Research Article |
Corresponding author: Yong Wang ( yongwangbis@aliyun.com ) Academic editor: Cecile Gueidan
© 2019 Chuan-Gen Lin, Darbhe J. Bhat, Jian-Kui Liu, Kevin D. Hyde, Yong Wang.
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:
Lin C-G, Bhat DJ, Liu J-K, Hyde KD, Wang Y (2019) The genus Castanediella. MycoKeys 51: 1-14. https://doi.org/10.3897/mycokeys.51.32272
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Two new species, Castanediella brevis and C. monoseptata, are described, illustrated and compared with other Castanediella taxa. Evidence for the new species is provided by morphological comparison and sequence data analyses. Castanediella brevis can be distinguished from other Castanediella species by the short hyaline conidiophores and fusiform, aseptate hyaline conidia, while C. monoseptata differs from other Castanediella species by its unbranched conidiophores and fusiform, curved, 0–1-sepatate, hyaline conidia. Phylogenetic analysis of combined ITS and LSU sequence data was carried out to determine the phylogenetic placement of the species. A synopsis of hitherto described Castanediella species is provided. In addition, Castanediella is also compared with morphologically similar-looking genera such as Idriella, Idriellopsis, Microdochium, Neoidriella, Paraidriella and Selenodriella.
new taxa, Castanediellaceae, hyphomycetes, phylogeny, Sordariomycetes
The genus Castanediella was established by
During a survey of hyphomycetes in Thailand, two hyaline-spored hyphomycetes were collected. They were shown to belong to the genus Castanediella based on morphology and phylogeny analyses of ITS and LSU sequence data. The new species C. brevis and C. monoseptata are introduced.
Dead leaves from a variety of plants in two forests (Lampang province and Chiang Mai province) were collected in 2016 in Thailand. Samples were taken to the laboratory in Zip-lock plastic bags for examination. The specimens were incubated in sterile moist chambers and examined using a Motic SMZ 168 series microscope. Fungi were removed with a needle and placed in a drop of distilled water on a slide for morphological study. Photomicrographs of fungal structures were captured with a Canon 600D digital camera attached to a Nikon ECLIPSE Ni compound microscope. All measurements were made by the Tarosoft (R) Image FrameWork program. Photo-plates were made with Adobe Photoshop CS3 (Adobe Systems, USA). Isolation of the fungi on to potato dextrose agar (PDA) was performed by the single spore isolation method (
Genomic DNA was extracted from fungal mycelium grown on PDA or malt extract agar (MEA) at room temperature using the Fungal gDNA Kit (BioMIGA, USA) according to the manufacturer’s instructions. The internal transcribed spacer region of ribosomal DNA (ITS) and large subunit nuclear ribosomal DNA (LSU) genes were amplified via polymerase chain reaction (PCR) using the following primers: ITS5 and ITS4 (
Original sequences were checked using BioEdit version 7.0.5.3 (
Taxa | Isolatea | ITS | LSU |
---|---|---|---|
Castanediella acaciae | CPC 24869, CBS 139896 | NR_137985 | KR476763 |
Castanediella brevis | KUMCC 18-0132 | MH806361 | MH806358 |
Castanediella cagnizarii | MUCL 41095 | KC775732 | KC775707 |
Castanediella cagnizarii | CBS 101043 | KP859051 | KP858988 |
Castanediella cagnizarii | CBS 542.96 | KP859054 | KP858991 |
Castanediella camelliae | CNUFC-DLHBS5-1 | MF926620 | MF926614 |
Castanediella camelliae | CNUFC-DLHBS5-2 | MF926621 | MF926615 |
Castanediella communis | CPC 27631 | KY173393 | – |
Castanediella couratarii | CBS 579.71 | NR_145250 | KP858987 |
Castanediella eucalypti | CPC 24746, CBS 139897 | NR_137981 | KR476758 |
Castanediella eucalypticola | CPC 26539 | NR_145254 | KX228317 |
Castanediella eucalyptigena | CBS 143178, CPC 32055 | MG386036 | MG386089 |
Castanediella hyalopenicillata | CPC 25873 | KX306751 | KX306780 |
Castanediella malaysiana | CPC 24918 | NR_154810 | KX306781 |
Castanediella monoseptata | KUMCC 18-0133 | MH806360 | MH806357 |
Castanediella ramosa | MUCL 39857 | KC775736 | KC775711 |
Subsessila turbinata | MFLUCC 15-0831 | KX762288 | KX762289 |
The best models of evolution for each gene region were determined using Akaike information criterion (AIC) as implemented in MrModeltest v2 (
MP analyses were performed in PAUP*4.0b10 (
ML analyses were carried out in raxmlGUI v 1.5b1 (
For BI analysis, Posterior probabilities (PP) (
Phylogenetic trees were drawn with TreeView 1.6.6 (
The aligned sequence matrix comprises LSU and ITS sequence data for 16 taxa (ingroup) and one outgroup taxon with a total of 1438 characters after alignment including the gaps, of which 120 were parsimony informative, 77 parsimony-uninformative, and 1241 characters constant. The dataset consists of thirteen species within the genus. The tree was rooted with Subsessila turbinata (MFLUCC 15-0831). Maximum parsimony analysis resulted in two trees with TL = 391, CI = 0.657, RI = 0.642, RC = 0.422, HI = 0.343. For the Bayesian analysis, two parallel runs with six chains were run for 1,000,000 generations and trees were sampled every 100th generation, resulting in 20002 trees from two runs of which 15002 trees were used to calculate the posterior probabilities (each run resulted in 10001 trees of which 7501 trees were sampled).The MP and ML (lnL = -4041.301739) analyses based on combined LSU and ITS sequence data provided similar tree topologies, and the result of MP analysis is shown in Fig.
Phylogenetic tree generated from MP analysis based on combined LSU and ITS sequence data for the genus Castanediella. Bootstrap support values for maximum parsimony (MP, first set) and maximum likelihood (ML, second set) greater than 50% are indicated above or below the nodes. Ex-type strains are in bold, the new isolates are in red. The tree is rooted with Subsessila turbinata (MFLUCC 15-0831).
The novelty of the species, Castanediella brevis and C. monoseptata, described in this study are supported by sequence data analyses as belonging to the genus Castanediella, but with low bootstrap support values. Isolates of Castanediella brevis and C. monoseptata formed separate clades in the phylogenetic inference, respectively. Castanediella brevis is sister to C. malaysiana and C. ramosa, while C. monoseptata shows close phylogenetic relationship to C. couratarii and C. malaysiana. Both the new taxa can be recognized as phylogenetically distinct species and are clearly novel based on the recommendations for molecular data (
MP, ML and BI were used for phylogenetic analyses in this study. The tree topologies of MP and ML resulted from the combined LSU and ITS sequence data are similar, but most of the nodes are in low bootstrap support (Fig.
THAILAND. Lampang: Amphoe Mueang Pan, Tambon Chae Son, on decaying leaves, 24 September 2016, Chuangen Lin, LCG 10-1 (MFLU 18-1695, holotype; HKAS 102198, isotype), ex-type living cultures KUMCC 18-0132.
In reference to the short conidiophores.
Saprobic on plant host. Asexual morph: Colonies on substrate effuse, white. Mycelium partly superficial, composed of septate, branched, smooth, hyaline to subhyaline hyphae. Conidiophores macronematous, mononematous, solitary, erect, unbranched, straight or flexuous, short, 0–1-septate, hyaline, subcylindrical, ampulliform, smooth, often reduced to conidiogenous cells. Conidiogenous cells holoblastic, polyblastic, sympodial, integrated, terminal, subcylindrical, ampulliform, hyaline, denticulate, with 2–4 tiny protuberant denticles, 3–14 × 1.5–5.5 μm. Conidia solitary, dry, acropleurogenous, smooth, fusiform, curved, aseptate, hyaline, 12.5–21.7 × 1.2–3 μm (av. 16.95 × 2.2 μm, n = 60). Sexual morph: Undetermined.
Culture characteristics: Conidia germinating on PDA within 24 h. Colonies on PDA effuse, greyish white to dark from above and below, reaching a diam. of 5–7 cm in 30 days at 25 °C.
Based on a megablast search of the NCBI nucleotide database using the ITS sequence of the ex-type culture, the highest similarities found were with Castanediella malaysiana (GenBank NR_154810; identities = 526/537(98%), gaps = 1/537(0%)) and C. couratarii (GenBank KX960789; identities = 521/538(97%), gaps = 3/538(0%)). Castanediella brevis differs from these two species by its conidiophore morphology. Castanediella couratarii has pale brown conidiophores and longer conidiogenous cells (10.5–37 × 2–3.5 μm) whereas C. malysiana has pale brown and longer conidiophores (76–157 × 2.5–3 μm).
Among the species that produce more or less falcate and aseptate conidia, Castanediella communis, C. eucalypti, C. eucalypticola and C. eucalyptigena are most similar to C. brevis. However, Castanediella brevis differs from these species by its short, unbranched and 0–1-septate conidiophores.
THAILAND. Chiang Mai: on decaying leaves, 24 August 2016, Chuangen Lin, MRC 3-1 (MFLU 18-1696, holotype; HKAS 102199, isotype), ex-type living cultures KUMCC 18-0133.
In reference to the 0–1-septate conidia
Saprobic on plant host. Asexual morph: Colonies on substrate effuse, white. Mycelium partly superficial, composed of septate, branched, hyaline to subhyaline, smooth hyphae. Conidiophores macronematous, mononematous, solitary, erect, unbranched, straight or flexuous, septate, hyaline, subcylindrical, smooth, 8–29 × 2–4 μm. Conidiogenous cells polyblastic, integrated, sympodial, subcylindrical, hyaline, with several scars. Conidia solitary, dry, acropleurogenous, smooth, fusiform, curved, 0–1-sepatate, hyaline, 15.4–25.8 × 1.5–2.3 μm (av. 23.03 × 1.98 μm, n = 45). Sexual morph: Undetermined.
Culture characteristics: Conidia germinating on PDA within 24 h. Colonies on PDA effuse, grayish white to dark from above and below, reaching a diam. of 5–7 cm in 30 days at 25 °C.
A megablast search of the NCBI nucleotide database using the ITS sequence of the ex-type culture showed the highest similarities with uncultured Sordariales fungi (GenBank GQ268569; identities = 518/539(96%), gaps = 3/539(0%)) and Castanediella couratarii (GenBank KX960789; identities = 516/540(96%), gaps = 4/540(0%)).
Five Castanediella species, C. cagnizarii, C. diversispora, C. hyalopenicillata, C. malaysiana and C. ramosa, were reported to produce 1-septate conidia. Castanediella monoseptata can be distinguished from these species by its unbranched conidiophores and falcate and 15.4–25.8 × 1.5–2.3 μm conidia. Castanediella monoseptata is phylogenetically closely related to C. couratarii and C. ramosa, but differs from both species by its conidial morphology. Castanediella couratarii has shorter conidia (9.5–19 × 2–3 μm) are aseptate and C. ramosa has larger conidia (26–44 × 2–3 μm) that are 0–3-septate.
In this study, two new Castanediella species, C. brevis and C. monoseptata, were identified from decaying leaves in Thailand and a synopsis of hitherto described Castanediella species is provided (Table
Taxa | Conidiophores | Conidiogenous cells | Conidia | |||
---|---|---|---|---|---|---|
Shape | Size (μm) | Septa | Colour | |||
C. acaciae | Subcylindrical, medium brown, 40–80 × 2–3 μm. | Polyblastic, ampulliform, pale brown, 10–15 × 2–3 μm. | Falcate with subobtuse ends | (8–)10–11(–12) × 1.5(–2) | 0 | Hyaline |
C. brevis | Subcylindrical, ampulliform, hyaline, often reduced to conidiogenous cells | Polyblastic, cylindrical, hyaline, 3–14 × 1.5–5.5 μm | Fusiform, curved | 12.5–21.7 × 1.2–3.0 | 0 | Hyaline |
C. cagnizarii | Cylindrical, brown at the base, subhyaline towards the apex, up to 45 μm long. | Polyblastic, sympodial, subhyaline, 5–22 × 3–4 μm. | Cylindrical to fusiform, curved at the ends | Two sizes, 10–15 × 2 or 20–26 × 2 | Hyaline | |
C. camelliae | Conidiophores reduced to conidiogenous cell. | Cylindrical, ampulliform, globose to subglobose, or irregularly-shaped, 5.5–20.5 × 2–4.5 μm. | Straight to slightly curved, sometimes swollen in the middle part | 18.5–51.5 × 1.6–2.5 | Septum indistinct | Hyaline |
C. communis | Subcylindrical, medium brown, 20–60 × 3–4 μm. | Polyblastic, subcylindrical to ampulliform, pale brown, 10–35 × 2–4 μm. | Falcate with subobtuse ends | (13–)17–20(–22) × (2–)2.5(–3) | 0 | Hyaline |
C. couratarii | Pale brown | Lageniform to cylindrical, hyaline to pale brown, 10.5–37 × 2–3.5 μm | Lunate | 9.5–19 × 2–3 | 0 | Hyaline |
C. diversispora | Pale brown to brown | Polyblastic, sympodial, pale brown to brown, 4–9 × 2–3.5 μm. | Type i) cylindrical, slightly uncinate at the ends, straight | Type i) 11.5–16 × 2 | Type i) 1-septate | Hyaline |
Type ii) cylindrical to slightly subacerose, slightly uncinate at the apex, abruptly attenuated at the base, straight | Type ii) 19.5–25 × 1.5–2 | Type ii) 1-septate | ||||
Type iii) long filiform, obtuse or rounded at the apex attenuated at the base, straight or curved | Type iii) 28.5–47 × 1 | Type iii) 1–3-septate | ||||
C. eucalypti | Subcylindrical, medium brown, 10–30 × 3–4 μm. | Polyblastic, subcylindrical to ampulliform, pale brown, 8–25 × 2.5–4 μm. | Falcate, slightly curved, widest in middle with subobtuse ends | (15–)18–21(–23) × 2–3 | 0 | Hyaline |
C. eucalypticola | Subcylindrical, medium brown, 5–30 × 3–5 μm. | Polyblastic, subcylindrical to ampulliform or lanceolate, pale brown, 5–20 × 3–3.5 μm. | Falcate, straight to curved, widest in the middle, apex subobtusely rounded, base truncate, 0.5 μm diam | (15–)20–26(–30) × (2.5–)3 | 0 | Hyaline |
C. eucalyptigena | Subcylindrical, hyaline, frequently reduced to conidiogenous loci on hyphae, up to 15 μm tall, 3–5 μm diam. | Polyblastic, hyaline, ampulliform or subcylindrical, 2–10 × 2–5 μm | Falcate, tapering to acute ends that are subobtusely rounded | (13–)18–24(–30) × 2(–2.5) | 0 | Hyaline |
C. hyalopenicillata | Cylindrical, penicillate, mono-, bi-, and terverticillate, hyaline, 24–69 × 1.5–3 μm. | Mono- and polyblastic, short cylindrical, ampulliform, hyaline, 6.5–14 × 2–4 μm | Fusiform, base pointed, apex obtuse | 14–24 × 2–3 | 0–1 | Hyaline |
C. malaysiana | Cylindrical, biverticillate, pale brown, 76–157 × 2.5–3 μm. | Polyblastic, cylindrical, subcylindrical, hyaline, 19–28 × 2.5–3.5 μm. | Fusiform, curved, apex acuminate, and base acuminate or slightly flattened | 18–30 × 2–3 | 0–1 | Hyaline |
C. monoseptata | Subcylindrical, unbranched, hyaline, 8–29 × 2–4 μm | Polyblastic, cylindrical, hyaline | Fusiform, curved | 15.4–25.8 × 1.5–2.3 | 0–1 | Hyaline |
C. ramosa | Cylindrical, penicillate, brown at the base, subhyaline at the apex, up to 70 μm long | Polyblastic, subhyaline, 10–20 x 2.5–3.5 μm | Falcate | 26–44 × 2.2–3 | (0–) 1 (–3) | Hyaline |
Presently, the genus Castanediella contains 14 species, and is shown to be diverse in its habitats. Most of Castanediella species have been collected from plant leaves. Castanediella acaciae, C. camelliae, C. communis, C. eucalypti, and C. eucalypticola were isolated from disease symptoms on different host plant leaves (
The genus Castanediella is morphologically similar to Idriella, Idriellopsis, Microdochium, Neoidriella, Paraidriella, Selenodriella (
Genera | Conidiophores | Conidiogenous cells | Conidia | Chlamydospores |
---|---|---|---|---|
Castanediella | Branched, pale brown to brown at the base and subhyaline at the apex. | Sympodial, small denticles or scars, subhyaline. | 0–1-sepate, falcate, lunate, cylindrical or fusiform, hyaline | Not observed. |
Idriella | Brown, mostly reduced to conidiogenous cells | Denticulate, sympodial | Aseptate, lunate, curved, hyaline | Brown, uni- or pluricellular. |
Idriellopsis | Unbranched, brown at the base, almost hyaline at the apex, mostly reduced to conidiogenous cells | Terminal, denticulate | 0–1-septate, falcate, curved, hyaline | Not observed |
Microdochium | More or less verticillate, reduced to conidiogenous cells, hyaline | Hyaline, sympodial or percurrent, sometimes denticulate | Aseptate or multiseptate, lunate, falcate, fusiform, filiform, obovoid or subpyriform, straight or curved, hyaline | Terminal or intercalary, solitary, in chains or grouped in clusters, brown. |
Neoidriella | Mostly unbranched, pale brown, mostly reduced to conidiogenous cells | Sympodial, denticulate, terminal. | Aseptate, cylindrical to obovoid, hyaline | Intercalary or terminal, pale brown. |
Paraidriella | Unbranched, pale brown, mostly reduced to conidiogenous cells. | Sympodial, denticulate, terminal. | Aseptate, cylindrical to oblong, hyaline | Not observed. |
Selenodriella | Unbranched or verticillate, brown. | Sympodial, denticulate, terminal and intercalary. | Aseptate, falcate, hyaline | Not observed |
We would like to thank Dr. Shaun Pennycook (Landcare Research Manaaki Whenua, New Zealand) for advising on the fungal names. The research is supported by the Thailand Research grants entitled The future of specialist fungi in a changing climate: baseline data for generalist and specialist fungi associated with ants, Rhododendron species and Dracaena species (grant no: DBG6080013), Impact of climate change on fungal diversity and biogeography in the Greater Mekong Subregion (grant no: RDG6130001). Y. Wang would like to thank the projects of the National Natural Science Foundation of China (No. 31560489, 31500451), Talent project of Guizhou science and technology cooperation platform ([2017]5788-5), Bijie Science and Technology Project ([2016]19) and Guizhou science and technology department international cooperation base project ([2018]5806). J.K. Liu would like to thank the National Natural Science Foundation of China (NSFC 31600032) and the Science and Technology Foundation of Guizhou Province (LH [2015]7061).