Epichloë species are fungal symbionts (endophytes) of grasses, six European and four North American biological species in genus Epichloë have been described in previous researches. In this study we describe a new Epichloë species, Epichloë yangzii Li et Wang, found in natural symbioses with Roegneria kamoji native to China. We investigated the host specificity, morphology, interfertility tests and molecular phylogenetic evidences of this new species. The results indicated that E. yangzii is host specific and seedborne. Most morphological characteristics of this new species are typical in the genus. However differences are evident in several features including size of perithecia, asci and ascospores. In mating tests E. yangzii was not interfertile with E. elymi isolates from related hosts in genera Elymus. Phylogenetic relationships based on sequences of beta-tubulin gene (tub2) introns and translation elongation factor 1-alpha gene (tef1) introns showed that members of the new species grouped into exclusive clades with high bootstrap value.
In this study we describe a new Neotyphodium species, Neotyphodium sinicum, found in natural symbiosis with asymptomatic Roegneria spp. (Elymus spp.) native to China. We investigated the host specificity, morphology, mating compatibility and molecular phylogenetic evidence for recognition of this new species. N. sinicum is host specific and seedborne and widely distributed in China. Most morphological characteristics of this new species are typical of other Neotyphodium species, but the growth on PDA plates tends to be more rapid. Three of five N. sinicum strains examined had double alleles of tefA and two of the five strains had double alleles of both tubB and tefA. Phylogenetic relationships based on sequences of tubB introns and tefA introns revealed that the allele-1 of tefA and tubB clustered together in a distinct subclade in the Epichloe bromicola/E. yangzii clade. The allele-2 grouped in the Epichlok" typhina clade (ETC). N. sinicum therefore appears to be an interspecific hybrid related to both E. yangzii and members of ETC. In mating tests N. sinicum failed to hybridize with E. yangzii.
We describe a new stromata-producing Neotyphodium species symbiotic with clonal Calamagrostis epigeios (L.) Roth. Stromata on the grass, 47.5-186 mm long, occurred frequently, but neither perithecium nor mature ascus was observed. Morphology of fungal isolates obtained from symptomatic and asymptomatic tillers were identical to each other and similar to those of epichloë endophytes. In phylogenetic analysis all selected five fungal isolates clustered into a significantly distinct clade based on sequences of beta-tubulin gene (tubB) introns and translation elongation factor 1-alpha gene (tefA) introns with bootstrap values of 99%, supporting erection of a new species. Concerning the production of extremely long stromata on the host plants and absence of sexual spores, we propose the name Neotyphodium stromatolongum Y. Ji, L. Zhan et Z. Wang, sp. nov.
We describe a new Epichloë species found in symbiosis with Poa pratensis ssp. pratensis in Liyang, China. Stromata characteristic of Epichloë spp. were present on some of the reproductive tillers of individual host grasses. Only three of the 98 stromata observed on field plants became orange and produced perithecia. Phylogenetic analyses based on sequences of tubB and tefA indicated that this Epichloë sp. was an interspecific hybrid related to both E. yangzii and members in the E. typhina complex clade (ETC). Allele-1 of tefA and tubB grouped in the E. bromicola/E. yangzii clade; allele-2 of these two genes clustered in a distinct subclade in the ETC. This is the first report of an Epichloë species that has interspecific hybrid origins. We propose the name Epichloë liyangensis Z. Wang, Y. Kang et H. Miao, sp. nov. for this species.
The fungus Neotyphodium sinicum forms an asymptomatic symbiosis with Roegneria grass species which are widely distributed in China. We collected 1,809 asymptomatic specimens of Roegneria, belonging to six species, from around 14 cities in nine provinces. 45.7% of the 1,809 asymptomatic Roegneria plants were endophyteinfected. Of 100 Neotyphodium isolates subsequently obtained, 31 were tested for cultural characteristics. These isolates showed morphological diversity and varying growth rates as well as differences in colony appearance and in conidial lengths. While conidial length of most of the isolates was typical of interspecific hybrid Neotyphodium species, conidial length of some isolates was smaller, similar to that of haploid species. A phylogenetic study was conducted of the tubB and tefA genes in 21 Neotyphodium isolates. The isolates were phylogenetically clustered into three types: 15/ 21 isolates had two divergent alleles of both tubB and tefA; 3/21 had two alleles of only tubB and 3/21 had two alleles of only tefA. In the tubB NJ tree, alleles-1 formed two distinct sub-clades in the Epichloë bromicola/E. yangzii clade (EBY); allele-2 was located in three sub-clades widely distributed within the E. typhina clade (ETC). The allele-1 of the tefA gene was also located within the EBY clade and formed two sub-clades; allele-2 was located in 3 sub-clades within the ETC clade. No relationship was observed between phylogenetic relationships and morphology in the N. sinicum isolates. The phylogenetic characteristics had little relationship to the host species or site of origin of the isolates. An explanation consistent with the phylogenetic findings is that N. sinicum is a complex of species that has resulted from multiple independent hybridization events.
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