Flow of Botrytis cinerea inoculum between lettuce crop and soil

Botrytis cinerea causes grey mould, a disease common on many economically important crops. Although much attention is paid to the airborne inoculum of this fungus, as it sporulates abundantly in favourable conditions, knowledge on the abundance and genetic characteristics of soilborne inoculum could help improve control strategies. In this study, the soilborne inoculum of B. cinerea was quantified in two greenhouses at different times before and after the cultivation of four successive lettuce crops. Between 0 and 1177 colony‐forming units (CFU) of B. cinerea per gram of soil were recorded. There was no significant correlation between abundance of soilborne inoculum and subsequent disease incidence on lettuce (P = 0·11). Sixty‐five isolates collected from diseased plants and 66 isolates collected from the soil were investigated for their genetic diversity. The soil strains showed lower genetic diversity than the lettuce strains when considering the unbiased gene diversity within the nine microsatellite loci, the mean number of alleles per locus and the haplotypic diversity. The genetic differentiation between lettuce and soil strains decreased over three successive lettuce crops. At the same time, the genetic structure of the two groups of strains tended to become similar. These results are consistent with the hypothesis of a flow of inoculum between the lettuce crop and the soil, and vice versa. The study shows that grey mould management should pay more attention to the inoculum of B. cinerea present in the soil.     

Genetic diversity and structure of Hemileia vastatrix populations on Coffea spp.

Coffee leaf rust is the most limiting disease for coffee cultivation in Brazil. Despite its importance, relatively little is known about the genetic diversity of Hemileia vastatrix, the rust causal agent. In this work, the DNA from 112 monopustule isolates from different geographic locations and coffee genotypes were analysed by amplified fragment length polymorphisms (AFLP). The objectives were to assess the influence of the host and geographic origin on the diversity and population differentiation in H. vastatrix. The fungal population showed a low level of genotypic diversity. Gene diversity (h) was 0·027 and the hypothesis of random mating in the total population was rejected, but evidence for recombination was found for two subpopulations (São Paulo and Paraná). The analysis of molecular variance revealed that 90% of the genetic distribution of the pathogen occurs among isolates within the subpopulation (states or host of origin). There was no correlation between geographic and genetic distance (r = ?0·024, P = 0·74), which together with the high number of migrants and the low degree of differentiation in populations of H. vastatrix, is consistent with the fact that the inoculum is probably easily dispersed by wind over long distances, allowing dispersal of the pathogen among coffee growing areas in Brazil. Therefore, it is difficult to predict the durability of resistant sources to coffee rust. The recommendation for the breeding programmes is thus to incorporate multigenic resistance as a control strategy.     

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.     

Evidence of low levels of genetic diversity for the Phytophthora austrocedrae population in Patagonia,Argentina

Phytophthora austrocedrae is a recently discovered pathogen that causes severe mortality of Austrocedrus chilensis in Patagonia. The high level of susceptibility of the host tree, together with the distribution pattern of the pathogen, have led to the hypothesis that P. austrocedrae was introduced into Argentina. The aim of this study was to assess the population structure of P. austrocedrae isolates from Argentina in order to gain an understanding of the origin and spread of the pathogen. Genetic diversity was determined based on amplified fragment length polymorphisms (AFLPs). In total, 48 isolates of P. austrocedrae were obtained from infected A. chilensis trees, representing the geographical range of the host. Four primer combinations were used for the AFLP analysis. Of the 332 scored bands, 12% were polymorphic. Gene diversity (h) ranged from 0·01 to 0·03; the Shannon index (I) ranged from 0·01 to 0·04. A high degree of genetic similarity was observed among the isolates (pairwise S values = 0·958–1; 0·993 ± 0·009, mean ± SD). A frequency histogram showed that most of the isolate pairs were identical. Principal coordinate analysis using three‐dimensional plots did not group any of the isolates based on their geographical origin. The low genetic diversity (within and between sites) and absence of population structure linked to geographic origin, together with the aggressiveness of the pathogen and the disease progression pattern, suggest that P. austrocedrae might have been introduced into Argentina.     

SSR assessment of Phytophthora infestans populations on tomato and potato in British gardens demonstrates high diversity but no evidence for host specialization

Phytophthora infestans populations can differ in composition as a result of host specialization on tomato and potato hosts. In Great Britain many amateur gardeners grow outdoor tomatoes but there is little or no commercial tomato production outdoors. This study analysed isolates of P. infestans from British gardens with 12 multiplexed simple sequence repeat markers that are used to monitor the disease on commercial potato crops. Samples of P. infestans from tomato hosts were collected in 3 years and from potato in 1 year from across Great Britain. Seven previously unreported clonal lineages were detected in garden populations and higher frequencies of unique clonal lineages (28–40%) were present compared with populations from British commercial potato crops reported elsewhere. Garden populations had a lower proportion (11–48% less) of the most common lineages (13_A2 and 6_A1) that together made up at least 86% of the commercial potato populations during the sampling period. Host species accounted for only 2·0% of molecular variance detected between garden potato‐ and tomato‐hosted samples. No significant difference in clonal lineage composition was found between host species in Great Britain and this could be due to the whole P. infestans population overwintering on potato. British garden populations on both hosts were much more diverse than those on commercial potato crops; this finding may be influenced by less frequent fungicide use by gardeners and a higher diversity of unsprayed susceptible potato cultivars, enabling metalaxyl‐sensitive and less aggressive genotypes to survive in gardens.     

Genotypic diversity and migration of clonal lineages of Botrytis cinerea from chickpea fields of Bangladesh inferred by microsatellite markers

An analysis of allelic diversity at nine microsatellite loci provided an insight into the population structure of Botrytis cinerea from four fields (sampled in 2003 and 2004) that represented important regional locations for chickpea production in Bangladesh. Although three populations were limited by sample size after clone‐correction, a total of 51 alleles were amplified among 146 B. cinerea isolates from Bangladesh, which revealed a high amount of within‐population and overall genetic diversity (H_S = 0·48 and H_T = 0·54, respectively). The percentage of maximal genotypic diversity (G) ranged between populations (G = 23–40), with a total of 69 haplotypes detected (G = 25). Bayesian cluster analysis depicted two major clusters distributed among the four Bangladesh populations, indicating population admixture from two origins that have spread throughout these regions. Genotype flow between regions was detected and indicated the spread of clonal lineages, consistent with relatively low differentiation among the four populations (mean G_ST = 0·1, P < 0·05). These results highlighted the potential threat of host resistance breakdown as a result of considerable genetic diversity, genotype flow and the evolutionary potential of B. cinerea.     

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.     

Genetic analysis of the population structure of the rice false smut fungus,Villosiclava virens,in China using microsatellite markers mined from a genome assembly

Studies on population genetics of Villosiclava virens are limited because of the lack of polymorphic markers. Based on a draft genome sequence of isolate HWD‐2 produced in this study, 20 of 403 potential simple sequence repeats (SSR) loci showed polymorphisms in preliminary screening using eight diverse V. virens isolates. Among polymorphic loci, most of them with tetra‐ to hexanucleotide unit motifs showed higher levels of polymorphism than loci with smaller motifs. After testing with 20 polymorphic SSR markers, the 87 isolates of V. virens from eight populations in China showed a high level of genetic diversity, with each as a unique haplotype. This differs from some previous findings showing little to no genetic variation based on random amplified polymorphic DNA and amplified fragment length polymorphism analyses. Among eight populations from major rice production areas of China, the population from Guangxi province in south China showed the highest levels of polymorphism, which led to the speculation that it might be closer to the centre of origin of this pathogen. The northern, central and eastern populations (Jilin, Liaoning, Hubei, Hunan, Jiangxi and Zhejiang), when considered together as a group, showed significant molecular variation compared to the southern populations (Fujian and Guangxi) (Φ_PT = 0·043, P = 0·037). A significant relationship (Mantel test, P = 0·027) but with low correlation (R² = 0·23) was also found between geographic distance and genetic distance. The 20 polymorphic SSR primer pairs designed in this study provide a tool for further research on the population diversity of this emerging fungal pathogen of rice.     

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.     

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.     

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.     

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.     

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.     

Genetic diversity of Botrytis in New Zealand vineyards and the significance of its seasonal and regional variation

Species‐ and population‐specific differences in fungicide resistance and aggressiveness within Botrytis makes basic data on genetic diversity important for understanding disease caused by this fungus. Genetic diversity of Botrytis was surveyed between 2008 and 2012 from grapes from five New Zealand wine‐growing regions. A total of 1226 isolates were gathered from symptomless flower buds at the start of the growing season and 1331 isolates from diseased fruit at harvest. Two species were found, B. cinerea and B. pseudocinerea. Botrytis pseudocinerea was common in both Auckland vineyards sampled, and infrequent elsewhere. However, even in Auckland, it was rarely isolated from diseased fruit. The presence of the Boty and Flipper transposons was assessed. Isolates with all four transposon states (Boty only, Flipper only, both Boty and Flipper, no transposons) were found for both species. Both vineyards in the Auckland region had high numbers of Flipper‐only isolates at flowering; both vineyards from the Waipara region had high numbers of Boty‐only isolates at flowering. Most isolates from diseased fruit at harvest contained both transposons. These observations suggest that B. pseudocinerea, and isolates with one or both of the transposons missing, may be less aggressive than B. cinerea, or than isolates with both transposons present. Two clades were resolved within B. pseudocinerea, only one of which has been reported from European vineyards. Phylogenetic diversity within B. cinerea in New Zealand was similar to that known from Europe, including isolates that appear to match Botrytis ‘Group S’. The taxonomic implications of this genetic diversity are discussed.     

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.     

Genetic population structure and fungicide resistance of Botrytis cinerea in pear orchards in the Western Cape of South Africa

Botrytis cinerea isolates from pear blossoms (Pyrus communis) in South Africa were collected from four orchards in two production areas in the Western Cape. The cryptic species status based on vegetative‐incompatibility alleles of the Bc‐hch gene indicated that all the isolates belonged to B. cinerea. A microsatellite analysis of B. cinerea populations was performed to assess the genetic population structure. Total gene diversity (H) was high, with a mean of 0.69 across all populations. Some genotype flow was evident between orchards as indicated by the spread of microsatellite multilocus genotypes, in agreement with the moderate, but significant population differentiation among orchards (mean φ_PT = 0.118, P = 0.001). Index of association analyses (I_A and r?_d) suggest that the populations reproduce mostly asexually, even though mating type distribution did not differ significantly from a 1:1 ratio, suggesting frequency‐dependent selection. Isolates resistant to benomyl were evident in one orchard only. This orchard was also significantly differentiated from all other populations, suggesting infrequent localized selection for benomyl resistance. Overall, the findings of this study highlight the dangers of a mixed reproduction system, and stress the importance of regularly monitoring fungicide resistance levels towards developing more efficient management practices.     

Most Colletotrichum species associated with tree tomato (Solanum betaceum) and mango (Mangifera indica) crops are not host‐specific

An important constraint for crop production in Colombia is the high incidence of anthracnose caused by Colletotrichum species. Although several studies have focused on these fungi, the relationship between the different fungal species within the genus and their hosts and whether they display any host preference or host specificity has yet to be examined. In Colombia, diseases caused by Colletotrichum species are particularly severe in mango (Mangifera indica) and tree tomato (Solanum betaceum) crops. In a previous investigation, the Colletotrichum phylogenetic species attacking these crops were identified. The present study aimed to determine whether isolates collected from tree tomato and mango showed host preference or host specificity by assessing aggressiveness, spore density, latent period, and fitness of each strain on the two hosts. In the departments of Cundinamarca and Tolima, Colombia, isolates were collected from plants that presented typical anthracnose symptoms and were identified as C. acutatum, C. asianum, C. boninense, C. gloeosporioides, C. tamarilloi and C. theobromicola. Inoculation of conidia of each isolate onto both hosts showed isolates had no host preference and only the C. gloeosporioides isolate showed host specificity. However, in general, isolates produced a higher spore density when inoculated on the alternate host, which may indicate a difference in the degree of adaptation to each host. Statistical analyses of the assessed parameter values revealed that isolates use different infection strategies when infecting each host. In light of these results, the implications of using quantitative estimations of fitness when studying fungal pathogens are discussed.     

Genetic structure of Verticillium dahliae isolates infecting olive trees in Tunisia using AFLP,pathogenicity and PCR markers

Since 2006, verticillium wilt of olive induced by Verticillium dahliae has caused considerable economic losses in olive orchards in Tunisia. The genetic structure of V. dahliae isolates collected from different olive growing regions was investigated using virulence tests, vegetative compatibility grouping (VCG) and amplified fragment length polymorphism (AFLP) analyses. In total, 42 isolates of V. dahliae from diseased olive trees were tested. Cluster analysis and principal coordinate analysis revealed that geographic origin was the main factor determining the genetic structure of V. dahliae populations and both methods indicated a genetic separation between the central and coastal isolates. Isolates were divided into two major groups: the AFLP‐I group included all isolates from Sidi Bouzid, Kairouan, Kasserine and Sfax (centre of the country) and the AFLP‐II group included isolates from Monastir, Zaghouane, Sousse, Mahdia (coastal region), and two isolates from Sfax. Analysis of the molecular variance (amova ) indicated a significant level of genetic differentiation among (76%) and within (23%) the two populations. Analyses of both the defoliating (D) and non‐defoliating (ND) pathotypes and VCG markers indicated that most of the isolates belong to VCG 2A and 4B/ND pathotype. The disease severity was highly variable among the isolates tested (P < 0·05) with no evidence of association between aggressiveness and geographical origin of the isolates. Overall, results of this study revealed a clear association between the genetic diversity of the isolates and their geographic origin, but not between genetic diversity and virulence patterns.     

Heterogeneity of Pseudomonas savastanoi populations infecting Myrtus communis in Sardinia (Italy)

Genetic, phenotypic and host range diversity among Pseudomonas savastanoi isolates from Myrtus communis were investigated. Thirty‐one isolates from six Sardinian commercial myrtle orchards and three isolates from plants growing spontaneously on the island of Rhodes (Greece) were compared with reference strains of P. savastanoi from olive, oleander, ash and myrtle. Multilocus sequence analysis (MLSA) indicated the presence of a monomorphic population with a very low level of variability. Conversely, Biolog phenotypic fingerprinting and phytohormone production analyses showed a considerable metabolic diversity, as bacteria obtained from single infected tissue differed more than bacteria obtained from different orchards. When pathogenicity tests were carried out on myrtle plants, different types of symptoms were induced: knots, canker lesions with or without tissue proliferations and, occasionally, wilting of the inoculated twig, a symptom never reported before for P. savastanoi. Comparable symptoms were also observed in the natural environment both on spontaneous and cultivated plants. Moreover, the host range of the myrtle population was heterogeneous and not well defined. Some isolates showed a wide host range whilst others were pathogenic only to their natural host. Overall these findings suggest that the diversity of the P. savastanoi population from myrtle does not depend so much on the locality or the natural host and does not allow the Sardinian and Greek isolates, together with previously characterized myrtle strains, to be ascribed to a known pathovar of P. savastanoi, nor to propose their belonging, as a whole, to a new pathovar.