Hirooka Y et al. (2011), A morphological and phylogenetic revision of th...

ECB-ART-41983

Stud Mycol 2011 Jan 01;68:35-56. doi: 10.3114/sim.2011.68.02.

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A morphological and phylogenetic revision of the Nectria cinnabarina species complex.

Hirooka Y , Rossman AY , Chaverri P .

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The genus Nectria is typified by N. cinnabarina, a wood-inhabiting fungus common in temperate regions of the Northern Hemisphere. To determine the diversity within N. cinnabarina, specimens and cultures from Asia, Europe, and North America were obtained and examined. Their phylogeny was determined using sequences of multiple loci, specifically act, ITS, LSU, rpb1, tef1, and tub. Based on these observations, four species are recognised within the N. cinnabarina complex. Each species is delimited based on DNA sequence analyses and described and illustrated from specimens and cultures. The basionym for N. cinnabarina, Sphaeria cinnabarina, is lectotypified based on an illustration that is part of the protologue, and an epitype specimen is designated. Nectria cinnabarinas. str. is recircumscribed as having 2-septate ascospores and long stipitate sporodochia. Nectria dematiosa, previously considered a synonym of N. cinnabarina, has up to 2-septate ascospores and sessile sporodochia or no anamorph on the natural substrate. A third species, Nectria nigrescens, has up to 3-septate ascospores and short to long stipitate sporodochia. One newly described species, Nectria asiatica with a distribution restricted to Asia, has (0-)1-septate ascospores and short stipitate sporodochia. Young and mature conidia developing on SNA were observed for each species. Mature conidia of N. asiatica, N. cinnabarina, and N. nigrescens but not N. dematiosa bud when the mature conidia are crowded. On PDA the optimal temperature for growth for N. dematiosa is 20 °C, while for the other three species it is 25 °C. Based on our phylogenetic analyses, three subclades are evident within N. dematiosa. Although subtle culture and geographical differences exist, these subclades are not recognised as distinct species because the number of samples is small and the few specimens are insufficient to determine if morphological differences exist in the natural environment.

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Genes referenced: LOC583794 LOC590297 snai2

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	Fig. 1. Members of the genus Nectria. Combined act, tub, rpb1, ITS, LSU, tef1 Bayesian cladogram (Ln â€“21514.704). BI posterior probabilities/ML bootstrap values indicated at branches.
	Fig. 2. Members of the Nectria cinnabarina species complex (NCSC). Combined act, tub, rpb1, ITS, LSU, tef1 Bayesian cladogram (Ln â€“11408.155). BI posterior probabilities/ML bootstrap values indicated at branches. T: Teleomorph observed in the natural environment; A: Anamorph observed in the natural environment; X: no holomorph observed in the natural environment.
	Fig. 3. Graphs of 95 % confidence intervals of length to width ratios of ascospores and conidia.
	Fig. 4. Mycelial growth of NCSC at different temperatures on PDA.
	Fig. 5. Aâ€“R. Nectria asiatica. A. Perithecia and short stipitate sporodochia in the natural environment. B. Perithecia on nature. C. Median section of perithecium. D. Median section of perithecial wall. E. Ascus. F. 0â€“1 septates ascospores. G. Short stipitate sporodochium in the natural environment. H. Median section of short stipitate sporodochium. I. Edge of short stipitate sprodochium. J. Acropleurogenous conidiophores in the natural environment. K. Conidia in the natural environment. L. Aerial conidiophores and conidial mass on SNA. M. Lateral phialidic pegs and conidia on SNA. N. Short aerial conidiophores and conidia on SNA. O. Densely blanched aerial conidiophores and conidia on SNA. P. Mature conidia and young conidia on SNA. Q. Budding mature conidia on SNA. R. Budding and germinating mature conidia (arrow) that were streaked onto SNA. Scale bars: A, L = 1 mm; B, C, G, H = 300 Î¼m; D, I = 100 Î¼m; E, J, K, M, R = 30 Î¼m; F, N, O, P, Q = 15 Î¼m.
	Fig. 6. Aâ€“R. Nectria cinnabarina. A. Perithecia and long stipitate sporodochia in the natural environment. B. Peritheica in the natural environment. C. Median section of perithecium. D. Median section of perithecial wall. E. Ascus. F. 0â€“2 septates ascospores. G. Long stipitate sporodochia in the natural environment. H. Median section of long stipitate sporodochia. I. Conidia in the natural environment. J. Acropleurogenous conidiophore in the natural environment. K. Aerial conidiophores and conidial mass on SNA. L. Lateral phialidic pegs on SNA. M. Aerial conidiophores and young conidia. N. Densely blanched aerial conidiophores and young conidia. O. Mature conidia on SNA. P. Budding mature conidia and secondly conidia on SNA. Q. Slimy head of young and mature conidia on lateral phialidic peg on SNA. R. Budding and germinating mature conidia (arrow) that were streaked onto SNA. Scale bars: A = 500 Î¼m; C = 300 Î¼m; D, = 100 Î¼m; E, J, L, M, N, P, R = 30 Î¼m; F, I, O, Q = 15 Î¼m; B, G, H, K = 1 mm.
	Fig. 7. Aâ€“W. Nectria dematiosa species complex. A. Perithecia in the natural environment. B. Median section of perithecium. C. Median section of perithecial wall. D. Ascus. E. 1â€“2 septates ascospores. F. Astipitate sporodochium in the natural environment. G. Median section of stipitate sporodochium. H. Acropleurogenous conidiophore in the natural environment. I. Conidia in the natural environment. J. Aerial conidiophores and conidial mass on SNA. K. Young conidia on SNA. L. Lateral phialidic pegs and young conidia on SNA. M. Short aerial conidiophores and conidia on SNA. N. Densely blanched aerial conidiophores on SNA. O. Mature conidia and young conidia of N. dematiosa subclade A. P. Mature conidia and young conidia of N. dematiosa subclade B. Q. Mature conidia and young conidia of N. dematiosa subclade C. R. Germinating mature conidia (arrows) of N. dematiosa subclade A on SNA. S. Germinating mature conidia (arrows) of N. dematiosa subclade B on SNA. T. Germinating mature conidia (arrow) of N. dematiosa subclade C on SNA. U. Germinating mature conidia of N. dematiosa subclade A that were streaked onto SNA. V. Germinating mature conidia of N. dematiosa subclade B that were streaked onto SNA. W. Germinating mature conidia of N. dematiosa subclade C that were streaked onto SNA. Scale bars: A, J = 1 mm; B = 300 Î¼m; C, F, G = 100 Î¼m; D, H, I, R = 30 Î¼m; E, K, Lâ€“W = 15 Î¼m.
	Fig. 8. Aâ€“S. Nectria nigrescens. A. Perithecia and short stipitate sporodochia in the natural environment. B. Peritheica in the natural environment. C. Median section of perithecia. D. Median section of perithecial wall. E. Ascus. F. One and three septatas ascospores. G. Long stipitate sporodochia in the natural environment. H. Median section of long stipitate sporodochium. I. Edge of long stipitate sporodochium. J. Acropleurogenous conidiophore in the natural environment. K. Conidia in the natural environment. L. Aerial conidiophores and conidial mass on SNA. M. Young conidia on SNA. N. Lateral phialidic pegs on SNA. O. Short and densely blanched aerial conidiophores, and conidia on SNA. P. Mature conidia and young conidia on SNA. Q, R. Budding mature conidia on SNA. S. Germinating mature conidia that were streaked onto SNA. Scale bars: A, G, H, L = 1 mm; B, C = 300 Î¼m; D, I = 100 Î¼m; E, J, K, O, R = 30 Î¼m; F, M, N, P, Q, S = 15 Î¼m.

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