Pathogenicity of Meloidogyne incognita and M. javanica on recombinant inbred lines from a crossing of Cucurbita pepo subsp. pepo × C. pepo subsp. ovifera

The response of recombinant inbred lines (RILs) from a cross of zucchini × scallop (Cucurbita pepo subsp. pepo ‘Murcia MU-CU-16’ × C. pepo subsp. ovifera ‘Scallop UPV-196’) to Meloidogyne incognita and M. javanica was determined after completion of a nematode reproduction cycle in experiments carried out in a growth chamber. The nematode differentiated the C. pepo genotypes at the subspecies level due to lower egg mass production on subspecies pepo than ovifera, and thus subspecies pepo was a poorer host than ovifera. In addition, Murcia MU-CU-16 discriminated M. incognita from M. javanica in terms of egg masses (EM), eggs per gram of root and reproduction factor (Rf), whereas Scallop UPV-196 did so in eggs per gram of root and Rf. The RILs differed in gall formation and EM production depending on the nematode × line combination. Comparisons between nematode isolates resulted in four significant combinations for pathogenic potential (galls/initial population (Pi) × 100), seven for parasitic success (egg masses/Pi × 100), and nine for host efficiency (egg masses/galls per root system × 100) which included all the lines tested against both isolates. Lines that restricted nematode development by at least 90% were considered as having intermediate resistance to M. incognita based on the definition of the International Seed Federation. They included lines 28-1, 35A, 107A, 110-3 and 153-2. All the RILs were susceptible hosts for M. javanica. The information presented here will be helpful for nematode management and also for plant breeders working on pathogen resistance on C. pepo.     

Colonization of rose by Sphaerotheca pannosa (Wallr.) Lév. var. rosae Wor.

Colonization of rose by powdery mildew (Sphaerotheca pannosa) is described in terms of mycelium growth, conidiophore production and sporulation in time. The data used are gathered during different years, put together and treated by means of graphic models. Colonies could be separated into fast and slow growing colonies. Colonies initiated on leaves of increasing age showed a decreasing growth rate. Production of conidiophores and conidia started on the same day, and the relative activity of conidiophore production reached its maximum 6 days after the end of the latency period, followed 1 day later by the maximum activity of conidium production. Both conidiophore and conidium production continued for a long time at a low level. The effect of leaf age on conidiophore production found expression in differences in production rate during the first days of colony development and in final production levels. Observations on naturally infected leaves in an outdoor experiment showed a rapid decrease of sporulation on leaves of 10 days and older. Highest percentages of sporulating leaf area were observed on leaves between 7 and 10 days old.     

Response to the letter on “Climatic distribution of citrus black spot caused by <Emphasis Type="Italic">Phyllosticta citricarpa.</Emphasis> A historical analysis of disease spread in South Africa” by Fourie et al. (2017)

In a previous study, Martínez-Minaya et al. (European Journal of Plant Pathology 143, 69–83, 2015) performed an analysis of climate-based distribution of citrus black spot (CBS) in South Africa. It was found that CBS was initially confined to humid areas with summer rainfall, but later spread to arid steppe and even desert climates. A strong spatial autocorrelation of CBS distribution was found. Fourie et al. (European Journal of Plant Pathology, 2017) take a critical view of our study, but without presenting any analysis of results to refute our findings. Furthermore, Fourie et al. (European Journal of Plant Pathology, 2017) appear to have misunderstood our work, since many of their criticisms relate to the potential distribution of CBS in Europe, which is beyond the scope of our original study. Fourie et al. (European Journal of Plant Pathology, 2017) highlight the limitations of climate classifications in species distribution modelling. However, this was made explicit in our study, indicating that it was a preparatory work and further advanced modelling studies, including spatial effects, will be needed. Fourie et al. (European Journal of Plant Pathology, 2017) incorrectly assume that we used all of South Africa as the background in the spatial autocorrelation analysis. However, only citrus areas were used and a strong spatial autocorrelation was detected at all distances evaluated. Contrary to what Fourie et al. (European Journal of Plant Pathology, 2017) suggest, similar climate distributions of CBS were obtained at 5′ and 30′ resolution, and also with the national land-cover map of South Africa. The figure comparison presented by Fourie et al. (European Journal of Plant Pathology, 2017) appears to ignore the fact that the maps we used were grid cells of 10 × 10 km and not the line polygons they suggest. Therefore, we consider the conclusions from the Martínez-Minaya et al. (European Journal of Plant Pathology 143, 69–83, 2015) remain entirely valid.     

PCR-mediated Detection of <Emphasis Type="Italic">Xanthomonas oryzae </Emphasis>pv. <Emphasis Type="Italic">oryzae </Emphasis>by Amplification of the 16S–23S rDNA Spacer Region Sequence

A detection method specific for Xanthomonas oryzae pv. oryzae, the pathogen responsible for bacterial blight of rice, was based on the polymerase chain reaction (PCR) and designed by amplifying the 16S–23S rDNA spacer region from this bacterium. The nucleotide sequence of the spacer region between the 16S and 23S rDNA, consisting of approximately 580-bp, from X. oryzae pv. oryzae, X. campestris pv. alfalfae, X. campestris pv. campestris, X. campestris pv. cannabis, X. campestris pv. citri, X. campestris pv. cucurbitae, X. campestris pv. pisi, X. campestris pv. pruni and X. campestris pv. vitians, was determined. The determined sequences had more than 95% identity. Therefore, a pair of primers, XOR-F (5′-GCATGACGTCATCGTCCTGT-3′) and XOR-R2 (5′-CTCGGAGCTATATGCCGTGC-3′) was designed and found to specifically amplify a 470-bp fragment from all strains of X. oryzae pv. oryzae isolated from diverse regions in Japan. No PCR product was amplified from X. campestris pathovars alfalfae, campestris, cannabis, carotae, cucurbitae, dieffenbachiae, glycines, pisi, pruni, vitians or zantedeschiae, except for pathovars citri, incanae and zinniae. The method could also detect the pathogen in infected rice leaves within 3 hr, at a detection limit of 4×10¹ cfu/ml. Received 17 December 1999/ Accepted in revised form 10 April 2000     

Virulence structure of the <Emphasis Type="Italic">Blumeria graminis</Emphasis> DC.f. sp. <Emphasis Type="Italic">avenae</Emphasis> populations occurring in Poland across 2010–2013

The aim of the present study was to determine the virulence structure of powdery mildew of oats (Blumeria graminis DC.f. sp. avena) in Poland in the years 2010–2013. For this purpose, powdery mildew isolates were collected from three experimental stations in Poland. To assess the virulence of the isolates, eight oat varieties with different responses to the pathogen were used. The results showed that a significant proportion of powdery mildew isolates found in Poland overcame the resistance genes of varieties Bruno (Pm6), Jumbo (Pm1) and Mostyn (Pm3). In contrast, lines Av1860 (Pm4), Am27 (Pm5) and Cc3678 (Pm2) were completely resistant to all pathogen isolates involved in the experiment. Changes constantly occurring in the powdery mildew population perfectly reflect diversity indexes, which were the smallest in the first year of observation, where in the following years these parameters were significantly higher. It is worth noting that the presence of powdery mildew is seasonal and local, which is reflected in the prevalence of the disease in a defined area of the country.     

Black band of Jew’s marrow caused by <Emphasis Type="Italic">Lasiodiplodia theobromae</Emphasis>

Stem rot and wilt of Jew’s marrow (nalta jute, Corchorus olitorius) were found on Is. Okinawa, Okinawa Prefecture, Japan, in March 2000. An anamorphic fungus, Lasiodiplodia theobromae was isolated repeatedly from the diseased plants and demonstrated to cause the disease. We coined the Japanese name “kurogare-byô” of Jew’s marrow for the present disease because it was new to Japan, although it had already been reported in India and Bangladesh as black band of the plant.     

Involvement of the β-cinnamomin elicitin in infection and colonisation of cork oak roots by <Emphasis Type="Italic">Phytophthora cinnamomi</Emphasis>

The virulence of two wild type (PA45 and PA37) and two genetically modified (13C: hygromycin resistant; FATSS: hygromycin resistant and β-cin knock-down) Phytophthora cinnamomi strains towards cork oak (Quercus suber) was assessed via a quantitative evaluation of disease symptoms arising from a soil infestation assay, and by a histological analysis of root colonization. Comparison of virulence, as expressed by symptom severity, resulted in the following ranking: highly virulent (wild type strains), medium virulence (strain 13C) and weakly virulent (FATSS). Both transgenic strains were compromised in their virulence, as expressed by symptom severity, but strain 13C was much less affected than FATSS. Microscopic observation showed that the FATSS strain was unable to effectively invade the root, while 13C and the two wild type strains were all able to rapidly colonize the whole root, including the vascular tissue. These results strengthen the notion that elicitins are associated, either directly or indirectly, with the infection process of Phytophthora.     

Bois noir affects the yield and wine quality of <Emphasis Type="Italic">Vitis vinifera</Emphasis> L. cv. ‘Chardonnay’

The Bois noir (BN) disease induced by ‘Candidatus Phytoplasma solani’ (CPs) is common in European vineyards. Its damage has not been fully investigated, especially with regards to wine attributes. The impact of BN on yield, berry composition and wine characteristics of Vitis vinifera L. cv. ‘Chardonnay’ was therefore comprehensively characterized in a 3-year field experiment in Hungary, Eger winegrowing region. Additionally, the bindweed-related tuf-b1 genotype was identified to be involved in the BN pathosystem in the experimental vineyard. Infection of CPs tuf-b1 genotype resulted in severe yield loss, the average decrease in number of bunches and total yield per vine was 56.7% and 68.4%, respectively. Analyses of wines produced from grapes of BN infected vines revealed decreased alcohol, epicatechin and iron contents; and increased organic acids, titratable acidity, catechin and calcium contents. Sensory evaluation of these wines confirmed unfavourable characteristics, i.e. higher acidity, bitterness, and usually pinkish discolouration. Negative impact on berry composition and wine quality were pronounced in the vintage with favourable weather conditions for grapevine production, whereas the negative effects of BN infection were less prominent, even masked, in the vintages with unfavourable weather (wet and cool). To reduce BN-caused damage, the need for improved preventative and curative measures for BN disease is highlighted.     

Chromosome constitution of hybrid strains constructed by protoplast fusion between the tomato and strawberry pathotypes of <Emphasis Type="Italic">Alternaria alternata</Emphasis>

To analyze the genetics of host-specific toxin production and its relation to the specific pathogenicity of a mitosporic fungus Alternaria alternata, we developed a protoplast fusion system. Protoplasts of drug-resistant transformants of the A. alternata tomato pathotype (AAL-toxin producer) and A. alternata strawberry pathotype (AF-toxin producer) were fused by electrofusion. Of five fusion strains examined, two strains were pathogenic on both tomato and strawberry host plants, whereas the rest of the fusion strains were pathogenic only on tomato. Pulsed-field gel electrophoresis analysis demonstrated that the hybrid strains pathogenic on both tomato and strawberry carry 1.0- and 1.05-Mb conditionally dispensable (CD) chromosomes derived, respectively, from the parental strains of the tomato and strawberry pathotypes. On the other hand, the fusion strains appeared to maintain only a single homologous chromosome derived from one of the parental strain in the case of essential chromosomes (A chromosomes). The results suggest that fusion strains between two different pathotypes of A. alternata might be haploid resulting from the deletion of extra sets of essential chromosomes in the fused nuclei, whereas the CD chromosomes derived from each parental strain could be maintained stably in a new genetic background with an expanded range of pathogenicity. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank database under the accession numbers AB469331to AB469354.     

Studies on the induction of rat uterine ornithine decarboxylase by DDT analogs. I. Comparison with estradiol-17β activity

The effect of DDT analogs and estradiol-17β on uterine ornithine decarboxylase activity in the immature intact and ovariectomized rat was studied. Pretreatment with various doses of o,p′DDT [1-(o-chlorophenyl)-1-(p-chlorophenyl)-2,2,2-trichloroethane] or estradiol-17β caused a marked increase in the specific activity of ornithine decarboxylase in the 20,000g supernatant fraction of uterine homogenates but not in liver homogenates. Doses as low as 0.5 mg of o,p′DDT or 0.002 μg of estradiol-17β stimulated uterine ornithine decarboxylase activity in the ovariectomized rat. The peaks of activity after treatment with o,p′ DDT and estradiol-17β occurred at 6 and 5 hr, respectively. The level of ornithine decarboxylase activity in untreated groups was consistently lower in ovariectomized rats than in intact immature animals. Treatment with o,p′ DDT (10 mg/100 g body weight) of ovariectomized and intact immature rats demonstrated at 131-fold and an about 20-fold increase in uterine ornithine decarboxylase activity, respectively. Treatment of ovariectomized rats with cycloheximide or actinomycin D effectively blocked the increase in ornithine decarboxylase caused by o,p′ DDT. Similar results were obtained with cycloheximide in the intact immature rat. Animals subjected to both adrenalectomy and ovariectomy demonstrated an increase in ornithine decarboxylase activity when treated with either estradiol-17β or o,p′ DDT. Dose-response curves obtained for estradiol-17β and o,p′ DDT suggest a similar mechanism of action for the two compounds. Graphic analysis of the dose-response curves for estradiol-17β and o,p′ DDT demonstrated an ED₅₀ of 0.038 μg/100 g body weight and 1.8 mg/100 g body weight, respectively. The examination of various DDT analogs in intact and ovariectomized animals showed that o,p′ DDT was the most potent inducer of ornithine decarboxylase. The order of decreasing potency of DDT analogs was o,p′ DDT, o,p′ DDD. p,p′ DDT, p,p′ DDD, and p,p′ DDE.     

Identification of a novel point mutation in the <Emphasis Type="Italic">β</Emphasis>-tubulin gene of <Emphasis Type="Italic">Botrytis cinerea</Emphasis> and detection of benzimidazole resistance by a diagnostic PCR-RFLP assay

The molecular basis of resistance to benzimidazole fungicides with laboratory and field mutant isolates of Botrytis cinerea was investigated. After chemical mutagenesis with N-methyl-N-nitrosogouanidine (NMNG) two different benzimidazole-resistant phenotypes were isolated on media containing carbendazim or a mixture of carbendazim and diethofencarb. The mutant isolates from the fungicide-mixture-containing medium were moderately resistant to carbendazim with wild-type tolerance to diethofencarb while mutant isolates from carbendazim-containing medium were highly resistant to carbendazim but sensitive to diethofencarb. The studied field isolates were highly resistant to benzimidazoles and sensitive to diethofencarb. Study of fitness characteristics of benzimidazole highly-resistant isolates showed that the resistance mutation(s) had no apparent effect on fitness-determining parameters. Contrary to this, the moderately benzimidazole-resistant strains, with no increased diethofencarb sensitivity, had a significant reduction in certain ecological fitness-determining characteristics. Analysis of the sequence of the β-tubulin gene revealed two amino acid replacements in the highly benzimidazole-resistant mutants compared to that of the wild-type parent strain. One was the glutamic acid (GAG) to alanine (GCG) change at position 198 (E198A), identified in both laboratory and field highly benzimidazole-resistant isolates, a mutation previously implicated in benzimidazole resistance. The second was a novel benzimidazole resistance mutation of glutamic acid (GAG) to glycine (GGG) substitution at the same position 198 (E198G), identified in a highly benzimidazole-resistant laboratory mutant strain. Molecular analysis of the moderately benzimidazole-resistant strains revealed no mutations at the β-tubulin gene. A novel diagnostic PCR-RFLP assay utilising a BsaI restriction site present in the benzimidazole-sensitive (E198) but absent in both resistant genotypes (E198G and E198A) was developed for the detection of both amino acid replacements at the β-tubulin gene.     

Quantification of viable cells of Clavibacter michiganensis subsp. sepedonicus in digester material after heat treatment by TaqMan® BIO-PCR

Journal of Plant Diseases and Protection - With bacterial ring rot affected potato lots must be decontaminated under official control without causing a risk for the dissemination of the disease....     

Rapid and sensitive detection of “<Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter asiaticus” by cycleave isothermal and chimeric primer-initiated amplification of nucleic acids

We developed a detection method for “Candidatus Liberibacter asiaticus”, causal agent of citrus huanglongbing, using isothermal and chimeric primer-initiated amplification of nucleic acids combined with cycling probe technology (Cycleave ICAN). With Cycleave ICAN, the reaction was done in one tube in 1 h without the need for electrophoresis, and false positives were not generated. In addition, Cycleave ICAN method was more sensitive than the conventional PCR method. Cycleave ICAN helps shorten the time for the large-scale detection needed to manage huanglongbing.     

Molecular diversity of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma mali’ and ‘<Emphasis Type="Italic">Ca</Emphasis>. P. prunorum’ in orchards in Slovenia

The plant parasitic nematode Longidorus poessneckensis found in Austria, the Czech Republic, Germany, Poland and the Slovak Republic was molecularly characterized. Mitochondrial genes encoding cytochrome c oxidase subunit I (COI) and nicotinamide dehydrogenase subunit 4 (nad4), the D2 and D3 expansion segments of 28S rRNA and Internal transcribed spacer 1 (ITS1) rRNA were sequenced for 16 L. poessneckensis populations. Six haplotypes of COI and five haplotypes of nad4 were determined. Nucleotide intraspecific variation was up to 17.1% for the partial sequenced COI gene and up to 17.7% for the partial sequenced nad4 gene, the latter being the highest up to date known intraspecific variation in this genus. The analyses of multiple amino acid sequence alignments of mitochondrial genes revealed low variability (0–2.4%) for COI gene and high divergence (0–7.6%) for nad4 gene. Intraspecific sequence diversity for the D2-D3 of 28S rRNA gene was up to 1.2% and for ITS1 rRNA gene was up to 1.6%. It has been hypothesized, that during the Last Glacial Maximum, L. poessneckensis populations probably persisted in refuge areas in the Carpathian Mountains and subsequently expanding from these areas and colonizing other European regions.     

Elimination of in vitro bacterial contaminants in shoot cultures of ‘MRS 2/5’ plum hybrid by the use of <Emphasis Type="Italic">Melia azedarach</Emphasis> extracts

The antimicrobial activity of leaf and callus extracts of Melia azedarach was tested on in vitro shoot cultures of the peach rootstoch ‘MRS 2/5’ (Prunus cerasifera × Prunus spinosa) that were heavily contaminated with Sphingomonas paucimobilis (Sp) and Bacillus circulans (Bc). The extracts were filter-sterilised and added at 0%, 1%, 5%, 10% and 20% to a modified Murashige and Skoog proliferation medium previously autoclave-sterilised. Up to about 17% shoots died with 10–20% extract, except for Sp-contaminated shoots, whose survival was reduced to 50% after treatment with 20% extract. No shoots died with 1% to 5% supplement. The undiluted leaf extract showed bactericidal activity on plated Sp and Bc isolates. The homogenates of shoots randomly collected from treated cultures were processed for bacterial colony counting. Thus the 10% supplement was the best treatment for ridding Bc-contaminated cultures of bacteria (although 5% had a similar bactericidal effect), and allowing shoot growth and proliferation comparable to controls at the fifth subculture on a standard medium, while 20% extract was needed to eliminate Sp, and could induce higher growth and proliferation rates in surviving shoots than in untreated cultures. Callus extract was ineffective. The bactericidal activity of the leaf extract seemed attributable to a synergistic effect of azadirachtin with other unidentified compounds present in the extract.     

Efficacy and dose–response relationship in biocontrol of Fusarium disease in maize by Streptomyces spp.

Two isolates of Streptomyces spp. DAUFPE 11470 and DAUFPE 14632 were evaluated to determine the antagonist–pathogen inoculum concentration relationship under greenhouse conditions. Pathogen and antagonist concentration, significantly (P < 0.05) affected development of Fusarium disease in maize with a significant interaction between pathogen and antagonist concentration. Dose–response relationship also differed significantly (P < 0.05) between the two isolates, but both isolates demonstrated effective control of Fusarium disease, regardless of pathogen concentration. The isolate DAUFPE 11470 provided the most effective control. The lowest value for disease incidence occurred at low pathogen (10³ chlamydospore g⁻¹ soil) and high antagonist concentration (10⁶ cfu ml⁻¹) for both isolates. The disease incidence for control plants ranged from 19% to 76%. However, in relation to control the lowest disease reduction occurred at low pathogen (10³ chlamydospore g⁻¹ soil) and high antagonist concentrations (10⁶ cfu ml⁻¹). These reductions were 10.6% and 13% for DAUFPE 14632 and DAUFPE 11470, respectively. The highest disease reductions, in relation to control plants, occurred at high pathogen (10⁶ chlamydospore g⁻¹ soil) and antagonist (10⁶ cfu ml⁻¹) concentrations for both isolates. These values were 55% and 62.2% for DAUFPE 14632 and DAUFPE 11470, respectively. The chlamydospore germination of Fusarium moniliforme was affected by glucose addition, antagonist isolates and type of inoculum. The lowest chlamydospore germination was observed with bacterial suspensions of the isolates, for all glucose additions. The results suggested that both Streptomyces spp. isolates were effective at different doses as biocontrol agents against F. moniliforme. Also, there was evidence for at least two mechanisms of biocontrol and that apparently, both isolates showed the same mechanisms of biocontrol action related to production of bioactive compounds and competition for carbon. Further studies will be developed to improve the level and effectiveness of control by these isolates.     

Natural parasitism of<Emphasis Type="Italic">Chrysoperla carnea</Emphasis> by hymenopterous parasitoids in cotton-growing areas of Çukurova,Turkey

A 4-year study of parasitoids attackingChrysoperla carnea (Stephens) in the cotton fields of Çukurova, Turkey, revealed the activities of three principal taxa:Telenomus sp. nr.suvae attacks the eggs ofC. carnea: Catolaccus sp. andBaryscapus sp. are larval and pupal parasitoids, respectively. In general, percent egg parasitism increased starting from the second half of June and into July, then decreased until the beginning of August before rising again at the end of growing season. During the early season, weekly parasitism was found to be as high as 94%. Overall, seasonal egg parasitism varied between 13.6% and 62.0% among all study sites. According to the results of a one-year field study at Hac?ali, total larval and pupal parasitism was found to be 12.5% and 55.6%, respectively. Therefore, different factors influencing the effectiveness of the generalist predatorC. carnea against cotton pests, and the differential effects of cotton treatments on both host and parasitoids, need to be evaluated.     

Diversity of defence mechanisms in plant–oomycete interactions: a case study of <Emphasis Type="Italic">Lactuca</Emphasis> spp. and <Emphasis Type="Italic">Bremia lactucae</Emphasis>

Plant pathogenic oomycetes, including biotrophic downy mildews and hemibiotrophs/necrotrophs such as Phytophthora and Pythium, cause enormous economic losses on cultivated crops. Lettuce breeders and growers face the threat of Bremia lactucae, the causal agent of lettuce downy mildew. This pathogen damages leaf tissues and lettuce heads and is also frequent on wild Asteraceae plants. The interactions of Lactuca spp. with B. lactucae (abbr. lettuce–Bremia) display extreme variability, due to a long co-evolutionary history. For this reason, during the last 30 years, the lettuce–Bremia pathosystem has been used as a model for many studies at the population, individual, organ, tissue, cellular, physiological and molecular levels, as well as on genetic variability and the genetics of host–parasite interactions. The first part of this review summarizes recent data on host–parasite specificity, host variability, resistance mechanisms and genetics of lettuce–Bremia interactions. The second part focuses on the development infection structures. Phenotypic expression of infection, behaviour of B. lactucae on leaf surfaces, the process of penetration, development of primary infection structures, hyphae and haustoria are discussed in relation to different resistance mechanisms. In the third part, the components of host resistance and the variability of defence responses are analysed. The role of reactive oxygen species (ROS), antioxidant enzymes, nitric oxide (NO), phenolic compounds, reorganization of cytoskeleton, electrolyte leakage, membrane damage, cell wall disruption, hypersensitive reaction and plant energetics are discussed in relation to defence responses. In general, the extreme variability of interactions between lettuce and Bremia, and their phenotypic expression, results from diversity of the genetic background. Different mechanisms of resistance are conditioned by an orchestra of defence responses at the tissue, cell, and molecular levels. The various events responsible for defence involve a complex interaction of the processes and reactions mentioned above. This review also provides an overview on the timing of pathogen development, host pathological anatomy, cytology and physiology of lettuce–Bremia associations. The significance of these factors on the expression of different resistance mechanisms (non-host and host resistance, race-specific and race non-specific resistance, field resistance) is discussed.     

Influence of Stigmatic Morphology on Flower Colonization by <Emphasis Type="Italic">Erwinia amylovora</Emphasis> and <Emphasis Type="Italic">Pantoea agglomerans</Emphasis>

The morphology of apple and pear stigma was investigated with confocal laser scanning microscopy and scanning electron microscopy. The floral colonization process by Erwinia amylovora was studied with gfp-labelled bacteria and confocal laser scanning microscopy to allow the in vivo observation of the pathogen colonization on intact, viable plant tissues without any kind of staining of the specimens. The interaction on the stigma between Erwinia amylovora and Pantoea agglomerans, both labelled with genes encoding for fluorescent proteins (DsRed-GFP), was also investigated. A stylar groove, covered by papillae and dwelling from the stigma along the style, was visualized. In laboratory conditions, this groove was shown to be an important way for E. amylovora migration towards the nectarthodes. Due to its anatomical structure the groove can sustain bacterial multiplication and thus may play an important role on the interactions between the pathogen and the bacterial antagonist P. agglomerans.