New Endophytic Fusarium spp. from Fraxinus excelsior Leaves in Slovakia

Abstract

Fraxinus excelsior is currently suffering from ash dieback disease caused by the fungus Hymenoscyphus fraxineus. Co-occurrence of large numbers of other fungi with endophytic as well as pathogenic lifestyles has been observed on F. excelsior. In this study, new endophytic Fusarium spp. isolate Fraxinus excelsior Fusarium from Slovak territory, associated with F. excelsior, was characterised. Using morphology-based approaches, the fungus was assigned to the genus Fusarium. The isolate formed thick-walled hyaline, fusiform, slightly curved macroconidia, produced from monophialides in sporodochia, pointed at the tip, mostly 3 – 5 septate, occasionally 6 septate, and 40 – 68 × 3.8 – 5.0 µm in size with basal pedicellate cells. For more accurate taxonomic affiliation, molecular sequence analyses of the internal transcribed spacer, translation elongation factor 1-alpha sequences, and partial β-tubulin gene were performed. While the internal transcribed spacer sequence of the isolate showed the highest similarity to the Fraxinus oxysporum species, translation elongation factor 1-alpha and partial β-tubulin sequences were distantly related to the Fraxinus avenaceum species. Based on these data, the Fraxinus excelsior Fusarium isolate could probably be considered a new species of the Fusarium genus.

Key words

Oleaceae - Fusarium - endophyte - morphology - new species - phylogenetic analyses

Publication History

Received: 07 October 2019

Accepted after revision: 06 June 2020

Article published online:
24 June 2020

• References

• 1 Hinsinger DD, Basak J, Gaudeul M, Cruaud C, Bertolino P, Frascaria-Lacoste N, Bousquet J. The phylogeny and biogeographic history of ashes (Fraxinus, Oleaceae) highlight the roles of migration and vicariance in the diversification of temperate trees. PLoS One 2013; 8: e80431
• 2 Kostova I, Iossifova T. Chemical components of Fraxinus species. Fitoterapia 2007; 78: 85-106
• 3 Sarfraz I, Rasul A, Jabeen F, Younis T, Zahoor MK, Arshad M, Ali M. Fraxinus: A plant with versatile pharmacological and biological activities. Evid Based Complement Alternat Med 2017; 2017: 4269868
• 4 Drenkhan R, Adamson K, Hanso M. Fraxinus sogdiana, a Central Asian ash species, is susceptible to Hymenoscyphus fraxineus . Plant Protect Sci 2015; 51: 150-152
• 5 Gross A, Holdenrieder O. Pathogenicity of Hymenoscyphus fraxineus and Hymenoscyphus albidus towards Fraxinus mandshurica var. japonica . For Pathol 2015; 45: 172-174
• 6 Kowalski T, Bilański P, Kraj W. Pathogenicity of fungi associated with ash dieback towards Fraxinus excelsior . Plant Pathol 2017; 66: 1228-1238
• 7 Gouda S, Das G, Sen SK, Shin HS, Patra JK. Endophytes: a treasure house of bioactive compounds of medicinal importance. Front Microbiol 2016; 7: 1538
• 8 Power MW, Hopkins AJ, Chen J, Bengtsson SB, Vasaitis R, Cleary MR. European Fraxinus species introduced into New Zealand retain many of their native endophytic fungi. Balt For 2017; 23: 74-81
• 9 Sieber TN. Endophytic fungi in forest trees: are they mutualists?. Fungal Biol Rev 2007; 21.2 – 3: 75-89
• 10 Ibrahim M, Sieber TN, Schlegel M. Communities of fungal endophytes in leaves of Fraxinus ornus are highly diverse. Fungal Ecol 2017; 29: 10-19
• 11 Bamisile BS, Dash CK, Akutse KS, Keppanan R, Wang L. Fungal endophytes: beyond herbivore management. Front Microbiol 2018; 9: 544
• 12 Bakys R, Vasaitis R, Barklund P, Thomsen IM, Stenlid J. Occurrence and pathogenicity of fungi in necrotic and non-symptomatic shoots of declining common ash (Fraxinus excelsior) in Sweden. Eur J For Res 2009; 128: 51-60
• 13 Kowalski T, Kraj W, Bednarz B. Fungi on stems and twigs in initial and advanced stages of dieback of European ash (Fraxinus excelsior) in Poland. Eur J For Res 2016; 135: 565-579
• 14 Smith IM, Dunez J, Phillips DH, Lelliott RA, Archer SA. European Handbook of Plant Diseases. Oxford: Blackwell Scientific Publications; 1988
• 15 Leslie JF, Summerell BA, Bullock S. The Fusarium Laboratory Manual. Ames, Iowa: Wiley-Blackwell; 2006
• 16 Schoch CL, Seifert KA, Huhndorf S, Robert V, Spouge JL, Levesque CA, Chen W. Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. Proc Nat Acad Sci 2012; 109: 6241-6246
• 17 Nitschke E, Nihlgard M, Varrelmann M. Differentiation of eleven Fusarium spp. isolated from sugar beet, using restriction fragment analysis of a polymerase chain reaction-amplified translation elongation factor 1α gene fragment. Phytopathology 2009; 99: 921-929
• 18 Divakara ST, Santosh P, Aiyaz M, Ramana MV, Hariprasad P, Nayaka SC, Niranjana SR. Molecular identification and characterization of Fusarium spp. associated with sorghum seeds. J Sci Food Agric 2014; 94: 1132-1139
• 19 OʼDonnell K, Ward TJ, Robert VA, Crous PW, Geiser DM, Kang S. DNA sequence-based identification of Fusarium: current status and future directions. Phytoparasitica 2015; 43: 583-595
• 20 Akbar A, Hussain S, Ullah K, Fahim M, Ali GS. Detection, virulence and genetic diversity of Fusarium species infecting tomato in Northern Pakistan. PLoS One 2018; 13: e0203613
• 21 Yli-Mattila T, Paavanen-Huhtala S, Bulat SA, Alekhina IA, Nirenberg HI. Molecular, morphological and phylogenetic analysis of the Fusarium avenaceum/F. arthrosporioides/F. tricinctum species complex – a polyphasic approach. Mycol Res 2002; 106: 655-669
• 22 Kosawang C, Amby DB, Bussaban B, McKinney LV, Xu J, Kjær ED, Collinge DB, Nielsen LR. Fungal communities associated with species of Fraxinus tolerant to ash dieback, and their potential for biological control. Fun Biol 2018; 122: 110-120
• 23 Kowalski T, Kehr RD. Endophytic fungal colonization of branch bases in several forest tree species. Sydowia 1992; 44: 137-168
• 24 Chen J. Fungal Community Survey of Fraxinus excelior in New Zealand [dissertation]. Uppsala: Swedish University of Agricultural Sciences; 2012
• 25 Goodwin DC, Lee SB. Microwave miniprep of total genomic DNA from fungi, plants, protists and animals for PCR. Biotechniques 1993; 15: 438 441–442, 444
• 26 White TJ, Bruns T, Lee S, Taylor JW. Amplification and direct Sequencing of fungal ribosomal RNA Genes for Phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ. eds. PCR Protocols: a Guide to Methods and Applications. San Diego: Academic Press; 1990: 315-322
• 27 Pristas P, Gaperova S, Gaper J, Judova J. Genetic variability in Fomes fomentarius reconfirmed by translation elongation factor 1-α DNA sequences and 25S LSU rRNA sequences. Biologia 2013; 68: 816-820
• 28 Nosratabadi M, Kachuei R, Rezaie S, Harchegani AB. Beta-tubulin gene in the differentiation of Fusarium species by PCR-RFLP analysis. Infez Med 2018; 26: 52-60
• 29 Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 2013; 30: 2725-2729