Research Article |
Corresponding author: Bao-Ju Li ( libaoju@caas.cn ) Academic editor: Marco Thines
© 2019 Ying-Ming Li, Roger G. Shivas, Bao-Ju Li, Lei Cai.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Li Y-M, Shivas RG, Li B-J, Cai L (2019) Diversity of Moesziomyces (Ustilaginales, Ustilaginomycotina) on Echinochloa and Leersia (Poaceae). MycoKeys 52: 1-16. https://doi.org/10.3897/mycokeys.52.30461
|
A combined ecological, morphological, and molecular approach was used to examine 26 herbarium specimens and eight strains of Moesziomyces. The phylogenetic analysis resolved eight well-supported clades, of which three contained type specimens of known species of Moesziomyces. One clade contained two specimens that produced a teleomorph in the flowers of Echinochloa kimberleyensis in Australia. The name Moesziomyces kimberleyensis is proposed for this smut fungus. Another clade contained specimens that produced sori in the flowers of Leersia hexandra. The name Thecaphora globuligera (now Moesziomyces globuligerus) is available for this species, which is lectotypified. The teleomorph of Moesziomyces antarcticus, previously known only from Japan, is found for the first time in China, on Echinochloa crus-galli.
Ecology, plant pathogens, phylogeny, Ustilaginaceae, Ustilaginomycotina
The genus Moesziomyces (Ustilaginales, Ustilaginaceae) was established by
The teleomorphs of Ustilaginaceae are mostly host specific (
Specimens borrowed from several herbaria were examined by light microscopy (Table
Collection details for Moesziomyces specimens newly sequenced in this study.
Species | Specimen/strain no.1 | Host | Source | Location | Year of collection | ITS GenBank accession number2 |
---|---|---|---|---|---|---|
Moesziomyces antarcticus | HMAS 248025 | Echinochloa crus-galli | Sorus | China | 2017 | MK027038 |
M. antarcticus | HMAS 248026 | E. crus-galli | Sorus | China | 2017 | MK027039 |
M. antarcticus | HMAS 60130 | E. crus-galli | Sorus | China | 1989 | MK027043 |
M. bullatus | HMAS 146471 | E. crus-galli | Sorus | China | 2003 | MK027040 |
M. bullatus | HMAS 50052 | E. crus-galli | Sorus | China | 1985 | MK027041 |
M. bullatus | LC-CLS58-3-2 | Setaria faberii | Leaf surface | China | 2017 | MK024201 |
M. bullatus | LC-CLS58-3-21 | S. faberii | Leaf surface | China | 2017 | MK024202 |
M. bullatus | LC-CLS58-3-22 | S. faberii | Leaf surface | China | 2017 | MK024203 |
M. bullatus | LC-CLS60-2-22 | Pennisetum sp. | Leaf surface | China | 2017 | MK024204 |
M. bullatus | LC-CLS60-2-4 | Pennisetum sp. | Leaf surface | China | 2017 | MK024205 |
M. bullatus | LC-SY1-2-11 | Digitaria sp. | Leaf surface | China | 2017 | MK024206 |
M. bullatus | LC-SY1-2-21 | Digitaria sp. | Leaf surface | China | 2017 | MK024207 |
M. bullatus | LC-SY1-2-22 | Digitaria sp. | Leaf surface | China | 2017 | MK024208 |
M. bullatus | HMAS 50454 | E. crus-galli | Sorus | Japan | 1985 | MK027042 |
M. bullatus | HMAS 70876 | E. crus-galli | Sorus | China | 1991 | MK027045 |
M. bullatus | HMAS 73871 | E. crus-galli | Sorus | China | 1996 | MK027046 |
M. bullatus | HUV 2442* | E. crus-galli | Sorus | Poland | 1869 | MK027047 |
M. bullatus | HUV 305 | E. crus-galli | Sorus | Germany | 1905 | MK027050 |
M. globuligerus | BRIP 27384 | Leersia hexandra | Sorus | Australia | 1998 | MK027025 |
M. globuligerus | BRIP 44301 | L. hexandra | Sorus | Australia | 2004 | MK027029 |
M. globuligerus | BRIP 44569 | L. hexandra | Sorus | Australia | 2004 | MK027030 |
M. globuligerus | BRIP 47767 | L. hexandra | Sorus | Thailand | 2005 | MK027031 |
M. globuligerus | BRIP 47768 | L. hexandra | Sorus | Thailand | 2005 | MK027032 |
M. globuligerus | BRIP 51872 | L. hexandra | Sorus | Australia | 2008 | MK027035 |
M. globuligerus | HMAS 248027 | L. hexandra | Sorus | China | 2017 | MK027037 |
M. kimberleyensis | BRIP 51843* | E. kimberleyensis | Sorus | Australia | 2008 | MK027034 |
M. kimberleyensis | BRIP 52498 | E. kimberleyensis | Sorus | Australia | 2009 | MK027036 |
M. penicillariae | HUV 2487 | Pe. glaucum | Sorus | Gambia | 1973 | MK027048 |
M. penicillariae | HUV 2488 | Pe. glaucum | Sorus | India | 1912 | MK027049 |
M. verrucosus | BRIP 39886 | Paspalum distichum | Sorus | Australia | 2003 | MK027026 |
M. verrucosus | BRIP 43727 | Pa. distichum | Sorus | Australia | 2004 | MK027027 |
M. verrucosus | BRIP 43735 | Pa. distichum | Sorus | Australia | 2004 | MK027028 |
M. verrucosus | BRIP 51772 | Pa. distichum | Sorus | India | 1992 | MK027033 |
M. verrucosus | HMAS 66437 | Pa. distichum | Sorus | India | 1992 | MK027044 |
Sori were carefully removed from herbarium specimens, up to 149 years old, with a fine needle, sterilized by dipping in 75% ethanol for 30 s, air-dried on sterilized filter paper, and deposited in cell lysis solution (CTAB). Pure yeast colonies grown on yeast extract peptone dextrose (YPD) plates were transferred to cell lysis solution directly. Genomic DNA was extracted following the protocol of
PCR amplifications were carried out in 25 μl reactions containing 1 μl of genomic DNA template, 9.5 μl distilled water, 12.5 μl of 2 X Taq Plus Master Mix (Nanjing Vazyme Biotech Co. Ltd, Nanjing, China) and 1 μl of each primer (10 μM). Amplification reactions were run as follows: initial denaturation of 95 °C for 5 min followed by 35 cycles at 95 °C for 30 s, 45 s at 58 °C (annealing temperature) and 1 min at 72 °C with a final extension of 10 min at 72 °C. PCR products were sent to Tianyihuiyuan (Beijing, China) for sequencing with the forward and reverse primers indicated above. AB1 sequence traces were assembled with Sequencher version 5 (Genecodes, Ann Arbor, USA).
The sequences included in this study (Tables
For BI, MrBayes was used with a Markov Chain Monte Carlo algorithm incorporating four runs, each consisting of four chains, until the standard deviation of split frequencies was reached. The cold chain was heated at a temperature of 0.25. Substitution model parameters were sampled every 50 generations and trees were saved every 5000 generations. Convergence of the Bayesian analysis was confirmed using AWTY (
The ITS dataset comprised the newly sequenced Moesziomyces specimens and strains (Table
Phylogram obtained from a ML analysis based on the ITS sequence alignment. Values above the branches represent ML bootstrap values (> 70%) from RaxML and PhyML analysis respectively. Thickened branches represent Bayesian posterior probabilities (> 0.95). The scale bar indicates 0.03 expected substitutions per site. * indicates type specimens or type strains.
List of Moesziomyces, Triodiomyces, and Ustilago sequences taken from GenBank and used in the phylogenetic analysis.
Species | Source | ITS GenBank accession number | Reference |
---|---|---|---|
Moesziomyces antarcticus | – | JX094775 | Gujjari et al. (unpubl.) |
– | JN942669 | An (unpubl.) | |
unpolished Japanese rice | AB089360 |
|
|
Antarctica sediment | AF294698 |
|
|
Albizia julibrissin flower | AY641557 |
|
|
lake sediment | AB089358 |
|
|
tomato rhizosphere | KF493994 | Johnston-Monje et al. (unpubl.) | |
Echinochloa crus-galli sorus | LC368624 | ||
Echinochloa crus-galli sorus | LC368624 | ||
Echinochloa crus-galli sorus | LC368624 | ||
Echinochloa crus-galli sorus | LC368624 | ||
Echinochloa crus-galli sorus | LC368624 | ||
Echinochloa crus-galli sorus | LC368624 | ||
Echinochloa crus-galli sorus | LC368625 | ||
Moesziomyces bullatus | human preterm low birth weight infant | KF926673 |
|
– | DQ831013 |
|
|
Japanese pear fruit | AB204896 |
|
|
Saccharum officinarum | AB704889 |
|
|
Leucaena glauca | HQ662536 |
|
|
human | EU105207 |
|
|
human blood | AB089362 |
|
|
human | HQ848933 | Xie et al. unpubl. | |
Fallopia japonica | KC282385 | Wang & Liu (unpubl.) | |
human blood | KM610219 | Bosco-Borgeat & Taverna (unpubl.) | |
Leucaena glauca | HQ647299 |
|
|
Saccharum officinarum | AB704890 |
|
|
poplar leaf | KM268868 | Sun & Yan (unpubl.) | |
Forcipomyia taiwana | KM555221 | Chen (unpubl.) | |
seaweed | KP269028 | Wang et al. (unpubl.) | |
aphid secretion | AF294699 |
|
|
Neoreglia cruenta | FN424100 | Garcia et al. (unpubl.) | |
Saccharum officinarum | AB704878 |
|
|
giant panda secretion | KF973199 | Li et al. (unpubl.) | |
Camellia sinensis leaf lesions | HQ832804 | Li et al. (unpubl.) | |
Echinochloa crus-galli | GU390690 | Hamayun & Ahmad (unpubl.) | |
aphid secretion on Solanum pseudocapsicum | JN942666 | An (unpubl.) | |
Citrus leaf | JQ425372 | Soliman (unpubl.) | |
– | JN942667 | An (unpubl.) | |
mouldy Zea mays leaf | AB089370 |
|
|
plant leaf | HE650886 | ||
ex-leaf of corn | AF294697 |
|
|
Hyoscyamus muticus | AB500693 | Abdel-Motaal & Itu (unpubl.) | |
Coffea arabica | EU002890 | Vega et al. (unpubl.) | |
Coffea arabica | DQ778919 |
|
|
Saccharum officinarum leaf | LC053989 | Surussawadee & Limtong (unpubl.) | |
marine environment | DQ178645 |
|
|
Helicoverpa armigera larva gut | AM160637 | Molnar & Prillinger (unpubl.) | |
Moesziomyces bullatus | marine sediment | KC834821 | Qu et al. (unpubl.) |
– | KR047769 | Wang et al. (unpubl.) | |
pharmaceutical effluent | KF922220 | Selvi & Das (unpubl.) | |
barley kernels and leaf | HG532070 |
|
|
Ericaceae roots | HQ260042 |
|
|
cleaned rice | AB235999 |
|
|
Arabidopsis thaliana infected with Albugo laibachii | KY930224 |
|
|
Echinochloa crus-galli | KY424439 |
|
|
Echinochloa crus-galli | KY424428 |
|
|
Echinochloa crus-galli | KY424429 |
|
|
Echinochloa muricata | KY424430 |
|
|
Echinochloa muricata | KY424431 |
|
|
Echinochloa muricata | KY424432 |
|
|
Echinochloa muricata | KY424433 |
|
|
Echinochloa crus-galli | KY424434 |
|
|
Echinochloa crus-galli | KY424435 |
|
|
Echinochloa crus-galli | KY424436 |
|
|
Echinochloa crus-galli | KY424437 |
|
|
Echinochloa crus-galli | KY424427 |
|
|
Echinochloa crus-galli | KY424438 |
|
|
shoot of tip pepper | GU975792 | Sim et al. (unpubl.) | |
Moesziomyces eriocauli | Eriocaulon cinereum | AY740041 |
|
Moesziomyces parantarcticus | – | KP132543 |
|
human blood | AB089356 |
|
|
– | NR130693 | An (unpubl.) | |
– | JN544036 | Chen (unpubl.) | |
yam tuber steep water | KF619567 |
|
|
Axonopus compressus soil | HQ436080 | Kee & Chia (unpubl.) | |
Moesziomyces penicillariae | Pennisetum glaucum | KY424440 |
|
Moesziomyces verrucosus | Paspalum distichum | AY740153 |
|
Triodiomyces altilis | Triodia pungens | AY740166 |
|
Ustilago echinata | Phalaris arundinacea | AY345001 |
|
Based on the phylogenetic analysis and the hosts of the teleomorphs, a new species of Moesziomyces is described and another species resurrected. Additionally, the teleomorph of M. antarcticus is reported for the first time from China.
Sporobolomyces antarcticus Goto, Sugiyama & Iizuka, Mycologia 61: 759 (1969). [Basionym]
Candida antarctica (Goto, Sugiyama & Iizuka) Kurtzman et al. Yeasts: 86 (1983).
Vanrija antarctica (Goto, Sugiyama & Iizuka) R.T. Moore, Bibltheca Mycol. 108: 167 (1987).
Pseudozyma antarctica (Goto, Sugiyama & Iizuka) Boekhout, J. Gen. Appl. Microbiol. 41: 364 (1995).
Trichosporon oryzae H. Ito, Iizuka & T. Sato, Agric. Biol. Chem. 38: 1599 (1974). (synonymy by Q.M. Wang, Begerow, F.Y. Bai and Boekhout).
Sori in scattered ovaries, sometimes deciduous, globose to ovoid, 2–3 mm in length, covered by a smooth green membrane of host tissue origin that becomes brown and ruptures irregularly to expose a granular, black to dark brown mass of spore balls; columella absent. Spore balls variable in shape and size, globose, subglobose, ovoid, elongate to irregular, 130–200 μm in diameter, dark brown, composed of up to several hundred spores, united firmly by fungal sterile cells and spore meshes and wings. Spore globose, ovoid to irregular, slightly polyhedral, (8–) 8.5–9.5 (–10) × (6–) 7–9 (–10) μm, usually with well-developed meshes and wings, subhyaline to pale yellowish-brown; wall 0.5 μm thick, smooth. Some of the sterile cells empty at maturity, thin-walled, with irregular meshes or wings on the spore surface when the spores separates; other sterile cells, globose, ovoid to irregular, slightly polyhedral, (8–) 8.5–9.5 (–10) × (6–) 7–9 (–10) μm, subhyaline to pale yellowish brown; wall 1–1.5 μm thick, smooth.
CHINA, Sichuan, Chengdu, on Echinochloa crus-galli, 15 Sept. 1989, L. Guo leg., HMAS 60130; Guangxi, on E. crus-galli, Oct. 2017, R.G. Shivas, M.D.E. Shivas & Y.-M. Li leg., HMAS 208025; Guangxi, on E. crus-galli, Oct. 2017, R.G. Shivas, M.D.E. Shivas & Y.-M. Li leg., HMAS 208026.
The teleomorph of Moesziomyces antarcticus was previously reported from Japan, on Echinochloa crus-galli (
Thecaphora globuligera Berk. & Broome, Trans. Linn. Soc. London, Bot., Ser. 2, 1: 407 (1880). — Type: AUSTRALIA, Queensland, Brisbane, on Leersia hexandra, F.M. Bailey, No. 86 (K(M) 252436, lectotype designated here, MBT 385180, not seen; K(M) 252437, syntype). [Basionym]
Tolyposporium globuligerum (Berk. & Broome) Ricker, J. Mycol. 11:112 (1905).
Testicularia leersiae Cornu, Ann. Sci. Nat. Bot., Sér. 6, 15: 275 (1883).
Sori in some of the ovaries, often deciduous, ellipsoidal to oval, 2.5–4 × 1.5–3 mm, green at first, later brown, smooth, ruptures irregularly to reveal a granular, dark brown mass of spore balls; columella absent. Spore balls subglobose, ellipsoidal or irregular, 75–150 µm in diameter, yellowish brown, composed of up to several hundred spores that separate by moderate pressure. Spores subglobose, ovoid to irregularly polyhedral, (8–) 8.5–11 (–13) × (6–) 7–9 (–10) μm (xˉ = 9.6 ± 1.2 × 7.9 ± 0.9 μm, n = 50), subhyaline to pale yellowish brown, attached together by multiple narrow cylindrical protuberances about 2 μm wide and 1–2 μm long; wall with irregular meshes and wings, less than 0.5 μm thick, smooth. (Based on specimen BRIP 27384).
AUSTRALIA, Queensland, Willowbank, on Leersia hexandra, 9 Mar. 1998, C. Vánky & K. Vánky leg., BRIP 27384; Queensland, Mareeba, on L. hexandra, 1 May 2004, M.D.E. Shivas & R.G. Shivas leg., BRIP 44301; Queensland, Mt Garnet, on L. hexandra, 5 May 2005, T.S. Marney & R.G. Shivas leg., BRIP 44569; Northern Territory, Darwin, on L. hexandra, 15 Apr. 2008, J. Ray, A.A. Mitchell, A.R. McTaggart & R.G. Shivas leg., BRIP 51872. CHINA, Guangxi province, on L. hexandra, Oct. 2017, R.G. Shivas, M.D.E. Shivas, Y.-M. Li, P. Zhao & X.-H. Qi leg., HMAS 248027. THAILAND, Kanchanaburi, on L. hexandra, 16 Dec. 2005, R.G. Shivas & M.D.E. Shivas leg., BRIP 47767; Chiang Mai, on L. hexandra, 26 Dec. 2005, R.G. Shivas & M.D.E. Shivas leg., BRIP 47768.
AUSTRALIA, Western Australia, Kununurra, Mulligan’s Lagoon Road, on Echinochloa kimberleyensis, 9 Apr. 2008, A.R. McTaggart, V.L. Challinor, A.D.W. Geering, M.D.E. Shivas & R.G. Shivas leg. (holotype: BRIP 51843).
Named after the Kimberley region of northern Western Australia from where it was collected.
Sori in some of the ovaries, often deciduous, globose to ovoid, 3–6 × 2–4 mm, green at first, later brown, smooth, ruptures irregularly to reveal a granular, dark brown mass of spore balls; columella absent. Spore balls subglobose, ovoid, elongate or irregular, 275–100 µm diam, dark brown, composed of up to several hundred spores, separated by moderate pressure. Spore globose, ovoid to irregular, slightly polyhedral, (9–) 9.5–12 (–14.5) × (8–) 8.5–9.5 (–10) μm (xˉ = 10.5 ± 1.2 × 8.9 ± 0.7 μm, n = 50), subhyaline to yellowish brown, attached together by multiple narrow cylindrical protuberances about 2 μm wide and 1–2 μm long; wall with irregular meshes and wings, 0.5 μm thick, smooth.
AUSTRALIA, Western Australia, Kununurra, Mulligan’s Lagoon Road, on E. kimberleyensis, 7 May 2009, A.R. McTaggart, M.J. Ryley, M.D.E. Shivas & R.G. Shivas leg. (BRIP 52498).
Moesziomyces kimberleyensis was shown in the phylogenetic analysis to reside in a well-supported clade sister to M. bullatus. Moesziomyces kimberleyensis is only known from the teleomorph, which forms sori in flowers of E. kimberleyensis, and thereby differs from M. bullatus by host association. Moesziomyces kimberleyensis is only known from one location in Western Australia on E. kimberleyensis, which is an endemic grass in the tropical and subtropical woodlands of northern Australia.
The phylogenetic analyses in this study supported the host specificity of the teleomorphic stage of six species of Moesziomyces, specifically, M. antarcticus on Echinochloa crus-galli, M. bullatus on E. crus-galli and E. muricata, M. globuligerus on Leersia hexandra, M. kimberleyensis on E. kimberleyensis, M. penicillariae on Pennisetum glaucum, and M. verrucosus on Paspalum distichum. The teleomorph of M. eriocauli may be specific to Eriocaulon spp., although this cannot be ascertained from the sequence data of one specimen. Specimens that have been assigned to M. bullatus were not well resolved and formed a number of smaller clades with varying degrees of support (Fig.
The anamorphs of Moesziomyces, together with most members of the Ustilaginales, have a dimorphic lifecycle comprised of a parasitic dikaryotic phase characterized by teliospores, together with a saprobic yeast-like haploid phase (
We thank Dr Begoña Aguirre-Hudson (Royal Botanic Gardens, Kew) for providing information about the syntypes of Thecaphora globuligera. We are also grateful to Dr Julia Kruse (University of Southern Queensland) for helpful comments about the manuscript. Marjan Shivas, Peng Zhao, Fang Liu, and Xiao-Hua Qi are thanked for assistance with specimen collection. This study was financially supported by CAS-QYZDB-SSW-SMC044 and CAAS-ASTIP-IVFCAAS.