Identification of endophytic fungi from Livistona chinensis based on morphology and rDNA sequences

Abstract

A survey of the endophytic fungi in fronds of Livistona chinensis was carried out in Hong Kong. The endophyte assemblages identified using morphological characters consisted of 16 named species and 19 ' morphospecies ', the latter grouped based on cultural morphology and growth rates. Arrangement of taxa into morphospecies does not reflect species phylogeny, and therefore selected morphospecies were further identified based on ribosomal DNA (rDNA) sequence analysis. The 5n8S gene and flanking internal transcribed spacers (ITS1 and ITS2) regions of rDNA from 19 representative morphospecies were amplified by the polymerase chain reaction and sequenced. Phylogenetic analysis based on 5n8S gene sequences showed that these morphospecies were filamentous Ascomycota, belonging in the Loculoascomycetes and Pyrenomycetes. Further identification was conducted by means of sequence comparison and phylogenetic analysis of both the ITS and 5n8S regions. Results showed that MS704 belonged to the genus Diaporthe and its anamorph Phomopsis of the Valsaceae. MS594 was inferred to be Mycosphaerella and its anamorph Cladosporium of the Mycosphaerellaceae. MS339, MS366, MS370, MS395, MS1033, MS1083 and MS1092 were placed in the genus Xylaria of the Xylariaceae. MS194, MS375 and MS1028 were close to the Clypeosphaeriaceae. MS191 and MS316 were closely related to the Pleosporaceae within the Dothideales. The other 5 morphospecies, MS786, MS1043, MS1065, MS1076 and MS1095, probably belong in the Xylariales. The value of using DNA sequence analysis in the identification of endophytes is discussed.

Community structure of endophytic fungi of four mangrove species in Southern China

“…For example, Phomopsis fungi were widely isolated from mangroves (Suryanarayanan et al 1998; Suryanarayanan and Kumaresan 2000; Pang et al 2008; Xing et al 2011; Xing and Guo 2011) and non-mangrove plants (Taylor et al 1999; Guo et al 2000, 2008; Cannon and Simmons 2002; Murali et al 2006; Sun et al 2011, 2012). In addition, Pestalotiopsis as generalist fungi were common mangrove endophytes and widely occurred on plants across Palmae, Rhizophoraceae, Planchonellae, and Podocarpaceae in China (Liu et al 2010a, 2012; Xing and Guo 2011; Gong et al 2014).…”

“…In addition, Pestalotiopsis as generalist fungi were common mangrove endophytes and widely occurred on plants across Palmae, Rhizophoraceae, Planchonellae, and Podocarpaceae in China (Liu et al 2010a, 2012; Xing and Guo 2011; Gong et al 2014). Xylaria , as endophytic fungi were dominant in subtropical and tropical ecosystems (Bayman et al 1998; Guo et al 2000). Phyllosticta taxa were the most frequently isolated fungi of 10 mangrove plants in Thailand (Chaeprasert et al (2010).…”

“…Surface sterilisation followed the method of Guo et al (2000). Segments were surface sterilised by consecutive immersion for 1 min in 75% ethanol, 3 min in 3.25% sodium hypochlorite, and 30 s in 75% ethanol.…”

“…The non-sporulated cultures were designated as M ycelia sterilia , which were divided into different “morphotypes” according to cultural characteristics such as colony colour, texture, and extension rate on MEA (Guo et al 2000). One representative strain of each morphotype or sporulated isolate was selected for further molecular identification.…”

“…Genomic DNA was extracted from fresh cultures following the protocol of Guo et al (2000). Fresh fungal mycelia ( c .…”