Abstract
The haustorium is a specific organ for penetration of host tissue in parasitic plants. To better understand the molecular mechanisms of the establishment of host-parasite relationships using gene expression profiles, early stage haustorium germination from mature mistletoe seeds was examined. We have generated a cDNA library that is an excellent source of expressed sequence tags (ESTs) specifically related to haustorium development and host penetration. We analyzed 4,771 ESTs derived from a mistletoe (Viscum album) haustorium cDNA library. Cluster analysis identified 727 contigs (23.88%) and 2,317 singletons (76.12%), yielding a total of 3,044 unique genes. Among these genes, 1,760 clones (57.8%) showed significant similarity to known genes; the remaining 1,284 clones were novel. Annotation of genes with Gene Ontology indicated that the most abundant ESTs reveals that defense against biotic / abiotic stresses, primary metabolic processes, cell wall loosening and modification is critical for haustorium development and establishment of the host-parasite connection. Especially, it is likely that xyloglucan endotransglycosylases (XETs), glucanase, expansins and other cell wall hydrolases cooperate in cell wall modification during the stages of host-parasite connection. Functional expression of full-length forms corresponding to target ESTs was applied to characterize haustorium genes. This EST information will help gene discovery and characterization and provide promising targets for genetic engineering. The EST analysis presented here represents the first reported transcriptome data for V. album var. coloratum. Although the number of ESTs analyzed may not be sufficient to completely elucidate the mechanism of the host-parasite connection, the integrated approaches reported here represent an essential step toward understanding haustorium development in parasitic plants and provide a valuable resource for defining molecular mechanisms in the host-parasite connection.
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Literature Cited
Albert, M., M. Werner, P. Proksch, S.C. Fry, and R. Kaldenhoff. 2004. The cell wall-modifying xyloglucan endotransglycosylase/hydrolase lexth1 is expressed during the defense reaction of tomato against the plant parasite Cuscuta reflexa. Plant Biol. 6:402–407.
Albrecht, H., J.I. Yoder, and D.A. Phillips. 1999. Flavonoids promote haustoria formation in the root parasite Triphysaria versicolor. Plant Physiol. 119:585–591.
Borsics, T. and M. Lados. 2002. Dodder infection induces the expression of a pathogenesis-related gene of the family PR-10 in alfalfa. J. Expt. Bot. 53:1831–1832.
Bourquin, V., N. Nishikubo, H. Abe, H. Brumer, S. Denman, M. Eklund, M. Christiernin, T.T. Teeri, B. Sundberg, and E.J. Mellerowic. 2002. Xyloglucan endotransglycosylases have a function during the formation of secondary cell walls of vascular tissues. Plant Cell 14:3073–3088.
Braun, J.M., H.L. Ko, and J.M. Schierholz. 2002. Standardized mistletoe extract augments immune response and down regulates local and metastatic tumor growth in murine models. Anticancer Res. 22:4187–4190.
Burger, A.M., U. Mengs, J.B. Schuler. 2001. Anticancer activity of an aqueous mistletoe extract (AME) in syngeneic murine tumor models. Anticancer Res. 21:1965–1968.
Büssing, A. 2000. Mistletoe, the genus Viscum. Harwood Academic Publishers, Amsterdam, The Netherlands. p. 83–85.
Campbell, P. and J. Braam. 1999. Xyloglucan endotransglycosylases: Diversity of genes, enzymes and potential wall-modifying functions. Trends Plant Sci. 4:361–366.
Catala, C., J.K.C. Rose, and A.B. Bennett. 2000. Auxin-regulated genes encoding cell wall-modifying proteins are expressed during early tomato fruit growth. Plant Physiol. 122:527–534.
Chang, S., J. Puryear, and J. Cairney. 1993. Simple and efficient method for isolating RNA from Pine trees. Plant Mol. Biol. Rep. 11:113–116.
Chen, F.H., H. Nonogaki, and K.J. Bradford. 2002. A gibberellin-regulated xyloglucan endotransglycosylase gene is expressed in the endosperm cap during tomato seed germination. J. Expt. Bot. 53:215–223.
Choi, D.W., J.D. Jung, Y.I. Ha, H.W. Park, D.S. In, H.J. Chung, and J.R. Liu. 2005. Analysis of transcripts in methyl jasmonate-treated ginseng hairy roots to identify genes involved in the biosynthesis of ginsenosides and other secondary metabolites. Plant Cell Rep. 23:557–566.
Chou, H.H. and H.H. Holmes 2001. DNA sequence quality trimming and EST vector removal. Bioinformatics 17:1093–1104.
Cosgrove, D.J. 1999. Enzymes and other agents that enhance cell wall extensibility. Annu. Rev. Plant Physiol. Plant Mol. Biol. 50:391–417.
Delseny, M., R. Cooke, M. Raynal, and F. Grellet. 1997. The Arabidopsis thaliana cDNA sequencing projects. FEBS Lett. 405:129–132.
Dos Santos, C.V., P. Letousey, P. Delavault, and P. Thalouarn. 2003. Defense gene expression analysis of Arabidopsis thaliana parasitized by Orobanche ramose. Phytopathology 93:451–457.
Elluru, S., J.P.D. van Huyen, B. Wootla, S. Delignat, F. Prost, V.S. Negi., and S.V. Kaveri. 2007. Tumor regressive effects of Viscum album preparations. Medicina 67:85–89.
Emani, C., J.M. Garcia, E. Lopata-Finch, M.J. Pozo, P. Uribe, D.J. Kim, G. Sunilkumar, D. R. Cook, C.M. Kenerley, and K.S. Rathore. 2003. Enhanced fungal resistance in transgenic cotton expressing an endochitinase gene from Trichoderma virens. Plant Biotechnol. J. 1:321–336.
Epple, P., K. Apel, and H. Bohlmann. 1997. Overexpression of an endogenous thionin enhances resistance of Arabidopsis against Fusarium oxysporum. Plant Cell 9:509–520.
Ewing, B., L. Hillier, M.C. Wendl, and P. Green. 1998. Base-calling of automated sequencer traces using Phred. Genome Res. 8:175–185.
Fukunaga, T., K. Nishiya, and I. Kajikawa. 1989. Studies on the constituents of Japanese mistletoes from different host trees and their antimicrobial and hypotensive properties. Chem. Pharm. Bull. (Tokyo) 37:1543–1546.
Guan, B., G.Q. Jiang, Y.X. Wang, Z.C. Wang, Y. Haxim, Q.A. Bao, Y.Z. Hu, F.C. Zhang, and Y. Wang. 2010. Identification of differentially expressed transcripts involved in the salt-stress response of Salsola ferganica by suppression subtractive hybridization. Plant Cell Tiss. Org. Cult. 103:343–352.
Groot, S.P.C., B. Kieliszewska-Rokicka, E. Vermeer, and C.M. Karssen. 1988. Gibberellin-induced hydrolysis of endosperm cell walls in gibberellin-deficient tomato seed prior to radicle protrusion. Planta 174:500–504
Huang, X. and A. Madan. 1999. CAP3: A DNA sequence assembly program. Genome Res. 9:868–877.
Jain, S., D. Kumar, M. Jain, P. Chaudhary, R. Deswal, and N.B. Sarin. 2012. Ectopic overexpression of a salt stress-induced pathogenesis-related class 10 protein (PR10) gene from peanut (Arachis hypogaea L.) affords broad spectrum abiotic stress tolerance in transgenic tobacco. Plant Cell Tiss. Org. Cult. 109:19–31.
Jin, T., Q. Chang, W. Li, D. Yin, Z. Li, D. Wang, B. Liu, and L. Liu. 2010. Stress-inducible expression of GmDREB1 conferred salt tolerance in transgenic alfalfa. Plant Cell Tiss. Org. Cult. 100:219–227.
Jung, H.Y., A.N. Lee, S.J. Kim, S.M. Park, T.J. Song, B.W. Ko, Y.H. Kim, H.S. An, Y.C. Yoo, B.R. Min, and J.B. Kim. 2008. Effect of Korean mistletoe (Viscum album coloratum) extract (KME) on the extensions of life span in silkworm moth, Bombyx mori. Korean J. Pharm. 39:310–315.
Kagi, J.H.R. and A. Schaffer. 1988. Biochemistry of metallothionein. Biochemistry 27:8509–8515.
Kebdani, N., L. Pieuchot, E. Deleury, F. Panabieres, J.Y. Le Berre, and M. Gourgues. 2010. Cellular and molecular characterization of Phytophthora parasitica appressorium-mediated penetration. New Phytol. 185:248–257.
Kim, D., R. Kocz, L. Boone, W.J. Keyes, and D.G. Lynn. 1998. On becoming a parasite: Evaluating the role of wall oxidases in parasitic plant development. Chem. Biol. 5:103–117.
Kim, S.W., S.M. Ko, and J.R. Liu. 2008. In vitro seed germination and callus formation on flower bud of Korean mistletoe (Viscum album L. var. cololatum [Kom.] Ohwi). J. Plant Biotechnol. 35:47–53.
Kuijt, J. 1977. Haustoria of phanerogamic parasites. Annu. Rev. Phytophathol. 17:91–118.
Kwak, J.M., S.A. Kim, S.W. Hong, and H.G. Nam. 1997. Evaluation of 515 expressed sequence tags obtained from guard cells of Brassica campestris. Planta 202:9–17.
Lange, B.M., M.R. Wildung, E.J. Stauber, C. Sanchez, D. Pouchnik, and R. Croteau. 2000. Probing essential oil biosynthesis and secretion by functional evaluation of expressed sequence tags from mint glandular trichomes. Proc. Natl. Acad. Sci. USA 97:2934–2939.
Liu, J., C. Hara, M. Umeda, Y. Zhao, T.W. Okita, and H. Uchimiya. 1995. Analysis of randomly isolated cDNAs from developing endosperms of rice (Oryza sativa L.): Evaluation of expressed sequence tags, and expression levels of mRNAs. Plant Mol. Biol. 29:685–689.
Liu, J.R., D.W. Choi, H.J. Chung, and S.S. Woo. 2002. Production of useful secondary metabolites in plants: Functional genomics approaches. J. Plant Biol. 45:1–6.
Löffler, B. 1923. Contributions to the development history of the female flower, berry and haustorium of mistletoe (Viscum album L.). Tharandter Forstliches Jahrbuch 74:49–62.
Maiti, S., D. Chakraborty, S. Kundu, S. Paul, S. Sengupta, and A. Pal. 2011. Developmentally regulated temporal expression and differential acid invertase activity in differentiating cotyledonary explants of mungbean [Vigna radiata (L.) Wilczek]. Plant Cell Tiss. Org. Cult. 107:417–425.
Matvienko, M., A. Wojtowicz, R. Wrobe, D. Jamison, Y. Goldwasser, and J.I. Yoder. 2001a. Quinone oxidoreductase message levels are differentially regulated in parasitic and non-parasitic plants exposed to allelopathic quinines. Plant J. 25:375–387.
Matvienko, M., M.J. Torres, and J.I. Yoder. 2001b. Transcriptional responses in the hemiparasitic plant Triphysaria versicolor to host plant signals. Plant Physiol. 127:272–282.
McCann, M.C., B. Wells, and K. Roberts. 1990. Direct visualization of cross-links in the primary cell wall. J. Cell Sci. 96:323–334.
Menossi, M., M.C. Silva-Filho, M. Vincentz, M.A. Van-Sluys, and G.M. Souza. 2008. Sugarcane functional genomics: Gene discovery for agronomic trait development. Int. J. Plant Genomics 2008:1–11.
Mir, G., J. Domènech, G. Huguet, W.J. Guo, P. Goldsbrough, S. Atrian, and M. Molinas. 2004. A plant type 2 metallothionein (MT) from cork block responds to oxidative stress. J. Expt. Bot. 55:2483–2493.
Nickrent, D.L., V. Malecot, R. Vidal-Russell, and J.P. Der. 2010. A revised classification of Santalales. Taxon 59:538–558.
Ochocka, R.J. and A. Piotrowski. 2002. Biologically active compounds from European mistletoe (Viscum album L.). Can. J. Plant Pathol. 24:21–28.
Palmieri, D.A., V.M. Novelli, M. Bastianel, M. Cristofani, G.A. Monge, E.F. Carlos, A.C. Oliveira, and M.A. Machado. 2007. Frequency and distribution of microsatellites from ESTs of Citrus. Genet. Mol. Biol. 30:1009–1018.
Park, J.H., C.K. Hyun, and H.K. Shin. 1999. Cytotoxic effects of the components in heat-treated mistletoe (Viscum album). Cancer Lett. 139:207–213.
Park, W.B., S.Y. Lyu, and J.H. Kim. 2001. Inhibition of tumor growth and metastasis by Korean mistletoe lectin is associated with apoptosis and antiangiogenesis. Cancer Biother. Radiopharm. 16:439–447.
Pelegrini, P.B. and O.L. Franco. 2005. Plant γ-thionins: Novel insights on the mechanism of action of a multi-functional class of defense proteins. Int. J. Biochem. Cell Biol. 37:2239–2253.
Ribereau-Gayon, G., M.L. Jung, and M. Franz. 1997. Modulation of cytotoxicity and enhancement of cytokine release induced by Viscum album L. extracts or mistletoe lectins. Anti-Cancer Drugs 8:3–8.
Riopel, J.L. and M.P. Timko. 1995. Haustorial initiation and differentiation, p. 39–79. In: M.C. Press and J.D. Graves (eds.). Parasitic plants. Chapman and Hall, London.
Sallé, G. 1983. Germination and establishment of Viscum album L., p. 145–159. In: D.M. Calder and P. Bernhardt (eds.). The biology of mistletoes. Academic Press, London.
Selitrennikoff, C.P. 2001. Antifungal proteins. Appl. Environ. Microbiol. 67:2883–2894.
Stewart, G.R. and M.C. Press. 1990. The physiology and biochemistry of parasitic angiosperms. Annu. Rev. Plant Physiol. Plant Mol. Biol. 41:127–151.
Tang, Z., J.H. Choi, C. Hemmerich, A. Sarangi, J.K. Colbourne, and Q. Dong. 2009. ESTPiper-a web-based analysis pipeline for expressed sequence tags. BMC Genomics 10:174.
Thoday, D. 1951. The haustorial system of Viscum album. J. Expt. Bot. 2:1–19
Torres, M.J., A.A. Tomilov, N. Tomilova, R.L. Reagan, and J.I. Yoder. 2005. Pscroph, a parasitic plant EST database enriched for parasite associated transcripts. BMC Plant Biol. 5:24.
Wang, K., L. Gan, E. Jeffery, M. Gayle, A.M. Gown, M. Skelly, P.S. Nelson, W.V. Ng, M. Schummer, L. Hood, and J. Mulligan. 1999. Monitoring gene expression profile changes in ovarian carcinomas using cDNA microarray. Gene 229:102–108.
Wang, A., Y. Wang, Z. Gu, S. Li, Y. Shi, and X. Guo. 2011. Development of expressed sequence tags from the pearl oyster, Pinctada martensii Dunker. Mar. Biotechnol. 13:275–283.
Whitney, S.E.C., M.G.E. Gothard, J.T. Mitchell, and M.J. Gidley. 1999. Roles of cellulose and xyloglucan in determining the mechanical properties of primary cell walls. Plant Physiol. 121:657–663.
Yamamoto, K. and T. Sasaki. 1997. Large-scale EST sequencing in rice. Plant Mol. Biol. 35:135–144.
Yoshida, S., J.K. Ishida, N.M. Kamal, A.M. Ali, S. Namba, and K. Shirasu. 2010. A full-length enriched cDNA library and expressed sequence tag analysis of the parasitic weed, Striga hermonthica. BMC Plant Biol. 10:55.
Zasloff, M. 2002. Antimicrobial peptides of multicellular organisms. Nature 415:389–395.
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Ko, S.M., Kwon, Y.K., Kim, J.H. et al. Transcriptome analysis of mistletoe (Viscum album) haustorium development. Hortic. Environ. Biotechnol. 55, 352–361 (2014). https://doi.org/10.1007/s13580-014-0033-6
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DOI: https://doi.org/10.1007/s13580-014-0033-6