Research Article |
Corresponding author: Hai-Xia Wu ( aileen2008haixia@gmail.com ) Academic editor: Nalin Wijayawardene
© 2020 Xinhao Li, Hai-Xia Wu, Jinchen Li, Hang Chen, Wei 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:
Li X, Wu H-X, Li J, Chen H, Wang W (2020) The insights into the evolutionary history of Translucidithyrium: based on a newly-discovered species. MycoKeys 76: 1-16. https://doi.org/10.3897/mycokeys.76.58628
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During the field studies, a Translucidithyrium-like taxon was collected in Xishuangbanna of Yunnan Province, during an investigation into the diversity of microfungi in the southwest of China. Morphological observations and phylogenetic analysis of combined LSU and ITS sequences revealed that the new taxon is a member of the genus Translucidithyrium and it is distinct from other species. Therefore, Translucidithyrium chinense sp. nov. is introduced here. The Maximum Clade Credibility (MCC) tree from LSU rDNA of Translucidithyrium and related species indicated the divergence time of existing and new species of Translucidithyrium was crown age at 16 (4–33) Mya. Combining the estimated divergence time, paleoecology and plate tectonic movements with the corresponding geological time scale, we proposed a hypothesis that the speciation (estimated divergence time) of T. chinense was earlier than T. thailandicum. Our findings provided new insights into the species of Translucidithyrium about ecological adaptation and speciation in two separate areas.
Divergence time, morphological characteristics, new species, Phaeothecoidiellaceae, phylogeny, speciation, taxonomy
The sooty blotch and flyspeck fungi are widespread species and commonly occur on the surface of leaves, stems and fruits in tropical and subtropical zones (
Phaeothecoidiellaceae K.D. Hyde & Hongsanan was introduced by
Translucidithyrium X.Y. Zeng & K.D. Hyde (2018) was introduced as a monotypic genus in Phaeothecoidiellaceae, which is represented by T. thailandicum X.Y. Zeng & K.D. Hyde (2018). It was characterised by epiphytes on the reverse of living leaves, semi-transparent ascomata, globose to subglobose asci and fusiform ascospores with verrucose and appendages. Ascospores germinated on MEA (Malt Extract Agar Medium) within 24 h. The colonies slowly grow on media, white to grey, circular and villiform (
In this study, we collected an extraordinary new species of Translucidithyrium in Xishuangbanna, Yunnan Province, China. We described the morphological characteristics and built a phylogenetic tree to determine the classification of the new taxon. We compared and analysed the estimated divergence time of Translucidithyrium with the environmental changes around the corresponding time range to propose the evolutional history hypothesis of Translucidithyrium distributed in two different regions (China and Thailand).
Fresh living leaves with olivaceous dots were collected at Xishuangbanna, China 21°55'51"N, 101°15'08"E, 540 m alt.) and delivered to the laboratory for observation. According to
According to the manufacturer’s instructions, genomic DNA was extracted from mycelium growing on PDA at room temperature by using the Forensic DNA Kit (OMEGA, USA). The primer pair LR0R and LR5 was used to amplify the large subunit (LSU) rDNA (
BioEdit version 7.0.5.3 (
Maximum Likelihood (ML) analysis was conducted by using RAxMLGUI v.1.0 (
For Bayesian analysis, MrModeltest 2.3 (
The fossil Protographum luttrellii (
Divergence time analysis was carried out using BEAST v1.8.4 (
The phylogenetic tree and MCC tree were visualized in FigTree v.1.4.3 (
The dataset of combined LSU and ITS sequences comprised 1350 characters after alignment. Bayesian Inference, in total, generated 20,001 trees and the average standard deviation of split frequencies reached 0.0096. A total of 15,001 trees were finally used to calculate posterior probabilities. Phylogenetic analysis showed that the new collection clusters with T. thailandicum with 100% Maximum Likelihood bootstrap support and 1.00 posterior probabilities (Fig.
The topology shows family relationships of Capnodiales, based on combined LSU and ITS dataset analysis. Bootstrap values of Maximum Likelihood higher than 60% are shown on the left, while values of Bayesian posterior probabilities above 80% are shown on the right. New species is given in bold. Clades of the key species or family are given in bold. The tree is rooted with Dothidea sambuci (Dothideaceae, Dothideales).
Selected taxa in this study with their corresponding GenBank accession numbers. The newly-generated sequences are shown in bold.
No. | Species | Vouncher /strain no. | LSU | ITS |
---|---|---|---|---|
1 | Acidomyces acidophilus | MH1085 | JQ172741 | JQ172741 |
2 | Asterina phenacis | TH 589 | GU586217 | – |
3 | Asterotexiaceae sp. | VUL.535 | MG844162 | – |
4 | Aulographum sp. | VUL.457 | MG844158 | – |
5 | Batcheloromyces proteae | CBS 110696 | JF746163 | JF746163 |
6 | Baudoinia compniacensis | CBS 123031 | GQ852580 | – |
7 | Brunneosphaerella protearum | CPC 16338 | GU214397 | GU214626 |
8 | Buelliella minimula | Lendemer 42237(NY) | KX244961 | – |
9 | Camarosporula persooniae | CBS 116258 | JF770461 | JF770449 |
10 | Capnobotryella renispora | CBS 214.90 | GU214398 | AY220612 |
11 | Capnodium coffeae | CBS 147.52 | GU214400 | DQ491515 |
12 | Catenulostroma protearum | CPC 15368 | GU214402 | GU214628 |
13 | Chaetothyrina guttulata | MFLUCC15–1080 | KU358917 | KX372277 |
14 | Chaetothyrina guttulata | MFLUCC15–1081 | KU358914 | KX372276 |
15 | Chaetothyrina musarum | MFLUCC 15–0383 | KU710171 | – |
16 | Cladosporium herbarum | CBS 121621 | KJ564331 | EF679363 |
17 | Cladosporium hillianum | CBS 125988 | KJ564334 | HM148097 |
18 | Cladosporium ramotenellum | CBS 170.54 | DQ678057 | AY213640 |
19 | Colletogloeum sp. | NY1_3.2F1c | FJ031986 | FJ425193 |
20 | Conidiocarpus (Phragmocapnias) betle | MFLUCC 10–0050 | JN832605 | – |
21 | Devriesia staurophora | ATCC 200934 | KF901963 | AF393723 |
22 | Dissoconium aciculare | CBS 204.89 | GU214419 | AY725520 |
23 | Dothidea sambuci | AFTOL-ID 274 | AY544681 | DQ491505 |
24 | Dothistroma pini | CBS 121011 | JX901821 | JX901734 |
25 | Elasticomyces elasticus | CCFEE 5547 | KF309991 | – |
26 | Exopassalora zambiae | YHJN13 | GQ433631 | GQ433628 |
27 | Extremus adstrictus | TRN96 | KF310022 | – |
28 | Friedmanniomyces endolithicus | CCFEE 5199 | KF310007 | JN885547 |
29 | Hispidoconidioma alpinum | L2–1/2 | FJ997286 | FJ997285 |
30 | Hortaea werneckii | CBS 100496 | GU301817 | AY128703 |
31 | Houjia yanglingensis | YHJN13 | GQ433631 | GQ433628 |
32 | Lecanosticta pini | CBS 871.95 | GQ852598 | – |
33 | Lembosia albersii | MFLUCC 13–0377 | KM386982 | – |
34 | Lembosina sp. | VUL.644 | MG844165 | – |
35 | Leptoxyphium cacuminum | MFLUCC 10–0049 | JN832602 | – |
36 | Melanodothis caricis | CBS 860.72 | GU214431 | GU214638 |
37 | Microcyclosporella mali | CPC 16171 | GU570545 | GU570528 |
38 | Microxyphium citri | CBS 451.66 | KF902094 | – |
39 | Morenoina calamicola | MFLUCC 14–1162 | NG059779 | NR154210 |
40 | Mycosphaerella pneumatophorae | AFTOL-ID 762 | KJ176856 | – |
41 | Neodevriesia coryneliae | CPC 23534 | KJ869211 | KJ869154 |
42 | Neodevriesia hilliana | CPC 15382 | GU214414 | GU214633 |
43 | Neodevriesia xanthorrhoeae | CBS 128219 | HQ599606 | HQ599605 |
44 | Neopseudocercosporella capsellae | CBS 127.29 | KF251830 | KF251326 |
45 | Nowamyces globulus | CBS 144598 | MN162196 | MN161935 |
46 | Nowamyces piperitae | CBS 143490 | MN162200 | MN161944 |
47 | Parapenidiella tasmaniensis | CBS 124991 | KF901844 | KF901522 |
48 | Passalora eucalypti | CBS 111318 | KF901938 | KF901613 |
49 | Penidiella columbiana | CBS 486.80 | EU019274 | KF901630 |
50 | Periconiella velutina | CBS 101950 | EU041840 | EU041783 |
51 | Petrophila incerta | TRN 77 | GU323963 | – |
52 | Phaeophleospora eugeniae | CPC 15159 | KF902095 | KF901742 |
53 | Phaeothecoidea eucalypti | CBS 120831 | KF901848 | KF901526 |
54 | Phaeothecoidiella illinoisensis | CBS 125223 | GU117901 | GU117897 |
55 | Phaeothecoidiella missouriensis | CBS 125222 | AY598917 | AY598878 |
56 | Phloeospora maculans | CBS 115123 | GU214670 | GU214670 |
57 | Piedraia hortae | CBS 480.64 | GU214466 | GU214647 |
58 | Piedraia quintanilhae | CBS 327.63 | GU214468 | – |
59 | Pseudocercospora vitis | CPC 11595 | GU214483 | GU269829 |
60 | Pseudoramichloridium henryi | CBS 124775 | KF442561 | KF442521 |
61 | Pseudotaeniolina globosa | CCFEE 5734 | KF310010 | KF309976 |
62 | Pseudoveronaea obclavata | CBS 132086 | JQ622102 | – |
63 | Racodium rupestre | L346 | EU048583 | GU067666 |
64 | Racodium rupestre | L424 | EU048582 | GU067669 |
65 | Ramichloridium apiculatum | CBS 156.59 | EU041848 | EU041791 |
66 | Ramularia endophylla | CBS 113265 | AY490776 | AY490763 |
67 | Ramularia pusilla | CBS 124973 | KP894141 | KP894248 |
68 | Ramulispora sorghi | CBS 110578 | GQ852653 | - |
69 | Readeriella mirabilis | CBS 125000 | KF251836 | KF251332 |
70 | Recurvomyces mirabilis | CBS 119434 | GU250372 | FJ415477 |
71 | Repetophragma zygopetali | VIC42946 | KT732418 | |
72 | Schizothyrium pomi | CBS 486.50 | EF134948 | EF134948 |
73 | Scolecostigmina mangiferae | CBS 125467 | GU253877 | GU269870 |
74 | Scorias spongiosa | CBS 325.33 | GU214696 | GU214696 |
75 | Septoria cytisi | USO 378994 | JF700954 | JF700932 |
76 | Septoria lysimachiae | CBS 123794 | KF251972 | KF251468 |
77 | Sonderhenia eucalyptorum | CBS 120220 | KF901822 | KF901505 |
78 | Sphaerulina myriadea | CBS 124646 | JF770468 | JF770455 |
79 | Sporidesmajira pennsylvaniensis | CBS 125229 | MH874965 | MF951287 |
80 | Stenella araguata | CBS 105.75 | EU019250 | EU019250 |
81 | Teratoramularia kirschneriana | CBS 113093 | GU214669 | GU214669 |
82 | Teratosphaeria fibrillosa | CBS 1217.07 | GU323213 | KF901728 |
83 | Toxicocladosporium irritans | CBS 185.58 | EU040243 | EU040243 |
84 | Toxicocladosporium rubrigenum | CBS 124158 | FJ790305 | FJ790287 |
85 | Translucidithyrium chinense | IFRDCC 3000 | MT659404 | MT659671 |
86 | Translucidithyrium thailandicum | MFLUCC 16–0362 | MG993048 | MG993045 |
87 | Tripospermum myrti | CBS 437.68 | GU323216 | – |
88 | Trochophora simplex | CBS 124744 | GU253880 | GU269872 |
89 | Uwebraunia communis | CBS 114238 | EU019267 | AY725541 |
90 | Vermiconia foris | CCFEE 5459 | GU250390 | KF309981 |
91 | Xenoconiothyrium catenatum | CMW 22113 | JN712570 | JN712502 |
92 | Zasmidium cellare | CBS 146.36 | EU041878 | EU041821 |
93 | Zygophiala cryptogama | OH4_1A1a | FJ147157 | FJ425208 |
94 | Zygophiala tardicrescens | MWA1a | EF164901 | AY598856 |
95 | Zygophiala wisconsinensis | OH4_9A1c | FJ147158 | FJ425209 |
Refer to the location of species, China.
IFRD9208
Epiphytic on living leaves, ascomata with papillate. Superficial hyphae absent. Sexual morph: Ascomata solitary or scattered, 480–870 μm diam. (x̄ = 741 μm, n = 6), 65–82 µm high (x̄ = 72 μm, n = 8), olivaceous to brown, slightly semi-transparent under highlighted background, circular to suborbicular, with slightly prominent papilla, membranous, without ostiole (Fig.
Translucidithyrium chinense (IFRD 9208, holotype) A plant leaves B acscoma on leaves surface C squash of ascoma at 20 times amplification D cross section of ascoma in blue cotton at 20 times amplification E, F cross section of ascoma in blue cotton at 40 times amplification G asci at 100 times amplification H–K asci in blue cotton at 100 times amplification L ascospore at 100 times amplification M–P ascospore in blue cotton at 100 times amplification. Scale bars: 200 µm (B); 100 µm (C, D); 50 µm (E, F); 20 µm (G–K); 10 µm (L–P). We slightly adjusted the contrast, saturation and hue of images and removed the contaminants around main object in images in PS software without obscuration, erasure or distortion of any information existing in the original document.
Ascospores germinating on MEA at 36 h after spore-isolation, germinating on PDA at 48 h after spore-isolation. Colonies slow growing on MEA and PDA, irregular, villiform, convex, white on surface, yellow to brown at base. After a long period of growth, the pigments produced by culture discolour the medium, roots generate at the bottom (Fig.
China, Yunnan Province, Xishuangbanna Dai Autonomous Prefecture, Xishuangbanna Botanical Garden; 21°55'51"N, 101°15'08"E, 540 m alt.; 21 Apr 2019; Haixia Wu and Xinhao Li leg; collected on living leaves of Alpinia blepharocalyx (IFRD 9208, holotype), ex-type living culture (IFRDCC 3000).
This new species is morphologically similar to Translucidithyrium thailandicum in having semi-transparent and largish ascomata, globose asci and hyaline ascospores with 1-septate. However, Translucidithyrium chinense has a slightly papilla thyriothecium with weaker transmittance and ascospores with guttules at both ends, while T. thailandicum has a flattened thyriothecium with higher transmittance and ascospores with appendages at both ends; besides, the size of ascomata and asci of T. chinense are slightly larger than those of T. thailandicum (795 μm vs. 621 μm; 77 μm vs. 64 μm). The culture characteristics of both species are different: the culture of T. chinense grows more slowly, has roots inserting into medium and turn the bottom brown. Phylogenetically, T. chinense clusters with T. thailandicum as a distinct clade with high support (100% ML / 1.00 PP, Fig.
The Maximum Clade Credibility (MCC) tree was similar to the major lineages in the Bayesian and ML trees. The crown age of Translucidithyrium showed 16 Mya (4–33), which was earlier than the divergence time of most genera in Phaeothecoidiellaceae. The estimated divergence time of Phaeothecoidiellaceae from
Translucidithyrium thailandicum was found in the north of Thailand (
The ascomata of Translucidithyrium are different from related genera of Phaeothecoidiellaceae: Nowamyces has immersed ascomata, Chaetothyrina has ascomata with setae and Rivilata has subcuticular ascomata (
The MCC tree with divergence times estimates of Phaeothecoidiellaceae obtained from a Bayesian approach (BEAST). Numbers at nodes indicate posterior probabilities (pp) for node support; bars correspond to the 95% highest posterior density (HPD) intervals. The key species are given in blue.
Historical events amongst different biological groups could then be compared with the dates of plate tectonic movements and paleoecology, according to the corresponding geological time scale (
Funds for research were provided by the Grant for Essential Scientific Research of National Nonprofit Institute (no. CAFYBB2019QB005), the Yunnan Province Ten Thousand Plan of Youth Top Talent Project (no. YNWR-QNBJ-2018-267) and the Yunnan Fundamental Research Projects (grant NO. 202001AT070014). The authors are deeply grateful to Prof. K.D. Hyde (Mae Fah Luang University, Thailand, MFU) for editing the English language of the manuscript, to Dr. Xiang-Yu Zeng and Dr. Nawaz Haider for revising this manuscript and to Dr. Rungtiwa Phookamsak for guiding experiment operation.