Taxonomy and Multigene Phylogeny of Diaporthales in Guizhou Province, China

In a study of fungi isolated from plant material in Guizhou Province, China, we identified 23 strains of Diaporthales belonging to nine species. These are identified from multigene phylogenetic analyses of ITS, LSU, rpb2, tef1, and tub2 gene sequence data coupled with morphological studies. The fungi include a new genus (Pseudomastigosporella) in Foliocryphiaceae isolated from Acer palmatum and Hypericum patulum, a new species of Chrysofolia isolated from Coriaria nepalensis, and five new species of Diaporthe isolated from Juglans regia, Eucommia ulmoides, and Hypericum patulum. Gnomoniopsis rosae and Coniella quercicola are newly recorded species for China.


Introduction
Diaporthales is an important and species-rich ascomycetous order in the subclass Diaporthomycetidae (Sordariomycetes). Despite its cosmopolitan distribution and high diversity with distinctive morphology, this order has received relatively little attention. Currently, the existing classification lists 31 accepted families within the order Diaporthales [1], including Foliocryphiaceae, Diaporthaceae, Gnomoniaceae, and Schizoparmaceae. Members of Diaporthales have a wide range of ecological habitats and numerous modes of nutrition [2]. Excepting the members of Tirisporellaceae, most taxa in Diaporthales occur in terrestrial habitats. Species in Diaporthales form solitary or aggregated, immersed to erumpent, rarely superficial, and orange, brown, or black perithecial ascomata, with short or long necks that are located in stromatic tissues or substrates and with a lack of hamathecium or with few paraphyses [2][3][4][5]. Their asci are unitunicate with a conspicuous refractive ring [5,6]. Their ascospores are diverse in shape, size, and color. The asexual morphs of Diaporthales are generally coelomycetous [6], producing acervuli, pycnidial, or synnematal conidiomata and with or without a well-developed stroma. Conidiogenesis is phialidic or rarely annellidic, and conidia are usually unicellular or one-septate [6].
In China, the first monograph for Diaporthales referred to Phomopsis (=Diaporthe), which introduced 133 morphological taxa (including two specialized forma) isolated from 74 familial plants [7]. One diaporthalean pathogen that causes a devastating wilt disease for Cyathea lepifera was reported in Taiwan [8]. Pustulomyces accommodated in Diaporthaceae was revealed by morphology and molecular analyses [9]. Two novel families, Melansporellaceae and Diaporthosporellaceae, were introduced to accommodate the monotypic genera, Melanosporella and Diaporthosporella, based on both holomorphic morphology and phylogenetic analysis [10,11], and Foliocryphiaceae was established by Jiang et al. [1] to retain

Species Strain Number
GenBank Accession Number Phylogenetic analyses were carried out by maximum likelihood (ML), maximum parsimony (MP), and Bayesian inference (BI). The ML analysis was performed using RAxML-HPC BlackBox(8.2.12) [42] partial and general time reversible model (GTR) using the discrete gamma distribution as the evolution model by CIPRES Science Gateway version 3.3 [43]. Non-parametric bootstrap analysis was implemented with 1000 iterations. The resulting duplicates were plotted onto the best-scoring tree previously obtained.
Maximum parsimony (MP) analyses were performed with PAUP on XSEDE (4.a168) on CIPRES Science Gateway v. 3.3 using the heuristic search option with 1000 random sequence addition replicates and tree bisection and reconnection (TBR) with reconnection limit (=8) as the branch-swapping algorithm. Maxtrees were set to 5000 (and not increased). Branches were collapsed, creating polytomies if maximum branch length was zero. Tree length (TL), consistency index (CI), retention index (RI), rescaled consistency index (RC), and homoplasy index (HI) were calculated for each tree generated.
Bayesian inference (BI) analysis was performed by MrBayes 3.2.7a [44] in the CIPRES Science Gateway version 3.3. The optimal substitution model with gamma rates and dirichlet base frequencies for ITS, LSU, rpb2, tef1, and tub2 sequences was decided by modelGUI for each locus [45]. The Markov chain Monte Carlo (MCMC) sampling approach was used to calculate posterior probabilities (PP) [46]. Six simultaneous Markov chains were run for 50 million generations and trees were sampled every 1000th generation; thus, 50,000 trees were obtained. The first 25% 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.
The phylogenetic trees were viewed with FigTree v. 1.4.3 [47] and processed with Adobe Illustrator CS5. ML bootstrap support (MLBS) and MP bootstrap support (PBS) equal or greater than 70% [48] and Bayesian posterior probabilities (PP) equal or greater than 0.95 [49] are displayed in the first, second, and third positions on the edited phylogenetic tree, respectively.

Genealogical Phylogenetic Species Recognition (GCPSR) Analysis
Morphologically and phylogenetically related species were analyzed using GCPSR as described by Taylor et al. [17] by the pairwise homogeneity index test (PHI) [50]. The PHI tests were performed in SplitsTree v. 4.17.1 [18,51] as described by Quaedvlieg et al. [52] to determine the level of recombination within phylogenetically closely related species. This test determines the null hypothesis probability (p-value) of no recombination within the dataset. When the p-value is less than 0.05, we reject the null hypothesis and accept the alternate hypothesis that there is evidence of the presence of recombination. The results were visualized by constructing a split graph using LogDet conversion and Splits options.

Phylogenetic Analyses
To reveal the phylogenetic position of the family Foliocryphiaceae, genera Diaporthe, Gnomoniopsis, and Coniella, within the order Diaporthales, phylogenetic analyses were performed with ITS, LSU, rpb2, tef1, and tub2 sequence data.
The first sequence dataset of ITS, LSU, rpb2, tef1, and tub2 was analyzed to focus on Cryphonectriaceae and Foliocryphiaceae. The alignment included 43 taxa, including representatives of Cryphonectriaceae and Foliocryphiaceae and outgroup sequences of Dwiroopa lythri (CBS 109755, ex-type strain) and Dw. punicae (CBS 143163, ex-type strain) ( Table 2). The aligned five-locus datasets comprised 3596 characters of the family Foliocryphiaceae, viz. ITS: 1-769, LSU: 770-1617, rpb2: 1618-2442, tef1: 2443-2952, and tub2: 2953-3596. Of these, 2145 characters were constant, 1131 characters were parsimony-informative, and 320 were parsimony-uninformative (gaps were treated as missing). The parameter settings used are shown in Table 6. A RAxML tree was selected to show the topology (Figure 1), and MP and Bayesian analyses resulted in similar topology to ML.   Two new strains of Chrysofolia coriariae sp. nov. (GUCC 416.4, ex-type strain and GUCC 416.14) collected during this study in Guizhou Province shared the same branch length with 100% MLBS/99% MPBS/1 PP support and were grouped with the type strains of Ch. colombiana (CPC 24986) and Ch. barringtoniae (TBRC 5647) with high statistical support, being (76% MLBS/99% MPBS)/(100% MLBS/99% MPBS/1 PP) (Figure 1), respectively. A comparison of the DNA base composition (Table 7) indicated that between our two strains and Ch. colombiana (CPC 24986), there were seven different bases in the ITS region, two different bases in the LSU region, and 149 different bases in the tef1 region. Between GUCC 416.4, GUCC 416.14, and Ch. barringtoniae (TBRC 5647), there were 31 different bases in the ITS region and four different bases in the LSU region. Unfortunately, Ch. colombiana did not have rpb2 or tub2 sequence data, and Ch. barringtoniae did not have rpb2, tef1, or tub2 sequence data.
Two new strains of Diaporthe hyperici sp. nov. (GUCC 414.4, ex-type strain and GUCC 414.41) formed a high-support subclade (100% ML, 100% MP, 1.00 PP) with D. caulivora (CBS 127268, ex-type strain, Dip1, and Dpc11). There were 17 base pair differences in ITS, 30 base pair differences in tef1, and 11 base pair differences in the tub2 from our strains based on a DNA base comparison ( Table 7). The PHI test (Figure 2f) did not find any statistically significant evidence of recombination (p-value = 1.0) between our two strains (GUCC 414.4, GUCC 414.41) and strains of D. caulivora.
The fourth sequence dataset of ITS, LSU, and tef1 was analyzed in combination to infer the interspecific relationships within Coniella. The alignment included 32 taxa, including the outgroup sequences of C. fragariae (CBS 172.49, ex-type strain) and C. nigra (CBS 165.60, extype strain) ( Table 5). The aligned three-locus datasets comprised 2165 characters of Coniella, viz. ITS: 1-595, LSU: 596-1767, and tef1: 1768-2165. Of these, 1717 characters were constant, 385 characters were parsimony-informative, and 63 were parsimony-uninformative (gaps were treated as missing). The parameter settings that were used are shown in Table 6. A RAxML tree was selected to show the topology (Figure 4) 16), there were identical sequences in the ITS and LSU regions, but 1/5/10/10/18/7/6 bases were different in the tef1 region. The PHI test (Figure 2h) did not find statistically significant evidence of recombination (p-value = 0.2264) between our five strains and related taxa C. quercicola (CBS 904.69, ex-type strain, CBS 283.76, and CPC 12133).

Taxonomy
Pseudomastigosporella S.Y. Wang, Yong Wang bis, and Y. Li, gen. nov. MycoBank Number: MB846026 Etymology: In reference to Mastigosporella, to which this genus is morphologically similar. Classification: Foliocryphiaceae, Diaporthales, Sordariomycetes. Description: Life style: Parasitic, leaves of Hypericum patulum and Acer palmatum. Asexual morph: Conidiomata pycnidial, globose or subglobose, base immersed, separate to aggregated, mycelium superficial, fluffy, granular, white or gray-white to pale yellow, exuding light brown-orange to medium brown-orange to deep brown-orange conidial masses, bright yellow or light orange in lactic acid, 2-5 wall layers of olive to gray-green textura angularis. Conidiophores reduced to conidiogenous cells. Conidiogenous cells arising from base, central cushion of hyaline cells, densely aggregated, slightly thicker, cylindrical to ampulliform, simple, lining the inner cavity of base, mostly hyaline, sometimes pale olive, smooth, cylindrical to ampulliform, straight to curved, sometimes wider at the base. Conidia solitary, hyaline, smooth, guttulate, fusoid to ellipsoidal, sometimes long bubbleshaped, straight to curved, aseptate, base tapering with flattened scar, apex with 1 tubular appendage. Sexual morph: Unknown.
Type species: Pseudomastigosporella guizhouensis S.Y. Wang, Yong Wang bis & Y. Li. Notes: In Foliocryphiaceae, the important morphological characters of asexual morph was to produce dimorphic conidia. The microconidia were minute, cylindrical, aseptate, hyaline to pale brown; macroconidia were fusoid, aseptate, hyaline [1]. However, Pseudomastigosporella only had macroconidia but like species in Mastigosporellaceae with an apical appendage developing as continuation of conidium body. This feature contradicted the root of "key to genera in Cryphonectriaceae, Foliocryphiaceae, and Mastigosporellaceae" provided by Jiang et al. [1]. However, following our phylogenetic analyses we still proposed that Pseudomastigosporella should be placed in Foliocryphiaceae family.
Pseudomastigosporella guizhouensis S.Y. Wang, Yong Wang bis, and Y. Li, sp. nov. MycoBank Number: MB846027, Figure 5.     Description: Life style: Parasitic, leaves of Hypericum patulum and Acer palmatum. Asexual morph: Conidiomata pycnidial, globose or subglobose, base immersed, separate to aggregated, mycelium superficial, fluffy, granular, white or gray-white to pale yellow, producing light brown-orange to medium brown-orange to deep brown-orange conidial masses, up to 570 µm diam., bright yellow or light orange in lactic acid, 2-5 wall layers of olive to gray-green textura angularis, 50-570 µm diam. Conidiophores reduced to conidiogenous cells. Conidiogenous cells arising from base, central cushion of hyaline cells, densely aggregated, slightly thicker, cylindrical to ampulliform, simple, lining the inner cavity of base, mostly hyaline, sometimes pale olive, smooth, cylindrical to ampulliform, straight to curved, sometimes wider at the base, 5-20 × 1. Culture characteristics: Colonies covering 9 cm Petri dish after 2 weeks at 25 • C and under a 12 h light/dark regime. On PDA, white or gray-white, fluffy, granular, effuse surface, reverse white or beige; on OA, white or gray-white to pale yellow, fluffy, granular, effuse surface, exuding light brown-orange to medium brown-orange to deep brown-orange conidial masses, reverse white or beige to pale yellow.
(2-)2.5(-3) μm [54]. The results of the DNA base comparisons (Table 7) showed that there were striking differences in each gene among our four Pseudomastigosporella strains and adjacent genera. Based on its distinct morphological characteristics, DNA phylogeny, DNA base differences, and pairwise homoplasy index (PHI) test results, Pseudomastigosporella was described here as a new genus in Foliocryphiaceae with Ps. guizhouensis as the type species.
Chrysofolia coriariae S.Y. Wang, Yong Wang bis, and Y. Li, sp. nov. MycoBank Number: MB845958, Figure 6.  Description: Life style: Parasitic, leaves of Coriaria nepalensis. Asexual morph: Conidiomata pycnidial, globose or subglobose, separate to aggregated, mycelium superficial and immersed, exuding yellow to bright orange to brown-orange wet conidial masses, greenbrown in lactic acid, but bright yellow or light orange in sterile water, 2-6 wall layers of green-brown to brown textura angularis, 50-400 μm diam.; neck 15-60 μm long, 50-200 Culture characteristics: Colonies culturing under a controlled temperature light incubator at 25 • C and under a 12 h light/dark regime for 2 weeks. Colonies on PDA 75-90 mm diam. after 2 weeks at 25 • C, light brown to white or gray-white, felty, effuse surface, with white fluffy even mycelium margin, reverse brown to light brown to white edge. Colonies on OA 65-85 mm diam. after 2 weeks at 25 • C, light brown to white or gray-white, flat surface, exuding orange or brown conidial masses, reverse light brown to white or gray-white.
Culture characteristics: Colonies covering 9 cm Petri dish after 2 weeks at 25 • C and under a 12 h light/dark regime; spreading with uneven aerial mycelium. On PDA, surface with abundant white to pale yellow, uneven zonated aerial mycelium and margin, distinctly imbricated like a flower; reverse with pale yellow to light brown and pale pink, uneven zonated aerial mycelium and margin, exuding abundant dark green to black spots with age. On OA surface with uneven white to olive aerial mycelium, forming black conidial masses surrounded by white or gray-white mycelium; reverse white to olive, irregular. On pine needles with irregular dark green to black subglobose conidial masses surrounded by thick gray-white mycelium.
Culture characteristics: Colonies covering 9 cm diam. Petri dish after 2 weeks at 25 • C under a 12 h light/dark regime. On PDA surface with thick aerial mycelium, flat, velvet, white and beige; reverse white to pale yellow. On OA surface with white or pale white thin aerial mycelium, exuding black conidial masses, surrounded by white mycelium; reverse white or beige. On pine needles, irregular, black, globose conidial masses surrounded by thick white mycelium.
Notes: The conidiomata of D. dejiangensis (2 mm diam.) are larger than those of D. cotoneastri (1.5 mm diam.) [58], while its alpha conidia (6-8.5 × 1.5-3 µm) are smaller than those of D. cotoneastri (6-10 × 2-3 µm). Neither beta nor gamma conidia were observed for D. dejiangensis, while D. cotoneastri produced beta conidia (18-25 × 1 µm). Diaporthe dejiangensis was phylogenetically distinct from the species presently known based on the DNA data ( Figure 3). The results of the DNA base comparisons are shown in Table 7 and indicate that there were many base differences among three genes. Based on its distinct morphological characteristics, DNA phylogeny, DNA base differences, and pairwise homoplasy index (PHI) test results, D. dejiangensis was described here as a new species.
Culture characteristics: Colonies covering 9 cm diam. Petri dish after 2 weeks at 25 °C under a 12 h light/dark regime. On PDA surface with thick aerial mycelium, flat, velvet, white and beige; reverse white to pale yellow. On OA surface with white or pale white Culture characteristics: Colonies covering 9 cm diam. Petri dish after 2 weeks at 25 • C and a 12 h light/dark regime; spreading with aerial mycelium and uneven zonation. On PDA, surface with abundant aerial mycelium, with white uneven zonated aerial mycelium in the middle; reverse with white to pale yellow to light brown, uneven zonated aerial mycelium. On OA, surface with white or pale white, thin aerial mycelium, exuding black conidial masses surrounded by white mycelium; reverse white or beige. On pine needles, irregular black subglobose conidial masses surrounded by white mycelium.
Notes: The conidiomata of D. hyperici (3 mm diam.) are larger than those of D. caulivora (230-310 µm diam.) [60], but the alpha conidia of D. hyperici (5-9.5 × 1.5-3 µm) are shorter than those of D. caulivora (8.9-9.2 × 2.4-2.5 µm). Diaporthe caulivora produces a sexual morph with unitunicate asci, while D. hyperici has no known sexual morph. Diaporthe hyperici was phylogenetically distinct from the presently known species based on the DNA data ( Figure 3). A comparison of the DNA bases (Table 7)  sexual morph with unitunicate asci, while D. hyperici has no known sexual morph. D aporthe hyperici was phylogenetically distinct from the presently known species based the DNA data ( Figure 3). A comparison of the DNA bases (  Figure 12. . Asexual morph: Con iomata erumpent, separated, immersed or superficial, globose to depressed, initially a pearing deep brown to black, slowly oozing transparent white or pale-yellow oily sphe with age, up to 600 μm diam., 5-7 wall layers of olive brown to brown textura angula Conidiophores reduced to conidiogenous cells. Conidiogenous cells lining the inner cavi hyaline, smooth, subcylindrical, branched at base or not, frequently branched above, si ple, tapering, hyaline, smooth, subcylindrical, tapering towards apex, 6-15 × 1-3.5 μm = 11 × 2 μm; n = 20). Conidia solitary, aseptate, fusoid, hyaline, asymmetrical, guttula smooth-walled, rounded to acute apex, 6-12 × 2-4 μm ( ̅ = 8.5 × 3 μm; n = 30).   Culture characteristics: Colonies cultured at 25 • C and a 12 h light/dark regime for 2 weeks on PDA 60-85 mm diam., forming a circle of transparent mycelium in the center, followed by a circle of white or gray-white thick ridges, then uneven zonated aerial mycelium, slightly imbricated, thick, initially appearing white to pale yellow, slowly turning olive-gray with age outside the two concentric rings, with an uneven edge; reverse transparent to white or olive and white uneven imbricated zonated to white or light brown uneven edge. Colonies on OA covering the whole dish, pale white or light gray-white, flat surface, exuding deep brown to black conidial masses, slowly oozing transparent white or pale-yellow oily spheres with age, reverse pale white or light gray-white.
Notes: Gnomoniopsis represented a genus of mostly host-specific fungi [61,62]. Gnomoniopsis rosae (GUCC 408.7 and GUCC 408.17) was phylogenetically identical to the ex-type strain (CBS 145085) isolated by Crous et al. [31] in ITS, LSU, and rpb2, and we also supplemented the DNA sequences of this species with tef1 and tub2 genes. The DNA base comparison results are shown in Table 7; there were no DNA base differences among several genes. The isolates of G. rosae were newly recorded for China based on their morphological characteristics, DNA phylogeny, DNA base differences, and pairwise homoplasy index (PHI) test results.
Notes: Coniella quercicola was originally described as Macroplodia quercicola on the leaves of Quercus robur collected in the Netherlands. It was described as having pale-brown, cylindrical conidia, 24 [64] in ITS, LSU, and tef1 genes ( Figure 14). According to the results of the DNA base comparison (Table 7), we note that base differences almost only occur in the tef1 region. The identification of C. quercicola was based on its morphological characteristics, DNA phylogeny, DNA base differences, and pairwise homoplasy index (PHI) test results. eral genes. The isolates of G. rosae were newly recorded for China based on their morp logical characteristics, DNA phylogeny, DNA base differences, and pairwise homopl index (PHI) test results.
MycoBank Number: MB 817831, Figure 13. x FOR PEER REVIEW 31 of 36 Figure 14. Phylogram generated from RAxML analysis of a concatenated ITS-LSU-tef1 sequence dataset to represent the phylogenetic relationships of taxa in Coniella. The tree was rooted with C. fragariae (CBS 172.49, ex-type strain) and C. nigra (CBS 165.60, ex-type strain). Bootstrap support values for ML and MP equal to or greater than 70% and the Bayesian posterior probabilities equal to or higher than 0.95 PP are indicated above the nodes as ML/MP/PP. Support values lower than 70% ML/MP and 0.95 PP are indicated by a hyphen (-). The newly generated sequences are indicated in red.

Discussion and Conclusions
Families, genera, and species within Diaporthales are now characterized and separated based on a combination of morphology and molecular data [12,29,30,[64][65][66][67][68][69][70][71][72]. The present study described and illustrated nine species (within five genera) of Diaporthales isolated from various host plants in Guizhou Province, China, including Gnomoniopsis mostly as host-specific fungi [61,62,65,73]. Based on their unique morphological characteristics, DNA phylogeny, DNA base differences, and pairwise homoplasy index (PHI) Figure 14. Phylogram generated from RAxML analysis of a concatenated ITS-LSU-tef1 sequence dataset to represent the phylogenetic relationships of taxa in Coniella. The tree was rooted with C. fragariae (CBS 172.49, ex-type strain) and C. nigra (CBS 165.60, ex-type strain). Bootstrap support values for ML and MP equal to or greater than 70% and the Bayesian posterior probabilities equal to or higher than 0.95 PP are indicated above the nodes as ML/MP/PP. Support values lower than 70% ML/MP and 0.95 PP are indicated by a hyphen (-). The newly generated sequences are indicated in red.

Discussion and Conclusions
Families, genera, and species within Diaporthales are now characterized and separated based on a combination of morphology and molecular data [12,29,30,[64][65][66][67][68][69][70][71][72]. The present study described and illustrated nine species (within five genera) of Diaporthales isolated from various host plants in Guizhou Province, China, including Gnomoniopsis mostly as host-specific fungi [61,62,65,73]. Based on their unique morphological characteristics, DNA phylogeny, DNA base differences, and pairwise homoplasy index (PHI) test evaluations, we described one new genus, seven new species, and two new fungal records for China. Only asexual morphology was observed for all the taxa described in this paper.
Foliocryphiaceae (Diaporthales) was established by Jiang et al. [1] based on the type genus Foliocryphia [54] and two allied genera, Chrysofolia [53] and Neocryphonectria [1]. Chrysofolia and Foliocryphia were originally placed in the family Cryphonectriaceae but they were transferred to Foliocryphiaceae by Jiang et al. [1]. Species of Chrysofolia usually exude a yellow slimy mass of conidia from a globose pycnidium with an immersed base. Only two species are listed in MycoBank (www.mycobank.org; accessed on 8 October 2022), Ch. colombiana [53], a pathogen of Eucalyptus urophylla from Colombia, and Ch. barringtoniae [55], an endophyte of Barringtonia acutangula from Thailand. Chrysofolia coriariae sp. nov. observed in the present study represents the first taxon of Chrysofolia in China.
Diaporthe is a large genus in Diaporthaceae with 1168 epithets listed in Index Fungorum (http://www.indexfungorum.org/; accessed on 4 July 2022) but only one-fifth of these taxa have been studied with molecular data [73][74][75]. The sexual morph of Diaporthe is characterized by immersed perithecial ascomata and an erumpent pseudostroma with more or less elongated perithecial necks; unitunicate clavate to cylindrical asci; and fusoid, ellipsoid to cylindrical, septate or aseptate, hyaline ascospores, which are biseriately to uniseriately arranged in the ascus, sometimes having appendages [29,30,76]. The asexual morph is characterized by ostiolate conidiomata, with cylindrical phialides producing three types of hyaline, aseptate conidia [76,77]. Type I α-conidia are hyaline, fusiform, straight, guttulate, or eguttulate; aseptate; and smooth-walled. Type II β-conidia are hyaline, filiform, straight or hamate, aseptate, smooth-walled, and eguttulate. Type III γ-conidia are rarely produced, and are hyaline, multiguttulate, and fusiform to subcylindrical with an acute or rounded apex, while the bases are sometimes truncate. Five new taxa of Diaporthe were introduced, which indicates that more potential novel and known taxa in this genus could be discovered because of the rich biodiversity in Guizhou Province.
Gnomoniaceae is a large family within Diaporthales, containing 38 accepted genera [65,[78][79][80][81]. Among them, Gnomoniopsis is a well-delimited genus inhabiting the leaves, branches, and fruits of hosts in three families: Fagaceae, Onagraceae, and Rosaceae [62,65,73]. The sexual morph of Gnomoniaceae is characterized by ascomata that are generally immersed, solitary, or aggregated in an undeveloped stroma [6,61]. The perithecia are dark brown to black and pseudoparenchymatous with central, eccentric, or lateral necks [6,61]. The asci usually have an inconspicuous or distinct apical ring. Ascospores are generally small, hyaline, and uniseptate. The asexual morph is characterized by acervular or pycnidial conidiomata, phialidic conidiogenous cells, and non-septate conidia [82]. Gnomoniopsis rosae in our study was isolated as asexual morph from Rosa sp. and was newly recorded for China.
The family Schizoparmeaceae (Diaporthales) was introduced by Rossman et al. [6]. Historically, the family consisted of three genera, two of which only produce asexual morphs (Coniella and Pilidiella), while one can produce sexual morphs (Schizparme) [6]. This family was reassessed by Alvarez et al. [64], who proposed that Pilidiella is a taxonomic synonym of Coniella. Coniella was erected by Höhnel [83] and typified by C. pulchella, [84] who separated the genus into Euconiella (with dark conidia) and Pseudoconiella (with pale conidia) [64]. The key characteristics of Coniella are erumpent, brown, or black ascomata or conidiomata that later become superficial and an irregularly thickened peridium with plate-like ornamentation and one-celled ascospores, initially hyaline and later becoming pale to dark brown [30]. The present isolates of C. quercicola represent a new country record for China and new host records.
The molecular data provided evidence that our new genus belongs to Foliocryphiaceae, although in morphology it is similar to Mastigosporella in Mastigosporellaceae. In this molecular era, morphological conclusions are increasingly being reduced to a subordinate or even insignificant position. Thus, we accepted the phylogenetic conclusion to create the monotypic genus Pseudomastigosporella. Despite this, we still require additional strains of Diaporthales in order to compare the genome-wide information of members in this order due to the high level of similarities in morphology but measurable differences in molecular data. Currently, there are too few data for the adequate comparison of fungi.