The pitch canker pathogen Fusarium circinatum was first found to cause damage in nurseries and pine plantations in northern Spain in 2004. Since then, establishment of pine plantations in the region has decreased as a result of the prohibitions placed on planting Pinus spp. and Pseudotsuga menziesii in areas affected by the disease. However, although most pine species have been found to be susceptible to the pathogen under nursery conditions, little is known about how the fungus affects the trees in the field. Furthermore, it is not known whether some of the native or exotic species commonly planted in the area are also susceptible to F. circinatum. The aim of this study was to evaluate the susceptibility of several conifer species commonly planted in northern Spain to the pitch canker pathogen. For this purpose, two different trials were carried out, one under controlled laboratory conditions and the other in the field. Although most of the conifers were affected by the pathogen in the laboratory tests, only Pinus radiata, Pinus nigra, Pinus pinaster and Pinus uncinata were susceptible to the pathogen in the field.
Fusarium circinatum, a fungus that causes pitch canker disease, has been present in Europe since at least 2003, when it was detected in northern Spain and found to be producing severe damage in tree nurseries and pine plantations. In this study, we tested a method of biological control of the disease with Trichoderma viride, a fungal species successfully used against many other pathogens. In vitro and in vivo assays were carried out to test the efficacy of this antagonist in controlling F. circinatum. The T. viride isolate exerted a significant effect on the growth of F. circinatum in the in vitro assay, reducing the length of the pathogen colony by half. However, although we tested three different concentrations of the T. viride spore solution, no clear conclusions were obtained with regard to the effects on the Pinus radiata seedlings. To our knowledge, this is the first study carried out with the aim of using Trichoderma spp. to control pitch canker disease.
Fusarium circinatum is a highly virulent invasive pathogen that causes the disease commonly known as pine pitch canker (PPC). On mature trees, the most common symptoms of PPC include resin bleeding, resinous cankers, wilting of needles and dieback. In nurseries the main symptoms are yellowish needles, and wilting of foliage and shoots. PPC is considered one of the most important diseases of conifers globally and at least 60 species of Pinus along with Pseudotsuga menziesii are known to be susceptible. Quarantine regulations are crucial to minimize the risk of new introductions into disease‐free countries. However, if these measures fail, implementation of environmentally friendly control methods is currently encouraged in an integrated management approach that minimizes the use of chemicals in forests. This review therefore summarizes current knowledge of biological control using endophytic fungi, bacteria and viruses, and other environmentally friendly methods to control PPC, including thermotherapy, natural products with fungicidal effects, promoters of innate host resistance mechanisms and breeding for resistance. Key research gaps to be addressed for improvement of effective management of this disease are discussed.
Pitch canker is a serious disease of pines caused by the ascomycete fungus Gibberella circinata (anamorph = Fusarium circinatum). Three distinct mitovirus strains have been described in this fungus: Fusarium circinatum mitovirus 1 (FcMV1), FcMV2-1 and FcMV2-2. Here, we investigated the frequency and population variation of these viruses and closely related sequence variants in northern Spain using RT-PCR and sequencing. Each virus strain and similar sequence variants shared >95 % sequence identity and were collectively designated as virus types. All virus types were relatively common in Spain, with estimated prevalence of 18.5 %, 8.9 % and 16.3 % for FcMV1, FcMV2-1 and FcMV2-2, respectively.
Laccase enzymes (benzenediol: oxygen oxidoreductase, EC 1.10.3.2) play a major role in the degradation of phenolic compounds such as lignin. They are common in fungi and have been suggested to participate in host colonization by pathogenic fungi. Putative mycoviruses have recently been isolated from the causal agent of pine pitch canker disease, Fusarium circinatum Nirenberg & O´Donell. In this study, the effects of single and double mycoviral infections on laccase activity, growth rate and pathogenicity were investigated in fourteen F. circinatum strains. Extracellular laccase activity was analyzed by the Bavendamm test, image processing and a spectrophotometric method. Mycelial growth, in vivo pathogenicity and seedling survival probability were also determined in Monterrey pine (Pinus radiata D. Don) seedlings. The findings showed that (i) mycelial growth of isolates from the same fungal population was homogeneous, (ii) the presence of mycovirus appears to increase the virulence of fungal isolates, (iii) co-infection (with two mycoviruses) caused cryptic effects in fungal isolates, and (iv) laccases embody a possible auxiliary tool in fungal infection. The prospects for biocontrol, the adaptive role of F. circinatum mycoviruses and the importance of laccase enzymes in host colonization are discussed.
Fusarium circinatum
is a harmful pathogenic fungus mostly attacking
Pinus
species and also
Pseudotsuga menziesii
, causing cankers in trees of all ages, damping-off in seedlings, and mortality in cuttings and mother plants for clonal production. This fungus is listed as a quarantine pest in several parts of the world and the trade of potentially contaminated pine material such as cuttings, seedlings or seeds is restricted in order to prevent its spread to disease-free areas. Inspection of plant material often relies on DNA testing and several conventional or real-time PCR based tests targeting
F
.
circinatum
are available in the literature. In this work, an international collaborative study joined 23 partners to assess the transferability and the performance of nine molecular protocols, using a wide panel of DNA from 71 representative strains of
F
.
circinatum
and related
Fusarium
species. Diagnostic sensitivity, specificity and accuracy of the nine protocols all reached values >80%, and the diagnostic specificity was the only parameter differing significantly between protocols. The rates of false positives and of false negatives were computed and only the false positive rates differed significantly, ranging from 3.0% to 17.3%. The difference between protocols for some of the performance values were mainly due to cross-reactions with DNA from non-target species, which were either not tested or documented in the original articles. Considering that participating laboratories were free to use their own reagents and equipment, this study demonstrated that the diagnostic protocols for
F
.
circinatum
were not easily transferable to end-users. More generally, our results suggest that the use of protocols using conventional or real-time PCR outside their initial development and validation conditions should require careful characterization of the performance data prior to use under modified conditions (i.e. reagents and equipment). Suggestions to improve the transfer are proposed.
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