Taxonomy
Phyllosticta chiangmaiensis D. Gomdola & K.D. Hyde, sp. nov. Figure 4
Index Fungorum number: IF 557882, Facesoffungi number: FoF 12965
Etymology – The specific epithet refers to Chiang Mai province, where the specimen was collected.
Holotype – MFLU22-0176
Saprobic on fallen leaves of Musa sp. (Musaceae). Sexual morph: Not observed. Asexual morph: Coelomycetous. Conidiomata 50–160 × 50–160 µm (x̄=90 × 95 µm, n = 20), solitary, uniloculate, globose to sub-globose, scattered or gregarious, semi-immersed, conspicuous on host surface, black. Pycnidial wall 2.75–14.5 µm wide (x̄=7.7 µm, n = 30), comprising 1–2 layers of thick-walled textura angularis cells, outer layer dark brown to black, inner layer pale brown. Conidiophores reduced to conidiogenous cells. Conidiogenous cells 6–14 × 4.5–9 µm (x̄=9.8 × 6.4 µm, n = 25), sub-globose or ellipsoidal or ovoid, guttulate, aseptate, hyaline, smooth. Conidia 13–18 × 8.5–12.5 µm (x̄=15.5 × 10.0 µm, n = 60), solitary, ellipsoidal to obovoid, guttulate, verruculose or with a single large central guttule, aseptate, hyaline, smooth-walled, tapering towards a narrowly truncate base and broader apex, surrounded by a mucilaginous sheath, thicker on both sides, 1.2–4.7 µm thick (x̄=2.6 µm, n = 60), thinner at the apex and base, 0.2–2.0 µm thick (x̄=1.0 µm, n = 25), with a hyaline, apical mucoid appendage. Appendages 4.7–11 × 0.8–1.2 µm (x̄=7.4 × 1.0 µm, n = 10), flexuous, unbranched, straight to curved, tapering towards an acutely rounded tip.
Material examined – Thailand, Chiang Mai Province, forests around the Mushroom Research Center, on fallen dead leaves of Musa sp. (Musaceae), 1 April 2021, D. Gomdola, (MFLU22-0176, holotype).
Distribution – Thailand.
GenBank accession numbers: ITS = OP693476, LSU = OP693472, TEF-1α = OQ189918, ACT = OQ189922.
Notes – Phyllosticta chiangmaiensis is sister to P. musaechinensis (GZAAS 6.1247 and GZAAS 6.1384) with strong support (100%ML, 100%MP, 1.00PP) (Fig. 3). Characters of the conidia match the species concept of Phyllosticta. Conidial length of P. chiangmaiensis ranges from 13–18 µm and that of P. musaechinensis (GZAAS 6.1247) is similar, ranging from 14–18 µm. However, conidiomata diameter and sheath thickness of P. chiangmaiensis are larger than that of the phylogenetically closely related taxa (Table 6). Other differences and similarities between P. chiangmaiensis and sister taxa are given (Table 6). Even though P. chiangmaiensis grouped with other species (P. musaechinensis, P. musarum, P. maculata, P. cavendishii) that were also isolated from Musa sp., it formed distinct lineages. Excluding gaps, in pairwise nucleotide comparisons of the type species of P. musaechinensis and P. chiangmaiensis (MFLU22-0176), there are 10 nucleotide base pair (bp) differences across ITS (567 nucleotides), 1 bp difference across ACT (195 nucleotides), and 1 bp difference in TEF-1α (316 nucleotides). In the ITS region of the type species of P. musarum, 14 nucleotide bp differences were observed across 433 nucleotides, excluding gaps. Hence, based on the recommendations provided by Chethana et al. (2021c), Jayawardena et al. (2021), Manawasinghe et al. (2021), Pem et al. (2021), we introduce P. chiangmaiensis as a new species.
Table 6
Morphological comparison of P. chiangmaiensis and sister taxa
|
Species
|
Species characters
|
P. chiangmaiensis MFLU22-0176
|
P. musaechinensis GZAAS 6.1247
|
P. musarum BRIP 55434
|
P. maculata CPC 18347
|
Morphological features
|
Conidiomata
|
Size (µm)
|
50–160 diam., 50–160 high
|
45–145 diam.
|
69–118 diam., 52–80 high
|
84–137 diam., 68–132 high
|
Shape and colour
|
Globose to sub-globose, black.
Ostiole not observed
|
Globose or subglobose, black, shiny, with a rounded central ostiole
|
Pycnidial, ostiolate
|
Pycnidial, ostiolate
|
Conidia
|
Size (µm)
|
13–18 × 8.5–12.5
|
14–18 × 8–12
|
(12)13–16(20) × (7)9–10(11)
|
(15)16–19(21) × (9)10–12(13)
|
Shape
|
Ellipsoidal to obovoid, coarsely guttulate, smooth-walled, broad apex and narrowly truncate base
|
Ellipsoidal or clavate, coarsely guttulate, thin- and smooth-walled
|
Oblong, obovoid or ellipsoidal, coarsely guttulate, thin- and smooth-walled, broad rounded or obtuse apex, truncate base becoming obtuse with age
|
Oblong or obovoid to subclavate, coarsely guttulate, smooth-walled, broad rounded or obtuse apex, truncate base becoming obtuse with age
|
Mucilaginous sheath
|
Thickness (µm)
|
1.2–4.7
|
0.5–3.5
|
1–3
|
2–4(6)
|
Appendage
|
Length (µm)
|
4.7–11
|
4–18.5
|
(12)14–18(20)
|
(12)15–26(37)
|
Shape
|
Straight to curved, tapering towards acutely rounded apex
|
Straight to curved
|
Straight to curved, tapering towards acute apex
|
Straight to curved, tapering towards acute apex
|
Reported morph
|
|
Asexual
|
Asexual
|
Asexual and sexual
|
Asexual and sexual
|
Symptoms
|
|
Saprobic
|
Weak pathogen, causing symptom similar to freckle disease
|
Leaf spots
|
Leaf spots
|
Hosts
|
|
Musa sp.
|
Musa sp.
|
Musa paradisiaca
|
Musa sp.
|
Gene region(s)
|
|
ITS, LSU, ACT, TEF-1α
|
ITS, ACT, TEF-1α, GAPDH
|
ITS
|
ITS, LSU, ACT, TEF-1α
|
References
|
|
This study
|
Wu et al. (2014)
|
Wong et al. (2012)
|
Wong et al. (2012)
|
Phyllosticta citribrasiliensis O.L. Pereira, Glienke & Crous (2011) Fig. 5
Index Fungorum number: IF831482, Facesoffungi number: FoF 12964
Associated with leaf spots of Laburnum anagyroides Medik. (Fabaceae). Sexual morph: Not observed. Asexual morph: Coelomycetous. Conidiomata 100–160 × 80–110 µm (x̄=111 × 97.5 µm, n = 15), solitary, uniloculate, globose to sub-globose, scattered, semi-immersed, conspicuous on host surface, black. Pycnidial walls 13.7–27 µm wide (x̄=18.6 µm, n = 15), comprising several layers of textura angularis cells, outer layers dark brown to black, inner layers pale brown to hyaline. Ostiole single, central, 12.5–17.5 µm wide (x̄=14.8 µm, n = 5). Conidiophores reduced to conidiogenous cells. Conidiogenous cells 5.6–15 × 1.3–2.6 µm (x̄=10.1 × 2.3 µm, n = 15), enteroblastic, phialidic, integrated, truncate to cylindrical to ampulliform, hyaline, formed from the inner layer of pycnidial wall. Conidia 7–9.8 × 4.5–6.6 µm (x̄=8.5 × 5.7 µm, n = 30), solitary, ellipsoidal to obovoid, coarsely guttulate, aseptate, hyaline, smooth-walled, with narrowly truncate base, surrounded by a mucilaginous sheath, thicker on both sides, 1.3–2 µm thick (x̄=1.6 µm, n = 20), thinner at apex and base, 0.4–0.9 µm thick (x̄=0.6 µm, n = 20), with a hyaline, apical mucoid appendage. Appendages 2.9–27 × 0.7–1.8 µm (x̄=8.3 × 1.1 µm, n = 20), flexuous, unbranched, tapering towards acutely rounded tip.
Material examined – Russia, Krasnodar Region, Sochi, Khostinsky City District, M.V. Frunze Health Care Resort, park, on senescing leaves of Laburnum anagyroides (Fabaceae), 15 October 2018, Timur S. Bulgakov, T-7583 (MFLU22-0175).
Distribution – Brazil, Russia (Farr and Rossman 2023).
GenBank accession numbers: LSU = OP684319, TEF-1α = OQ189917, ACT = OQ189921.
Notes – Our strain clusters with the type strain (CBS 100098) as well as other strains of P. citribrasiliensis (LGMF09, LGMF08, CPC 17466, CPC 17465, CPC 17464) with strong support (88%ML, 83%MP, 1.00PP) (Fig. 3). Excluding gaps, in pairwise nucleotide comparisons of the type strain of P. citribrasiliensis and our isolate (MFLU22-0175), there is no nucleotide base pair (bp) difference across LSU (761 nucleotides) and ACT (181 nucleotides), and 1 bp difference across TEF-1α (226 nucleotides). Morphological differences and similarities are also given (Table 7). Based on multigene phylogenetic analyses and morphological characters, we establish our strain as P. citribrasiliensis, a new host and country record. This is the first time P. citribrasiliensis is reported from Laburnum anagyroides in Russia (Farr and Rossman 2023).
Table 7
Comparison of P. citribrasiliensis (type strain) and strain obtained in this study
Species characters
|
MFLU22-0175
|
CBS 100098
|
Morphological features
|
Conidiomata
|
Size (µm)
|
100–160 diam., 80–110 high
|
Up to 250 diam.
|
Shape and color
|
Solitary, globose to sub-globose, black
|
Solitary, globose, black, erumpent, exuding colourless to opaque conidial masses
|
Central ostiole
|
Width (µm)
|
12.5–17.5
|
Up to 30 µm
|
Conidia
|
Size (µm)
|
7–9.8 × 4.5–6.6
|
(8)10–12(13) × 6–7(8)
|
Shape
|
Solitary, ellipsoidal to obovoid, coarsely guttulate, smooth-walled, with narrowly truncate base
|
Solitary, ellipsoid to obovoid, coarsely guttulate, thin- and smooth-walled, tapering toward a narrowly truncate base
|
Mucilaginous sheath
|
Thickness (µm)
|
1.3–2
|
2–4
|
Appendage
|
Length (µm)
|
2.9–27 × 0.7–1.8
|
7–15 × 1.5–2
|
Shape
|
Straight to curved
|
Straight to flexuous, unbranched
|
Reported morph
|
|
Asexual
|
Asexual
|
Hosts
|
|
Laburnum anagyroides
|
Citrus limon
|
Gene region(s)
|
|
LSU, ACT, TEF-1α
|
ITS, LSU, ACT, TEF-1α, GAPDH, RPB2
|
References
|
|
This study
|
Glienke et al. (2011)
|
Phyllosticta capitalensis Henn. (1908) Fig. 6
Index Fungorum number: IF168326, Facesoffungi number: FoF06888
For morphological description, see Glienke et al. (2011) and Wikee et al. (2013b).
Material examined – Thailand, Chiang Rai market, on fruit of Phyllanthus emblica (Phyllanthaceae), 3 March 2021, N. Huanraluek, FUA 03/3 (MFLU22-0177, dry culture), living culture (MFLUCC 22–0115); Thailand, Chiang Mai, Doi Inthanon National park, on fallen leaves of Morus alba (Moraceae), 18 October 2021, D. Gomdola, DGO2-L2A (MFLU22-0178, dry culture), living culture (MFLUCC 22–0116); Thailand, Chiang Mai, Doi Inthanon National park, on fallen fruit of Ficus auriculata (Moraceae), 18 October 2021, D. Gomdola, DGD1-L1-N3B (MFLU22-0179, dry culture), living culture (MFLUCC 22–0117).
GenBank accession numbers: (MFLU22-0177: ITS = OP686473, LSU = OP686475, ACT = OQ189919, TEF-1α = OQ189916, RPB2 = OQ189923), (MFLU22-0178: RPB2 = OQ189924), (MFLU22-0179: ITS = OP688119, LSU = OP688118, ACT = OQ189920, RPB2 = OQ189925)
Notes – In our multi-locus phylogenetic analyses, MFLU22-0177, MFLU22-0178 and MFLU22-0179 are in the P. capitalensis species complex (Fig. 3). Our isolates of P. capitalensis produced pycnidia on PDA. However, no conidia or other morphological characters were observed. Nonetheless, culture characteristics of our isolates are consistent with those described by Wikee et al. (2013b), also shown herein (Fig. 6).
We report our three strains of P. capitalensis as new host records in northern Thailand. This is the first time P. capitalensis is isolated from Phyllanthus emblica, Morus alba, and Ficus auriculata. Phyllosticta capitalensis has previously been reported on other Ficus species such as F. benjamina in Thailand and F. macrophylla in Iran (Wikee et al. 2013a, b; Sabahi et al. 2022; Farr and Rossman 2023). Phyllosticta capitalensis (MFLU22-0177 and MFLU22-0179) are established based on multigene phylogenetic analyses. However, P. capitalensis (MFLU22-0178) is reported solely based on the RPB2 gene region.
Excluding gaps, in pairwise nucleotide comparisons of the type species of P. capitalensis and our isolate (MFLU22-0177), there is no nucleotide base pair (bp) difference in ITS (523 nucleotides), ACT (201 nucleotides), TEF-1α (178 nucleotides) and RPB2 (749 nucleotides), and 1 nucleotide base pair (bp) difference across LSU (763 nucleotides). There is no nucleotide bp difference across RPB2 (749 nucleotides) when compared with P. capitalensis (MFLU22-0178). There is no nucleotide bp difference across ITS (534 nucleotides), LSU (763 nucleotides) and ACT (127 nucleotides), but 2 bp differences across RPB2 (749 nucleotides) when compared with P. capitalensis (MFLU22-0179). Since there are insignificant differences across each gene region, we determine these three strains as P. capitalensis.