New reports of Bursaphelenchus species associated with conifer trees in Romania.

Summary The 2013 annual monitoring programme for the pinewood nematode, Bursaphelenchus xylophilus, analysed a total of 267 wood samplescollected from declining or symptomatic coniferous trees distributed among national forests, gardens, public parks, distribution centres,wood-processing industries, as well as 104 samples collected from wood packing material originated from several other countries. From atotal of eight species found, five have been previously reported from Romania, while three other species (Bursaphelenchus abietinus, Bursa-phelenchus fraudulentus and Bursaphelenchus fuchsi) represent new findings for Romania. In addition, B. mucronatus was found in packingwood originated from Russia. Herein, we provide a morphological, morphometric and molecular characterization for the new species foundfor Romania. Bursaphelenchus xylophilus was not detected. 1 Introduction The pinewood nematode (PWN), Bursaphelenchus xylophilus (Steiner and Buhrer, 1934) Nickle, 1970, is considered one ofthe most important pests and pathogens of conifer forests worldwide. The general fear of establishment of the PWN, causalagent of the pine wilt disease (PWD) into new areas, assumes huge relevance due to the devastating damage that this spe-cies can cause to pine forests (Vicente et al. 2012). As the first report of this species in Europe (Mota et al. 1999), all Euro-pean Union members have adopted recommended regulations and have performed national surveys for monitoring thepotential entry of this quarantine species.In the past years, Romania has been following EU regulations, such as the 2012/535/UE decision on emergency mea-sures to prevent the introduction and spread of the PWN, thereby annual national surveys have been conducted under theMinistry of Agriculture and Rural Development. The total forest area in Romania consists of over 6.3 million ha, corres-ponding to 27% of the total land area of the country. Romanian forests are diverse, with 31% represented by conifers,including several Pinus species (e.g. Pinus cembra, P. mugo, P. nigra and P. sylvestris) (Donita et al. 1990; Borlea et al.2006). Although the current phytosanitary measures and monitoring programme in Romania aim to minimizes the risk ofentry of the PWN into the country, the favourable climatic conditions, associated with the presence of highly susceptiblepine trees (P. nigra and P. sylvestris) and distribution of Monochamus species (Ruicanescu 2007), which can mediate thetransportation and transmission of the PWN, impose a continuous challenge to protect the national pine forests.This quarantine programme has not only allowed monitoring of the national pine forests and several critical areas wherethe PWN could entry/establish (entry ports and wood factories), but also has provided the opportunity to identify andcharacterize other species belonging to the genus Bursaphelenchus. The first report of the genus in Romania was given in2013 (Calin et al. 2013), followed by the identification and characterization of five species of this genus, namely B. hofman-ni Braasch, 1998; B. poligraphi Fuchs, 1937; B. vallesianus Braasch, Sh€onfeld, Polomski & Burgermeister, 2004; B. willibaldiShonfeld, Braasch & Burgermeister, 2006, including the€ first description of B. andrassyi Dayi, Calin, Akbulut, Gu, Schr€oder,Vieira & Braasch, 2014 (Dayi et al. 2014).During 2013, the sampled areas throughout the country and the number of wood samples collected were intensified, aswell as the number of samples collected from wood originating from other countries. As a result, three additional Bursaphe-lenchus species, new for Romania, have been identified. In this context, we provide a morphological and molecular charac-terization of the newly found Bursaphelenchus species.