M. Uhlen, R.P. Bhalerao, O. Nilsson, G. Sandberg, J. Karlsson, J. Lundeberg, and S. Jansson. 2004. A Populus EST resource for plant functional genomics. P. Natl. Acad. Sci. USA. 101:13951–1396.
Suzuki, H., L. Achnine, R. Xu, S.P. Matsuda, and R.A. Dixon. 2002. A genomics approach to the early stages of triterpene saponin bio synthesis in Medicago truncatula. Plant J. 32:1033–1048.
Tomatsu, M., M. Ohnishi-Kameyama, and N. Shibamoto. 2003. Aralin, a new cytotoxic protein from Aralia elata, inducing apoptosis in human cancer cells. Cancer Lett. 199:19–25.
Wang, Y.C., C.P. Yang, G.F. Liu, J. Jiang, and J.H. Wu. 2006. Generation and analysis of expressed sequence tags from a cDNA library of Tamarix androssowii. Plant Sci. 170:28–36.
Yagi-Chaves, S.N., G. Liu, K. Yamashita, M. Manabe, S.J. Song, and H. Kodama. 2006. Effect of five triterpenoid compounds isolated from root bark of Aralia elata on stimulus-induced superoxide generation, tyrosyl or serine/threoninephosphorylation and translocation of p47 (phox), p67 (phox), and rac to cellmembrane in human neutrophils. Arch. Biochem. Biophys. 446:84–90.
Yang, Y., S. Cun, X. **e, J. Lin, J. Wei, W. Yang, C. Mou, C. Yu, L. Ye, Y. Lu, Z. Fu, and A. Xu. 2003. EST analysis of gene expression in the tentacle of Cyanea capillata. FEBS Lett. 538: 183–191.
Author information
Authors and Affiliations
Corresponding author
Additional information
These authors contributed equally to this work.
Rights and permissions
About this article
Cite this article
Boo, K.H., Lee, D., **, S.B. et al. A profile of expressed sequence tags in newly develo** leaves of Aralia elata seem. Hortic. Environ. Biotechnol. 53, 421–427 (2012). https://doi.org/10.1007/s13580-012-0741-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13580-012-0741-8