Biodiversity Data Journal :
Taxonomic Paper
|
Corresponding author: Yu-Lan Jiang (yljchsd@163.com), Guang-Can Tao (tgcan@gzata.cn)
Academic editor: Ning Jiang
Received: 13 Jun 2021 | Accepted: 13 Dec 2021 | Published: 23 Dec 2021
© 2021 Hong Zhang, Tian-Peng Wei, Yu-Tao Mao, Ming-Xia Ma, Kai Ma, Ying Shen, Mei-Juan Zheng, Wei-Yu Jia, Ming-Yan Luo, Yan Zeng, Yu-Lan Jiang, Guang-Can Tao
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:
Zhang H, Wei T-P, Mao Y-T, Ma M-X, Ma K, Shen Y, Zheng M-J, Jia W-Y, Luo M-Y, Zeng Y, Jiang Y-L, Tao G-C (2021) Ascodesmis rosicola sp. nov. and Talaromyces rosarhiza sp. nov., two endophytes from Rosa roxburghii in China. Biodiversity Data Journal 9: e70088. https://doi.org/10.3897/BDJ.9.e70088
|
Rosa roxburghii Tratt., a deciduous shrub of the family Rosaceae, is usually used as food and medicinal materials and also cultivated as an ornamental. Plant endophytic fungi are a large class of microbial resources not fully researched, with great potential applications. Two strains of Ascodesmis and Talaromyces were isolated during a survey of biodiversity on endophytic fungi of R. roxburghii in China. Multigene phylogenetic analyses showed that each of the two fungi formed a distinct lineage and separated from known congeneric species and they are proposed as two novel taxa.
Ascodesmis rosicola sp. nov. usually has one or two conspicuous simple or branched ridges extending to the majority of the ascospore surface and remarkably small asci, distinguishing it from the previously-described species in the genus Ascodesmis. Talaromyces rosarhiza sp. nov., of the section Talaromyces, is closely related to T. francoae. It differs from the latter by having both monoverticillate and biverticillate conidiophores, while those of T. francoae are biverticillate. Both novel endophytes are illustrated and described.
multigene phylogenetics, morphology, new taxa, taxonomy, endophytic fungi
Rosa roxburghii Tratt. is currently attracting significant attention for notably high vitamin C, superoxide dismutase and flavonoids (
Endophytic fungi are phylogenetically diverse microorganisms that can colonise asymptomatically in various parts of living and healthy plants, ranging from the roots, stems and leaves to the flowers, fruits and seeds. To date, endophytes have been obtained from almost all plants investigated. Woody plants, in particular, may contain hundreds or thousands of fungal endophytes (
Endophytic fungi belong to Ascomycota, Basidiomycota and Zygomycota, but Ascomycota is the predominant group compared to others (
The genus Talaromyces, within the order Eurotiales, was erected by
In this study, we introduce Ascodesmis rosicola and Talaromyces rosarhiza as two novel taxa. Both of them are confirmed by multigene phylogeny and morphological characters.
The wild-type R. roxburghii was collected from Guizhou Province, China (April and August 2020) (Table
Location charateristics and climate features of samples collection sites
Sampling site |
Latitude and longitude |
Altitude (m) |
Mean annual temperature (℃) |
Mean annual rainfall (mm) |
Mean annual sunshine hours (h) |
Sampling time |
Guiyang City, Guizhou Province |
|
1184 |
15.3 |
1130 |
1235 |
22 April 2020 |
Liupanshui City, Guizhou Province |
|
2047 |
15.2 |
1390 |
1593 |
4 August 2020 |
All tissues, especially roots, were cleaned under tap water for half an hour, rinsed with double-distilled water for 10 min,and dried under natural conditions. Then, they were cut into small pieces and transferred to the clean bench for surface disinfection. The margin of these pieces was trimmed off under aseptic conditions. All samples were surface-sterilised in 75% ethanol (1 min) and rinsed three times with sterile water. Subsequently, they were treated with 1% (w/v) aqueous sodium hyprochlorite (NaOCl) for several minutes (roots, 2 min; fruit, 1 min) and washed three times with sterile water again. After washing, the surface water of tissues was blotted with sterile filter papers. The potato dextrose agar (PDA, Shanghai Bio-way Technology Co., Ltd., China) medium had been supplemented with streptomycin sulphate (0.5 g/l) to avoid bacterial contamination. Each culture plate contained three segments. These plates were incubated at 28±1°C in a 12-h light:12-h dark photoperiod for 3 to 7 days. After several days, hyphal tips were subcultured into fresh PDA plates to obtain pure endophytic strains (
The effectiveness of surface sterilisation was monitored with three methods. First, the final rinsing water (0.1 ml) was coated on PDA plates (
Morphological identification was performed with PDA (Shanghai Bio-way Technology Co., Ltd., China), oatmeal agar (OA, Beijing Solarbio Science & Technology Co., Ltd., China), and malt-extract agar (MEA, Shanghai Bio-way Technology Co., Ltd., China). Macroscopic morphology, for example, growth rate, colony colour and pigmentation, were examined at 2-day intervals. Microscopic characteristics of ascospores or conidia were determined and recorded and spores of each strain were randomly selected for measurement (length and width) (
DNA was extracted from fresh mycelia grown on PDA for 7 to 10 days using the Fungal gDNA Isolation Kit (BW-GD2416, Biomiga, China), following the manufacturer’s instructions. The products were stored at -20°C until polymerase chain reaction (PCR) was performed. PCR amplifications were conducted on a T100 Thermal Cycler (BIO-RAD, USA). According to manufacturer’s instructions in a total volume of 25 μLl, including 12.5 µl 2X SanTaq PCR Mix (the mixture of MgCl2, dNTP, Taq DNA Polymerase, PCR buffer and PCR enhancer solution), 9.5 µl double-distilled water, 1.0 µl each of forward and reverse primers (0.05–1 µmol/l) and 1.0 µl DNA sample (1–10 ng/µl). The forward and reverse PCR reaction primers are shown in Table
Genus |
Phylogenetic analysis |
Model |
||||
ITS (ITS4/ITS5) |
LSU (LR0R/LR5) |
BenA (Bt2a/Bt2b) |
CaM (Cmd5/Cmd6) |
RPB2 (fRPB2-5F/ fRPB2-7cR) |
||
Ascodesmis |
ML analysis |
TNe+I+G4 |
TIM3+F+I+G4 |
- |
- |
TNe+I+G4 |
BI analysis |
GTR+F+I |
GTR+F+I+G4 |
- |
- |
SYM+I+G4 |
|
Talaromyces |
ML analysis |
TNe+R3 |
- |
TIM2e+I+G4 |
K2P+I+G4 |
K2P+I+G4 |
BI analysis |
GTR+F+I+G4 |
- |
GTR+F+I+G4 |
SYM+I+G4 |
SYM+I+G4 |
Forward and reverse sequences were utilised to create consensus sequences by BioEdit v. 7.0.9.0 (
Taxa used in this study and their corresponding GenBank accession numbers.
Species |
Strain no. |
GenBank accession no. | ||||
ITS |
LSU |
BenA |
CaM |
RPB2 |
||
Ascodesmis microscopica |
CBS 275.80 |
- |
- |
- |
||
A. nigricans |
CBS 389.68 T |
- |
- |
- |
||
A. sphaerospora |
CBS 125.61 T |
- |
- |
- |
||
A. rosicola sp. nov. |
GUCC 190035.1 T |
- |
- |
|||
A. rosicola sp. nov. |
GUCC 190204.1 |
- |
- |
|||
Boubovia nicholsonii |
KH.03.65 (FH) |
- |
- |
- |
||
Eleutherascus lectardii |
CBS 626.71 T |
- |
- |
- |
||
E. lectardii |
AFTOL-ID 933 |
- |
- |
- |
||
E. peruvianus |
CBS 101.75 T |
- |
- |
- |
||
E. peruvianus |
CBS 104.77 |
- |
- |
- |
||
Geopyxis vulcanalis |
KH.04.37 (FH, dupl. DBG) |
- |
- |
- |
||
Lasiobolus ciliatus |
KS-94-005 (C) |
- |
- |
- |
- |
|
L. papillatus |
KH.08.30 |
- |
- |
- |
||
Pseudocoprotus catenipilus |
ALV9774 |
- |
- |
|||
P. catenipilus |
- |
- |
- |
|||
Pseudombrophila theioleuca |
DHP 3498 (FH) |
- |
- |
- |
||
Pulvinula constellatio |
KH.03.64 (FH) |
- |
- |
- |
||
Pu. convexella |
KH.01.20 (C) |
- |
- |
- |
||
Talaromyces assiutensis |
CBS 147.78 T |
- |
||||
T. bacillisporus |
CBS 296.48 T= IMI 040045 = NRRL 1025 |
- |
||||
T. calidicanius |
CBS 112002 T |
- |
||||
T. cecidicola |
CBS 101419 T= DAOM 233329 |
- |
||||
T. cinnabarinus |
CBS 267.72 T= NHL 2673 |
- |
||||
T. coalescens |
CBS 103.83 T |
- |
||||
T. diversiformis |
CBS 141931 T= CGMCC3.18204 = DTO 317-E3 |
- |
||||
T. duclauxii |
CBS 322.48 T= IMI 040044 = MUCL 28672 = NRRL 1030 |
- |
||||
T. euchlorocarpius |
PF 1203 = DTO 176-I3 T= DTO 176-I4 |
- |
||||
T. flavus |
CBS 310.38 T= IMI 197477 = NRRL 2098 |
- |
||||
T. francoae |
CBS 113134 T= IBT 23221 = DTO 056-D9 |
- |
||||
T. helicus |
CBS 335.48 T= DSM 3705 = IMI 040593 = NRRL 2106 |
- |
||||
T. intermedius |
CBS 152.65 T= BDUN 267 = IFO 31752 = IMI 100874 |
- |
||||
T. islandicus |
CBS 338.48 T= IMI 040042 = MUCL 31324 = NRRL 1036 |
- |
||||
T. kendrickii |
CBS 136666 T = DTO 273-F4 = IBT 13593 |
- |
||||
T. mangshanicus |
CGMCC 3.18013 |
- |
||||
T. marneffei |
CBS 388.87 T = ATCC 18224= CBS 334.59 = IMI 068794ii = IMI 068794iii |
- |
||||
T. mimosinus |
CBS 659.80 T = FRR 1875 = IMI 223991 |
- |
||||
T. neorugulosus |
CBS 140623 T= CGMCC3.18215 = DTO 318-A8 |
- |
||||
T. panamensis |
CBS 128.89 T= IMI 297546 |
- |
||||
T. purpureus |
CBS 475.71 T= FRR 1731 = IMI 181546 |
- |
||||
T. purpurogenus |
CBS 286.36 T= IMI 091926 |
- |
||||
T. qii |
AS3.15414 T= CBS 139515 |
- |
||||
T. resinae |
AS 3.4387 = CBS 324.83 T= DTO 027-G5 |
- |
||||
T. reverso-olivaceus |
CBS 140672 T= CGMCC3.18195 = DTO 317-C3 |
- |
||||
T. rosarhiza sp. nov. |
GUCC 190040.1 T |
- |
||||
T. rosarhiza sp. nov. |
GUCC 197011.1 |
- |
||||
T. rugulosus |
CBS 371.48 T= IMI 040041 = MUCL 31201 = NRRL 1045 |
- |
||||
T. stipitatus |
CBS 375.48 T= NRRL 1006 = IMI 39805 |
- |
||||
T. subinflatus |
CBS 652.95 T= IBT 17520 |
- |
||||
T. tenuis |
CBS 141840 T = DTO 340-G9 |
- |
||||
T. thailandensis |
CBS 133147 T= KUFC 3399 |
- |
||||
T. trachyspermus |
CBS 373.48 T = IMI 040043 |
- |
||||
T. tzapotlensis |
NRRL 35203 T |
- |
||||
T. viridis |
CBS 114.72 T= ATCC 22467 = NRRL 5575 |
- |
||||
Trichocoma paradoxa |
CBS 247.57, CBS 103.73, CBS 788.83 T |
- |
New species are marked in bold; T: indicates type or ex-type strains.
Maximum Likelihood (ML) and Bayesian Inference (BI) methods were used to construct the phylogenetic trees. The best-fit partition models were inferred for the independent sequence datasets using ModelFinder (
Endophytic fungi of R. roxburghii. Asexual morph not observed. Sexual morph (Fig.
Cultural characteristics: Description based on GUCC 190035.1. On PDA, reaching 6.0–7.5 cm in diameter after 14 days of cultivation in dark at 28°C, superficial, margin irregular, white, with abundant aerial mycelium, uneven, centre and margin uplifted; reverse yellowish. On MEA, 28°C: medium sparse, circular, flat. Mycelium white, cottony, margin regular. On OA, 28°C: radial, aerial mycelium sparse, pale white, growth regular. On MEA and OA, reaching 85 mm in diameter under the same conditions.
The name refers to the host plant, Rosa roxburghii, from which this fungus was isolated.
Ascodesmis rosicola is introduced as a new species, based on morphological characteristic and phylogenetic analysis. The genus Ascodesmis, established by
Phylogram generated from Maximum Likelihood analysis, based on combined ITS, LSU and RPB2 sequence data. Geopyxis vulcanalis (KH.04.37 (FH, dupl. DBG)) is used as the outgroup taxa. Bootstrap support values for ML greater than 50% and Bayesian posterior probabilities greater than 0.90 are given near nodes (BS/PP), respectively. The novel taxon is indicated in bold and black. T: type or ex-type.
Endophyte of R. roxburghii. Sexual morph not observed. Asexual morph (Fig.
Cultural characteristics: Description based on GUCC 190040.1. On PDA, reaching 50 mm in diameter after 14 days of cultivation in dark at 28°C; moderately deep, slightly sulcate, flat; margin entire, mycelium white. On OA, reaching 42 mm in diameter under the same conditions, low, flat; margin low, entire; mycelium white; velvety; abundant sporulation; conidia en masse dark olive green. On MEA 28°C, 14 days: reaching 12 mm; raised; margin low, flat, entire; mycelium white; floccose to velvety; abundant sporulation, conidia en masse dull green; soluble pigments absent; exudates absent; reverse greyish orange.
The word "rosarhiza" originated from “rosa” referring to the host plant, Rosa roxburghii and “rhiza” referring to root, from which this fungus was isolated.
Talaromyces rosarhiza is described as a new species, based on morphology and phylogenetic analyses. Phylogenetic analysis (Fig.
Species |
Conidiophores pattern |
No. of metulae per verticil |
Metulae size (μm) |
No. of phialides per metulae |
Phialides size (μm) |
Conidia shape |
Conidial walls |
Conidia size (μm) |
T. rosarhiza sp. nov. |
Monoverticillate and biverticillate |
2–3 |
6.0–14.5 × 2.0–3.0 |
2-3 |
6.5–15.0 × 1.5–3.5 |
Ellipsoidal |
Smooth |
2.5–3.5 × 2.0 –3.0 |
T. francoae |
Biverticillate |
3-6 |
8–13 × 2.5–4.5 |
3–6 |
8.5–12 × 2.5–4 |
Globose |
Verrucose, rough |
2.5–4 × 2.5–4 |
T. kendrickii |
Biverticillate |
3–8 |
10–13 × 3–4 |
3–5 |
9–12 × 2.5– 3.5 |
Subglobose |
Roughened |
2.5–3 × 2.5–3 |
T. mangshanicus |
Biverticillate |
3–6 |
11–13.5 × 4–5 |
3–6 |
10.5–13.5 × 3.5–4.5 |
Subglobose to ellipsoidal |
Echinulate |
4.5–5.5 × 4–5 |
T. qii |
Biverticillate |
4–6 |
7–11 × 2.5–3 |
2–4 |
7–9×2–2.5 |
Ovoid to subglobose |
Echinulate |
3–3.5 |
T. thailandensis |
Biverticillate |
3–5 |
7.2–10.9 × 2.2–3.4 |
3–7 |
11.5–13.7 × 1.5–2.4 |
Subglobose to ellipsoidal |
Smooth |
1.8–2.4 × 1.7 –2.3 |
Phylogram generated from Maximum Likelihood analysis, based on combined ITS, BenA, CaM and RPB2 sequence data. Trichocoma paradoxa (CBS 788.83) is used as the outgroup taxa. Bootstrap support values for ML greater than 50% and Bayesian posterior probabilities greater than 0.90 are given near nodes (BS/PP), respectively. The new taxon is indicated in bold and black. T: type or ex-type.
Ascodesmis rosicola: For the genus Ascodesmis, only three species, A. microscopica, A. nigricans and A. sphaerospora, have available sequence data. Therefore, phylogenetic relationships were deduced using sequence data of Pezizales. Nineteen strains were included in the combined analyses (ITS, LSU and RPB2) which comprised 1999 characters (561 for ITS, 819 for LSU, 619 for RPB2). The tree topology of the ML analysis was similar to the BI analysis. Two new strains, GUCC 190035.1 and GUCC 190204.1, were related to Ascodesmis spp., clustered with good support (BS = 94, PP = 1) and formed an independent moderately-supported clade (BS = 61, PP = 0.92) (Fig.
Talaromyces rosarhiza: Phylogenetic relationships were investigated using sequence data of Talaromyces. Thirty-six strains were included in the combined analyses (ITS, BenA, CaM and RPB2) which comprised 2549 characters (541 for ITS, 570 for BenA, 647 for CaM, 791 for RPB2). The tree topology of ML analysis was similar to the BI analysis. Two new strains, GUCC 190040.1 and GUCC 197011.1, clustered with strong support (BS = 100, PP = 1) and formed a group with five species of Talaromyces that had been reported and belonged to section Talaromyces. Talaromyces rosarhiza was closely related to T. francoae, but placed in a distinct clade with high bootstrap support (BS = 90) and posterior probability (PP = 1) (Fig.
In this study, A. rosicola and T. rosarhiza, two new endophytes isolated from R. roxburghii in China, are proposed and described. Multigene phylogenetic analyses revealed that A. rosicola was phylogenetically close to A. nigricans and A. sphaerospora and was placed in a distinct clade with moderate support. However, combined with morphological characters, the novel species is confirmed. Similarly, based on morphology and phylogenetic analyses, T. rosarhiza is suggested as a new species.
Although there are 13 species of Ascodesmis listed in Mycobank (https://www.mycobank.org/), A. aurea and A. hawaiiensis are considered dubious species due to the lack of adequate description and material (
Talaromyces, another genus in this study, is important in biotechnology, medicine and the food industry (
In the present study, a total of 127 strains of endophytic fungi were successfully isolated from R. roxburghii and categorized into six classes (Sordariomycetes, Dothideomycetes, Eurotiomycetes, Pezizomycetes, Leotiomycetes and Agaricomycetes), of which strains GUCC 190035.1, GUCC 190204.1, GUCC 190040.1 and GUCC 197011.1 were identified as two new taxa. As known, besides agriculture and medicine, many endophytic fungi may play significant roles in shaping and maintaining the balance of microbial communities in plants (
This work is supported by the following projects: the National Natural Science Foundation of China (No. 32060009) and the Guizhou Provincial Science and Technology Projects ([2020]1Y043 and [2018]5404).