Abstract
Mitochondrial DNA (mtDNA) haplotypes were determined using restriction fragment length polymorphism (RFLP) for P. infestans sampled from 513 foliar lesions of late blight found on potato and tomato in different regions of Ethiopia. Among the four reported mitochondrial haplotypes of Phytophthora infestans, Ia, Ib and IIb were detected in 93 % of the samples analyzed but the vast majority of these were Ia. The remaining 7 % represented a previously unreported haplotype. DNA sequencing of this new haplotype also confirmed a single base nucleotide substitution that resulted in loss of EcoRI restriction site and gain of two additional MspI sites in cox1 and atp1 genes, respectively. There were 28 polymorphic sites among all nucleotide sequences including five reference isolates. Sites with alignment gaps were observed in P4 with one nucleotide deletion in 11 Ethiopian isolates. None of the reference sequence produced frame-shifts, with the exception of the 3-nucleotide deletion in the P4 region by Phytophthora andina, a feature that can be used to distinguish the new Ethiopian isolates from P. andina. While a distinguishing molecular data presented here clearly separated them from P. infestans, 7 % of the isolates that share this feature formed an important component of the late blight pathogen causing disease on Solanum tuberosum in Ethiopia. Thus, these Ethiopian isolates could represent a novel Phytophthora species reported for the first time here.
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References
Adler N, Erselius LJ, Chacon MG, Flier WG, Ordonez ME, Kroon LPNM, Forbes GA (2004) Genetic diversity of Phytophthora infestans sensu lato in Ecuador provides new insight into the origin of this important plant pathogen. Phytopatholology 94:154–162
Avila-Adame C, Gómez-Alpizar L, Zisman V, Jones K, Buell CR, Ristaino JB (2006) Mitochondrial genome sequences and molecular evolution of the Irish potato famine pathogen, Phytophthora infestans. Curr Genet 49:39–46
Berga L, Gebre-Medhin W, Teriessa J. Bereketsehai T (1992) Potato agronomy research. In: Herath E, Lemma D (eds) Horticulture research and development in Ethiopia, Proceedings of the 2nd National Horticultural Workshop of Ethiopia, Institute of Agricultural Research, Addis Ababa, 2–5 February, pp. 182–196
Blair JE, Coffey MD, Park SY, Geiser DM, Kang S (2008) A multi-locus phylogeny for Phytophthora utilizing markers derived from complete genome sequences. Fungal Gen Biol 45:266–277
Carter DA, Archer SA, Buck KW, Shaw DS, Shattock RC (1990) Restriction fragment length polzmorphism of mitocondrial DNA of Phytophthora infestans. Mycol Res 94:1123–1128
Chen JZ, Hebert PDN (1999) Intra-individual sequence diversity and hierarchical approach to the study of mitochondrial DNA mutations. Mutat Res 434:205–217
Chesnick JM, Goff M, Graham J, Ocampo C, Lang BF, Seif E, Burger G (2000) The mitochondrial genome of the stramenopile alga Chrysodidymus synuroideus. Complete sequence, gene content and genome organization. Nucleic Acid Res 28:2512–2518
Cooke DEL, Lees AK (2004) Markers, old and new, for examining Phytophthora infestans diversity. Plant Pathol 53:692–704
Cooke D, Lees A (2006) Eucablight protocol—Sampling of P. infestans from foliar epidemics. http://www.eucablight.org/Lib/Eucablight/Protocol/BlightSampling_V1.0.pdf. Accessed 07 March 2011
Cooke DEL, Young V, Birch PRJ, Toth R, Gourlay F, Day JP, Carnegie S, Duncan JM (2003) Phenotypic and genotypic diversity of Phytophthora infestans populations in Scotland (1995–97). Plant Pathol 52:181–192
Daniel S (2015) Diversity of Phytophthora infestans (Mont.) de Bary on Potato (Solanum tuberosum L.) and Tomato (Solanum lycopersicum Mill.) in Ethiopia. Dissertation, Haramaya University, Haramay, Ethiopia
Flier WG, Grünwald NJ, Kroon LPNM, Sturbaum AK, van den Bosch TBM, Garay-Serrano E, Lozoya-Saldaña H, Fry WE, Turkensteen LJ (2003) The population structure of Phytophthora infestans from the Toluca valley of Central Mexico suggests genetic differentiation between populations from cultivated potato and wild Solanum spp. Phytopathology 93:382–390
Fry WE, Goodwin SB, Dyer AT, Ma-tuszak JM, Drenth A, Tooley PW, Sujkowski LS, Koh YJ, Cohen BA, Spielman LJ, Deahl KL, Inglis DA, Sandlan KP (1993) Historical and recent migrations of Phytophthora infestans: chronology, pathways, and implications. Plant Dis 77:653–661
Fry WE, Grunwald NJ, Cooke DEL, Mcleod A, Forbes GA, Cao K (2009) Population genetics and population diversity of Phytophthora infestans. In: Lamour K, Kamoun S (eds) Oomycete genetics and genomics: diversity, interactions, and research tools, Wiley, pp 139–164
Galtier N, Nabholz B, Glémin S, Hurst GDD (2009) Mitochondrial DNA as a marker of molecular diversity: a reappraisal. Mol Ecol 18:4541–4550
Gardes M, Bruns TD (1993) ITS primers with enhanced specificity of basidiomycetes: application to the identification of mycorrhizae and rusts. Mol Ecol 2:113–118
Gavino PD, Fry W (2002) Diversity in and evidence for selection of the mitochondrial genome of Phytophthora infestans. Mycologia 94:781–793
Goodwin SB, Cohen BA, Fry WE (1994) Panglobal distribution of a single clonal lineage of the Irish potato famine fungus. Proc Natl Acad Sci 91:11591–11595
Gorfu D, Woldegiorgis G (2013) Pest risk analysis for potato importation into Ethiopia: a case of four source countries. In: Woldegiorgis G, Schulz S, Berihun B (eds) Seed potato tuber production and dissemination experiences, challenges and prospects. EIAR and ARARI, Bahir Dar, pp 7–20
Goss EM, Cardenas ME, Myers K, Forbes GA, Fry WE et al (2011) The plant pathogen Phytophthora andina emerged via hybridization of an unknown Phytophthora species and the Irish potato famine pathogen P. infestans. PLoS One 6(9):e24543. doi:10.1371/journal.pone.0024543
Griffith GW, Shaw DS (1998) Polymorphism in Phytophthora infestans: four mitochondrial haplotypes are detected after PCR amplification of DNA from pure cultures or from host lesions. Appl Envir Microbiol 64(10):4007–4014
Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis for Windows 95/98/NT. Nucl Acids Symp Ser 41:95–98
Hirpa A, Meuwissen PMM, Tesfaye A, Lommen JMW, Lansink AO, Tsegaye A, Struik PC (2010) Analysis of seed potato systems in Ethiopia. Am J Potato Res 87:537–552
Lang BF, Forget L (1993) The mitochondrial genome of Phytophthora infestans. In: O’Brien SJ (ed) Genetic maps: locus maps of complex genomes. Cold Spring Harbor Laboratory Press, pp 3.133–3.135
Lassiter ES, Russ C, Nusbaum C, Zeng Q, Hu CH, Thorne JL, Ristaino JB (2010) Inferring evolutionary relationships of species in the Phytophthora Ic clade using nuclear and mitochondrial genes. Phytopathology, S68
Lassiter ES, Russ C, Nusbaum C, Zeng Q, Saville A, Olarte R, Carbone I, Hu CH, Seguin-Orlando A, Samaniego JA, Thorne JF, Ristaino J (2015) Mitochondrial genome sequences reveal evolutionary relationships of the Phytophthora Ic clade species. Curr Genet 61(4):567–577
Laufer B (1938) The American Plant Migration. Part I: The Potato Archeology Series. Field Museum of National History. Vol 28, Number I. Publication 418. Chicago
Lees AK, Wattier R, Shaw DS, Sullivan L, Williams NA, Cooke DEL (2006) Novel microsatellite markers for the analysis of Phytophthora infestans populations. Plant Pathol 55:311–319
Librado P, Rozas J (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphic data. Bioinformatics 25:1451–1452
Martin FN (2008) Mitochondrial haplotype determination in the oomycete plant pathogen Phytophthora ramorum. Curr Genet 54:23–34
Martin FN, Zhang Y, Grünwald N, Coffey MD (2012) Expanded analysis of P. infestans mitochondrial haplotypes and correlation with nuclear genotype. Phytopathology 54:76
Mesfin T, Wondirad M, Bekele K (2009) Review of Research on Diseases of Root and Tuber Crops in Ethiopia. In: Abraham T (ed) Increasing crop production through improved plant protection -, vol II. Plant Protection Society of Ethiopia (PPSE) and EIAR, Addis Ababa, pp 169–202
Oliva RF, Chacón MG, Cooke DEL, Lees AK, Forbes GA (2007) Is Phytophthora infestans a good taxonomist? Host recognition and co-evolution in the Phytophthora/Solanum interaction. Acta Horticulturae 745:465–471
Oliva RF, Kroon LPNM, Chacón G, Flier WG, Ristaino JB, Forbes GA (2010) Phytophthora andina sp. nov., a newly identified heterothallic pathogen of solanaceous hosts in the Andean highlands. Plant Pathol 59:613–625
Ordoñez ME, Hohl HR, Velasco JA, Ramon MP, Oyarzun PJ, Smart CD, Fry WE, Forbes GA, Erselius LJ (2000) A novel population of Phytophthora, similar to P. infestans, attacks wild solanum species in Ecuador. Phytopathology 90(2):197–202
Paquin B, Laforest MJ, Forget L, Roewer I, Wang Z, Longcore J, Lang BF (1997) The fungal mitochondrial genome project: evolution of fungal mitochondrial genomes and their gene expression. Curr Genet 31:380–395
Schiessendoppler E, Molnar O (2002) Characterization of Phytophthora infestans populations in sub-Saharan Africa as a basis for simulation modeling and integrated disease management. In: Lizárraga C (ed) Late blight: managing the global threat. Proceedings of the global initiative on late blight conference, Hamburg, Germany, 11–13 July 2002, International Potato Centre, Lima, p 140
Spielman LJ, Drenth A, Davidse LC, Sujkowski LJ, Gu WK, Tooley PW, Fry WE (1991) A second world-wide migration and population displacement of Phytophthora infestans? Plant Pathol 40:422–430
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The Clustal X Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acid Res 24:4876–4882
Vega-Sánchez ME, Erselius LJ, Rodriguez AM, Bastidas O, Hohl HR, Ojiambo PS, Mukalazi J, Vermeulen T, Fry WE, Forbes GA (2000) Host adaptation to potato and tomato within the US–1 clonal lineage of Phytophthora infestans in Uganda and Kenya. Plant Pathol 49:531–539
Wattier RAM, Gathercole LL, Assinder SJ, Gliddon CJ, Deahl KL, Shaw DS, Mills DI (2003) Sequence variation of intergenic mitochondrial DNA spacers (mtDNA-IGS) of Phytophthora infestans (Oomycetes) and related species. Mol Ecol Notes 3:136–138
Yang ZH, Qi MX, Qin YX, Zhu JH, Gui XM, Tao B, Xu XH, Zhang FG (2013) Mitochondrial DNA polymorphisms in Phytophthora infestans: new haplotypes are identified and re-defined by PCR. J Microbiol Methods 95:117–121
Zhang Z, Schwartz S, Wagner L, Miller W (2000) A greedy algorithm for aligning DNA sequences. J Comput Biol 7:203–214
Acknowledgments
We thank Dr Bekele Kassa (Plant Pathology Department, Holeta Agricultural Research Centre, Ethiopia) for help with part of sample collection in 2011. We would like to express our thanks to Department of Forest Mycology and Plant Pathology, Swedish University of Agriculture Sciences (SLU) for help with molecular laboratory facility. A Swedish Government Initiative, the SLU Africa Food Security Research Program supports this work.
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Communicated by M. Kupiec.
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Shimelash, D., Hussien, T., Fininsa, C. et al. Mitochondrial DNA assessment of Phytophthora infestans isolates from potato and tomato in Ethiopia reveals unexpected diversity. Curr Genet 62, 657–667 (2016). https://doi.org/10.1007/s00294-016-0572-8
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DOI: https://doi.org/10.1007/s00294-016-0572-8