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Chaosia purpurea, a new mycorrhizal species associated with epiphytic orchids in tropical Asia

Takayuki Aokia, Takashi Yaguchib, Yoshitaka Onoc, Tsutomu Hattoria,*

a National Institute of Fungi, 1-1 XXX, Tsukuba, Ibaraki 305-1234, Japan

b University of YYY, ., Japan

c University of ZZZ, ., Japan

*Corresponding author. National Institute of Fungi, 1-1 XXX, Tsukuba, Ibaraki 305-1234, Japan.

E-mail address: abcd@efg.hi.jp (T. Hattori).

Text: 15 pages; tables: 2; figures: 3

Supplemental materials: 2 Supplementary Tables and 3 Supplementary Figures

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ABSTRACT(Full papers: 200 words or less. Short communications/Notes: 150 words or less as a standard format)

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Keywords: Atractiellales, Phalaenopsis, Phylogeny, Pucciniomycotina, TaxonomyComment by : Use a comma to delimit words

(up to 5 words or phrases; in alphabetical order; do not repeat words already present in the title; not capitalize initials of common words except for proper nouns)

1. Introduction(primary heading; in bold; hierarchically numbered; used ONLY for Full papers/Reviews)Comment by : In articles on taxonomy, cite authors of all species and infraspecific taxa only at the first use in the text. In abstract, however, authors of taxa are not cited except for special articles.Author names are written in full or abbreviated. In principle, abbreviation styles follow the Authors of Fungal Names [Index of Fungi Supplement, Kirk and Ansell (1992) or http://www.indexfungorum.org/AuthorsOfFungalNames.htm].Use italics for scientific names (generic, infrageneric, specific and infraspecific names).

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2. Materials and methods

2.1. Morphology[second-level heading; in italics; hierarchically numbered; you may use the second-level headings (but not hierarchically numbered) for Short communications/Notes, if necessary]

Start second-level headings at the left margin in boldface (but not italic). Authors are urged to deposit voucher specimens and cultures in internationally acknowledged herbaria and culture collections. The registered specimen numbers or strain numbers must be cited in the paper.

According to the recommendations in the International Code of Nomenclature for algae, fungi, and plants (ICNafp), authors describing new species or new infraspecific taxa are recommended to deposit a living culture (ex-type culture), whenever practicable, in at least two institutional culture or genetic resource collections, and cite these accession numbers in the paper (G. Okada, personal communication, August 19, 2017).

2.2. Molecular phylogeny

2.2.1. DNA extraction, amplification, sequencing and sequence alignment(third-level heading; in italics; hierarchically numbered)

Third-level headings are in italics; bold formatting is not used. Molecular sequence data must be deposited in a molecular sequence repository (DDBJ, http://www.ddbj.nig.ac.jp; EMBL, http://www.ebi.ac.uk; or GenBank, http://www.ncbi.nlm.nih.gov/Genbank/) and the accession number(s) must be cited in the paper.

2.2.2. Phylogenetic analyses

Third-level headings are italicized. Authors are requested to deposit sequence alignments in TreeBASE (http://treebase.org/treebase-web/) or in other internationally acknowledged databases and to indicate the accession number in the manuscript.

3. Results

3.1. Morphology

Start the first paragraph after main headings here. Do not start sentences with numbers, symbols, or abbreviated words (e.g., abbreviated taxon names). Do not forget to cite all figures in the text (e.g., Fig. 1; Fig. 1A, B; Fig. 1Aa, b, Ba, b; Figs. 1, 2; Figs. 1, 2, 4; Figs. 13).

3.2. Taxonomy

Chaosia purpurea S. Takam., G. Okada & Y. Ono, sp. nov. Figs 1, 2.Comment by : Add a dot for abbreviated names.Comment by : Use "&" or "et", preferably "&". Do not use "and".Comment by : not Figs. 12.

MycoBank no.: MB #####.

Diagnosis: to be described in English. Diagnosis is a brief statement composed of key words and phrases to clearly distinguish a proposed taxon from allied taxa. Diagnosis here is different from a technical Description below.

Type: COUNTRY, locality, host or substrate, date of isolation or collection, isolator or collector, registration numbers: e.g., MALAYSIA, Perlis State, Chuping, in roots of Phalaenopsis violacea Teijsm. & Binn., 1 Jan 2017, leg. Y. Ono (holotype, TNS-F-#####; isotype, TFM:FPH-#####; ex-type cultures, JCM ##### = CBS #####).

Gene sequences ex-holotype: AB ##### (18S), AB ##### (ITS), AB ##### (EF-1).

Etymology: Purpureus, from the color of orchid flower.

Start the description in English. Record measurements as length by width (or diameter). Place exceptional dimensions in parentheses. Indicate mean values separately.

Example: Conidia (10)1316(18.5) 78(9) m, 15.5 7.5 m on average. Comment by : Use en dash, not hyphen.Comment by : Use "", not a letter "x".

Chlamydospores 1012(13) m diam, finely warted.

Habitat and distribution: xxxxxxxxxx.

Additional specimens/cultures examined: xxxxxxxxxx.

3.3. Molecular phylogeny

4. Discussion

4.1. Taxonomy and phylogeny of the Chaosia species

4.2. Ecology of Chaosia purpurea

4.3. Key to species of Chaosia

1a. Mycelial layer tomentose ............................................................................................. 2

1b. Mycelial layer smooth .................................................................................................. 3

2a. Button spherical (0.81.2 cm) attached to a thick subiculum, expanded basidiome 1.42.5 cm wide, exoperidium not splitting in two layers, capillitial threads 3.45 m diam, branched and tapering tips ......................................................... C. purpurea

2b. Button ovate and bluntly pointed (25 cm) with a poorly developed subiculum, expanded basidioma 36 cm wide, exoperidium splitting in two layers, capillitial threads up to 11 m diam, often branched near the tapering tips ..........C. velutina

3a. Expanded basidiome 15 cm in diam, spores 3.25.5 m diam, ornamented with columnar processes up to 0.6 m high ......................................................... C. saccata

3b. Expanded basidiome 22.5 cm diam, spores 23 m diam, ornamented with low verrucae up to 0.2 m high ................................................................................... C. minutispora

Disclosure

The author(s) declare(s) no conflicts of interest. All the experiments undertaken in this study comply with the current laws of the country(-ies) where they were performed.

AcknowledgementsComment by : Acknowledegments, not Acknowledgments.

This study was supported, in part, by a Grant-in-Aid for Scientific Research (no. XXXXX) from the Japan Society for the Promotion of Sciences. We thank Dr. YYYYY, National Museum of Nature and Science, Tsukuba, for providing Phalaenopsis plants for inoculation experiments.

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Figure legendsComment by : Indicate the specimen/strain number(s).Fig. 1 Aspergillus niger (CBS 1234). A: xxxxx. B: xxxxx.orFig. 1 Aspergillus niger. A: xxxxx (CBS 1234). B: xxxxx (CBS 2345).

Fig. 1 Chaosia purpurea (TNS-F-#####). A: Light micrograph (LM) of a transverse section through the root of Phalaenopsis violacea. B: TEM image showing root colonization in P. violacea. C: Simple-septate basidia (LM). D: Basidiospores (LM). E: Basidiospores (SEM). Bars: A 20 m; B 1 m; CE 5 m.Comment by : Do not cite author name(s) in the legends.

Fig. 2 Neighbor-joining tree derived from the 18S rRNA gene sequences from the 50 fungi classified in the Atractiellales. The scale bar indicates one base change per 100 nucleotide positions. Bootstrap values were calculated from 1000 replications.

(Fig. 1 Sample figure 1.)Comment by Hattori Tsutomu: Figures should be submitted in separate files with enough high resolutions of at least 300 dpi at expected print sizes (see Guide for Authors).Comment by Hattori Tsutomu: Figure number and Figure Legends should not include on the figure, itself)Comment by : Pota et al. (2013). Mycoscience, 54, 1928.

Fig. 2 Sample figure 2.Comment by Hattori Tsutomu: Figure number and Figure Legends should not include on the figure, itself)Comment by : Zhao & Cui. (2013). Mycoscience 54: 231240.

(Fig. 3. Sample figure 3. (The font size should be adjusted as readable when printed in A4 size page; to be larger than that shown in this figure, as the minimal size but not recommended))Comment by : Figure number and Figure Legends should not include on the figure, itself)Comment by : Meeboon et al. (2013). Mycoscience, 54, 183187.

Table 1 Comment by : Please upload tables in separate files from the main body of text. Use the table function of Microsoft Word.Avoid vertical lines and unnecessary horizontal lines.

Comparison of Chaosia purpurea with similar species.

Species

Associated plant

Size of basidiospores (m)

Source

C. purpurea

Phalaenopsis violacea

1015 712

This study

C. viridi

P. appendiculata

1823 915

Tubaki (1950)

C. nigra

P. amboinensis

610 56.5

Kobayasi and Hiratsuka (1920)

Table 2

Taxa sequenced in this study.

Anamorph

Locality

Strain no.

DDBJ/GenBank/EMBL accession no.

18S

ITS

EF-1

Chaosia purpurea

Malaysia

ATCC ##### a

AB ######

AY ######

AF ######

C. purpurea

Thailand

CBS #####

AB ######

AY ######

AF ######

C. nigra

Indonesia

JCM #####

AB ######

AY ######

AF ######

Helicogloea lagerheimii

Sweden

MAFF #####b

AB ######

AY ######

AF ######

Atractiella solani

Poland

NBRC #####

AB ######

AY ######

AF ######

a Ex-holotype.

b Ex-neotype.

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