Botryosphaerialean fungi associated with woody oil plants cultivated in Sichuan Province, China

Wen-Li Li; Rui-Ru Liang; Asha J. Dissanayake; Jian-Kui Liu

doi:10.3897/mycokeys.97.103118

Research Article

Botryosphaerialean fungi associated with woody oil plants cultivated in Sichuan Province, China

Wen-Li Li^‡, Rui-Ru Liang^‡, Asha J. Dissanayake^‡, Jian-Kui Liu^‡

‡ Electronic Science and Technology University, Chengdu, China

Corresponding author: Jian-Kui Liu ( ljiankui@gmail.com )

Academic editor: Nalin Wijayawardene

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 W-L, Liang R-R, Dissanayake AJ, Liu J-K (2023) Botryosphaerialean fungi associated with woody oil plants cultivated in Sichuan Province, China. MycoKeys 97: 71-116. https://doi.org/10.3897/mycokeys.97.103118

Abstract

Woody oil plants are important economic trees which are widely cultivated and distributed throughout China. Surveys conducted during 2020 and 2021 on several woody oil plantations from five regions of Sichuan Province, China, revealed a high diversity of Botryosphaerialean fungi. The identification of 50 botryosphaeriaceous isolates was carried out based on both morphology and multi-gene phylogenetic analysis of internal transcribed spacer region (ITS), translation elongation factor 1-alpha gene (tef1) and β-tubulin gene (tub2). This allowed the identification of twelve previously known Botryosphaeriales species: Aplosporella prunicola, A. ginkgonis, Barriopsis tectonae, Botryosphaeria dothidea, Bo. fabicerciana, Diplodia mutila, Di. seriata, Dothiorella sarmentorum, Neofusicoccum parvum, Sardiniella guizhouensis, Sphaeropsis citrigena, and Sp. guizhouensis, and four novel species belonging to the genera Diplodia and Dothiorella, viz. Di. acerigena, Di. pistaciicola, Do. camelliae and Do. zanthoxyli. The dominant species isolated across the surveyed regions were Botryosphaeria dothidea, Sardiniella guizhouensis and Diplodia mutila, representing 20%, 14% and 12% of the total isolates, respectively. In addition, most isolates were obtained from Pistacia chinensis (14 isolates), followed by Camellia oleifera (10 isolates). The present study enhances the understanding of Botryosphaeriales species diversity on woody oil plants in Sichuan Province, China.

Keywords

Botryosphaeriales, diversity, new species, phylogeny, taxonomy

Introduction

Botryosphaeriaceae is a diverse group of fungi that includes endophytes, saprobes and plant pathogens (Phillips et al. 2013). They have broad host ranges, and are widely distributed in tropical and temperate regions (Batista et al. 2021). Botryosphaeriaceae was introduced by Theissen and Sydow (1918) to accommodate three genera Botryosphaeria, Dibotryon and Phaeobotryon. Botryosphaeriales was proposed to include the single family, Botryosphaeriaceae, based on multi-gene phylogeny (Schoch et al. 2006). Up to date, six families and 32 genera are accepted in Botryosphaeriales, while Botryosphaeriaceae is known to be the largest monophyletic family, including 22 genera and more than 200 species (https://botryosphaeriales.org/, accessed on 15^th April 2023).

The members of Botryosphaeriaceae have been taxonomically characterized based on both sexual and asexual morphs. The production of large, ovoid to oblong, typically hyaline, aseptate ascospores, which may become brown and septate with age, as well as bitunicate asci within unilocular or multilocular botryose ascomata known as pseudothecia is typical to the sexual state (Sivanesan 1984; Phillips et al. 2005). The asexual states of Botryosphaeriaceae exhibit a wide range of conidial morphologies; for example, its conidia can be thin-walled and hyaline, or thick-walled and pigmented, aseptate or 1–2-septate (Phillips et al. 2005). Additonaly, the spermatial states were also frequently observed in Botryosphaeriaceae species, which produced unicellular, hyaline, allantoid to rod-shaped spermatia on culture. Botryosphaeriaceae species are significantly different from other fungi in that the color of its aerial hyphae, changing from gray to black with age on 2% potato dextrose agar (PDA), which can be used for the rapid determination of botryosphaeriaceous fungi.

The geographic distribution and host range of botryosphaeriaceous taxa are diverse. Seven genera in Botryosphaeriaceae: Botryosphaeria, Diplodia, Dothiorella, Lasiodiplodia, Neodeightonia, Neofusicoccum and Phaeobotryon are common and frequently reported from various geographical regions (Batista et al. 2021), while Botryobambusa, Oblongocollomyces, Sakireeta and Sardiniella appear to be limited to a single region or country (Liu et al. 2012; Crous et al. 2015; Linaldeddu et al. 2016; Yang et al. 2017; Dissanayake et al. 2021). Many Botryosphaeriaceae species have wide host ranges (e. g. Botryosphaeria dothidea, Diplodia mutila, Dothiorella sarmentorum, Lasiodiplodia theobromae and Neofusicoccum parvum), while other species have narrower host ranges (e. g. Diplodia olivarum was reported on olive, oleaster, carob, grapevine, almond et al.) (Lazzizera et al. 2008; Granata et al. 2011; Linaldeddu et al. 2015; Olmo et al. 2016) or even in very specific hosts (e. g. Eutiarosporella darliae was only reported on infected wheat and wheat-stubble) (Thynne et al. 2015; Farr and Rossman 2021). Different species of Botryosphaeriaceae exhibit different environmental adaptations and host preferences (Braunsdorf et al. 2016). Botryosphaeriaceous taxa with narrow host ranges or limited geographic distribution will be more susceptible to climatic effects (Slippers et al. 2017; Li et al. 2020).

Woody oil plants are economically important as they are used for the production of cooking and industrial oil. Recently, many Botryosphaeriaceae species have been frequently reported on woody oil plants. Diplodia olivarum was first reported from rotting olive drupes in Italy (Lazzizera et al. 2008) and later it was reported as associated with declining Prunus dulcis trees in Spain (Gramaje et al. 2012). Diplodia insularis was isolated from branch canker of Pistacia lentiscus in Italy (Linaldeddu et al. 2015). Dothiorella gregaria was isolated from the stems with asymptomatic of Zanthoxylum bungeanum in China (Li et al. 2016b). Botryosphaeria dothidea, Diplodia mutila, Di. seriata, Dothiorella iberica, Do. omnivora, Do. sarmentorum, Lasiodiplodia citricola, L. pseudotheobromae, L. theobromae, Neofusicoccum mediterraneum, N. nonquaesitum, N. parvum, N. ribis, N. vitifusiforme, and Neoscytalidium dimidiatum have been reported as pathogens of English walnut (Juglans regia L.) in California (Chen et al. 2014), Chile (Jimenez Luna et al. 2022), China (Li et al. 2016a; Zhang et al. 2017), Iran (Abdollahzadeh et al. 2013; Panahandeh et al. 2019), South Africa (Cloete et al. 2011), Spain (Gramaje et al. 2012) and USA (Chen et al. 2014). However, very little is known about the Botryosphaerialean species occurring on native woody oil plants in China. Hence, the aim of this study was to gain a more comprehensive understanding of the diversity of Botryosphaeriaceae species associated with common woody oil plants grown in Sichuan Province, China.

Materials and methods

Isolates and morphology

The isolates in this study were collected from the woody oil tree plantations in Sichuan Province during the period of 2020 and 2021. The hosts include Acer truncatum, Camellia oleifera, Idesia polycarpa, Olea europaea, Paeonia suffruticosa, Pistacia chinensis, Vernicia fordii and Zanthoxylum bungeanum. The samples were collected from decayed stems, branches and twigs of woody oil trees. Mature fruiting bodies were selected for fungal isolation and for morphological observations under stereo microscope Motic SMZ 168 series. Measurements were made with Tarosoft Image Frame Work program v. 0.9.7 (Liu et al. 2010). Thirty conidia/ascospores were measured per isolate, and 10–30 measurements were taken of other morphological structures. At least 20 conidia/ascospores were used to calculate the average length/width ratio (L/W). Single spore isolation was conducted in accordance with the methods described in Chomnunti et al. (2014). Germinated spores were individually placed on PDA plates and grown at 25 °C in daylight.

Herbarium specimens were stored in the herbarium of Cryptogams Kunming Institute of Botany, Academia Sinica (KUN-HKAS) and duplicated at Herbarium, University of Electronic Science and Technology (HUEST), Chengdu, China. Living cultures were deposited at China General Microbiological Culture Collection Centre (CGMCC), Beijing, China and duplicated at the University of Electronic Science and Technology Culture Collection (UESTCC), Chengdu, China. MycoBank numbers were registered as outlined in MycoBank (http://www.MycoBank.org. Accessed on 11^th November 2022).

DNA extraction, PCR amplification and sequencing

The total genomic DNA was extracted from 7day-old isolates grown on 2% PDA median at 25 °C, using the EZ geneTM fungal gDNA kit (GD2416), following the manufacturer’s instructions and protocols. Partial gene sequences were determined for the internal transcribed spacer 1 and 2 including the intervening 5.8S nrDNA gene (ITS), the nuclear ribosomal 28s large subunit (LSU), the translation elongation factor 1-alphagene (tef1), and the β-tubulin gene (tub2). The primers used for ampliﬁcation are ITS5/ITS4 for ITS (White et al. 1990), LR0R/LR5 for LSU (Vilgalys and Hester 1990), EF1-728F/EF1-986R for tef1 (Carbone and Kohn 1999) and Bt2a/Bt2b for tub2 (Glass and Donaldson 1995). Polymerase chain reaction (PCR) amplification conditions were followed as of Dissanayake et al. (2021). PCR products were sent for sequencing at Beijing Tsingke Biological Engineering Technology and Services Co. Ltd. (Beijing, P.R. China). All newly generated sequences are deposited in GenBank, and the obtained accession numbers are listed in Table 1.

Table 1.

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All newly generated sequences in this study. Ex-type strains are indicated with *. N/A: Not available.

Taxon	Stain Number	GenBank Accession Number
Taxon	Stain Number	ITS	tef1	tub2
Aplosporella ginkgonis	UESTCC 22.0091	OQ190504	OQ241438	N/A
Aplosporella prunicola	UESTCC 22.0090	OQ190505	N/A	N/A
Barriopsis tectonae	UESTCC 22.0092	OQ190506	OQ241439	N/A
Botryosphaeria dothidea	UESTCC 22.0111	OQ190507	OQ241440	N/A
	UESTCC 22.0109	N/A	OQ241441	N/A
	UESTCC 22.0112	OQ190508	OQ241442	N/A
	UESTCC 22.0113	OQ190509	OQ241443	N/A
	UESTCC 22.0108	OQ190510	OQ241444	N/A
	UESTCC 22.0116	OQ190511	OQ241445	N/A
	UESTCC 22.0114	OQ190512	OQ241446	N/A
	UESTCC 22.0115	OQ190513	OQ241447	N/A
	UESTCC 22.0110	OQ190514	OQ241448	N/A
	UESTCC 22.0107	OQ190515	OQ241449	N/A
Botryosphaeria fabicerciana	UESTCC 22.0117	OQ190516	OQ241450	N/A
Botryosphaeria fabicerciana	UESTCC 22.0118	OQ190517	OQ241451	N/A
Diplodia acerigena*	CGMCC 3.24157	OQ190518	OQ241452	N/A
Diplodia acerigena	UESTCC 22.0074	OQ190519	OQ241453	OQ338163
Diplodia acerigena	UESTCC 22.0075	OQ190520	OQ241454	OQ338164
Diplodia mutila	UESTCC 22.0064	OQ190521	OQ241455	OQ338165
	UESTCC 22.0065	OQ190522	OQ241456	OQ338166
	UESTCC 22.0069	OQ190523	OQ241457	OQ338167
	UESTCC 22.0068	OQ190524	OQ241458	OQ338168
	UESTCC 22.0067	OQ190525	OQ241459	OQ338169
	UESTCC 22.0063	OQ190526	OQ241460	OQ338170
Diplodia pistaciicola *	CGMCC 3.24156	OQ190527	OQ241461	OQ338171
Diplodia pistaciicola	UESTCC 22.0071	OQ190528	OQ241462	OQ275062
Diplodia seriata	UESTCC 22.0072	OQ190529	OQ241463	N/A
Dothiorella camelliae	UESTCC 22.0080	OQ190530	N/A	OQ275063
Dothiorella camelliae *	CGMCC 3.24158	OQ190531	OQ241464	OQ275064
Dothiorella camelliae	UESTCC 22.0079	OQ190532	OQ241465	OQ275065
Dothiorella camelliae	UESTCC 22.0078	OQ190533	OQ241466	OQ275066
Dothiorella sarmentorum	UESTCC 22.0076	OQ190534	N/A	OQ275067
Dothiorella sarmentorum	UESTCC 22.0077	OQ190535	OQ241467	OQ275068
Dothiorella zanthoxyli *	CGMCC 3.24159	OQ190536	OQ241468	OQ275069
Dothiorella zanthoxyli	UESTCC 22.0083	OQ190537	OQ241469	OQ275070
Dothiorella zanthoxyli	UESTCC 22.0084	OQ190538	OQ241470	OQ275071
Neofusicoccum parvum	UESTCC 22.0096	OQ190539	OQ241471	N/A
	UESTCC 22.0094	OQ190540	N/A	N/A
	UESTCC 22.0093	OQ190541	N/A	N/A
	UESTCC 22.0095	OQ190542	N/A	N/A
Sardiniella guizhouensis	UESTCC 22.0100	OQ190543	OQ241472	N/A
	UESTCC 22.0101	OQ190544	OQ241473	N/A
	UESTCC 22.0099	OQ190545	OQ241474	N/A
	UESTCC 22.0097	OQ190546	OQ241475	N/A
	UESTCC 22.0098	OQ190547	OQ241476	N/A
	UESTCC 22.0102	OQ190548	OQ241477	N/A
	UESTCC 22.0103	OQ190549	OQ241478	N/A
Sphaeropsis citrigena	UESTCC 22.0106	OQ190550	OQ241479	N/A
Sphaeropsis citrigena	UESTCC 22.0105	OQ190551	OQ241480	N/A
Sphaeropsis guizhouensis	UESTCC 22.0104	OQ190552	OQ241481	N/A

Phylogenetic analyses

Sequence data for phylogenetic analyses were obtained from GenBank and from recent publications regarding Botryosphaeriaceae fungi (Dissanayake et al. 2021; Xiao et al. 2021; Zhang et al. 2021; Rathnayaka et al. 2022) (See Suppl. material 1). The single gene alignments were performed using MAFFT v.7.429 online service (https://mafft.cbrc.jp/alignment/server/, accessed on 15 October 2022) (Katoh et al. 2019) and ambiguous regions were excluded using TrimAI with the option “-automated1”, which trimmed sequences based on similarity statistics (Capella-Gutiérrez et al. 2009). Multi-gene sequences were concatenated by Sequence matrix software (Vaidya et al. 2011). Multi-gene phylogenetic analyses were obtained from maximum likelihood (ML) and Bayesian inference (BI) analyses following Dissanayake et al. (2020).

ML analyses was performed using RAxML (Stamatakis 2006). The tree search included 1,000 non-parametric bootstrap replicates and the best scoring tree was selected from suboptimal trees under the GTRGAMMA substitution model. Maximum likelihood bootstrap values equal or greater than 75% are marked near each node of the phylogenetic tree.

Bayesian analyses was performed in MrBayes 3.2.6 (Ronquist et al. 2012). The program MrModeltest 2.2 (Nylander 2004) was used to determine the best nucleotide substitution model for each data partition. The Markov Chain Monte Carlo (MCMC) sampling approach was used to calculate the posterior probabilities (PP) (Rannala and Yang 1996). Bayesian analyses of four simultaneous Markov chains were run for 10,000,000 generations with trees sampled every 1,000^th generations. The first 20% of trees, representing the burn-in phase of the analyses, were discarded, and the remaining trees were used for calculating posterior probabilities (PP) in the majority rule consensus tree. PP values equal or greater than 0.95 are marked near each node.

Trees were visualized with FigTree v1.4.0 (Rambaut 2006), and the layout was edited using Adobe Illustrator CS6 software (Adobe Systems, USA).

Results

Phylogenetic analyses

A concatenated dataset of ITS and tef1 was used to determine the phylogenetic position of Aplosporellaceae and Botryosphaeriaceae isolates obtained in this study. Combined sequences of ITS and tef1 were used for the analyses of Botryosphaeria, while ITS, tef1 and tub2 were used for the analyses of Diplodia and Dothiorella isolates. All details of the alignments are provided in Table 2.

Table 2.

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Alignment details and ML, BI analyses results of each phylogenetic tree constructed in this study.

Character		Overview phylogenetic tree	Botryosphaeria	Diplodia	Dothiorella
Number of base pairs in each gene region (including the gaps after alignment)		ITS (603 bp), tef1 (320 bp)	ITS (555 bp), tef1 (315 bp)	ITS (537 bp), tef1 (311 bp), tub2 (381 bp)	ITS (523 bp), tef 1 (294 bp), tub2 (427 bp)
Number of isolates obtained in this study		17	12	11	9
Number of taxa originated from GenBank		94	45	64	73
Outgroup taxa		Lecanosticta acicula (LNPV252)	Barriopsis iraniana (IRAN1448C) and Barriopsis iraniana (IRAN1449C)	Dothiorella dulcispinae (CMW 36460) and Dothiorella dulcispinae (CMW 36462)	Neofusicoccum luteum (CBS 562.92) and Neofusicoccum luteum (CMW 41365)
BI (model of each gene region)	ITS	GTR+I+G	SYM	K80+I+G	HKY+I+G
	tef1	GTR+I+G	HKY+G	GTR+G	GTR+G
	tub2	–	–	GTR+G	GTR+I+G

In an overview phylogenetic tree (Fig. 1), sixteen newly obtained isolates were nested with four genera of Botryosphaeriaceae, representing seven known species viz. Barriopsis tectonae, Neofusicoccum parvum, Sardiniella guizhouensis, Sphaeropsis guizhouensis and Sp. citrigena. Two isolates were clustered with the genus Aplosporella (Aplosporellaceae), and were identified as A. ginkgonis and A. prunicola.

Figure 1.

Phylogram generated from RAxML analysis based on combined ITS and tef1 sequence data of Botryosphaeriaceae and Aplosporellaceae isolates. The tree was rooted to Lecanosticta acicula (LNPV 252). The ML (≥ 75%) and BI (≥ 95%) bootstrap supports are given near the nodes, respectively. Isolates from this study are marked in red and ex-type strains are marked in bold.

Three individual phylogenetic trees were constructed for the genera Botryosphaeria, Diplodia and Dothiorella. Twelve isolates belonged to the genus Botryosphaeria and ten of them were nested with Bo. dothidea, while the remaining two isolates clustered with Bo. fabicerciana (Fig. 2). Another twelve isolates were treated in Diplodia and seven isolates were clustered with two known species of Diplodia (Di. mutila and Di. seriata, Fig. 3). The other five isolates did not cluster with any previously known Diplodia species, thus, two novel species were preliminarily identified based on phylogenetic evidence. Eight isolates were nested within Dothiorella and six isolates of them were occupied in the basal position of the Dothiorella tree and formed two well‐supported subclades, representing two new species. The other two isolates were nested within the Do. sarmentorum isolates (Fig. 4).

Figure 2.

Phylogram generated from RAxML analysis based on combined ITS and tef1 sequence data of Botryosphaeria isolates. The tree was rooted to Barriopsis iraniana (IRAN1448C and IRAN1449C). The ML (≥ 75%) and BI (≥ 95%) bootstrap supports are given near the nodes, respectively. Isolates from this study are marked in red and ex-type strains are marked in bold.

Figure 3.

Phylogram generated from RAxML analysis based on combined ITS, tef1 and tub2 sequence data of Diplodia isolates. The tree was rooted to Dothiorella dulcispinae (CMW 36460 and CMW 36462). The ML (≥ 75%) and BI (≥ 95%) bootstrap supports are given near the nodes, respectively. Isolates from this study are marked in red and ex-type strains are marked in bold.

Figure 4.

Phylogram generated from RAxML analysis based on combined ITS, tef1 and tub2 sequence data of Dothiorella isolates. The tree was rooted to Neofusicoccum luteum (CBS 562.92 and CMW 41365). The ML (≥ 75%) and BI (≥ 95%) bootstrap supports are given near the nodes, respectively. Isolates from this study are marked in red and ex-type strains are marked in bold.

Taxonomy

Aplosporella ginkgonis C.M. Tian, Z. Du & K.D. Hyde. Mycosphere 8(2): 1249 (2017).

MycoBank No: 552938

Fig. 5

Description

Saprobic on decaying branches of Zanthoxylum bungeanum. Sexual morph: Not observed. Asexual morph: Coelomycetous, Conidiomata 558–657 × 216–241 μm (x̄ = 235.5 × 228.5 μm, n = 10), immersed, partially erumpent when mature, multilocular, locules separated by pale brown cells of textura angularis. Peridium 65–106 μm wide, wall 6–10 cell-layers thick, outer layers composed of 3–4 layers of pale brown cells of textura globulosa, intermediate layers composed of dark brown cells of textura angularis, becoming pale brown towards the inner region. Ostiole 138–171 μm diam., central. Conidiophores reduced to conidiogenous cells. Conidiogenous cells 12–13 × 7.5–8 μm (x̄ = 12.5 × 8 μm, n = 20), holoblastic, hyaline, cylindrical to doliiform, smooth-walled. Conidia 17–20 × 6.5–7.5 μm (x̄ = 18.5 × 7 μm, n = 30), L/W ratio = 2.5, ellipsoidal to subcylindrical, with both ends rounded, initially hyaline, becoming dark brown, aseptate.

Figure 5.

Aplosporella ginkgonis (HUEST 22.0092, new host record) a–c appearance of conidiomata on natural substrate d vertical section of conidioma e section of peridium f conidiogenous cells and developing conidia g–k brown aseptate conidia l upper view of the colony on PDA after 14 d m reverse view of the colony on PDA after 14 d. Scale bars: 100 μm (d); 40 μm (e); 10 μm (f–k).

Culture characteristics

Colonies on PDA developing dense aerial mycelium with age, becoming white to gray-brown at the surface, and whitish to yellowish brown at the reverse, producing a brown pigment, with sinuate edges.

Material examined

China, Sichuan Province, Yaan City, Hanyuan County, 29°16'51"N, 102°37'48"E, elevation 1,689 m, on dead branches of Zanthoxylum bungeanum, 30^th October 2021, W.L. Li, HJ 511 (HUEST 22.0092), living culture UESTCC 22.0091.

Notes

Aplosporella ginkgonis was introduced by Du et al. (2017) and isolated from diseased branches of Ginkgo biloba and Morus alba from Gansu Province in China. One isolate (UESTCC 22.0091) obtained in this study from Zanthoxylum bungeanum is morphologically similar to the original description of Aplosporella ginkgonis, and the sequences data are identical to the previous data (99%–100%). We, thus, identify the new collection as Aplosporella ginkgonis and this is the first report from Zanthoxylum bungeanum.

Aplosporella prunicola Damm & Crous Fungal Diversity 27: 39 (2007).

MycoBank No: 504373

Fig. 6

Description

Saprobic on decaying branches of Zanthoxylum bungeanum. Sexual morph: Not observed. Asexual morph: Coelomycetous, Conidiomata 355–408 × 568.5–599 μm (x̄ = 381.5 × 584 μm, n = 10), immersed, partially erumpent when mature, multilocular, locules divided by pale brown cells of textura angularis. Peridium 107–122 μm wide, composed of 3–5 layers of pale brown cells of textura globulosa. Ostiole 70–88 μm diam., central. Conidiophores reduced to conidiogenous cells. Conidiogenous cells 6.5–10 × 2–3 μm (x̄ = 8 × 2.5 μm, n = 20), holoblastic, hyaline, cylindrical, smooth-walled. Conidia 20–23.5 × 12–13.5 μm (x̄ = 21.5 × 13 μm, n = 30), L/W ratio = 1.6, ellipsoidal to subcylindrical, with both ends broadly rounded, initially hyaline, becoming dark brown, aseptate, smooth.

Figure 6.

Aplosporella prunicola (HUEST 22.0091, new host record) a, b appearance of conidiomata on natural substrate c vertical section of multiloculate conidioma d, e section of peridium f–i conidiogenous cells and developing conidia j–n brown aseptate conidia. Scale bars: 100 μm (c); 20 μm (d, e); 10 μm (f–n).

Culture characteristics

Colonies on PDA after 7 d, becoming pale olivaceous-gray to olivaceous-black at the surface, and olivaceous black at the reverse, with irregular edges.

Material examined

China, Sichuan Province, Yaan City, Hanyuan County, 29°16'51"N, 102°37'48"E, elevation 1,689 m, on dead branches of Zanthoxylum bungeanum, 30^th October 2021, W.L Li, HJ 509 (HUEST 22.0091), living culture UESTCC 22.0090.

Notes

Our isolate UESTCC 22.0090 morphologically lines up with the description of Aplosporella prunicola provided by Damm et al. (2007) in having immersed to erumpent, multilocular conidiomata and brown, smooth‐walled, ovoid to oblong conidia. The strain UESTCC 22.0090 is phylogenetically and morphologically similar to A. yalgorensis and A. prunicola, however, A. yalgorensis can be distinguished from other Aplosporella species by its pitted conidial walls. Thus, the strain UESTCC 22.0090 was identified as A. prunicola based on current evidence. This is the first time A. prunicola is reported from Zanthoxylum bungeanum in China.

Diplodia acerigena L.W. Li & Jian K. Liu, sp. nov.

MycoBank No: 847163

Figs 7, 8

Etymology

The epithet ‘‘acerigena’’ refers to the host genus Acer, on which the holotype was collected.

Holotype

HKAS 125891.

Description

Saprobic on decaying branches of Acer truncatum. Sexual morph: Ascomata 304.5–321 × 217–260 (x̄ = 313 × 238.5 μm, n = 20), more or less subglobose, solitary or gregarious, semi-immersed, medium brown to dark brown, unilocular, papillate, ostiolate. Ostiole 101–115 μm diam., conical or circular, central, papillate, periphysate. Peridium 23–29 μm wide, composed of 3–5 layers of dark brown cells of textura angularis. Pseudoparaphyses 3.5–5 μm wide, hyaline, branched, septate. Asci 98–120 × 24–32.5 μm (x̄ = 109 × 28 μm, n = 30), (4–)8-spored, clavate, stipitate, irregularly bitunicate, apex rounded with an ocular chamber. Ascospores 24.5–31.5 × 13.5–16 μm (x̄ = 28 × 14.5 μm, n = 30), L/W ratio = 2, biseriate, broadly fusiform to oval, widest in the middle, both ends obtuse, hyaline, moderately thick-walled, smooth, becoming brown and 2-septate when aged. Asexual morph: Coelomycetous, pycnidia produced on mycelium in PDA. Conidiomata stromatic, mostly solitary, gray to black, globose to subglobose. Paraphyses 2–3.5 μm wide, hyaline, subcylindrical, branched, septate. Conidiophores absent. Conidiogenous cells 9–12 × 3.5–5 μm (x̄ = 10.5 × 4.5 μm, n = 20), holoblastic, hyaline, cylindrical. Conidia 21–24 × 10–11 μm (x̄ = 22.5 × 10.5 μm, n = 30), L/W ratio = 2, aseptate, thick-walled, wall externally smooth, roughened on the inner surface, initially hyaline becoming dark brown, obovoid to ellipsoid, both ends broadly rounded. Spermatogenous cells 7–9.5 × 2.5–3.5 μm (x̄ = 8 × 3 μm, n = 20), discrete or integrated, hyaline, smooth, cylindrical, holoblastic or proliferating via. determinate phialides with periclinal thickening. Spermatia 7–11.5 × 3–4 μm (x̄ = 9 × 3.5 μm, n = 30), hyaline, smooth, aseptate, rod-shaped with rounded ends.

Figure 7.

The sexual morph of Diplodia acerigena (HKAS 125891, holotype) a, b appearance of ascomata on natural substrate c vertical section of ascoma d ostiole e section of peridium f–h asci with hyaline ascospores i asci with brown 2-sepatate ascospores j, k hyaline immature aseptate ascospores l–n mature brown 2-septate ascospores o germinated ascospore p upper view of the colony on PDA after 14 d q reverse view of the colony on PDA after 14 d. Scale bars: 100 μm (c); 10 μm (d, e, j–o); 20 μm (f–i).

Culture characteristics

Ascospores germinating on PDA within 12 h. Colonies growing on PDA, reaching a diam. of 4 cm after five days at 25 °C, effuse, velvety, with entire to slightly undulate edge. Surface initially white and later turning dark olivaceous from the surrounding of the colony and dark gray in reverse.

Figure 8.

The asexual morph of Diplodia acerigena (HKAS 125891, holotype) a–d appearance of conidiomata on PDA e–i conidiogenous cells and developing conidia j, k hyaline immature conidia l–o mature brown aseptate conidia p–t Spermatogenous cells and Spermatia. Scale bars: 10 μm (e–o, t); 5 μm (p–s).

Material examined

China, Sichuan Province, Chengdu City, Pidu District, 30°19'57"N, 103°59'47"E, elevation 442 m, on dead branches of Acer truncatum (Anacardiaceae), 19^th March 2021, W.L Li, YBF 96 (HKAS 125891, holotype), ex-type living culture UESTCC 22.0073 = CGMCC 3.24157; ibid., YBF103 (HUEST 22.0075, paratype), living culture UESTCC 22.0074. Additional sequences: LSU: OQ164827 (CGMCC 3.24157), OQ164828 (UESTCC 22.0074).

Notes

Three isolates of Diplodia acerigena clustered closer to Di. pseudoseriata (CBS 124906) with high bootstrap support (ML/BI 100%/1). The asexual morph of Diplodia pseudoseriata was introduced by Pérez et al. (2010), collected and isolated from the Blepharocalyx salicifolius in Uruguay and its sexual morph has not been reported. The asexual morph of Diplodia acerigena differs from Di. pseudoseriata in having conidia which become 1-septate when aged. Diplodia acerigena shares similar sexual morph characters as of other Diplodia species by having immersed to semi-immersed pseudothecia, clavate asci, broadly fusiform to ovoid and hyaline ascospores. However, conidia of Diplodia acerigena become brown and septate when aged, which is rarely observed in any other sexual morph species of this genus.

Diplodia mutila (Fr.) Mont., Ann. Sci. nat., sér. 2, 1: 302. 1834.

MycoBank No: 201741

Fig. 9

Sphaeria mutila Fr., Syst. Mycol. (Lundae) 2: 424. 1823. Basionym.

≡ Physalospora mutila (Fr.) N.E. Stevens, Mycologia 28: 333. 1936.

= Botryosphaeria stevensii Shoemaker, Canad. J. Bot. 42: 1299. 1964.

Description

Saprobic on decaying branches of Camellia oleifera. Sexual morph: Not observed. Asexual morph: Coelomycetous, Conidiomata 330–394 × 215–230 μm (x̄ = 362 × 223 μm, n = 10), immersed, erumpent, gregarious, dark brown to black, subglobose, unilocular. Ostiole 48.5–67 μm diam., central. Peridium 29–38 μm wide, thick-walled, outer and inner layers composed of 1–2 layers dark brown textura angularis, intermediate layers composed of 3–5 layers of hyaline cells of textura angularis. Conidiophores reduced to conidiogenous cells. Conidiogenous cells 8.5–12 × 3–5 μm (x̄ = 10 × 4 μm, n = 20), cylindrical, thin-walled, hyaline, holoblastic, indeterminate, proliferating at the same level to produce periclinal thickenings, or proliferating percurrently giving rise to 2–3 indistinct annellations. Conidia 19–21 × 9.5–11 μm (x̄ = 20 × 10.5 μm, n = 30), L/W ratio = 2, oblong, with broadly rounded apex and truncate base, thick-walled, wall externally smooth, roughened on the inner surface, hyaline, aseptate, becoming dark brown when aged.

Figure 9.

Diplodia mutila (HUEST 22.0069, new host record) a, b appearance of conidiomata on natural substrate c vertical section of conidioma d ostiole e section of peridium f–i conidiogenous cells and developing conidia j hyaline immature conidium k–n mature brown conidia. Scale bars: 40 μm (c); 20 μm (d, e); 10 μm (f–n).

Culture characteristics

Colonies on PDA initially olivaceous buff in the center of the colony and white at the edge, becoming olivaceous within 7 d on the surface, with smooth edge.

Materials examined

China, Sichuan Province, Jiangyou City, Shuanghe County, 31°54'10"N, 104°55'57"E, elevation 657 m, on dead branches of Camellia oleifera, 11^th July 2021, W.L Li, 286 (HUEST 22.0069), living culture UESTCC 22.0068; ibid., 289 (HUEST 22.0068), living culture UESTCC 22.0067; ibid., Guangyuan city, Qingchuan County, 32°40'38"N, 105°28'57"E, elevation 634 m, on dead branches of Olea europaea, 20^th April 2021, W.L Li, 188 (HUEST 22.0065), living culture UESTCC 22.0064; ibid., 257 (HUEST 22.0070), living culture UESTCC 22.0069; ibid., on dead branches of Vernicia fordii, 20^th April 2021, W.L Li, 238 (HUEST 22.0066), living culture UESTCC 22.0065; ibid., Chengdu City, Pidu District, 30°49'27"N, 103°47'42"E, elevation 442 m, on dead branches of Pistacia chinensis, 5^th March 2021, W.L Li, A61 (HUEST 22.0064), living culture UESTCC 22.0063. Additional sequences: LSU: OQ164832 (UESTCC 22.0063), OQ164830 (UESTCC 22.0064), OQ164831 (UESTCC 22.0065).

Notes

The phylogenetic tree show that six strains isolated from Camellia oleifera, Olea europaea and Vernicia fordii nested with Diplodia mutila (CBS 112553) with a moderate bootstrap support (ML/BI 86%/1). Diplodia mutila, the type of the genus, is a well-known and most commonly reported species. It has been recorded mainly from woody substrates, and it is known from more than 50 hosts (Batista et al. 2021). Morphologically, one of the isolates obtained in this study UESTCC 22.0068 shares similar conidia shape and size with Di. mutila, but hardly observed the mature conidia with septa. We identify these taxa as Di. mutila based on morphology and phylogeny evidences. This is the first report of Di. mutila, isolated from Camellia oleifera, Olea europaea and Vernicia fordii.

Diplodia pistaciicola L.W. Li & Jian K. Liu, sp. nov.

MycoBank No: 847166

Fig. 10

Etymology

The epithet ‘‘pistaciicola’’ refers to the host genus Pistacia, on which the holotype was collected.

Holotype

HKAS 125890.

Description

Saprobic on decaying branches of Pistacia chinensis. Sexual morph: Not observed. Asexual morph: Coelomycetous, Conidiomata 353–441 × 274.5–316 μm (x̄ = 397 × 295 μm, n = 10), immersed, forming split-like opening on the host, solitary or gregarious, globose to subglobose, dark brown to black, unilocular, papillate, ostiolate. Ostiole 38–49.5 μm diam., conical or circular, centrically located. Peridium 42–60 μm wide, composed of thick walled, dark brown to hyaline cells of textura angularis. Conidiophores reduced to conidiogenous cells. Conidiogenous cells 10–14 × 3–4 μm (x̄ = 12 × 3.5 μm, n = 20), holoblastic, discrete, cylindrical, hyaline, smooth, indeterminate, arising from the inner cavity of the conidiomata. Conidia 24.5–27 × 11–13 μm (x̄ = 25.5 × 12 μm, n = 30), L/W ratio = 2.2, ellipsoid to obovoid, aseptate, hyaline, thick-walled, guttulate.

Figure 10.

Diplodia pistaciicola (HKAS 125890, holotype) a–c appearance of conidiomata on natural substrate d, e vertical section of conidiomata/conidioma f ostiole g section of peridium h–l conidiogenous cells and developing conidia m–q hyaline aseptate conidia. Scale bars: 200 μm (d); 50 μm (e); 20 μm (f, g); 10 μm (h–q).

Culture characteristics

Conidia germinate on PDA within 12 h. Colonies growing on PDA, reaching a diameter of 4 cm after five days at 25 °C, effuse, velvety, with entire to slightly undulate edge. The early stage of the colony is white, later turning dark olivaceous and dark gray in reverse.

Material examined

China, Sichuan Province, Chengdu City, Pidu District, 30°49'27"N, 103°47'42"E, elevation 442 m, on dead branches of Pistacia chinensis (Anacardiaceae), 5^th March 2021, W.L Li, 049 (HKAS 125890, holotype), ex-type living culture UESTCC 22.0070 = CGMCC 3.24156; ibid., 049B (HUEST 22.0072 isotype), ex-isotype living culture UESTCC 22.0071. Additional sequence: LSU: OQ164833 (CGMCC 3.24156).

Notes

Phylogenetic analyses showed that two strains of Diplodia pistaciicola isolated from Pistacia chinensis are distinct but closely related to Di. agrifolia (CBS 124.30). The comparison of ITS, tef1 and tub2 of these two species indicate 5 bp (502), 3bp (224), 9 bp (425) differances, respectively. Morphologically, Di. agrifolia differs from Di. pistaciicola in producing two to three times larger ascomata than that of Di. pistaciicola (721–836 vs. 274.5–316 μm) and possessing smaller conidia (27–36.5 × 14.5–17.8 μm vs. 24.5–27 × 11–13 μm). In addition, conidia of Di. pistaciicola are hyaline, aseptate, rarely becoming pale brown and uniseptate with age, whereas conidia of Di. agrifolia are mostly dark brown and uniseptate before discharge from pycnidia.

Diplodia seriata De Not., Micr. Ital. Dec. 4: 6. (1942).

MycoBank No: 180468

Fig. 11

Description

Saprobic on decaying branches of Camellia oleifera. Sexual morph: Ascomata 301–343 × 293–340 (x̄ = 322 × 316 μm, n = 10), more or less subglobose, solitary or gregarious, semi-immersed, medium brown to dark brown, unilocular, papillate, ostiolate. Ostiole 72–78 μm diam., conical or circular, central, papillate, periphysate. Peridium 33–44 μm wide, composed of dark brown, 4–6 layers of textura angularis. Pseudoparaphyses 2–2.5 μm wide, hyaline, branched, septate. Asci 112–141 × 27.5–30 μm (x̄ = 126 × 28.5 μm, n = 30), clavate, stipitate, bitunicate, containing eight, biseriate ascospores. Ascospores 31.5–32.5 × 12–13.5 μm (x̄ = 32 × 13 μm, n = 30), L/W ratio = 2.5, broadly fusiform to oval, widest in the middle, both ends obtuse, hyaline, moderately thick-walled, smooth, becoming brown when aged. Asexual morph: Not observed.

Figure 11.

Diplodia seriata (HUEST 22.0073, new host record) a–c appearance of ascomata on natural substrate d vertical section of ascomata e section of peridium f–i asci j–n ascospores. Scale bars: 100 μm (d); 10 μm (e, j–n); 20 μm (f–i).

Culture characteristics

Ascospores germinate on PDA within 12 h. Colonies growing on PDA, reaching a diameter of 4 cm after five days at 25 °C, effuse, velvety, with entire to slightly undulate edge.

Material examined

China, Sichuan Province, Jiangyou City, shuanghe County, 31°54'10"N, 104°55'57"E, elevation 656 m, on dead branches of Camellia oleifera, 10^th June 2021, W.L Li, 288 (HUEST 22.0073), living culture UESTCC 22.0072.

Notes

The morphology of the taxa isolated from decaying woody oil plants is similar to Diplodia seriata. In the multi-gene phylogenetic analysis, our new collection clustered with the ex-type strain of Di. seriata (CBS 112555) with strong bootstrap support. Diplodia seriata has been isolated from a wide range of hosts (121 species) and has a worldwide distribution (reported in 46 countries) (Batista et al. 2021). This is the first report of Di. seriata isolated from Camellia oleifera.

Dothiorella camelliae L.W. Li & Jian K. Liu, sp. nov.

MycoBank No: 847167

Fig. 12

Etymology

The epithet ‘‘camelliae’’ refers to the host genus Camellia, on which the holotype was collected.

Holotype

HKAS 125892.

Description

Saprobic on decaying branches of Camellia oleifera. Sexual morph: Ascomata 199–222 × 237–269 μm (x̄ = 210.5 × 253 μm, n = 10), submerged in the substrate, partly erumpent at maturity, solitary or gregarious, dark brown to black, subglobose, multilocular or unilocular. Ostiole 17–37 μm diam., central. Peridium 35–43 μm wide, thick-walled, outer layers composed of 1–2 layers dark brown cells of textura angularis, becoming hyaline towards the inner region. Pseudoparaphyses 3–4 μm wide, hyaline, frequently aseptate. Asci 80–96 × 22–25 μm (x̄ = 88 × 23.5 μm, n = 30), stipitate, clavate, thick-walled, bitunicate, (6–)8-spored, irregularly biseriate. Ascospores 21–25 × 9.5–12 μm (x̄ = 23 × 10.5 μm, n = 30), L/W ratio = 2, oblong, ovate to sub-clavate, (0–)1-septate, slightly constricted at the septum, hyaline to dark brown, moderately thick-walled, straight or inequilateral, basal cell tapering towards the acute end. Asexual morph: Not observed.

Figure 12.

Dothiorella camelliae (HKAS 125892, holotype) a, b appearance of ascomata on natural substrate c vertical section of ascoma d section of peridium e–g asci h–m ascospores n germinated ascospore. Scale bars: 50 μm (c); 10 μm (d, h–n); 20 μm (e–g).

Culture characteristics

Ascospores germinate on PDA within 12 h. Colonies growing on PDA, reaching a diameter of 4 cm after five days at 25 °C, effuse, velvety, with entire to slightly undulate edge. Surface initially white and later turning dark olivaceous from the surrounding of the colony and dark gray in reverse.

Materials examined

China, Sichuan Province, Leshan City, Wutongqiao District, 29°22'28"N, 103°45'49"E, elevation 383 m, on dead branches of Camellia oleifera (Theaceae), 23^th July 2021, Z.P Liu, 351 (HKAS 125892, holotype), ex-type living culture UESTCC 22.0081 = CGMCC 3.24158; ibid., 347 (HUEST 22.0081), living culture UESTCC 22.0080; ibid., Shizhong District, 29°42'13"N, 103°52'25"E, elevation 356 m, on dead branches of Paeonia suffruticosa, 23^th July 2021, W.L Li, A240 (HUEST 22.0080), living culture UESTCC 22.0079; ibid., A234 (HUEST 22.0079), living culture UESTCC 22.0078. Additional sequences: LSU: OQ164834 (CGMCC 3.24158), OQ164835 (UESTCC 22.0079), OQ164836 (UESTCC 22.0078).

Notes

Four strains isolated from Vernicia fordii and Camellia oleifera occupy a basal position in the Dothiorella phylogenetic tree by forming a well‐supported subclade sister to Do. zanthoxyli (ML/BI 97%/1, Fig. 4). The BLASTn searches of the ITS sequence of Dothiorella zanthoxyli resulted in 97% matches with Neofusicoccum vitifusiforme BRIP64010, the tef1 showed 91.23% matches with Do. symphoricarposicola BL158, and the tub2 BLASTn results indicated 96.53% similarity with Do. uruguayensis CBS 124908 and Do. viticola B116-3. Dothiorella camelliae can be distinguished from Do. zanthoxyli in the size of ascomata, ascus and L/W ratio of ascospores (Table 3). Dothiorella camelliae resembles the sexual morph of Do. sarmentorum in producing immersed to sub-immersed ascomata, clavate asci and ovate to sub-clavate, hyaline to brown conidia with (0–)1-septate. However, Do. sarmentorum morphologically can be distinguished from Do. camelliae in having larger ascomata (350–400 μm vs. 237–269 μm), thicker peridium (50–75 μm vs. 35–43 μm), and longer asci (140–210 μm vs. 80–96 μm) (Table 3). Phylogenetically, these two species reside in two distinct clades.

Table 3.

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A morphological comparison of the sexual morph of three Dothiorella species.

Taxa	Ascomata (μm)	Asci (μm)	Peridium (μm)	Ascospores
Taxa	Ascomata (μm)	Asci (μm)	Peridium (μm)	Size(μm)	Color	L/W ratio
Dothiorella camelliae	199–222 × 237–269	80–96 × 22–25	35–43	21–25 × 9.5–12	Hyaline to dark brown	2
Dothiorella sarmentorum	350–400	140–210 × 17–24	50–75	24.5–25.5 × 11.5–12.5	Dark brown	2.4
Dothiorella zanthoxyli	258–280 × 170–174	63.5–77 × 20–24.5	35–40	22.5–25 × 9.5–11	Hyaline to dark brown	2.6

Dothiorella sarmentorum (Fr.) A.J.L. Phillips, J. Luque & A. Alves, Mycologia 97: 522. (2005).

MycoBank No: 501403

Fig. 13

Sphaeria sarmentorum Fr., K. svenska Vetensk-Acad. Handl. 39: 107. 1818. Basionym.

≡ Diplodia sarmentorum (Fr.) Fr., Summ. veg. Scand. (Stockholm) 2: 417. 1849.

= Diplodia pruni Fuckel, Jahrb. Nassauischen Vereins Naturk., 23–24: 169. 1870 [1869].

= Botryosphaeria sarmentorum A.J.L. Phillips, J. Luque & A. Alves, Mycologia 97: 522. 2005.

Description

Saprobic on decaying branches of Pistacia chinensis. Sexual morph: Not observed. Asexual morph: Conidiomata 278–338 × 240–280 μm (x̄ = 308 × 260 μm, n = 10), immersed, erumpent, forming split-like opening on the host, gregarious, globose to subglobose, dark brown to black, unilocular or multilocular, papillate, ostiolate. Ostiole 52–57 μm diam., conical or circular, centrically located. Peridium 28.5–44 μm, comprising 5–8 layers of thick‐walled, dark brown to hyaline cells arranged in a textura angularis. Conidiophores reduced to conidiogenous cells. Conidiogenous cells 2.5–3.5 × 6–9 μm (x̄ = 3 × 7.5 μm, n = 20), holoblastic, discrete, cylindrical, hyaline, smooth, indeterminate, proliferating at the same level giving rise to periclinal thickenings, or rarely proliferating percurrently to form one or two close, indistinct annellations. Conidia 21.5–24 × 9–10 μm (x̄ = 22.5 × 9.5 μm, n = 30), L/W ratio = 2.4, ellipsoid to obovoid, with rounded ends, initially hyaline and aseptate becoming pigmented brown and 1-septate often while still attached to conidiogenous cell, brown walled, slightly constricted at the septum.

Figure 13.

Dothiorella sarmentorum (HUEST 22.0077, new host record) a, b appearance of conidiomata on natural substrate c vertical section of conidioma d–g conidiogenous cells and developing conidia h–l brown conidia. Scale bars: 50 μm (c); 10 μm (d–l).

Culture characteristics

Conidia germinate on PDA within 12 h. Colonies growing on PDA, reaching a diameter of 4 cm after three days at 25 °C, effuse, velvety, with entire to slightly undulate edge. Surface initially white and later turning dark olivaceous from the surrounding of the colony and dark gray in reverse.

Materials examined

China, Sichuan Province, Chengdu City, Pidu District, 30°19'57"N, 103°59'47"E, elevation 442 m, on dead branches of Pistacia chinensis, 19^th March 2021, W.L Li, 072 (HUEST 22.0077), living culture UESTCC 22.0076; ibid., Guangyuan City, Qingchuan County, 32°40'38"N, 105°28'57"E, elevation 638 m, 20^th April 2021, W.L Li, A189 (HUEST 22.0078), living culture UESTCC 22.0077. Additional sequences: LSU: OQ164837 (UESTCC 22.0076), OQ164838 (UESTCC 22.0077).

Notes

Dothiorella sarmentorum was introduced by Phillips et al (2005) with both asexual and sexual morphs. Recently, nine Dothiorella species (Do. californica, Do. iberica, Do. italica, Do. guttulata, Do. omnivora, Do. parva, Do. sempervirentis, Do. symphoricarpicola, Do. vidmadera) were synonymized under Do. Sarmentorum by Zhang et al. (2021) based on phylogenetic analyses. Two isolates obtained in the present study clustered with the group of Do. sarmentorum taxa in the phylogenetic analyses (Fig. 4).

Dothiorella zanthoxyli L.W. Li & Jian K. Liu, sp. nov.

MycoBank No: 847168

Fig. 14

Etymology

The epithet ‘‘zanthoxyli’’ refers to the host genus Zanthoxylum, on which the holotype was collected.

Holotype

HKAS 125893.

Description

Saprobic on decaying branches of Zanthoxylum bungeanum. Sexual morph: Ascomata 258–280 × 170–174 μm (x̄ = 269 × 172 μm, n = 10), submerged in the substrate, partly erumpent at maturity, solitary or gregarious, dark brown to black, subglobose, unilocular. Ostiole 42–44 μm diam., central. Peridium 35–40 μm wide, thick-walled, outer layers composed of 3–5 layers dark brown cells of textura angularis, becoming hyaline towards the inner region. Pseudoparaphyses 3–4.5 μm wide, hyaline, frequently aseptate. Asci 63.5–77 × 20–24.5 μm (x̄ = 70 × 22.5 μm, n = 30), short stipe, clavate, thick-walled, bitunicate, 8-spored, irregularly biseriate. Ascospores 22.5–25 × 9.5–11 μm (x̄ = 24 × 10 μm, n = 30), L/W ratio = 2.6, oblong, ovate to sub-clavate, (0–)1-septate, slightly constricted at the septum, hyaline to dark brown, moderately thick-walled, straight or inequilateral, basal cell tapering towards the acute end. Asexual morph: Not observed.

Figure 14.

Dothiorella zanthoxyli (HKAS 125893, holotype) a, b appearance of ascomata on natural substrate c vertical section of ascoma d ostiole e section of peridium f, k–n asci g–j brown ascospores. Scale bars: 50 μm (c); 20 μm (d–f); 10 μm (g–n).

Culture characteristics

Materials examined

China, Sichuan Province, Yanan City, Hanyuan County, 29°16'51"N, 102°37'48"E, elevation 1,689 m, on dead branches of Zanthoxylum bungeanum (Rutaceae), 30^th October 2021, W.L Li, 504 (HKAS 125893, holotype), ex-type living culture UESTCC 22.0082 = CGMCC 3.24159; ibid., 506 (HUEST 22.0084), living culture UESTCC 22.0083; ibid., 507 (HUEST 22.0085), living culture UESTCC 22.0084. Additional sequences: LSU: OQ164839 (CGMCC 3.24159), OQ164840 (UESTCC 22.0083), OQ164841 (UESTCC 22.0084).

Notes

Three strains of Dothiorella zanthoxyli isolated from Zanthoxylum bungeanum correspond well with sexual morph of Dothiorella described by Phillips et al. (2013), but morphologically differ from other species (Do. camelliae, Do. iberica and Do. sarmentorum) in the size of ascomata and asci (Table 3). A comparison of ITS and tef1 nucleotides shows that Do. zanthoxyli is significantly different from its sister species, Do. camelliae by 4/550 bp (0.72%) in ITS and 14/242 bp (5.8%) in tef1. In the phylogenetic analysis, these two species formed two distinct clades in Dothiorella (Fig. 4).

Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips, Stud. Mycol. 55: 248. (2006).

MycoBank No: 500879

Fig. 15

Fusicoccum parvum Pennycook & Samuels, Mycotaxon 24: 455. 1985. Basionym.

= Botryosphaeria parva Pennycook & Samuels, Mycotaxon 24: 455. 1985.

Description

Saprobic on decaying branches of Idesia polycarpa. Sexual morph: Ascomata 284–321 × 129–223 μm (x̄ = 302.5 × 176 μm, n = 10), pseudothecial, forming a botryose aggregation of up to 30, solitary or gregarious, stromatic, immersed, partially erumpent when mature, dark brown to black, more or less circular, multiloculate, individual locules 143.5–161 μm diam, thick-walled. Peridium 59–78 μm diam., composed of several layers of thick-walled, pale brown cells of textura angularis. Ostiole 43.5–58 μm wide, circular, central, papillate. Asci 95–99 × 20–21.5 μm (x̄ = 97 × 20.5 μm, n = 30), (6–)8‐spored, bitunicate, fissitunicate, cylindrical to clavate, apex rounded with an ocular chamber, sometimes short pedicellate. Ascospores 18.5–23 × 7–10.5 μm (x̄ = 20.5 × 9 μm, n = 30), L/W ratio = 3, fusoid to ovoid, with tapered ends and appearing spindle-shape, hyaline, aseptate, externally smooth, internally finely verruculose, biseriate in ascus. Asexual morph: Not observed.

Figure 15.

Neofusicoccum parvum (HUEST 22.0097, new host record) a, b appearance of ascomata on natural substrate c, d vertical section of ascomata e ostiole f section of peridium g–j ascospores k, l immature asci m, n mature asci. Scale bars: 100 μm (d); 25 μm (e, f); 10 μm (g–j); 20 μm (k–n).

Culture characteristics

Ascospores germinate on PDA within 12 h. Colonies growing on PDA, reaching a diam., of 7 cm after five days at 25 °C, effuse, velvety, with entire to slightly undulate edge. Surface initially white and later turning dark olivaceous from the surrounding of the colony and dark gray in reverse.

Materials examined

China, Sichuan Province, Leshan City, Jingyan County, 29°30'27"N, 103°57'14"E, elevation 682 m, on dead branches of Idesia polycarpa, 23^th July 2021, W.L Li, STZ 327 (HUEST 22.0095), living culture UESTCC 22.0094; ibid., STZ 359 (HUEST 22.0094), living culture UESTCC 22.0093; ibid., Leshan City, Shizhong Distinct, 29°42'13"N, 103°52'25"E, elevation 356 m, on dead branches of Paeonia suffruticosa, 23^th July 2021, W.L Li, YMD 366 (HUEST 22.0096), living culture UESTCC 22.0095; ibid., Guangyuan City, Qingchuan County, 32°40'38"N, 105°28'57"E, elevation 638 m, on dead branches of Vernicia fordii, 20^th April 2021, W.L. Li, YT 175 (HUEST 22.0097), living culture UESTCC 22.0096.

Notes

The morphology of our collections obtained from decaying woody oil plants are similar to the original description of Neofusicoccum parvum (Crous et al. 2006). In the multi-gene phylogenetic analysis, these four isolates clustered together (ML/BI 75%/0.99) with the ex-type of N. parvum. Neofusicoccum parvum has a wide range of hosts and has a worldwide distribution (Phillips et al. 2013). This is the first report of N. parvum on Idesia polycarpa.

Sardiniella guizhouensis Y.Y. Chen & Jian K. Liu. Phytotaxa 508 (2): 190. (2021).

MycoBank No: 558352

Fig. 16

Description

Saprobic on decaying branches of Pistacia chinensis. Sexual morph: Not observed. Asexual morph: Conidiomata 223–232 × 150–176 μm (x̄ = 227.5 × 163 μm, n = 10), dark brown to black, globose, submerged in the substrate, partially erumpent at maturity, ostiolate. Ostiole 28.5–45 μm diam., circular, central. Peridium 21–30 μm thick, composed of dark brown thick-walled cells of textura angularis, becoming thin-walled and hyaline towards the inner region. Conidiophores reduced to conidiogenous cells. Conidiogenous cells 6–9.5 × 3.5–5 μm (x̄ = 7.5 × 4 μm, n = 20), hyaline, short obpyriform to subcylindrical, holoblastic, indeterminate. Conidia 20.5–24 × 11.5–14 μm (x̄ = 22 × 13 μm, n = 30), L/W ratio = 1.6, ellipsoid to ovoid with both ends rounded, hyaline, aseptate, externally smooth, internally finely verruculose.

Figure 16.

Sardiniella guizhouensis (HUEST 22.0100, new host record) a–c appearance of conidiomata on natural substrate d vertical section of conidioma e section of peridium f ostiole g–j conidiogenous cells and developing conidia k–n conidia. Scale bars: 40 μm (d); 20 μm (e, f); 10 μm (g–n).

Culture characteristics

Conidia germinate on PDA within 12 h. Colonies growing on PDA, reaching a diameter of 7 cm after five days at 25 °C, effuse, velvety, with entire to slightly undulate edge. Surface initially white and later turning dark olivaceous from the surrounding of the colony and dark gray in reverse.

Material examined

China, Sichuan Province, Chengdu City, Pidu District, 29°16'50.70"N, 102°37'47.53"E, elevation 442 m, on dead branches of Pistacia chinensis, 19^th March 2021, W.L Li, 047 (HUEST 22.0101), living culture UESTCC 22.0100; ibid., 070 (HUEST 22.0102), living culture UESTCC 22.0101; ibid., 071 (HUEST 22.0100), living culture UESTCC 22.0099; ibid., 150 (HUEST 22.0098), living culture UESTCC 22.0097; ibid., 151 (HUEST 22.0099), living culture UESTCC 22.0098; ibid., A39 (HUEST 22.0103), living culture UESTCC 22.0102; ibid., A40 (HUEST 22.0104), living culture UESTCC 22.0103. Additional sequences: LSU: OQ164842 (UESTCC 22.0100), OQ164843 (UESTCC 22.0101), OQ164844 (UESTCC 22.0099), OQ164845 (UESTCC 22.0097), OQ164846 (UESTCC 22.0098), OQ164847 (UESTCC 22.0102).

Notes

Seven isolates of our collection are morphologically similar to the original description of Sardiniella guizhouensis (Chen et al. 2021). The multi-gene phylogenetic analysis showed that the newly obtained isolates clustered together with ex-type of Sa. guizhouensis (CGMCC 3.19222) and this is the first report of Sa. guizhouensis from Pistacia chinensis.

Sphaeropsis citrigena (A.J.L. Phillips, P.R. Johnst. & Pennycook) A.J.L. Phillips & A. Alves. Stud. Mycol. 76, 157. (2013).

MycoBank No: 805463

Fig. 17

Description

Saprobic on decaying branches of Camellia oleifera. Sexual morph: Ascomata 219–252 × 216–241 μm (x̄ = 235.5 × 228.5 μm, n = 10), brown to black, solitary or aggregated, immersed, becoming erumpent, ostiolate. Ostiole 71–92 μm, central, relatively broad. Peridium 37.5–45 μm diam., composed of several layers of dark brown cells of textura angularis. Pseudoparaphyses 1.5–2 μm wide, hyaline, smooth, septate. Asci 93.5–107 × 28.5–33 μm (x̄ = 100 × 30.5 μm, n = 30), bitunicate, 8-spored, stipitate, thick-walled, with well-developed apical chamber. Ascospores 29–35 × 13–15 μm (x̄ = 32 × 14 μm, n = 30), L/W ratio = 2.3, yellowish brown to dark brown, ellipsoid to ovoid with both ends rounded, with an apiculus at either end, aseptate, externally smooth, internally finely verruculose, widest in middle to upper third. Asexual morph: Not observed.

Figure 17.

Sphaeropsis citrigena (HUEST 22.0107, new host record) a, b appearance of ascomata on natural substrate c vertical section of ascomata d section of peridium e–h mature asci i–m dark brown ascospores. Scale bars: 100 μm (c); 20 μm (d–h); 10 μm (i–m).

Culture characteristics

Ascospores germinate on PDA within 12 h. Colonies growing on PDA, reaching a diam. of 7 cm after five days at 25 °C, effuse, velvety, with entire to slightly undulate edge. Surface initially white and later turning dark olivaceous from the surrounding of the colony and dark gray in reverse.

Materials examined

China, Sichuan Province, Chengdu City, Pidu District, 31°54'10"N, 104°55'57"E, 656 m, on dead branches of Camellia oleifera, 10^th June 2021, W.L Li, 285 (HUEST 22.0107), living culture UESTCC 22.0106; ibid., on dead branches of Acer truncatum, 30°19'57"N, 103°59'47"E, elevation 442 m, 19^th March 2021, W.L Li, A33 (HUEST 22.0106), living culture UESTCC 22.0105. Additional sequence: LSU: OQ164848 (UESTCC 22.0105).

Notes

The phylogenetic tree shows that two isolates of Sphaeropsis from our collection clustered together with the ex-type strain of Sp. citrigena (ICMP 16812) with high bootstrap support (ML/BI 100%/1). Sphaeropsis citrigena was first described as Phaeobotryosphaeria citrigena by Phillips et al. (2008), later transferred to Sphaeropsis based on morphological and phylogenetic analyses (Phillips et al. 2013). The new collection (UESTCC 22.0105) isolated from Camellia oleifera resembles Sp. citrigena isolated from Citrus sinensis (Phillips et al. 2013) in the shape of asci and ascospores, though their asci are somewhat smaller than those of Sp. citrigena (93.5–107 × 28.5–33 μm vs. 180–230 × 35–43 μm). In addition, there are no base pair differences in ITS and tef1 sequences of these two strains. We, thus, identify the new collection as Sp. citrigena and this is the first record of Sp. citrigena from Camellia oleifera.

Sphaeropsis guizhouensis Y.Y. Chen, A. J. Dissanayake & Jian K. Liu., J. Fungi 7, 893. (2021).

MycoBank No: 558475

Fig. 18

Description

Saprobic on decayed branched of Camellia oleifera. Sexual morph: Ascostromata 166–198 × 146.5–175 μm (x̄ = 182 × 160.5 μm, n = 20), initially immersed under host epidermis, becoming semi‐immersed to erumpent, solitary or gregarious, uniloculate, black, globose to subglobose, membraneous, ostiolate. Ostiole 75–80 μm wide, central, papillate, pale brown, relatively broad, periphysate. Peridium 23–27 μm wide, comprising 3–5 layers of relatively thick‐walled, dark brown to black‐walled cells arranged in a textura angularis. Pseudoparaphyses 2–2.5 μm diam., hyphae‐like, numerous, embedded in a gelatinous matrix. Asci 87.5–135 × 28.5–35 μm (x̄ = 111 × 32 μm, n = 20), 8‐spored, bitunicate, fissitunicate, cylindrical to clavate, sometimes short pedicellate, mostly long pedicellate, apex rounded with an ocular chamber. Ascospores 28.5–33 × 13–15 μm (x̄ = 30.5 × 14 μm, n = 20), overlapping uniseriate to biseriate, ellipsoidal to obovoid, pale brown to dark brown, septate, slightly wide at the center, minutely guttulate, smooth‐walled. Asexual morph: Not observed.

Figure 18.

Sphaeropsis guizhouensis (HUEST 22.0105, new host record) a, b appearance of ascomata on natural substrate c vertical section of ascoma d section of peridium e–h mature asci i–l brown ascospores. Scale bars: 20 μm (c–h); 5 μm (i–l).

Culture characteristics

Ascopores germinate on PDA within 12 h. Colonies growing on PDA, reaching a diam. of 7 cm after five days at 25 °C, effuse, velvety, with entire to slightly undulate edge. Surface initially white and later turning dark olivaceous from the surrounding of the colony and dark gray in reverse.

Material examined

China, Sichuan Province, Chengdu City, Pidu District, on dead branches of Pistacia chinensis, 30°19'57"N, 103°59'47"E, elevation 442 m, 24^th March 2021, W.L Li, 290 (HUEST 22.0105), living culture UESTCC 22.0104.

Notes

Sphaeropsis guizhouensis was introduced by Dissanayake et al. (2021) and isolated from an unknown host. One isolate obtained in the present study clustered with the ex-type isolate of Sp. guizhouensis (CGMCC 3.20352) in the phylogenetic analyses of combined ITS and tef1 sequence data with high bootstrap support. A comparison of ITS and tef1 shows that there are no base pair differences between the isolates of UESTCC 22.0104 and CGMCC 3.20352. The new collection is morphologically similar to Sp. guizhouensis, with immersed to erumpent, black ascostromata and biseriate, aseptate, ellipsoid to obovoid, thick‐walled conidia. In addition, ascospores become brown and septate when aged. Considering similar morphology and strong molecular evidence, we identify UESTCC 22.0104 as Sp. guizhouensis and this is the first record of Sp. guizhouensis on Camellia oleifera.

Diversity of Botryosphaerialean fungi collected in this study

Based on the phylogenetic and morphological analyses, 50 Botryosphaeriales isolates collected from the five regions (Chengdu, Guangyuan, Leshan, Mianyang and Yaan City) in Sichuan Province were identified as 16 species. Of these, Botryosphaeria dothidea was the most prevalent species (20%), followed by Sphaeropsis guizhouensis (14%) and Diplodia mutila (12%) (Fig. 19a). Aplosporella ginkgonis, Barriopsis tectonae and Sphaeropsis guizhouensis were identified only once. There are 14 isolates (28%) isolated from Pistacia chinensis, including Di. acerigena, Di. mutila, Di. pistaciicola, Dothiorella sarmentorum and Sardiniella guizhouensis. Ten isolates were from Camellia oleifera (20%), including Bo. dothidea, Bo. fabicerciana, Di. mutila, Do. camelliae, Sp. citrigena and Sp. guizhouensis. Nine isolates were from Olea europaea (18%), including Ba. tectonae, Bo. dothidea, Bo. fabicerciana, Di. mutila and Do. sarmentorum. Relatively few strains were found on Idesia polycarpa, Paeonia suffruticosa and Vernicia fordii, as each host presents two species, respectively. As of final conclusion, Bo. dothidea were isolated from five hosts, Di. mutila were isolated from four hosts, N. parvum were isolated from three hosts, Bo. fabicerciana, Di. acericola, Do. camelliae, Do. sarmentorum and Sp. citrigena were isolated from two hosts, but several fungal isolates were only isolated from one host species, such as A. prunicola, Sa. guizhouensis and Sp. guizhouensis (Fig. 19b).

Figure 19.

Botryosphaeriales species composition a the proportion of each species to the total number of isolates b the number of Botryosphaeriales fungi on each host and host distribution of species.

Discussion

In this study, 48 Botryosphaeriaceae isolates and two Aplosporellaceae isolates were obtained from woody oil plants in Sichuan Province, China, and they were identified as 16 species based on morphological characters and multi-gene phylogenetic analyses. These species included Aplosporella prunicola, A. ginkgonis, Barriopsis tectonae, Botryosphaeria dothidea, Bo. fabicerciana, Diplodia acerigena, Di. mutila, Di. pistaciicola, Di. seriata, Dothiorella camelliae, Do. sarmentorum, Do. zanthoxyli, Neofusicoccum parvum, Sardiniella guizhouensis, Sphaeropsis citrigena and Sp. guizhouensis. Of these, Di. acerigena, Di. pistaciicola, Do. camelliae and Do. zanthoxyli are introduced as novel species. Descriptions, illustrations and notes were provided for 13 species, and only sequences data were provided for the remaining three species viz. Barriopsis tectonae, Botryosphaeria dothidea and Bo. fabicerciana due to low specimen quality.

According to previous studies, Barriopsis tectonae, Sardiniella guizhouensis, Sphaeropsis citrigena and Sp. guizhouensis have limited geographical distribution. So far, Barriopsis tectonae has been reported from China, Thailand and South Africa (Doilom et al. 2014; Dissanayake et al. 2021). Sardiniella guizhouensis and Sphaeropsis guizhouensis were only found in China while Sp. citrigena was isolated from China, Colombia and New Zealand. It’s worth noting that most of the species obtained from this study were also reported previously from Guizhou province (Dissanayake et al. 2021). Earlier studies have shown that the distribution of Botryosphaeriaceae species is influenced by the climate condition (Úrbez-Torres et al. 2006; Pitt et al. 2010; Li et al. 2020; Vivas et al. 2021). Thus, we speculate that the adjacent geographical location and similar climatic conditions may be important reasons for the similarity of fungal species isolated from the Sichuan and Guizhou provinces.

The remaining Botryosphaeriaceae species identified in this study are all well-known and reported from various geographic regions. Botryosphaeria dothidea, Di. seriata and Ne. parvum are recognized to be globally distributed while Di. mutila and Do. sarmentorum are founded only in the temperate and Mediterranean areas. In addition, these species have a broad host range. Batista et al. (2021) reported Neofusicoccum parvum from 223 hosts, B. dothidea from 403 hosts and Di. seriata from 121 hosts. Diplodia mutila and Di. seriata have previously been reported on Olea europaea in Uruguay (Hernández-Rodríguez et al. 2022). Botryosphaeria dothidea was recently isolated from diseased Camellia oleifera in China (Hao et al. 2022). In this study, Bo. dothidea, Di. mutila and Ne. parvum occurred on most of the woody oil plants species we examined. However, some common genera e. g. Lasiodiplodia, Neodeightonia and Phaeobotryon have never been collected from this group of hosts (Fig. 19). The absence of these genera from there is likely a sampling effect.

Aplosporella (Aplosporellaceae) was introduced by Spegazzini (1880) with A. chlorostroma as the genetic type. In a previous study, Aplosporella represents anamorph lineage within the Botryosphaeriaceae. Slippers et al. (2013) later proposed the family Aplosporellaceae to accommodate Aplosporella and Bagnisiella. Aplosporella species are infrequently isolated in China. Aplosporella ginkgonis, isolated from Gansu Province, was first descripted by Du et al. (2017) while Aplosporella macropycnidia was reported in Yunnan Province. Subsequently, Jiang et al. (2021) isolated a new collection of A. prunicola. However, other species have not been recorded in China. Our study revealed new host records of A. ginkgonis and A. prunicola. Though the phylogenetic analyses indicated that A. yalgorensis and A. prunicola have a low genetic divergence (Taylor et al. 2008, in this study), A. yalgorensis is still considered as a different species as it differs from other Aplosporella species (including A. prunicola) by its pitted conidial walls.

Though there are more than 1,000 Diplodia epithets listed in Index fungorum (www. Index Fungorum. Accessed in November 2022), presently only 30 species are accepted in this genus based on phylogenetic analyses (Slippers et al. 2017; Wu et al. 2021). Holomorphic species in Diplodia are Di. tsugae, Di. seriata, Di. mutila and Di. sapinea. This study revealed two previously known Diplodia species, Di. mutila and Di. seriata, and two new species, Di. acerigena and Di. pistaciicola. Among them, Di. acerigena is a holomorphic species, as its sexual stage was observed on the dead branches of Acer truncatum, and the asexual stage produced on culture (PDA). However, the sexual morph of Di. mutila and Di. pistaciicola, as well as the asexual morph of Di. seriata have not been observed on woody oil plants.

Dothiorella was established by Saccardo with Do. pyrenophora as the type species (Saccardo 1880). Recently, Dothiorella encountered a series of revisions as many species in this genus have been reduced to synonymy, such as Do. americana, Do. eriobotryae and Do. iberica (Dissanayake et al. 2021; Zhang et al. 2021). So far, 31 species are valid in Dothiorella. Most of the species were reported as the asexual morph of Dothiorella and the sexual stage is rarely founded on nature (Dissanayake et al. 2016). Phillips et al. (2013) initiated a link of asexual-sexual morph for Do. sarmentorum, Do. iberica and Do. vidmadera. However, the latter two species were synonymized under Do. sarmentorum (Zhang et al. 2021). In this study, two new species Do. camelliae and Do. zanthoxyli are introduced based on their sexual morphs as well as strong molecular evidences. Besides, new collections of Do. sarmentorum is reported on Pistacia chinensis for the first time.

Multiple molecular systematic studies, mainly of pathogenic fungi of woody plants (Phillips et al. 2013; Slippers et al. 2013; Dissanayake et al. 2021; Zhang et al. 2021), have generated a robust phylogeny for Botryosphaeriaceae. However, the classification and identification of some species in this family remains a major challenge, due to the reasons 1) With the increase of the number of Botryosphaeriaceae species, morphological feature of inter-genera and inter-species is vague, 2) Some species occurred as asexual morph on nature and it is difficult to establish the link of asexual and sexual morph, 3) In general, Botryosphaeriaceae species do not show an obvious host specialization, while some populations displayed a certain degree of host association. Thus, the traditional host-based classification system made taxonomic position confusion of some species. Therefore, collection of more fresh specimens is very important for better understanding the life cycle of Botryosphaeriaceae species, their host range (e. g. native plants) and potential pathogenicity.

Acknowledgements

Wen-Li Li thanks Tian Zhang for her help with sample collections and Yan-Peng Chen for the fungal diversity analysis.

This study was supported by the Joint Fund of the National Natural Science Foundation of China and the Karst Science Research Center of Guizhou province (Grant No. U1812401).

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Supplementary material

Supplementary material 1

Taxa and GenBank accession numbers of sequences used in this study

Wen-Li Li, Rui-Ru Liang, Asha Dissanayake, Jian-Kui Liu

Data type: table (excel document)

This dataset is made available under the Open Database License (http://opendatacommons.org/licenses/odbl/1.0/). The Open Database License (ODbL) is a license agreement intended to allow users to freely share, modify, and use this Dataset while maintaining this same freedom for others, provided that the original source and author(s) are credited.

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﻿Abstract

Keywords

﻿Introduction

﻿Materials and methods

﻿Isolates and morphology

﻿DNA extraction, PCR amplification and sequencing

﻿Phylogenetic analyses

﻿Results

﻿Phylogenetic analyses

﻿Taxonomy

﻿ Aplosporella ginkgonis C.M. Tian, Z. Du & K.D. Hyde. Mycosphere 8(2): 1249 (2017).

Description

Culture characteristics

Material examined

Notes

﻿ Aplosporella prunicola Damm & Crous Fungal Diversity 27: 39 (2007).

Description

Culture characteristics

Material examined

Notes

﻿ Diplodia acerigena L.W. Li & Jian K. Liu, sp. nov.

Etymology

Holotype

Description

Culture characteristics

Material examined

Notes

﻿ Diplodia mutila (Fr.) Mont., Ann. Sci. nat., sér. 2, 1: 302. 1834.

Description

Culture characteristics

Materials examined

Notes

﻿ Diplodia pistaciicola L.W. Li & Jian K. Liu, sp. nov.

Etymology

Holotype

Description

Culture characteristics

Material examined

Notes

﻿ Diplodia seriata De Not., Micr. Ital. Dec. 4: 6. (1942).

Description

Culture characteristics

Material examined

Notes

﻿ Dothiorella camelliae L.W. Li & Jian K. Liu, sp. nov.

Etymology

Holotype

Description

Culture characteristics

Materials examined

Notes

﻿ Dothiorella sarmentorum (Fr.) A.J.L. Phillips, J. Luque & A. Alves, Mycologia 97: 522. (2005).

Description

Culture characteristics

Materials examined

Notes

﻿ Dothiorella zanthoxyli L.W. Li & Jian K. Liu, sp. nov.

Etymology

Holotype

Description

Culture characteristics

Materials examined

Notes

﻿ Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips, Stud. Mycol. 55: 248. (2006).

Description

Culture characteristics

Materials examined

Notes

﻿ Sardiniella guizhouensis Y.Y. Chen & Jian K. Liu. Phytotaxa 508 (2): 190. (2021).

Description

Culture characteristics

Material examined

Notes

﻿ Sphaeropsis citrigena (A.J.L. Phillips, P.R. Johnst. & Pennycook) A.J.L. Phillips & A. Alves. Stud. Mycol. 76, 157. (2013).

Description

Culture characteristics

Materials examined

Notes

﻿ Sphaeropsis guizhouensis Y.Y. Chen, A. J. Dissanayake & Jian K. Liu., J. Fungi 7, 893. (2021).

Description

Abstract

Introduction

Materials and methods

Isolates and morphology

DNA extraction, PCR amplification and sequencing

Phylogenetic analyses

Results

Phylogenetic analyses

Taxonomy

Aplosporella ginkgonis C.M. Tian, Z. Du & K.D. Hyde. Mycosphere 8(2): 1249 (2017).

Aplosporella prunicola Damm & Crous Fungal Diversity 27: 39 (2007).

Diplodia acerigena L.W. Li & Jian K. Liu, sp. nov.

Diplodia mutila (Fr.) Mont., Ann. Sci. nat., sér. 2, 1: 302. 1834.

Diplodia pistaciicola L.W. Li & Jian K. Liu, sp. nov.

Diplodia seriata De Not., Micr. Ital. Dec. 4: 6. (1942).

Dothiorella camelliae L.W. Li & Jian K. Liu, sp. nov.

Dothiorella sarmentorum (Fr.) A.J.L. Phillips, J. Luque & A. Alves, Mycologia 97: 522. (2005).

Dothiorella zanthoxyli L.W. Li & Jian K. Liu, sp. nov.

Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips, Stud. Mycol. 55: 248. (2006).

Sardiniella guizhouensis Y.Y. Chen & Jian K. Liu. Phytotaxa 508 (2): 190. (2021).

Sphaeropsis citrigena (A.J.L. Phillips, P.R. Johnst. & Pennycook) A.J.L. Phillips & A. Alves. Stud. Mycol. 76, 157. (2013).

Sphaeropsis guizhouensis Y.Y. Chen, A. J. Dissanayake & Jian K. Liu., J. Fungi 7, 893. (2021).

Diversity of Botryosphaerialean fungi collected in this study

Discussion

Acknowledgements

References