(1998) and subsequent authors (Bissett et al 2003; Atanasova et

(1998) and subsequent authors (Bissett et al. 2003; Atanasova et al. 2010) while revealing the existence of 12 undescribed phylogenetic species. In the present work we revise the taxonomy of the Longibrachiatum Clade of Trichoderma following the molecular phylogenetic analysis of Druzhinina et al. (2012). Materials and methods Trichoderma strains

were independently received by the Kubicek and Samuels labs from colleagues in several countries or from personal collecting. Hypocrea teleomorphs of Trichoderma species were collected in Australia, New Zealand, Sri Lanka, Canary Islands (La Palma) and Isle de la Réunion in the Indian Ocean; cultures derived from these collections were made by isolating solitary ascospores using a micromanipulator or a platinum buy SAR302503 needle on cornmeal agar Selleckchem STA-9090 (Difco or Sigma) + 2% dextrose (CMD). Strains described below as T. flagellatum were isolated from surface sterilized roots of Coffea arabica and T. solani originated in surface sterilized potato tubers. Growth rates were determined on PDA (potato dextrose agar, Difco) and SNA (Nirenberg 1976, without filter paper)

at 15, 20, 25, 30 and 35°C in Entinostat darkness (with intermittent light when they were measured at intervals of 24 h). To prepare inoculum, cultures were incubated at 25°C for a few days on cornmeal agar (Difco) with 2% glucose (CMD) or on SNA. The inoculum was placed at 10–15 mm distance from the edge of the plate. It should be noted that different brands of PDA can give different colony characteristics (Jaklitsch 2009). Measurements were made at intervals of 24 h until 96 h. Colony characters were taken from colonies incubated on PDA and SNA at 25°C with alternating cool white fluorescent light and darkness (12 h/12 h) after 7–10 day; these conditions are referred to in descriptions as ‘under light’. Typically else there is little intra-species variation. Measurements are reported as mean plus and minus standard deviation with extremes in brackets; the 95% confidence of the means (95% ci) is reported in cases of multiple collections for a species. Statistics were computed

using Systat 10© (Wilkinson 2000). Continuous measurements (dimensions of conidia, phialides etc.) and appearance of conidiophores and conidial pustules are determined from colonies incubated 7–10 day at 25°C under light conditions described above, usually from SNA but when conidia do not form on SNA, characters are taken from CMD, less frequently on cornmeal agar without added glucose. Thirty units of each character are measured from all available cultures of each species, except where noted. In some images Helicon Focus (http://​www.​heliconsoft.​com/​heliconfocus.​html) was used to provide depth of field. The present work derives from the phylogenetic analysis of Druzhinina et al. (2012). To facilitate the location of species in the phylogenetic context a modified version of their phylogenetic tree is given as Fig. 1.

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