No abstract

Pea plants grown in the field were used to study the natural incidence of endophytic bacteria in the stem. Eleven pea cultivars at the flowering stage were screened for the presence of endophytic bacteria using a printing technique with surface disinfested stem cross-sections on 5% Trypticase Soy Agar (TSA). Five cultivars showed colonization. Cultivar Twiggy showed the highest and most consistent colonization and was further investigated. Stems of cv. Twiggy at the pod stage were analyzed for endophytic bacterial types and populations. Cross-sections of surface disinfested stems were printed on 5% TSA. Endophytic bacterial populations decreased from the lower to the upper part of the stem. One section from the third and the fourth internode was surface disinfested, homogenized, and spiral plated on the media 5% TSA, R2A, and SC (Davis et al. 1980). Over a series of 30 samples, 5% TSA gave significantly better recovery of bacterial endophytes compared with R2A and SC media. For most stems, populations ranged from 10(4) to 10(5) CFU/g except in one of the field blocks in which endophyte populations were uniformly higher. Comparison of colony counts by spiral plating and printing showed a positive correlation. The most frequently recovered bacterial types were Pantoea agglomerans and Pseudomonas fluorescens. Less frequently isolated were Pseudomonas viridiflava and Bacillus megaterium.

Red band needle blight caused by Dothistroma septosporum and D. pini, and brown spot needle blight caused by Lecanosticta acicola provoke severe and premature defoliation in Pinus, and subsequent reduction of photosynthetic surfaces, vitality, and growth in young and adult trees. The recurrent damage results in branch and tree death. Until recently, pine needle blight diseases have had only minor impacts on native and exotic forest trees in the North of Spain, but in the past five years, these pathogen species have spread widely and caused severe defoliation and mortality in exotic and native plantations of Pinus in locations where they were not detected before. In an attempt to understand the main causes of this outbreak and to define the effectiveness of owners' management strategies, four research actions were implemented: a survey of the management activities implemented by the owners to reduce disease impact, the evaluation of specific symptoms and damage associated with infection, and the identification of the causative pathogenic species and their reproductive capacity. Morphological characteristics of the fungus and molecular identification were consistent with those of Lecanosticta acicola and Dothistroma spp., D. septosporum, D. Pini, and both mating types were present for the three identified pathogens. The local silvicultural management performed, mainly pruning and thinning, was not resulting in the expected improvement. The results of this study can be applied to establish guidelines for monitoring and controlling the spread of needle blight pathogens.

There is a concern on how emerging pests and diseases will affect the distribution range and adaptability of their host species, especially due to different conditions derived from climate change and growing globalization. Fusarium circinatum, which causes pitch canker disease in Pinus species, is an exotic pathogen of recent introduction in Spain that threatens its maritime pine (P. pinaster) stands. To predict the impact this disease will have on the species, we examine host resistance traits and their genetic architecture. Resistance phenotyping was done in a clonal provenance/progeny trial, using three-year-old cuttings artificially inoculated with the pathogen and maintained under controlled environmental conditions. A total number of 670 ramets were assessed, distributed in 10 populations, with a total of 47 families, 2 to 5 half-sibs per family, and 3–7 ramets per clone. High genetic variation was found at the three hierarchical levels studied: population, family and clone, being both additive and non-additive effects important. Narrow-sense and broad-sense heritability estimates were relatively high, with respective values of 0.43–0.58 and 0.51–0.8, depending on the resistance traits measured (lesion length, lesion length rate, time to wilting, and survival). These values suggest the species' high capacity of evolutionary response to the F. circinatum pathogen. A population originated in Northern Spain was the most resistant, while another from Morocco was the most susceptible. The total number of plants that did not show lesion development or presented a small lesion (length<30 mm) was 224 out of 670, indicating a high proportion of resistant trees in the offspring within the analyzed populations. We found large differences among populations and considerable genetic variation within populations, which should allow, through natural or artificial selection, the successful adaptation of maritime pine to 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.

Fusarium circinatum was recently detected as an endophyte in grasses causing no apparent damage. Our goal was to describe the endophytic colonization of herbaceous host plants growing in a plantation of Pinus radiata with symptoms of pitch canker disease, which may act as reservoir of inoculum. We detected the fungus in five species of dicots families (Asteraceae, Lamiaceae, Rosaceae), besides two species of Poaceae. It was found in the aerial part of nonsymptomatic hosts, so we describe F. circinatum as an endophyte mainly transmitted by spores through the air. However, vertical transmission occurs at least in Hypochaeris radicata. SSR markers analyses showed a unique haplotype regardless isolates origin, pine cankers or nonsymptomatic herbaceous plants. Thus, the same genotype can adopt a pathogenic or endophytic lifestyle. Non-symptomatic plants can act as reservoir of inoculum: pine seedlings can be infected from senescent tissue of non-symptomatic host colonized by the fungus.

Survival of Fusarium circinatum in colonized pine needles and wood pieces was measured. Naturally colonized branches and their needles were cut into small pieces and placed in mesh bags on the soil surface at two locations in northern Spain. Pieces were recovered periodically, cultured on a selective medium, and microscopically examined to identify the species. After 507 days, F. circinatum was recovered from 0 to 27% of the wood pieces and from none of the needles. After 858 days, F. circinatum was not recovered from any wood pieces but was found to be present on 1 out of 220 needle pieces analysed. Artificially infested pieces of wood and needles were placed on 5-mm sieved soil either in plastic boxes at controlled temperature or in mesh bags under field conditions. No survival was recorded after 794 days under field conditions and the decline over time occurred more rapidly in inoculated pieces under field conditions. Soil was also infested with conidia of F. circinatum and survival was estimated. No conidia were recovered after 224 days at 30°C, although at 20 and 5°C the respective populations were 20 and 3700 cfu/g soil. Fusarium circinatum was not recovered from 2-mm-sieved soil collected under pitch canker-infected pines. Results indicate that branch segments and needles naturally colonized by F. circinatum will not be a potential source of inoculum, and the fungus in soil is not likely to contribute to reinfection of new plantations after 2 years.

Pitch canker, caused by the fungus Fusarium circinatum, is a major disease of Pinus radiata currently controlled to some extent in nurseries by good hygiene and application of synthetic fungicides. The aim of this study was to evaluate alternative strategies to control fungal infections in nurseries and young pine plantations. The antagonistic effects of biocontrol bacteria and essential oils against F. circinatum in vitro and in young P. radiata trees were assessed. Pseudomonas fluorescens, Erwinia billingiae, and Bacillus simplex reduced the growth of the fungus in vitro by 17%-29%, and decreased the density of the mycelial mat. In young P. radiata trees, the length of F. circinatum lesions was reduced by 22%-25% by the same bacterial strains. Direct application of cinnamon and/or clove essential oils to wounds in stems of two-year-old P. radiata trees also limited the damage caused by F. circinatum. Lesion length was reduced by 51% following treatment with cinnamon oil (10% v/v), and by 45% following treatment with clove oil (15% v/v) or a combination of both oils. However, the oils were toxic to younger trees. The biocontrol bacteria and essential oils show promise as prophylactic treatments to reduce the devastating effects of F. circinatum on P. radiata.