Toward a natural classification of Botryosphaeriaceae : a study of the type specimens of Botryosphaeria sensu lato

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Authors

Zhang, Ying
Zhou, Yupei
Sun, Wei
Zhao, Lili
Pavlic-Zupanc, Draginja
Crous, Pedro W.
Slippers, Bernard
Dai, Yucheng

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Frontiers Media

Abstract

The genus Botryosphaeria includes more than 200 epithets, but only the type species, Botryosphaeria dothidea and a dozen or more other species have been identified based on DNA sequence data. The taxonomic status of the other species remains unconfirmed because they lack either morphological information or DNA sequence data. In this study, types or authentic specimens of 16 “Botryosphaeria” species are reassessed to clarify their identity and phylogenetic position. nuDNA sequences of four regions, ITS, LSU, tef1-a and tub2, are analyzed and considered in combination with morphological characteristics. Based on the multigene phylogeny and morphological characters, Botryosphaeria cruenta and Botryosphaeria hamamelidis are transferred to Neofusicoccum. The generic status of Botryosphaeria aterrima and Botryosphaeria mirabile is confirmed in Botryosphaeria. Botryosphaeria berengeriana var. weigeliae and B. berengeriana var. acerina are treated synonyms of B. dothidea. Botryosphaeria mucosa is transferred to Neodeightonia as Neodeightonia mucosa, and Botryosphaeria ferruginea to Nothophoma as Nothophoma ferruginea. Botryosphaeria foliicola is reduced to synonymy with Phyllachorella micheliae. Botryosphaeria abuensis, Botryosphaeria aesculi, Botryosphaeria dasylirii, and Botryosphaeria wisteriae are tentatively kept in Botryosphaeria sensu stricto until further phylogenetic analysis is carried out on verified specimens. The ordinal status of Botryosphaeria apocyni, Botryosphaeria gaubae, and Botryosphaeria smilacinina cannot be determined, and tentatively accommodate these species in Dothideomycetes incertae sedis. The study demonstrates the significance of a polyphasic approach in characterizing type specimens, including the importance of using of DNA sequence data.

Description

Supplementary Figure 1 | Botryosphaeria gaubae (W 1992-05937, holotype). (A,B) Ascomata erumpent through the lower side of the leaf. (C) Squash showing cylindrical or broadly cylindrical asci in cotton blue. (D) Part of the peridium. (E) Septate pseudoparaphyses in cotton blue. (F-H) Aseptate, fusiform to ellipsoid ascospores in cotton blue. Scale bars: (A) = 1 mm, (B) = 200 mm, (C) = 50 mm, (E) = 20 mm, (D,F-H) = 10 m m.
Supplementary Figure 2 | Laestadia apocyni (MICH 14281, isotype). (A) Ascomata erumpent through a piece of twig epidermis. (B) Released, hyaline, 1-septate ascospores. (C) Ascus in water. (D) Line drawing of ascus in water. Scale bars: (A) = 200 mm, (B-D) = 20 m m.
Supplementary Figure 3 | Sphaeria smilacinina (NYS f2818, holotype). (A) Ascomata erumpent through the twig epidermis. (B,C) Immature asci. (D) Released ascospores. (E) Line drawing of broadly clavate ascus. Scale bars: (A) = 500 mm, (B-D) = 20 mm, (E) = 40 m m.
Supplementary Figure 4 | One of the most parsimonious trees obtained from combined ITS, LSU, tub2, and tef1-a sequence data of Botryosphaeria spp. Outgroup taxa are Neofusicoccum luteum and Neofusicoccum parvum. Maximum parsimony (MP) support values above 70% and Bayesian posterior probabilities (PP) support above 80% are shown with MP bootstrap followed by Bayesian PP (MP/PP) values at the nodes. The species characterized in this study are in boldface.
Supplementary Figure 5 | One of the most parsimonious trees obtained based on combined ITS, tef1-a, and tub2 sequence data of Neofusicoccum spp. Outgroup taxon are Botryosphaeria dothidea and B. corticis. Maximum parsimony (MP) support values above 60% and Bayesian posterior probabilities (PP) support above 80% are shown with MP/PP, values at the nodes. The species characterized in this study are in boldface.
Supplementary Figure 6 | One of the most parsimonious trees obtained from LSU sequence dataset of Neofusicoccum spp. Outgroup taxa are Botryosphaeria corticis and B. dothidea. Maximum parsimony (MP) support values above 70% and Bayesian posterior probabilities (PP) support above 80% are shown with MP bootstrap followed by Bayesian PP (MP/PP) values at the nodes. The species characterized in this study are in boldface.
Supplementary Figure 7 | One of the most parsimonious trees obtained from ITS and LSU sequence dataset of Nothophoma spp. Outgroup taxa is Didymella calidophila. Maximum likelihood (ML) support values above 50%, Maximum parsimony (MP) support values above 50%, and Bayesian posterior probabilities (PP) support above 95% are shown with ML and MP bootstrap followed by Bayesian PP (MP/PP/ML) values at the nodes. The species characterized in this study are in boldface.
Supplementary Table 1 | Species, specimens and GenBank accession numbers of sequences used in this study (newly generated sequences are indicated in bold).

Keywords

Botryosphaeriales, Phylogeny, Sexual stage, Taxonomy, Type specimens

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Citation

Zhang, Y., Zhou, Y.P., Sun, W., Zhao, L.L., Pavlic-Zupanc, D., Crous, P.W., Slippers, B. & Dai, Y.C. (2021) Toward a Natural Classification of Botryosphaeriaceae: A Study of the Type Specimens of Botryosphaeria sensu lato. Frontiers in Microbiology 12:737541. DOI: 10.3389/fmicb.2021.737541