Phoma-like fungi associated with Convolvulaceae plants

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Phoma-like fungi is an extensive, non-taxonomic group of anamorphic ascomycetes, currently incorporating all micromycetes that were previously consumed as species of the genus Phoma. Didymellaceae is one of the largest families in the Pleosporales with a unique and underestimated biodiversity, encompassing main genera of Phoma-like fungi, namely Ascochyta, Didymella, Stagonosporopsis, etc. Several Convolvulaceae wild plants are widespread and one of the most harmful weeds, successful in many types of climates, are an exceptional source of biodiversity of Didymellaceae fungi. Phytosanitary examination of industrial fields, natural and ruderal areas has been carried out by authors from 1990 to the present. Leaves of Convolvulaceae plants (Calystegia inflata, Calystegia sepium, Calystegia sp., Convolvulus arvensis, Ipomoea purpurea) with symptoms of fungal etiology, namely leaf spots were collected in in different locations in Russia and neighboring countries (Kazakhstan, Kyrgyzstan). As a result of monitoring, a rich collection of fungal Phoma-like strains (at least 200) has been created and maintained. The aim of this study was to identify 28 Didymellaceae spp. strains isolated from Convolvulaceae plants according to the consolidated species concept (CSC) by their phylogenetic, micromorphological, and cultural features. Multilocus phylogenetic analysis inferred from nucleotide sequences of the internal transcribed spacer (ITS) and large subunit (28S) of ribosomal DNA, partial DNA-directed RNA polymerase II subunit (rpb2), and β-tubulin (tub2) genes revealed well-supported monophyletic clades corresponding to 18 Didymellaceae species. Among them: Ascochyta erotica, Didymella americana, D. bellidis, D. glomerata, D. macrostoma, D. pomorum, D. pseudomacrophylla, D. segeticola, D. sinensis, D. tanaceti, Epicoccum convolvulicola, E. pseudoplurivorum, Nothophoma brennandiae, N. gossypiicola, Phomatodes nebulosa, Stagonosporopsis caricae, S. heliopsidis, and S. inoxydabilis. Plants of the Convolvulaceae family were recorded as substrates for all identified fungal species for the first time. Seven species were revealed in Russia for the first time D. bellidis, D. segeticola, D. sinensis. D. tanaceti, Nothophoma brennandiae, Phomatodes nebulosa, and Stagonosporopsis caricae. Stagonosporopsis heliopsidis for the first time was found in the Kazakhstan. Additionally to detailed phylogenetic data, the manuscript is accompained with a detailed description of the cultural and micromorphological features of all species.

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作者简介

M. Gomzhina

All-Russian Institute of Plant Protection

编辑信件的主要联系方式.
Email: gomzhina91@mail.ru
俄罗斯联邦, St. Petersburg

E. Gasich

All-Russian Institute of Plant Protection

Email: elena_gasich@mail.ru
俄罗斯联邦, St. Petersburg

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2. Fig. 1. Combined maximum likelihood (ML) phylogenetic tree of Ascochyta, Didymella, Epicoccum, Nothophoma, Phomatodes, and Stagonosporopsis species based on the ITS, 28S, rpb2, and tub2 sequences. Bootstrap support and Bayesian probability (BPP, ≥ 0.7) are given at the branch nodes of the phylogram, respectively. Branches with ML=100 and BP=1 support are highlighted in bold. Numbers of type or representative strains are highlighted with T or R, respectively. Numbers of the studied strains are highlighted in blue.

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3. Fig. 2. Didymella americana MF 32.47 cultures (left half – upper part, right – reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days.

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4. Fig. 3. Didymella bellidis MF 32.65 cultures (left half – upper part, right – reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days; G–C – pycnidia; L–M – conidiogenous cells; N – conidia. Scale – 500 µm (G), 100 µm (G–C), 10 µm (L–M), 20 µm (N).

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5. Fig. 4. Didymella glomerata MF 32.38.1 cultures (left half – upper part, right – reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days; G–I – pycnidia; K–L – microsclerotia; M – conidia. Scale – 1 mm (G), 50 µm (G–L), 20 µm (M).

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6. Fig. 5. Didymella macrostoma MF 32.52.2 cultures (left half is the upper part, right half is the reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days; G–I – pycnidia; K–L – conidiogenous cells; M – conidia. Scale – 1 mm (G), 50 µm (H), 100 µm (I), 20 µm (K–M).

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7. Fig. 6. Didymella pomorum MF 32.67 cultures (left half is the upper part, right half is the reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days; G–H – pycnidia; I, K, M – multicellular chlamydospores; L – microsclerotia; N – unicellular chlamydospores; O – conidia. Scale bar – 1 mm (G), 50 µm (H, L), 20 µm (I, K, M–O).

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8. Fig. 7. Cultures of Didymella segeticola MF 32.69 (left half – upper part, right – reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days.

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9. Fig. 8. Didymella sinensis MF 32.66 cultures (left half – upper part, right – reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days.

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10. Fig. 9. Didymella tanaceti MF 17.86.4 cultures (left half – upper part, right – reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days; G–I – pycnidia; K – conidia. Scale – 1 mm (G), 200 µm (H), 100 µm (I), 20 µm (K).

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11. Fig. 10. Cultures of Nothophoma brennandiae MF 32.68 (left half is the upper part, right half is the reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days; G–H – pycnidia; I – conidiogenous cell; K – conidia. Scale – 1 mm (G), 100 µm (H), 10 µm (I), 20 µm (K).

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12. Fig. 11. Cultures of Nothophoma gossypiicola MF 52.4 (left half – upper part, right – reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA; 14 days; D – OA, 14 days; E – MEA, 14 days.

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13. Fig. 12. Phomatodes nebulosa MF 32.57.1 ​​cultures (left half – upper part, right – reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days.

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14. Fig. 13. Cultures of Stagonosporopsis caricae MF 32.73 (left half is the upper part, right half is the reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days; G–H – pycnidia; I – conidia. Scale – 500 µm (G), 50 µm (H), 20 µm (I).

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15. Fig. 14. Cultures of Stagonosporopsis heliopsidis MF 32.81 (left half is the upper part, right half is the reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days; G–H – pycnidia; I – conidiogenous cell; K – conidia. Scale – 1 mm (G), 50 µm (H), 20 µm (I, K).

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16. Fig. 15. Cultures of Stagonosporopsis inoxydabilis MF 9.239 (left half – upper part, right – reverse): A – KSA, 7 days; B – OA, 7 days; C – MEA, 7 days; G – KSA, 14 days; D – OA, 14 days; E – MEA, 14 days.

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