Increased bacterial endophyte populations in Nicotiana benthamiana with a branched‐chain alkane mixture

The aim of this study was to determine if treatment of soil with a branched‐chain alkane mixture known to induce resistance against Colletotrichum orbiculare also changes populations of bacterial endophytes from Nicotiana benthamiana. Eight culturable bacterial endophyte types matching six species of Bacillus and two species of Pseudomonas were found in roots, stem + petioles and/or leaves. Application of the branched‐chain alkane mixture resulted in significantly higher endophyte populations compared to the water or emulsifier controls for the Pseudomonas sp. LW3, Bacillus simplex LW4 and Bacillus pumilis LW5 colony types in roots and the B. simplex LW4 colony type in stem + petioles. The Pseudomonas sp. LW3 and B. simplex LW4 colony types also had higher populations in pure cultures under in vitro conditions with the branched‐chain alkane mixture compared to the controls. Inoculation with each of the eight colony types increased their population in the plant and induced resistance against C. orbiculare, with the most effective being Pseudomonas sp. LW3 and Pseudomonas alcaligenes SW1. Most of the endophytes could inhibit C. orbiculare growth in vitro, but the level of resistance in planta was not correlated to the ability of the colony type to inhibit C. orbiculare in culture. Thus, a branched‐chain alkane mixture can selectively affect the biomass of a subset of bacterial endophytes, demonstrating that it is a novel in planta endophyte growth promoter.     

Nematicidal activity of Ochradenus baccatus against the root‐knot nematode Meloidogyne javanica

Ochradenus baccatus is a widely distributed shrub in desert regions of the Middle East and North Africa. This plant's nematicidal activity against the root‐knot nematode Meloidogyne javanica was evaluated because it has been found to contain exceptionally high levels of glucosinolates. In in vitro assays with aqueous extracts of the plant, 100% of second‐stage juveniles were immobilized after exposure to 4% root‐core extract for 48 h; 8% root‐core extract suppressed their hatching by 87%, whereas stem, flower and root bark showed lower activity. Incorporation of root core or bark into the soil, as fresh or dry powder at 1 and 0·5% (w/w), respectively, reduced the number of nematodes recovered from the soil by 95–100%, whereas the flower and stem were much less effective. Results from further pot experiments indicated that only the root bark consistently contains nematicidal compounds which are effective in soil, whereas the nematicidal activity of the root core in soil was inconsistent. The presence of non‐volatile lipophilic and lipophobic nematicidal compounds in the root bark was suggested by extraction with different polar solvents, but these compounds do not seem to be isothiocyanates – glucosinolate‐hydrolysed compounds with nematicidal activity. Very poor host status of Ochradenus baccatus to M. javanica, M. incognita and M. hapla, but with root‐penetration rates of juveniles similar to those in tomato roots, suggest that this plant may be used as a cover plant or trap plant to reduce nematode populations in the soil.     

The influence of formulation on Trichoderma biological activity and frosty pod rot management in Theobroma cacao

Frosty pod rot (FPR), caused by Moniliophthora roreri, is responsible for significant losses in Theobroma cacao. Due to limited options for FPR management, biological control methods using Trichoderma are being studied. Combinations of three formulations and two Trichoderma isolates were studied between May 2009 and April 2011. The formulations were 0·3 mL L^?1 of the surfactant BreakThru 100SL (BT), a mixture of 1% w/v Sure‐Jell (source of pectin) and 1% w/v potato dextrose broth (PDB) (PP), and an invert oil emulsion of 50% v/v corn oil/2·5% w/v lecithin/0·5% w/v PDB (COP). Water and fungicide, copper oxychloride, were included as controls. Humidity chamber studies indicated that Trichoderma conidia germinated in all formulations if free water was maintained, while only the COP formulation supported germination under drying conditions. In the field, Trichoderma ovalisporum DIS‐70a and Trichoderma harzianum DIS‐219f were applied monthly in each of the three formulations at a rate of 180 mL per tree, 2·46 × 10⁷ conidia per mL. The COP/DIS‐70a formulation provided the largest yield increase compared to all other treatments, including the fungicide control. Averaged over the 2 years, the COP formulation increased yield to 30·7% healthy pods compared to 9·7% healthy pods in the water control. Although the formulation/isolate combinations did not consistently increase endophytic colonization, the PP/DIS‐219f, COP/DIS‐219f and COP/DIS‐70a combinations increased total endophytic/epiphytic colonization by Trichoderma. The invert corn oil formulation of DIS‐70a significantly enhanced yield of healthy cacao pods over 2 years providing a promising model for optimizing Trichoderma‐based biocontrol strategies.     

Temporal development and relationship amongst brown rot blossom blight,fruit blight and fruit rot in integrated and organic sour cherry orchards

The aim of this 4‐year study was to characterize temporal development of brown rot blossom blight and fruit blight (caused by Monilinia spp.) and their sporulating areas in sour cherry orchards; and to determine the relationships amongst incidence and sporulating area of blossom blight, fruit blight and fruit rot. The study was performed in integrated and organic orchard blocks on two cultivars (Újfehértói fürtös and Érdi b?term?). On both cultivars, disease progress on flowers and fruits was 2–10 times slower in the integrated than in the organic management system. The peak incidence values were 9 and 31 days after petal fall for blossom blight and fruit blight, respectively. After these dates, no new blight symptoms on flowers and/or fruits appeared and the disease was levelling off. Final blossom blight incidence ranged from 1 to 5% and from 12 to 34%, and fruit rot incidence from 2 to 6% and from 11 to 26% in the integrated and the organic orchards, respectively. The sum of fruit blight incidence ranged from 9 to 22% for the organic system, but was below 5% for the integrated system, while the final sporulating area was 5–16 mm² and <3 mm², respectively. Among the five highest Pearson's correlation coefficients, relationships between blossom blight and early fruit blight stage (r = 0·845, P = 0·0087 integrated; r = 0·901, P = 0·0015 organic), and between sporulating area and fruit rot (r = 0791, P = 0·0199 integrated; r = 0·874, P = 0·0039 organic) were the most significant relationships from an epidemic standpoint as they indicated a connection between different brown rot symptom types.     

Magnesium oxide nanoparticles induce systemic resistance in tomato against bacterial wilt disease

Bacterial wilt is a serious problem affecting many important food crops. Recent studies have indicated that treatment with biotic or abiotic stress factors may increase the resistance of plants to bacterial infection. This study investigated the effects of magnesium oxide nanoparticles (MgO NP) on disease resistance in tomato plants against Ralstonia solanacearum, as well as its antibacterial activity. The roots of tomato seedlings were inoculated with R. solanacearum and then immediately treated with MgO NP; the treated plants showed very little inhibition of bacterial wilt. In contrast, when roots were drenched with a MgO NP suspension prior to inoculation with the pathogen, the incidence of disease was significantly reduced. Rapid generation of reactive oxygen species such as O₂^?˙ radicals was observed in tomato roots treated with MgO NP. Further O₂^?˙ was rapidly generated when tomato plant extracts or polyphenols were added to the MgO NP suspension, suggesting that the generation of O₂^?˙ in tomato roots might be due to a reaction between MgO NP and polyphenols present in the roots. Salicylic acid‐inducible PR1, jasmonic acid‐inducible LoxA, ethylene‐inducible Osm, and systemic resistance‐related GluA were up‐regulated in both the roots and hypocotyls of tomato plants after treatment of the plant roots with MgO NP. Histochemical analyses showed that β‐1,3‐glucanase and tyloses accumulated in the xylem and apoplast of pith tissues of the hypocotyls after MgO NP treatment. These results indicate that MgO NP induces systemic resistance in tomato plants against R. solanacearum.     

Effect of defoliation by livestock on stem canker caused by Leptosphaeria maculans in Brassica napus

Brassica napus (canola, oilseed rape), an important break crop for cereals across the Australian wheat belt, is being rapidly adopted as a dual‐purpose (forage and grain) crop in mixed farming systems. Stem canker caused by the fungus Leptosphaeria maculans is the most important disease of B. napus in Australia. The primary source of inoculum is airborne ascospores released during autumn/winter which coincides with the grazing of dual‐purpose crops. Field experiments were defoliated by sheep to determine the effect of grazing on blackleg stem canker severity at plant maturity in B. napus cultivars differing in their resistance level and grazed at different times. One cultivar was sown on different dates to investigate the impact of grazing at the same time, but at different growth stages. Defoliation by mowing was compared to defoliation by livestock. Similar amounts of dry matter remained after defoliation by machinery (0·66 t ha^?1) or livestock (0·52 t ha^?1). However, stem canker severity was higher in the grazed (40% of crown cross‐section diseased) compared with the mown (25%) treatment, which was higher than the ungrazed control (9%). Stem canker severity generally increased with grazing, but the increase was eliminated or reduced in cultivars with good resistance. Grazing during vegetative plant growth minimized the increase in stem canker severity compared with grazing during reproductive growth. Currently, cultivars with good L. maculans resistance are recommended in high disease situations. To avoid excessive yield loss in dual‐purpose B. napus crops due to L. maculans it is recommended that such cultivars are grown even in low‐moderate disease situations.     

Development and validation of a set of standard area diagrams to aid in estimation of spot blotch severity on wheat leaves

This study aimed to develop and validate a standard area diagram (SAD) set to quantify the severity of spot blotch, caused by Bipolaris sorokiniana, on wheat leaves. The proposed SAD set includes images of leaves with 11 distinct disease severities (0·1, 1, 5, 10, 20, 30, 40, 50, 60, 70 and 83%). The SAD set was validated by 12 raters without experience in evaluating plant disease. Lin's concordance correlation analysis of estimated versus actual disease severity (based on image analysis) showed that precision and accuracy improved for all raters using the SAD set in contrast to assessments made without it. The SAD set improved accuracy (coefficient of bias, C_b = 0·88 and 0·99, without and with the SAD set, respectively) and agreement (Lin's concordance correlation coefficient, ρ_c = 0·81 and 0·96 without and with the SAD set, respectively) of the estimates of severity. The severity estimates were also more reliable when using the SAD set (coefficient of determination, R² = 0·76 unaided and R² = 0·92 with the SAD set, and intra‐class correlation ρ = 0·79 without the SAD set and ρ = 0·95 using the SAD set). The SAD set proposed in this study will improve the accuracy and reliability of estimates of spot blotch severity on wheat leaves.     

Evaluation of different methods for the characterization of carrot resistance to the alternaria leaf blight pathogen (Alternaria dauci) revealed two qualitatively different resistances

Alternaria leaf blight (ALB), caused by Alternaria dauci, is one of the most damaging foliar diseases of carrot worldwide. The aim of this study was to compare different methods for evaluating levels of carrot resistance to ALB. Three techniques were investigated by comparison with a visual disease assessment control: in vivo conidial germination, a bioassay based on a drop‐inoculation method, and in planta quantification of fungal biomass by quantitative PCR (Q‐PCR). Three carrot cultivars showing different degrees of resistance to A. dauci were used, i.e. a susceptible cultivar (Presto) and two partially resistant genotypes (Texto and Bolero), challenged with an aggressive or a very aggressive isolate of A. dauci. Both partially resistant genotypes produced a higher mean number of germ tubes per conidium (up to 3·42±0·35) than the susceptible one (1·26±0·18). The drop‐inoculation results allowed one of the partially resistant genotypes (Bolero, log₁₀(S+1) = 1·34±0·13) to be distinguished from the susceptible one (1·90±0·13). By contrast, fungal growth measured by Q‐PCR clearly differentiated the two partially resistant genotypes with log₁₀(I) values of 2·77±0·13 compared to the susceptible cultivar (3·65±0·13) at 15 days post‐inoculation. This result was strongly correlated (r² = 0·91) with the disease severity index scored at the same date. Data obtained with the different assessment methods strongly suggest that the Texto and Bolero genotypes have different genetic resistance sources.     

The effect of environmental factors on infection of blueberry fruit by Colletotrichum acutatum

Anthracnose fruit rot of blueberries caused by Colletotrichum acutatum is a serious problem in humid blueberry‐growing regions of North America. In order to develop a disease prediction model, environmental factors that affect mycelial growth, conidial germination, appressorium formation and fruit infection by C. acutatum were investigated. Variables included temperature, wetness duration, wetness interruption and relative humidity. The optimal temperature for mycelial growth was 26°C, and little or no growth was observed at 5 and 35°C. The development of melanized appressoria was studied on Parafilm‐covered glass slides and infection was evaluated in immature and mature blueberry fruits. In all three assays, the optimal temperature for infection was identified as 25°C, and infections increased up to a wetness duration of 48 h. Three‐dimensional Gaussian equations were used to assess the effect of temperature and wetness duration on the development of melanized appressoria (R² = 0·89) on Parafilm‐covered glass slides and on infection incidence in immature (R² = 0·86) and mature (R² = 0·90) blueberry fruits. Interrupted wetness periods of different durations were investigated and models were fitted to the response of melanized appressoria (R² = 0·95) and infection incidence in immature (R² = 0·90) and mature (R² = 0·78) blueberry fruits. Additionally, the development of melanized appressoria and fruit infection incidence were modelled in relation to relative humidity (R² = 0·99 and 0·97, respectively). Three comprehensive equations were then developed that incorporate the aforementioned variables. The results lay the groundwork for a disease prediction model for anthracnose fruit rot in blueberries.     

Intraspecific variation in susceptibility to dothistroma needle blight within native Scottish Pinus sylvestris

Dothistroma needle blight (DNB), caused by Dothistroma septosporum, is the most important disease currently affecting pine plantations in Britain. Intraspecific variation in susceptibility to DNB has been observed in several pine species, but it is not clear if similar variation occurs in Pinus sylvestris (Scots pine), Britain's only native pine. In three separate experiments 2‐ and 3‐year‐old Scots pine saplings from six native Scottish populations were artificially inoculated with D. septosporum conidial suspensions and incubated under conditions optimal for disease development. Conidial suspensions were produced using a single isolate from northeast Scotland. In one experiment, plants were also treated with various spore suspension concentrations to assess the impact of inoculum load on disease severity. There were no significant interactions between host population, plant height, and experiment/inoculum load (anova , P > 0·05), but population, height and inoculum load all significantly affected disease severity (anova , P < 0·05). Among the 2‐year‐old trees, those from Amat were less susceptible than those from Glen Loyne and Glen Cannich (anova , P < 0·05). Among the 3‐year‐old trees, those from Beinn Eighe were less susceptible than those from Abernethy. Plant height and DNB susceptibility had a slightly negative relationship. The use of a spore suspension with a concentration of 1·6 × 10⁶ spores mL^?1 was optimum for disease development. In an in vitro experiment, production of conidia was greater when cultures were incubated in darkness. This paper is the first to report intraspecific variation in DNB susceptibility within Scots pine.     

Species composition,toxigenic potential and pathogenicity of Fusarium graminearum species complex isolates from southern Brazilian rice

This study aimed to assess the extent and distribution of Fusarium graminearum species complex (FGSC) diversity in rice seeds produced in southern Brazil. Four species and two trichothecene genotypes were detected among 89 FGSC isolates, based on a multilocus genotyping assay: F. asiaticum (69·6%) with the nivalenol (NIV) genotype, F. graminearum (14·6%) with the 15‐acetyldeoxynivalenol (ADON) genotype, and F. cortaderiae (14·6%) and F. meridionale (1·1%), both with the NIV genotype. Seven selected F. asiaticum isolates from rice produced NIV in rice‐based substrate in vitro, at levels ranging from 4·7 to 84·1 μg g^?1. Similarly, two F. graminearum isolates from rice produced mainly 15‐ADON (c. 15–41 μg g^?1) and a smaller amount of 3‐ADON (c. 6–12 μg g^?1). One F. meridionale and two F. cortaderiae isolates did not produce detectable levels of trichothecenes. Two F. asiaticum isolates from rice and two from wheat (from a previous study), and one F. graminearum isolate from wheat, were pathogenic to both crops at various levels of aggressiveness based on measures of disease severity in wheat spikes and rice kernel infection in a greenhouse assay. Fusarium asiaticum and the reference F. graminearum isolate from wheat produced NIV, and deoxynivalenol and acetylates, respectively, in the kernels of inoculated wheat heads. No trichothecene was produced in kernels from inoculated rice panicles by any of the isolates. These findings constitute the first report of FGSC composition in rice outside Asia, and confirm the dominance of F. asiaticum in rice agroecosystems.     

Stem rot of jewel orchids caused by a new forma specialis,Fusarium oxysporum f. sp. anoectochili in Taiwan

Stem rot of Anoectochilus formosanus (Af) caused by Fusarium oxysporum (Fo) is a major limiting factor to jewel orchid production in Taiwan. Fo causes discoloration in vascular tissues. However, some newly collected Fo isolates from Af stem rot do not cause vascular discoloration, suggesting changes may have occurred in the pathogen. Among recent Fo isolates from Af there are two colony types, the cottony alba (CA) and the sporodochial (S). In order to confirm that both colony types cause Af stem rot, 200 isolates were obtained from diseased stems in Nantou County and characterized by colony type and whether or not the infected plants had vascular discoloration. Isolates of both the CA and S types caused stem rot of Af; some isolates in each colony type caused vascular discoloration whilst others did not. Pathogenicity tests with 22 isolates resulted in stem rot disease severity ratings on Af of 3·1–4·0 and 2·1–4·0 with CA and S type colonies, respectively. The same isolates failed to cause disease on Cattleya, Dendrobium or Phalaenopsis plants. Phylogenetic analysis of partial intergenic spacer sequences showed that these isolates were distinguishable from other formae speciales of Fo and could be separated into two groups correlated with the CA or S type colonies with high bootstrap. Based on pathogenic, morphological and molecular characterizations, the Fo that causes stem rot of Af is proposed to be a new forma specialis, F. oxysporum f. sp. anoectochili, with different pathotypes.     

Mechanisms of resistance in Brassica carinata,B. napus and B. juncea to Pseudocercosporella capsellae

Studies were undertaken to compare susceptible and resistant host responses to Pseudocercosporella capsellae in cotyledons of Brassica carinata, B. juncea and B. napus in order to define the mechanisms of resistance in these three species. On both resistant and susceptible hosts, hyphal penetration was always through stomatal openings and without infection pegs or appressoria. On resistant B. carinata ATC94129P, up to 72% of spores disintegrated and, generally, germination (<22%) and germ tube lengths (<25 μm) were comparatively low. Resistant B. napus Hyola 42 had the lowest germination (8%) and susceptible B. carinata UWA#012 had the highest (51%). On resistant B. carinata ATC94129P, germ tube extension was impeded across 24–60 h post‐inoculation (hpi) and percentage stomatal penetration lower (4%) at 60 hpi compared with susceptible B. carinata UWA#012 (26%). Stomatal densities (stomata/14 757 μm²) on resistant B. juncea Dune (2·12) and B. napus Hyola 42 (1·62) were lower than for susceptible B. juncea Vardan (2·40) and B. napus Trilogy (2·03). Resistant B. carinata ATC94129P had greater stomatal density (1·89) than susceptible B. carinata UWA#012 (1·58). Overall, B. juncea had greater stomatal density (2·26) compared with B. napus (1·83) and B. carinata (1·74). In resistant B. carinata ATC94129P, P. capsellae induced 28% stomata to close, while in susceptible B. carinata UWA#012 no such closure was induced. Epicuticular wax crystalloids were present only on resistant B. carinata ATC94129P and probably also contribute towards resistance.     

The influence of flavonoids in maize pericarp on fusarium ear rot symptoms and fumonisin accumulation under field conditions

Breeding efforts have been undertaken to increase resistance of maize to fusarium ear rot (FER) and to fumonisin accumulation. Flavonoids in the pericarp of the kernels are considered particularly able to reduce the fumonisin accumulation. The aim of this 2‐year field study was to assess the effect of flavonoids on FER symptoms and fumonisin contamination in maize kernels using two isogenic hybrids, one providing pigmentation in the pericarp (P1‐rr) and the other without it (P1‐wr). FER incidence (FERi), FER severity (FERs), the incidence of infections caused by Fusarium spp. in symptomless kernels (FF) and fumonisin contamination (FUM) were assessed in both hybrids. Significant differences between the two hybrids were detected mainly in 2012 trials where P1‐rr showed lower FERi (P < 0·01), FF (P < 0·05) and FUM (P < 0·1) than P1‐wr. Site, characterized by local temperature and precipitation, played a relevant role in modelling all the measured variables, as its effect was highly significant in both years, whether they were considered individually or altogether. The interaction of hybrid with location was a significant (P < 0·001) source of variation only for FF. FF, together with FERi, was also significantly (P < 0·001) influenced by the interaction of hybrid with year. In general, FUM was more influenced by year and location parameters, such as temperatures during late ripening, than by flavonoid presence in kernel pericarp. The results indicate that flavonoid pigments alone may not be an important component in the resistance of maize to fumonisin accumulation.     

Sulphate and sulphurous acid alter the relative susceptibility of wheat to Phaeosphaeria nodorum and Mycosphaerella graminicola

The relative abundances of DNA of Mycosphaerella graminicola and Phaeosphaeria nodorum in archived wheat samples are closely correlated with UK anthropogenic emissions of oxidized sulphur over the last 160 years. To test whether this could be a causal relationship, possible modes of action of sulphur on the two fungi were examined. Mycelial growth of the two fungi in solutions of sulphurous acid was similar. Sulphurous acid at pH 4 reduced percentage germination of P. nodorum conidia more strongly than M. graminicola conidia. In spray inoculations of wheat cv. Squarehead's Master, Cappelle Desprez and Riband with water or sulphurous acid (pH 4), the ratio of leaves infected by P. nodorum to leaves infected by M. graminicola was increased by factors of 2·5, 2·1 and 0·6, respectively at pH 4. The same three cultivars of wheat were grown in sand and vermiculite and fertilized with nutrient solution containing 2·5 or 0·5 mm sulphate. Both pathogens infected less frequently at 2·5 mm sulphate, by a factor of about 2. The severity of infection by M. graminicola was reduced on all three cultivars by a factor of about 4·5 at 2·5 mm sulphate, but severity of P. nodorum was reduced only by a factor of about 2. Both elevated free sulphate concentrations in soil and sulphite in rainwater could therefore increase the prevalence of P. nodorum relative to M. graminicola, which is consistent with the historical changes in abundance.     

Interactions between Trichoderma harzianum and defoliating Verticillium dahliae in resistant and susceptible wild olive clones

Verticillium wilt (VW) in olive is best managed by an integrated disease management strategy, of which use of host resistance is a key element. The widespread occurrence of a highly virulent defoliating (D) Verticillium dahliae pathotype has jeopardized the use of commercial olive cultivars lacking sufficient resistance to this pathogen. However, the combined use of resistant wild olive rootstocks and Trichoderma spp. effective in the biocontrol of VW can improve the management of VW in olive. In vivo interactions between D V. dahliae and Trichoderma harzianum were studied in olive and wild olive plants displaying different degrees of resistance against this pathogen using confocal microscopy. This multitrophic system included wild olive clones Ac‐4 and Ac‐15, olive cv. Picual, and the fungal fluorescent transformants T. harzianum GFP22 and V. dahliae V138I‐YFP, the latter being obtained in this study. In planta observations indicated that V138I‐YFP colonizes the roots and stems of the olive and wild olive genotypes, and that GFP22 grows endophytically within the roots of them all. YFP fluorescence signal quantifications showed that: (i) the degree of root and stem colonization by the pathogen varied depending upon the susceptibility of the tested wild olive genotype, being higher in Ac‐15 than in Ac‐4 plants; and (ii) treatment with T. harzianum GFP22 reduced the extent of pathogen growth in both clones. Moreover, root colonization by strain GFP22 reduced the percentage of pathogen colonies recovered from stems of olive and wild olive plants.     

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.     

Testing and modelling the effects of climate on the incidence of the emergent nut rot agent of chestnut Gnomoniopsis castanea

Gnomoniopsis castanea is an emerging fungal pathogen causing nut rot of sweet chestnut, Castanea sativa. This study was aimed at testing and modelling the effects of climate on disease incidence. Up to 120 ripe nuts were collected in 2011 from trees in each of 12 sites located in the northwest of Italy. The incidence of G. castanea in each site was expressed as the number of infected nuts out of the total number of nuts sampled (%), determined by combining the results of morphological identification of isolates obtained from nuts, and their typing through a newly developed taxon‐specific molecular assay. Disease incidence ranged from 20 to 93%, depending on site. Geostatistical analyses revealed that, despite the clustering of sites (P < 0·05), disease incidence was not spatially autocorrelated (P > 0·05). This finding suggests that the disease is influenced by site‐dependent factors whose scale (c. 7·5–15·6 km) is consistent with the climate variability throughout the sampling region. Multivariate analyses on maximum, mean and minimum temperatures and on rainfall showed that warmer temperatures were associated with higher levels of disease incidence. The temperatures of months before nut harvesting were selected as predictors for partial least squares regression (PLSR) models (GnoMods) of G. castanea incidence. External validation on data collected either on sites or in years not used for model fitting showed the good predictive abilities of the GnoMods (Spearman's ρ_obs/pred > 0·72, P < 0·05). The above findings support a relationship between climate and incidence of G. castanea, providing statistical tools to forecast disease incidence at site level.     

Corynelia uberata as a threat to regeneration of Podocarpus falcatus in Ethiopian forests: spatial pattern and temporal progress of the disease and germination studies

The order Coryneliales includes several fungi such as Corynelia spp. that are pathogenic to trees in the Podocarpaceae. The aim of this study was to assess the spatial pattern and temporal progress of disease caused by Corynelia uberata on Podocarpus falcatus in Ethiopian forests and to evaluate the germination potential of seed retrieved from fruit infected by C. uberata. Corynelia uberata was found on leaves, young stems and/or on fruit of P. falcatus in Ethiopian forests. Spatial analysis in the Adaba‐Dodola forest showed that disease intensity of C. uberata was significantly higher in non‐‘WAJIB’ blocks (disturbed forest) than ‘WAJIB’ blocks (sustainably managed forest) (P < 0·0001). In the temporal disease progress study, a significantly higher incidence and severity of disease on fruit was recorded during the wet season relative to dry season (P < 0·0001). The green milk stage of fruit exhibited significantly higher mean incidence (P < 0·0001) and severity (P < 0·0001) of disease compared to other growth stages of fruit. The disease incidence and severity in general, as well as on different fruit growth stages, were highly correlated (P < 0·0001, R² ≥ 0·95). Germination rate of seed decreased significantly with an increase in the level of fruit infection by C. uberata (P < 0·0001). Thus, C. uberata can apparently influence germination of seed and may pose a threat to the regeneration of P. falcatus from seeds in Ethiopian forests.     

Nematicidal activity of the leaf powder and extracts of Myrtus communis against the root‐knot nematode Meloidogyne javanica

Nematicidal activity of the leaf powder, leaf extracts and formulated leaf extracts of Myrtus communis, an evergreen shrub that is widely distributed in Israel and other Mediterranean countries, was evaluated using the root‐knot nematode Meloidogyne javanica in in vitro and pot experiments. Leaf powder added to sand at 0·1% (w/w) reduced the number of juveniles recovered from the sand by more than 50%. Reduction in galling index and number of nematode eggs on tomato roots was also observed by incorporating the leaf powder at 0·1–0·4% (w/w) in the soil in pot experiments. Leaf powder extracts with methanol or ethanol showed the highest nematicidal activity among all extracts tested. Emulsifiable concentrates of leaf‐paste extract at a concentration as low as 0·005% (a.i., w/w) reduced the number of juveniles recovered from treated sand and the gall index of cucumber seedlings. The extract paste at 26 g m^?2 was also effective in reducing the gall index of tomato plants in field‐plot experiments. The leaf powder at 0·2% and the formulated leaf‐paste extract at 0·02% were also nematicidal to Tylenchulus semipenetrans and Ditylenchus dipsaci, but not to Pratylenchus penetrans or Steinernema feltiae. At least three nematicidal compounds were found in the leaf extract upon fractionation by thin‐layer chromatography. The results suggest that the leaf powder and paste extract of M. communis are potential nematicides against root‐knot nematodes.