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A tale of three kingdoms: members of the Phylum Nematoda independently acquired the detoxifying enzyme cyanase through horizontal gene transfer from plants and bacteria

  • D. S. Zarlenga (a1), M. Mitreva (a2), P. Thompson (a1), R. Tyagi (a2), W. Tuo (a1) and E. P. Hoberg (a1)...


Horizontal gene transfer (HGT) has played an important role in the evolution of nematodes. Among candidate genes, cyanase, which is typically found only in plants, bacteria and fungi, is present in more than 35 members of the Phylum Nematoda, but absent from free-living and clade V organisms. Phylogenetic analyses showed that the cyanases of clade I organisms Trichinella spp., Trichuris spp. and Soboliphyme baturini (Subclass: Dorylaimia) represent a well-supported monophyletic clade with plant cyanases. In contrast, all cyanases found within the Subclass Chromadoria which encompasses filarioids, ascaridoids and strongyloids are homologous to those of bacteria. Western blots exhibited typical multimeric forms of the native molecule in protein extracts of Trichinella spiralis muscle larvae, where immunohistochemical staining localized the protein to the worm hypodermis and underlying muscle. Recombinant Trichinella cyanase was bioactive where gene transcription profiles support functional activity in vivo. Results suggest that: (1) independent HGT in parasitic nematodes originated from different Kingdoms; (2) cyanase acquired an active role in the biology of extant Trichinella; (3) acquisition occurred more than 400 million years ago (MYA), prior to the divergence of the Trichinellida and Dioctophymatida, and (4) early, free-living ancestors of the genus Trichinella had an association with terrestrial plants.


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Author for correspondence: D. S. Zarlenga, E-mail:


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Andersson, JO (2005) Lateral gene transfer in eukaryotes. Cellular and Molecular Life Sciences 62, 11821197.
Anderson, PM and Little, RM (1986) Kinetic properties of cyanase. Biochemistry 25, 16211626.
Anderson, IJ and Loftus, BJ (2005) Entamoeba histolytica: observations on metabolism based on the genome sequence. Experimental Parasitology 110, 173177.
Anderson, PM, Sung, Y-C and Fuchs, JA (1990) The cyanase operon and cyanate metabolism. FEMS Microbiology Reviews 7, 247252.
Blaxter, M (2007) Symbiont genes in host genomes: fragments with a future? Cell Host & Microbe 2, 211213.
Blaxter, M (2011) Nematodes: the worm and its relatives. PLoS Biology 9, e1001050.
Blaxter, M, Koutsovoulos, G, Jones, M, Kumar, S and Elsworth, B (2014) Phylogenomics of Nematoda. In Cotton, J, Hughes, J and Olson, P (eds), Next-generation Systematics. Cambridge, UK: Cambridge University Press, pp. 6283.
Bock, WJ (1959) Preadaptation and multiple evolutionary pathways. Evolution 13, 194211.
Brooks, DR and McLennan, DA (2002) The Nature of Diversity: An Evolutionary Voyage of Discovery. Chicago, USA: University of Chicago Press.
Crisp, A, Boschetti, C, Perry, M, Tunnacliffe, A and Micklem, G (2015) Expression of multiple horizontally acquired genes is a hallmark of both vertebrate and invertebrate genomes. Genome Biology 16, 50.
Dabert, M, Witalinski, W, Kazmierski, A, Olszanowski, Z and Dabert, J (2010) Molecular phylogeny of acariform mites (Acari, Arachnida): strong conflict between phylogenetic signal and long-branch attraction artifacts. Molecular Phylogenetics and Evolution 56, 222241.
Dagan, T, Artzy-Randrup, Y and Martin, W (2008) Modular networks and cumulative impact of lateral transfer in prokaryote genome evolution. Proceedings of the National Academy of Sciences, USA 105, 1003910044.
Dame, JB, Murrell, KD, Worley, DE and Schad, GA (1987) Trichinella spiralis: genetic evidence for synanthropic subspecies in sylvatic hosts. Experimental Parasitology 64, 195203.
Danchin, EGJ, Perfus-Barbeoch, L, Rancurel, C, Thorpe, P, Da Rocha, M, Bajew, S, Neilson, R, Guzeeva, ES, Da Silva, C, Guy, J, Labadie, K, Esmenjaud, D, Helder, J, Jones, JT and den Akker, SE (2017) The transcriptomes of Xiphinema index and Longidorus elongatus suggest independent acquisition of some plant parasitism genes by horizontal gene transfer in early-branching nematodes. Genes (Basel) 8, E287.
Dieterich, C and Sommer, RJ (2009) How to become a parasite – lessons from the genomes of nematodes. Trends in Genetics 25, 203209.
Dieterich, C, Clifton, SW, Schuster, LN, Chinwalla, A, Delehaunty, K, Dinkelacker, I, Fulton, L, Fulton, R, Godfrey, J, Minx, P, Mitreva, M, Roeseler, W, Tian, H, Witte, H, Yang, SP, Wilson, RK and Sommer, RJ (2008) The Pristionchus pacificus genome provides a unique perspective on nematode lifestyle and parasitism. Nature Genetics 40, 11931198.
Ebbs, S (2004) Biological degradation of cyanide compounds. Current Opinion in Biotechnology 15, 231236.
el Kouni, MH (2017) Pyrimidine metabolism in schistosomes: a comparison with other parasites and the search for potential chemotherapeutic targets. Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology 213, 5580.
Elleuche, S and Pöggeler, S (2008) A cyanase is transcriptionally regulated by arginine and involved in cyanate decomposition in Sordaria macrospora. Fungal Genetics and Biology 45, 14581469.
Elmore, MH, McGary, KL, Wisecaver, JH, Slot, JC, Geiser, DM, Sink, S, O'Donnell, K and Rokas, A (2015) Clustering of two genes putatively involved in cyanate detoxification evolved recently and independently in multiple fungal lineages. Genome Biology and Evolution 7, 789800.
Espie, GS, Jalali, F, Tong, T, Zacal, NJ and So, AK (2007) Involvement of the cynABDS operon and the CO2-concentrating mechanism in the light-dependent transport and metabolism of cyanate by cyanobacteria. Journal of Bacteriology 189, 10131024.
Gan, YH, Chua, KL, Chua, HH, Liu, B, Hii, CS, Chong, HL and Tan, P (2002) Characterization of Burkholderia pseudomallei infection and identification of novel virulence factors using a Caenorhabditis elegans host system. Molecular Microbiology 44, 11851197.
Gilbert, A, Castro, J, Ferguson, JD and Gorden, C (1973) Amine excretion in excysted larvae and adults of Trichinella spiralis. Comparative Biochemistry and Physiology. Part A, Molecular & Integrative Physiology 45, 819828.
Guilloton, M and Karst, F (1987) Cyanate specifically inhibits arginine biosynthesis in Escherichia coli K12: a case of by-product inhibition? Journal of General Microbiology 133, 655665.
Guindon, S, Dufayard, JF, Lefort, V, Anisimova, M, Hordijk, W and Gascuel, O (2010) New algorithms and methods to estimate Maximum-Likelihood phylogenies: assessing the performance of PhyML 3.0. Systematic Biology 59, 307321.
Haegeman, A, Jones, JT and Danchin, EGJ (2011) Horizontal gene transfer in nematodes: a catalyst for plant parasitism? Molecular Plant-microbe Interactions 24, 879887.
Haskins, WT and Weinstein, PP (1957) Nitrogenous excretory products of Trichinella spiralis larvae. Journal of Parasitology 43, 1924.
Hewagama, A, Guy, HI, Vickrey, JF and Evans, DR (1999) Functional linkage between the glutaminase and synthetase domains of carbamoyl-phosphate synthetase. Role of serine 44 in carbamoyl-phosphate synthetase-aspartate carbamoyltransferase-dihydroorotase (cad). The Journal of Biological Chemistry 274, 2824028245.
Holterman, M, van der Wurff, A, van den Elsen, S, van Megen, H, Bongers, T, Holovachov, O, Bakker, J and Helder, J (2006) Phylum-wide analysis of SSU rDNA reveals deep phylogenetic relationships among nematodes and accelerated evolution toward crown clades. Molecular Biology and Evolution 23, 17921800.
Kamennaya, NA and Post, AF (2013) Distribution and expression of the cyanate acquisition potential among cyanobacterial populations in oligotrophic marine waters. Limnology and Oceanography 58, 19591971.
Keeling, PJ and Palmer, JD (2008) Horizontal gene transfer in eukaryotic evolution. Nature Reviews. Genetics 9, 605618.
Keen, NT and Roberts, PA (1998) Plant parasitic nematodes: digesting a page from the microbe book. Proceedings of the National Academy of Sciences, USA 95, 47894790.
Kunz, DA and Nagappan, O (1989) Cyanase-mediated utilization of cyanate in Pseudomonas fluorescens. Applied and Environmental Microbiology 55, 256258.
Maeda, S and Omata, T (2009) Nitrite transport activity of the ABC-type cyanate transporter of the cyanobacterium Synechococcus elongatus. Journal of Bacteriology 191, 32653272.
Marti, HP, Murrell, KD and Gamble, HR (1987) Trichinella spiralis: immunization of pigs with newborn larval antigens. Experimental Parasitology 63, 6873.
McGill, LM, Fitzpatrick, DA, Pisani, D and Burnell, AM (2017) Estimation of phylogenetic divergence times in Panagrolaimidae and other nematodes using relaxed molecular clocks calibrated with insect and crustacean fossils. Nematology 19, 899913.
Mitreva, M, Smant, G and Helder, J (2009) Role of horizontal gene transfer in the evolution of plant parasitism among nematodes. Methods in Molecular Biology 532, 517535.
Mitreva, M, Jasmer, DP, Zarlenga, DS, Wang, Z, Abubucker, S, Martin, J, Taylor, CM, Yin, Y, Fulton, L, Minx, P, Yang, SP, Warren, WC, Fulton, RS, Bhonagiri, V, Zhang, X, Hallsworth-Pepin, K, Clifton, SW, McCarter, JP, Appleton, J, Mardis, ER and Wilson, RK (2011) The draft genome of the parasitic nematode Trichinella spiralis. Nature Genetics 43, 228235.
Mitreva-Dautova, M, Roze, E, Overmars, H, de Graaff, L, Schots, A, Helder, J, Goverse, A, Bakker, J and Smant, G (2006) A symbiont-independent endo-1,4-beta-xylanase from the plant-parasitic nematode Meloidogyne incognita. Molecular Plant-microbe Interactions 19, 521529.
Morris, JL, Puttick, MN, Clark, JW, Edwards, D, Kenrick, P, Pressel, S, Wellman, CH, Yang, Z, Schneider, H and Donoghue, PCJ (2018) The timescale of early land plant evolution. Proceedings of the National Academy of Sciences, USA 115, E2274E2283.
Nishikawa, BK, Fowlkes, DM and Kay, BK (1989) Convenient uses of polymerase chain reaction in analyzing recombinant cDNA clones. Biotechniques 7, 730735.
Opperman, CH, Bird, DM, Williamson, VM, Rokhsar, DS, Burke, M, Cohn, J, Cromer, J, Diener, S, Gajan, J, Graham, S, Houfek, TD, Liu, Q, Mitros, T, Schaff, J, Schaffer, R, Scholl, E, Sosinski, BR, Thomas, VP and Windham, E (2008) Sequence and genetic map of Meloidogyne hapla: a compact nematode genome for plant parasitism. Proceedings of the National Academy of Sciences, USA 105, 1480214807.
Polz, MF, Alm, EJ and Hanage, WP (2013) Horizontal gene transfer and the evolution of bacterial and archaeal population structure. Trends in Genetics 29, 170175.
Rensing, SA, Lang, D, Zimmer, AD, Terry, A, Salamov, A, Shapiro, H, Nishiyama, T, Perroud, PF, Lindquist, EA, Kamisugi, Y, Tanahashi, T, Sakakibara, K, Fujita, T, Oishi, K, Shin-I, T, Kuroki, Y, Toyoda, A, Suzuki, Y, Hashimoto, S, Yamaguchi, K, Sugano, S, Kohara, Y, Fujiyama, A, Anterola, A, Aoki, S, Ashton, N, Barbazuk, WB, Barker, E, Bennetzen, JL, Blankenship, R, Cho, SH, Dutcher, SK, Estelle, M, Fawcett, JA, Gundlach, H, Hanada, K, Heyl, A, Hicks, KA, Hughes, J, Lohr, M, Mayer, K, Melkozernov, A, Murata, T, Nelson, DR, Pils, B, Prigge, M, Reiss, B, Renner, T, Rombauts, S, Rushton, PJ, Sanderfoot, A, Schween, G, Shiu, SH, Stueber, K, Theodoulou, FL, Tu, H, Van de Peer, Y, Verrier, PJ, Waters, E, Wood, A, Yang, L, Cove, D, Cuming, AC, Hasebe, M, Lucas, S, Mishler, BD, Reski, R, Grigoriev, IV, Quatrano, RS and Boore, JL (2008) The Physcomitrella genome reveals evolutionary insights into the conquest of land by plants. Science 319, 6469.
Richards, TA, Dacks, JB, Jenkinson, JM, Thornton, CR and Talbot, NJ (2006) Evolution of filamentous plant pathogens: gene exchange across eukaryotic kingdoms. Current Biology 16, 18571864.
Rödelsperger, C and Sommer, RJ (2011) Computational archaeology of the Pristionchus pacificus genome reveals evidence of horizontal gene transfers from insects. BioMed Central Evolutionary Biology 11, 239.
Sanderson, MJ, Thorne, JL, Wikstrom, N and Bremer, K (2004) Molecular evidence on plant divergence times. American Journal of Botany 91, 16561665.
Schlachter, CR, Klapper, V, Wybouw, N, Radford, T, Van Leeuwen, T, Grbic, M and Chruszcz, M (2017) Structural characterization of a eukaryotic cyanase from Tetranychus urticae. Journal of Agricultural and Food Chemistry 65, 54535462.
Smant, G, Stokkermans, JP, Yan, Y, de Boer, JM, Baum, TJ, Wang, X, Hussey, RS, Gommers, FJ, Henrissat, B, Davis, EL, Helder, J, Schots, A and Bakker, J (1998) Endogenous cellulases in animals: isolation of beta-1, 4-endoglucanase genes from two species of plant-parasitic cyst nematodes. Proceedings of the National Academy of Sciences, USA 9, 49064911.
Walsh, MA, Otwinowski, Z, Perrakis, A, Anderson, PM and Joachimiak Andrzej, A (2000) Structure of cyanase reveals that a novel dimeric and decameric arrangement of subunits is required for formation of the enzyme active site. Structure 8, 505514.
Wang, Z, Zarlenga, D, Martin, J, Abubucker, S and Mitreva, M (2012) Exploring metazoan evolution through dynamic and holistic changes in protein families and domains. BioMed Central Evolutionary Biology 12, 138.
Wybouw, N, Balabanidou, V, Ballhorn, DJ, Dermauw, W, Grbić, M, Vontas, J and Van Leeuwen, T (2012) A horizontally transferred cyanase gene in the spider mite Tetranychus urticae is involved in cyanate metabolism and is differentially expressed upon host plant change. Insect Biochemistry and Molecular Biology 42, 881889.
Wybouw, N, Dermauw, W, Tirry, L, Stevens, C, Grbić, M, Feyereisen, R and Van Leeuwen, T (2014) A gene horizontally transferred from bacteria protects arthropods from host plant cyanide poisoning. eLIFE 3, e02365.
Wybouw, N, Van Leeuwen, T and Dermauw, W (2018) A massive incorporation of microbial genes into the genome of Tetranychus urticae, a polyphagous arthropod herbivore. Insect Molecular Biology 27, 333351.
Zarlenga, DS, Boyd, P, Lichtenfels, JR, Hill, D and Gamble, HR (2002) Identification and characterisation of a cDNA sequence encoding a glutamic acid-rich protein specifically transcribed in Trichinella spiralis newborn larvae and recognised by infected swine serum. International Journal for Parasitology 32, 13611370.
Zarlenga, DS, Nisbet, AJ, Gasbarre, LC and Garrett, WM (2011) A calcium-activated nucleotidase secreted from Ostertagia ostertagi 4th-stage larvae is a member of the novel salivary apyrases present in blood-feeding arthropods. Parasitology 138, 333343.
Zarlenga, D, Wang, Z and Mitreva, M (2016) Trichinella spiralis: adaptation and parasitism. Veterinary Parasitology 231, 821.


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