Pinewood nematode population growth in relation to pine phloem chemical composition

The pinewood nematode, Bursaphelenchus xylophilus, native to North America (NA), is now a major invasive pine pest in Eurasia. Pine species differ greatly in their susceptibility to B. xylophilus, and differences in susceptibility might be related to differences in phytochemistry. This study addressed two major questions: (i) which plant chemical profiles (including compounds that act as plant defences and those important for nutrition) favour pinewood nematode development; and (ii) are NA and Portuguese pinewood nematodes different in their capacity to subsist on plant tissues? For this, B. xylophilus isolates from NA and Portugal were tested on pine phloem and xylem of different pine species. It was found that unaltered pine phloem can sustain the population growth of B. xylophilus, but only in a few cases, and this was clearly related to distinct chemical profiles in some pine species. No evidence was found of evolutionary divergence between B. xylophilus populations in Portugal and NA. The pine species with phloem on which pinewood nematodes could grow tended to have low levels of total phenols, condensed tannins, flavonoids and lignin. Evidence was also found that nutrients such as N may be important.     

Sublethal infection of different pine species by the pinewood nematode

Bursaphelenchus xylophilus, the pinewood nematode and causal agent of pine wilt disease (PWD), is a globally destructive pine pest. PWD is favoured by high temperatures and drought, but B. xylophilus can also persist in symptomless hosts in areas not prone to PWD. We compared seedlings of different Pinus species that were inoculated with B. xylophilus when well-watered and under moderate temperatures, conditions that favour sublethal infections. We compared isolates of B. xylophilus in south-west Europe (Portugal) and south-east USA (Louisiana). P. pinea, P. pinaster, and P. radiata were challenged in both areas with inoculations of local B. xylophilus. P. sylvestris in Portugal and P. palustris in Louisiana were also tested. Seedling mortality was low, as expected, but nematode establishment was common. There were higher nematode densities and more damage in seedlings of P. sylvestris and P. radiata, followed by P. pinaster, with P. pinea and P. palustris proving to be more tolerant. Life history of hosts was a better predictor of resistance to B. xylophilus than geographical origin, even under sublethal conditions. Fast-growing species (P. radiata and P. pinaster) appeared most likely to be symptomless carriers in areas not prone to the development of PWD. Cold-adapted species (P. sylvestris) may still be suitable hosts for the pathogen, even at the sublethal conditions of their distribution areas. There were more nematodes per gram of host using nematode isolates from Portugal. Nonetheless, our results opposed the hypothesis that North American pinewood nematodes have such low virulence that they are obligatorily saprophytic.     

Alternative species to replace Monterey pine plantations affected by pitch canker caused by Fusarium circinatum in northern Spain

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.     

Occurrence and ITS diversity of wood‐associated Bursaphelenchus nematodes in Scots pine forests in Switzerland

The quarantine pathogen Bursaphelenchus xylophilus (pine wood nematode, PWN) represents a serious threat for Pinus species in Europe. To exclude its presence in Switzerland, in 2010 and 2011 a countrywide survey was conducted in 102 Pinus sylvestris stands, chosen according to whether they contained dying or dead trees or were located in areas at risk of PWN introduction. In total, 285 trees (1–5 per site) were sampled. Nematodes were extracted from wood chips using a standard procedure, and identified to species by internal transcribed spacer (ITS) sequencing. Bursaphelenchus species were present in 34% of the trees, but no B. xylophilus was identified, i.e. PWN is still not present in Switzerland. The nematodes found belonged to seven different species, with B. vallesianus the most frequent species, followed by B. sexdentati, B. mucronatus kolymensis and B. eggersi. Three other species (B. borealis, B. pinophilus, B. poligraphi) were each only present in one or two trees. Three groups of sequences could not be assigned to a species because of the lack of matching reference sequences. The species composition found in Switzerland suggests co‐existence of southern and central European Bursaphelenchus species. Intraspecific ITS variability differed considerably among the four most common species. Bursaphelenchus eggersi, B. mucronatus kolymensis and B. sexdentati had several variable sites in the ITS region, resulting in multiple ITS genotypes in each species. In contrast, all 99 B. vallesianus isolates had an identical ITS region. This could indicate a founder effect, and possibly that B. vallesianus is not native to the Alpine region.     

Susceptibility evaluation of Picea abies and Cupressus lusitanica to the pine wood nematode (Bursaphelenchus xylophilus)

Pine wilt disease (PWD), recently introduced into Europe, is caused by the pine wood nematode (PWN) Bursaphelenchus xylophilus and is a devastating illness that affects mainly pine trees. It is known that the PWN is capable of infecting other conifers; however, there is currently no information on which other plant species may be susceptible to PWD. In this study, the potential susceptibility of two common species of European forests, Picea abies and Cupressus lusitanica, to PWN was assessed through the monitoring of visual external symptoms, dimension and localization of the nematode population in stems, quantification of total chlorophyll, total soluble phenolics and lignin, at 7, 14, 21 and 28 days after inoculation. The degree of susceptibility was established through the comparison of symptoms with Pinus pinaster, a well‐known PWN host. Furthermore, the stem ultrastructure of P. abies, C. lusitanica and Pn. pinaster was analysed by scanning electron microscopy. The results suggest that P. abies and C. lusitanica are resistant to PWN, and that lignin biosynthesis in these species is affected at an early stage of the infestation. Nevertheless, P. abies seems to be a compatible host that could act as a repository for PWN.     

Growth environment and seedling age affect constitutive and inducible defence in radiata pine

There is increasing interest in induced resistance for crop protection. This study investigated effects of growth environment on inducible defences in developing radiata pine (Pinus radiata) seedlings. Seedlings were grown in the presence or absence of Trichoderma in either a temperature-controlled growth room or in a greenhouse. After 3, 5 or 7 months, seedlings were sprayed with 2.25 mm methyl jasmonate (MJ), 1 week before quantification of monoterpenes and total phenolics. The magnitude of induction of monoterpenes and total phenolics by MJ was inversely proportional to constitutive content, was greater in the greenhouse than the growth room and was greater at 3 months than at 7 months. This may indicate a cost-saving strategy to limit defence induction once constitutive defence reaches a certain threshold. Growth room seedlings expressed greater constitutive resistance to terminal crook (Colletotrichum acutatum) and to diplodia dieback (Diplodia sapinea) than greenhouse plants, consistent with the relative differences in constitutive defence chemistry. Indeed, while MJ induced resistance to diplodia dieback in both environments, there was no difference in terminal crook incidence (4.1%) between MJ-treated and untreated seedlings in the growth room. By the end of the study, growth room seedlings had c. 70% more β-pinene in stems and 140% more total phenolics in needles than greenhouse plants but were 40% smaller, indicating a defence–growth trade-off. Trichoderma did not have a significant effect on defence chemistry or on pathogen resistance. These results demonstrate the potential to manipulate defence in radiata pine by modification of the growth environment.     

Aleppo pine provenances vary in susceptibility and secondary chemical response to Gremmeniella abietina infection

Aleppo pine is a thermophilic species that has become a popular tree species in plantings even beyond its natural habitat. In 1999 the pathogenic fungus Gremmeniella abietina was isolated for the first time in Aleppo pine plantations in Spain. The main objective of this study was to analyse the variation in susceptibility to G. abietina (European race, biotype A) infections among trees from five Aleppo pine provenances. Artificial inoculations were performed at two different times in January 2012. The susceptibility of the trees was evaluated by recording disease severity and the length of internal stem necrosis produced by the pathogen. In addition, the concentrations of two flavanone compounds were quantified as putative indicators of resistance. Provenances and G. abietina isolates both significantly affected levels of necrosis although the results were dependent on the time of inoculation. There was a relationship between provenance and resistance, with trees of higher elevation provenances being more susceptible to disease than trees of lower elevation provenances. The amount of naringenin flavanone from trees was significantly different between provenances, suggesting that it is a possible indicator value for resistance. Conversely, no differences were found in the concentration of eriodictyol in trees between provenances. The results show that the provenance‐dependent variation in the susceptibility of Aleppo pine to G. abietina should be considered in selection of seed sources for restoration.     

Differences in MAT gene distribution and expression between Rhynchosporium species on grasses

Leaf blotch is a globally important disease of barley crops and other grasses that is caused by at least five host‐specialized species in the fungal genus Rhynchosporium. The pathogen R. commune (specialized to barley, brome‐grass and Italian ryegrass) has long been considered to reproduce only by asexual means, but there has been accumulating evidence for recombination and gene flow from population genetic studies and the detection of complementary MAT1‐1 and MAT1‐2 isolates in a c. 1:1 ratio in the field. Here, it is demonstrated that 28 isolates of the closely related species R. agropyri (on couch‐grass) and R. secalis (on rye and triticale), collected from Europe, were also either of MAT1‐1 or MAT1‐2 genotype and that the distribution of mating types did not deviate significantly from a 1:1 ratio. Evidence is then provided for MAT1‐1‐1 and MAT1‐2‐1 gene expression during mycelial growth for all three species. By contrast, 27 isolates of the more distantly related R. orthosporum (on cocksfoot) and R. lolii (on Italian and perennial ryegrasses) from Europe were exclusively of the MAT1‐1 genotype, and expression of the MAT1‐1‐1 gene could not be detected during mycelial growth. These data suggest that cryptic sexual cycles are more likely to exist for R. commune, R. agropyri and R. secalis than for either R. orthosporum or R. lolii. A phylogenetic analysis of partial MAT1‐1 idiomorph sequences resolved these five species into two distinct groups (R. commune, R. agropyri and R. secalis versus R. orthosporum and R. lolii) but provided only limited resolution within each group.     

Resistance to Sclerotinia sclerotiorum in wild Brassica species and the importance of Sclerotinia subarctica as a Brassica pathogen

Brassica crops are of global importance, with oilseed rape (Brassica napus) accounting for 13% of edible oil production. All Brassica species are susceptible to sclerotinia stem rot caused by Sclerotinia sclerotiorum, a generalist fungal pathogen causing disease in over 400 plant species. Generally, sources of plant resistance result in partial control of the pathogen although some studies have identified wild Brassica species that are highly resistant. The related pathogen S. subarctica has also been reported on Brassica but its aggressiveness in relation to S. sclerotiorum is unknown. In this study, detached leaf and petiole assays were used to identify new sources of resistance to S. sclerotiorum within a wild Brassica ‘C genome’ diversity set. High‐level resistance was observed in B. incana and B. cretica in petiole assays, whilst wild B. oleracea and B. incana lines were the most resistant in leaf assays. A B. bourgeai line showed both partial petiole and leaf resistance. Although there was no correlation between the two assays, resistance in the detached petiole assay was correlated with stem resistance in mature plants. When tested on commercial cultivars of B. napus, B. oleracea and B. rapa, selected isolates of S. subarctica exhibited aggressiveness comparable to S. sclerotiorum indicating it can be a significant pathogen of Brassica. This is the first study to identify B. cretica as a source of resistance to S. sclerotiorum and to report resistance in other wild Brassica species to a UK isolate, hence providing resources for breeding of resistant cultivars suitable for Europe.     

Substantial heritable variation for susceptibility to Dothistroma septosporum within populations of native British Scots pine (Pinus sylvestris)

The threat from pests and pathogens to native and commercially planted forest trees is unprecedented and expected to increase under climate change. The degree to which forests respond to threats from pathogens depends on their adaptive capacity, which is determined largely by genetically controlled variation in susceptibility of the individual trees within them and the heritability and evolvability of this trait. The most significant current threat to the economically and ecologically important species Scots pine (Pinus sylvestris) is dothistroma needle blight (DNB), caused by the foliar pathogen Dothistroma septosporum. A progeny‐population trial of 4‐year‐old Scots pine trees, comprising six populations from native Caledonian pinewoods each with three to five families in seven blocks, was artificially inoculated using a single isolate of D. septosporum. Susceptibility to D. septosporum, assessed as the percentage of non‐green needles, was measured regularly over a period of 61 days following inoculation, during which plants were maintained in conditions ideal for DNB development (warm; high humidity; high leaf wetness). There were significant differences in susceptibility to D. septosporum among families indicating that variation in this trait is heritable, with high estimates of narrow‐sense heritability (0.38–0.75) and evolvability (genetic coefficient of variation, 23.47). It is concluded that native Scots pine populations contain sufficient genetic diversity to evolve lower susceptibility to D. septosporum through natural selection in response to increased prevalence of this pathogen.     

Age‐related susceptibility of Eucalyptus species to Phytophthora boodjera

Phytophthora boodjera is a newly described pathogen causing damping off and mortality of Eucalyptus seedlings in Western Australian nurseries. This study evaluated the age‐related susceptibility of several taxa of mallee Eucalyptus to P. boodjera in sterilized washed river sand‐infestation pot trials. Phytophthora cinnamomi and P. arenaria were included for comparison. Seedlings of Eucalyptus taxa were inoculated at 0, 2, 4, 12 and 88 weeks with individual Phytophthora isolates. Pre‐emergent mortality in the presence of Phytophthora was almost 100%. Post‐emergent mortality was 50–100%, depending on isolate, compared to 0% for the control. Mortality was also high for inoculated 1 month‐old seedlings (46–68%) and root length of surviving seedlings was severely reduced. Death from root infection was not observed for seedlings inoculated at 12 and 88 weeks, but they developed root necrosis and reduced root dry weight compared to non‐inoculated controls. Phytophthora boodjera is a pre‐ and post‐emergent pathogen of mallee eucalypts. These eucalypts are susceptible to P. boodjera at all life stages tested, but the mortality rates declined with plant age. Similar results were obtained for P. cinnamomi and P. arenaria. The events leading to its recent appearance in the nurseries remain unknown and further investigations are underway to determine if this is an introduced or endemic pathogen. The approach used here to understand the impact of a Phytophthora species on multiple hosts at different seedling ages is novel and sets a benchmark for future work.     

Comparison of root and foliar applications of potassium silicate in potentiating post‐infection defences of melon against powdery mildew

The application of silicon to the roots or leaves reduces the severity of powdery mildew (Podosphaera xanthii) in melon but the latter treatment is less effective. This study compared key biochemical defence responses of melon triggered by P. xanthii after root or foliar treatment with potassium silicate (PS). Treatments consisted of pathogen‐inoculated or mock‐inoculated plants supplied with PS via roots or foliarly, as well as a non‐treated control. The activity of defence enzymes and the concentration of phenolic compounds, lignin and malondialdehyde were determined from leaf samples at different time points after inoculation. Pathogen‐inoculated plants irrigated with PS showed both an accumulation of silicon and primed defence responses in leaves that were not observed in pathogen‐inoculated plants either sprayed with PS or not treated. These responses included the anticipated activity of peroxidase and accumulation of soluble phenols, the activation of chitinase and repression of catalase, and the stronger activation of superoxide dismutase, peroxidase and β‐1,3‐glucanase. Moreover, the lignin concentration increased in response to inoculation, whereas the malondialdehyde concentration decreased. For the foliar treatment, however, only an increase in lignin deposition was observed compared with the control plants. The results show that silicon strongly plays an active role in modulating the defence responses of melon against P. xanthii when supplied to the roots as opposed to the foliage.     

Pathogenic variation of Mycosphaerella species infecting banana and plantain in Nigeria

Mycosphaerella species that cause the ‘Sigatoka disease complex’ account for significant yield losses in banana and plantain worldwide. Disease surveys were conducted in the humid forest (HF) and derived savanna (DS) agroecological zones from 2004 to 2006 to determine the distribution of the disease and variation among Mycosphaerella species in Nigeria. Disease prevalence and severity were higher in the HF than in the DS zone, but significant (P < 0·001) differences between agroecological zones were only observed for disease severity. A total of 85 isolates of M. fijiensis and 11 isolates of M. eumusae were collected during the survey and used to characterize the pathogenic structure of Mycosphaerella spp. using a putative host differential cultivar set consisting of Calcutta‐4 (resistant), Valery (intermediate) and Agbagba (highly susceptible). Area under disease progress curve (AUDPC) was higher on all cultivars when inoculated with M. eumusae than with M. fijiensis, but significant (P < 0·05) differences between the two species were only observed on Valery. Based on the rank‐sum method, 8·3% of the isolates were classified as highly aggressive and 46·9% were classified as aggressive. About 11·5% of all the isolates were classified as least aggressive, and all of these were M. fijiensis. The majority of M. eumusae isolates (seven out of 11; 64%) were classified as aggressive. A total of nine pathotype clusters were identified using cluster analysis of AUDPC. At least one M. fijiensis isolate was present in all the nine pathotype clusters, while isolates of M. eumusae were present in six of the nine clusters. Isolates in pathotype clusters III and V were the most aggressive, while those in cluster VIII were the least aggressive. Shannon’s index (H) revealed a more diverse Mycosphaerella collection in the DS zone (H = 1·81) than in the HF (H = 1·50) zone, with M. fijiensis being more diverse than M. eumusae. These results describe the current pathotype structure of Mycosphaerella in Nigeria and provide a useful resource that will facilitate screening of newly developed Musa genotypes for resistance against two important leaf spot diseases of banana and plantain.     

Relationships between morphological traits and resistance to pine wood nematode in two Japanese pines

It has been suggested that certain morphological traits of Japanese pines function as resistance factors against the pine wood nematode (PWN), Bursaphelenchus xylophilus. The aim of this study was to verify this concept, and to determine the relationships between traits and resistance at both the individual and family levels. Relationships between survival rate and morphological traits of Pinus densiflora and Pinus thunbergii were studied after inoculation with PWN. Morphometric parameters used were height, basal diameter of the axis, and the number of branches from the axis. Three indices of relationships were analysed: (1) among individuals within a family (survival difference between smaller and larger morphometric populations within each family); (2) among individuals (correlation between each morphometric average and survival rate in quintile populations of all subjects within each pine species based on the order of morphometric values); and (3) among families (correlation between each morphometric average and survival rate in the family). Both significant and non-significant differences were detected within a family, indicating a need for macro techniques. Among individuals, a thicker axis and more branches were correlated with increased survival, indicating that they are resistance factors. However, these correlations were not significant among different families. These results indicate that morphological traits cannot be used to evaluate resistance at a family level, even if they are significant at an individual level. Further studies on traits associated with resistance should be conducted at a family rather than an individual level, to breed for Japanese pines with resistance to PWN.

Testing and modelling the potential of three diploid plants in Poaceae as a new pathosystem to investigate the interactions between cereal hosts and cereal cyst nematode (Heterodera avenae)

Cereal cyst nematode (CCN), Heterodera avenae, is one of the most important pathogens of wheat worldwide, and causes significant yield losses. Research on CCN–wheat interactions is hampered by the lack of an effective model pathosystem. This study investigated the potential of the model cereal Brachypodium distachyon (Bd21‐3) and diploid wheat 2A (G1812) and 2D (AL8/78) as model hosts for CCN. Nematode infection analysis showed that although some CCN penetrated Bd21‐3 roots, these nematodes failed to develop to the later developmental stages or form cysts, indicating B. distachyon is not a host for CCN. A strong burst of reactive oxygen species (ROS) within Bd21‐3 roots infected with CCN was induced 3 days after infection and the expression of seven ROS‐producing genes was significantly increased. In contrast, CCN completed its life cycle in both diploid wheat 2A and 2D, and formed normal syncytia in these hosts. Although CCN developmental processes within both diploid wheat 2A and 2D were very similar to those in the susceptible control, the number of cysts formed on diploid wheat 2D was less than those formed on diploid wheat 2A and the susceptible control, indicating that diploid wheat 2A was a more suitable host for CCN than 2D. This is the first report of a potential new pathosystem for CCN–host interactions using diploid wheat.     

In vitro and in planta nematicidal activity of Fumaria parviflora (Fumariaceae) against the southern root‐knot nematode Meloidogyne incognita

Root and stem extracts of Fumaria parviflora showed strong nematicidal activity against Meloidogyne incognita in in vitro and in planta experiments. Phytochemical screening of F. parviflora revealed the presence of seven classes of bioactive compounds (alkaloids, flavonoids, glycosides, tannins, saponins, steroids and phenols). Quantitative determination of the plant extracts showed the highest percentages of alkaloids (0·9 ± 0·04) and saponins (1·3 ± 0·07) in the roots and total phenolic contents in the stem (16·75 ± 0·07 μg dry g^?1). The n‐hexane, chloroform, ethyl acetate and methanol extracts of roots and stems at concentrations of 3·12, 6·24, 12·5, 25·0 and 50·0 mg mL^?1, significantly inhibited hatching and increased mortality of second‐stage juveniles (J2s) compared with water controls. Percentage J2 mortality and hatch inhibition were directly related to exposure time. In pot trials with tomato cv. Rio Grande, root and stem extracts at concentrations of 1000, 2000 and 3000 ppm, applied as soil drenches, significantly reduced the number of galls, galling index, eggs masses, eggs and reproduction factor compared with the water control. Regardless of concentration, all the extracts significantly increased the host plant growth parameters studied. The n‐hexane extracts from the roots and stem were the most active, followed by the methanol ones, at all concentrations. The in vitro and in planta results suggest that extracts from the roots and stem of F. parviflora may be potential novel nematicides.     

Isolation and characterization of Streptomyces species from potato common scab lesions in Norway

The present survey was conducted to isolate and characterize Streptomyces species from common scab lesions of potato in Norway. Bacteria were isolated from scab lesions on tubers sampled in two consecutive years at different locations in Norway spanning ～1400 km from south to north. In total, 957 independent isolations from individual tubers were performed, with 223 putative pathogenic isolates obtained from 29 different potato cultivars and 130 different fields. Streptomyces europaeiscabiei was the most abundant species isolated from common scab lesions (69%), while 31% of the isolates obtained were S. turgidiscabies. Streptomyces scabies was not found. Pathogenicity of selected Streptomyces isolates was tested on potato. The ability of the bacterial isolates to infect potato was consistent with the presence of the txtAB operon. The results revealed no pattern in geographical distribution of S. europaeiscabiei and S. turgidiscabies; both could be found in the same field and even the same lesion. Four different pathogenicity island (PAI) genotypes were detected amongst the txtAB positive isolates: nec1+/tomA+, nec1–/tomA+, nec1+/tomA? and nec1?/tomA?. The current findings demonstrate that there is genetic variability within species and that the species are not spread solely by clonal expansion. This is thought to be the most comprehensive survey of Streptomyces species that cause common scab of potato in a European country.     

Virulence of Pectobacterium carotovorum subsp. brasiliense on potato compared with that of other Pectobacterium and Dickeya species under climatic conditions prevailing in the Netherlands

In western Europe, Pectobacterium carotovorum subsp. brasiliense is emerging as a causal agent of blackleg disease. In field experiments in the Netherlands, the virulence of this pathogen was compared with strains of other Dickeya and Pectobacterium species. In 2013 and 2014, seed potato tubers were vacuum infiltrated with high densities of bacteria (10⁶ CFU mL^?1) and planted in clay soil. Inoculation with P. carotovorum subsp. brasiliense and P. atrosepticum resulted in high disease incidences (75–95%), inoculation with D. solani and P. wasabiae led to incidences between 5% and 25%, but no significant disease development was observed in treatments with P. carotovorum subsp. carotovorum, D. dianthicola or the water control. Co‐inoculations of seed potatoes with P. carotovorum subsp. brasiliense and D. solani gave a similar disease incidence to inoculation with only P. carotovorum subsp. brasiliense. However, co‐inoculation of P. carotovorum subsp. brasiliense with P. wasabiae resulted in a decrease in disease incidence compared to inoculation with only P. carotovorum subsp. brasiliense. In 2015, seed potatoes were inoculated with increasing densities of P. carotovorum subsp. brasiliense, D. solani or P. atrosepticum (10³–10⁶ CFU mL^?1). After vacuum infiltration, even a low inoculum density resulted in high disease incidence. However, immersion without vacuum caused disease only at high bacterial densities. Specific TaqMan assays were evaluated and developed for detection of P. carotovorum subsp. brasiliense, P. wasabiae and P. atrosepticum and confirmed the presence of these pathogens in progeny tubers of plants derived from vacuum‐infiltrated seed tubers.     

Effect of Trichoderma viride pre-inoculation in pine species with different levels of susceptibility to Fusarium circinatum: physiological and hormonal responses

Pine pitch canker (PPC), caused by Fusarium circinatum, affects Pinus species worldwide. Although no effective solutions have yet been found to control it, there is a growing interest in using biological control agents (BCA) such as Trichoderma to avoid the application of chemical-based products. Using species with an increasing level of susceptibility to PPC (Pinus pinea, Pinus pinaster and Pinus radiata), this study aimed to evaluate the effect of Trichoderma viride pre-inoculation on disease development, assessing several physiological and hormonal parameters. A 2-week period elapsed between T. viride and F. circinatum inoculation. Sampling for each species was performed independently when at least 50% of the plants of one of the inoculated groups developed disease symptoms. Fusarium circinatum infection reduced water status and photosynthesis, but increased proline, abscisic acid and jasmonic acid concentrations in plants of P. radiata and P. pinaster with symptoms; while in P. pinea water relations were maintained and anthocyanin accumulation occurred in the presence of F. circinatum. In P. radiata, T. viride pre-inoculation accelerated disease progression, with some PPC-induced responses augmented (decreased water potential and photosynthesis; increased substomatal CO₂ concentration) and novel changes not found in seedlings inoculated exclusively with F. circinatum (increased electrolyte leakage and salicylic acid; decreased relative water content). This suggests that T. viride may be initially recognized as an invading organism, subverting the plant defence mechanisms for successful root colonization. If seedlings are not allowed to recover from this state, pathogen infection may thus be facilitated, highlighting the importance of application timing in BCA strategies.     

Fusarium species and chemotypes associated with fusarium head blight and fusarium root rot on wheat in Sardinia

Environmental conditions in Sardinia (Tyrrhenian Islands) are conducive to fusarium root rot (FRR) and fusarium head blight (FHB). A monitoring survey on wheat was carried out from 2001 to 2013, investigating relations among these diseases and their causal agents. FHB was more frequently encountered in the most recent years while FRR was constantly present throughout the monitored period. By assessing the population composition of the causal agents as well as their genetic chemotypes and EF‐1α polymorphisms, the study examined whether the two diseases could be differentially associated to a species or a population. Fusarium culmorum chemotypes caused both diseases and were detected at different abundances (88% 3‐ADON, 12% NIV). Fusarium graminearum (15‐ADON genetic chemotype) appeared only recently (2013) and in few areas as the causal agent of FHB. In F. culmorum, two haplotypes were identified based on an SNP mutation located 34 bp after the first exon of the EF‐1α partial sequence (60% adenine, 40% thymine); the two populations did not segregate with the chemotype but the A‐haplotype was significantly associated with FRR in the Sardinian data set (P = 0·001), suggesting a possible fitness advantage of the A‐haplotype in the establishment of FRR that was neither dependent on the sampling location nor the sampling year. The SNP determining the Sardinian haplotype is distributed worldwide. The question whether the A‐haplotype segregates with characters facilitating FRR establishment will require further validation on a specifically sampled international data set.