Joana Henriques

Efficacy of sulfuryl fluoride against the pinewood nematode, Bursaphelenchus xylophilus (Nematoda: Aphelenchidae), in Pinus pinaster boards

BACKGROUND The pinewood nematode (PWN) Bursaphelenchus xylophilus is an important conifer disease worldwide. It is the direct cause of the death of millions of pines in south-east Asia (mainly Japan, China and Korea) and has been established in Portugal since 1999. The phasing out of methyl bromide has created an urgent need for alternative treatment of wood packaging materials. The effect of sulfuryl fluoride (SF), a broad-spectrum fumigant used to control insects, was tested in Pinus pinaster boards naturally infested by PWN. RESULTS Boards were fumigated for 24 h at three different temperatures (15, 20 and 30 °C) with dosage ranges of 3169-4407, 1901-4051 and 1385-2141 gh m(-3) respectively. Treated wood was sampled for nematode identification and counting, before treatment and after 24 h, 72 h and 21 days. No survival was found in the 15 °C and 30 °C treatments, while at 20 °C the mortality ranged from 94.06 to 100%. Some reasons for the survival at 20 °C are presented. CONCLUSION Results confirm SF to be an effective quarantine treatment for PWN at 15 and 30 °C. Further studies are needed to obtain the most effective dosage at 20 °C, and to determine the toxicity of SF fumigation on B. xylophilus at other temperatures, especially at 25 °C.

Ambrosia fungi in the insect-fungi symbiosis in relation to cork oak decline

Ambrosia fungi live associated with beetles (Scolytidae and Platypodidae) in host trees and act as a food source for the insects. The symbiotic relation is important to the colonizing strategies of host trees by beetles. Ambrosia fungi are dimorphic: they grow as ambrosial form and as mycelium. The fungi are highly specialized, adapted to a specific beetle and to the biotope where they both live. In addition other fungi have been found such as tree pathogenic fungi that may play a role in insects host colonization success. Saprophytic fungi are also present in insects galleries. These may decompose cellulose and/or be antagonistic to other less beneficial fungi. This paper summarizes the importance of ambrosia fungi and the interaction with insects and hosts. The possibility of the transport of pathogenic fungi by Platypus cylindrus to cork oak thus contributing for its decline is discussed.

Diversity of Biscogniauxia mediterranea within Single Stromata on Cork Oak

Charcoal canker, caused by the fungus Biscogniauxia mediterranea, is one of the most frequent diseases of cork oak in Portugal. The pathogen has been considered a secondary invader that attacks only stressed hosts; however, in recent years, an increasing number of young trees exhibiting the disease symptoms have been recorded. A collection of monoascosporic cultures isolated from single stromata of B. mediterranea in cork oak from different locations was analyzed by means of microsatellite—Primed Polymerase Chain Reaction—using three microsatellite primers, in order to detect the genetic variation of the population thus discussing its plasticity and ability to adapt to different conditions. The results showed a high level of genetic variability among isolates obtained from the same stroma, being impossible to distinguish isolates from individual stromata neither from different geographical location.

Analysis of the genetic diversity and phylogenetic relationships of Biscogniauxia mediterranea isolates associated with cork oak

Biscogniauxia mediterranea is one of the most frequent fungal pathogens involved in cork oak decline in the Mediterranean Basin, causing charcoal canker. In Portugal, this disease is widespread on adult declining trees but nowadays it increasingly affects young trees and exhibits atypical symptoms, leading to the hypothesis that some change in the fungus may have occurred. In order to evaluate the genetic diversity and phylogenetic relationship of B. mediterranea associated with cork oak, 102 isolates were obtained from young and adult trees of Quercus suber and other hosts species with different disease expression, from several Mediterranean countries. The collection of isolates was analyzed by individual and multigene phylogenies using Maximum-Likelihood approach based on nucleotide sequences of the internal transcribed spacers of ribosomal DNA, translation elongation factor 1-α and β-tubulin genes, and by microsatellite-primed PCR profiles. Sequence analyses separated the Mediterranean isolates from those from other regions, while MSP-PCR analysis revealed relevant but unstructured diversity among the Mediterranean isolates under study, making this a monophyletic but diverse population. Considering the adaptive capacity of the fungus in the Mediterranean-climate ecosystems and the present climatic change scenario, all conditions are gathered to favor aggravation of the disease in cork oak stands.

Susceptibility of larvae and adults of Monochamus galloprovincialis to entomopathogenic fungi under controlled conditions

Six entomopathogenic fungal isolates were tested under controlled conditions, as biological control agents against Monochamus galloprovincialis (Coleoptera: Cerambycidae) (Olivier), the vector of the pine wood nematode Bursaphelenchus xylophilus (Nematoda: Parasitaphelenchidae) (Steiner and Buhrer). The fungi Beauveria bassiana (Bals.-Criv.) Vuill, Metarhizium anisopliae (Metchnikoff) Sorokin, Metarhizium sp. and Fusarium sp. were tested against larvae and adults of the insect, by comparing two exposure techniques: continuous contact and spraying. Regarding the larvae, only Fusarium sp. and Metarhizium sp. killed more than 40% of the individuals, while B. bassiana and M. anisopliae were found to kill 50% of the adults (LT50) within five days. Exposure to B. bassiana reduced adult longevity by 19 days, when compared to untreated beetles, and had a direct efficacy of 46%. Continuous contact with the fungi proved to be more effective than spraying. A larval disinfection protocol was developed to eliminate other fungal isolates from the wood-collected tested individuals. The implications of the results and possible applications of selected strains as bio-control agents against M. galloprovincialis are discussed.