Fusarium solani f. sp. passiflorae: a new forma specialis causing collar rot in yellow passion fruit

The aim of this study was to characterize a Fusarium population obtained from yellow passion fruit (YPF) with collar rot using pathogenicity, morphocultural characteristics and molecular tests. Pathogenicity and disease severity were assessed in six plant species: YPF, zucchini, tomato, bean, soya bean and cucumber. Potato dextrose agar medium (PDA) was used to determine mycelial growth at five temperatures (15–35°C). The colour produced by isolates was also determined on PDA at 25°C. Synthetic nutrient agar medium was used to evaluate: (i) type of mycelium and phialides; (ii) size, shape and number of septa from conidia; and (iii) production of chlamydospores and perithecia. Molecular tests consisted of sequencing the ITS–5·8S rDNA region and elongation factor 1α (EF‐1α) gene. The isolates caused large lesions on YPF, zucchini and tomato, with YPF having the highest mean disease severity and being the only one that showed wilt symptoms and death of the plant. Thus the isolates showed host specificity. Maximum mycelial growth occurred at 25°C and the predominant colour was bluish‐white. The isolates produced long phialides, dense aerial mycelium, oval microconidia with a mean size of 9·5 × 2·6 μm, macroconidia of 32·7 × 3·4 μm with 3·3 septa, and chlamydospores; only one isolate lacked perithecia. Phylogenetic trees of the ITS region and EF‐1α gene showed that isolates from YPF formed a distinct group within the F. solani group and the formae speciales of F. solani. It is proposed to name all isolates from YPF as F. solani f. sp. passiflorae.     

Variation in virulence of Greek isolates ofPhytophthora citrophthora as measured by their ability to cause crown rot on three peach rootstocks

The virulence ofPhytophthora citrophthora isolated from various host-plants on three peach rootstocks (GF677, PR204, KID I) was examined. There was no significant difference among the rootstocks with respect to their susceptibility to testedP. citrophthora isolates. The most virulent isolate originated from sycamore (Acer pseudoplatanus); isolates from pistachio trees (Pistacia vera) also showed high virulence but were significantly less virulent than the sycamore isolate. Isolates originating from plum (Prunus domestica), almond (Prunus amygdalus) and lemon (Citrus limon) trees were moderately virulent on peach rootstocks; those from cyclamen (Cyclamen persicum) showed the lowest virulence of those tested. There was thus great variation in virulence among the testedP. citrophthora isolates. It is possible that the isolates ofP. citrophthora from sycamore, pistachio, plum, almond and lemon trees are a threat to peach trees, whereas the low virulence of the isolates from cyclamen hosts suggests that these pathogens are not a serious threat to peach trees. http://www.phytoparasitica.org posting Jan. 3, 2002.     

Characterization of <Emphasis Type="Italic">Pectobacterium carotovorum</Emphasis> subsp. <Emphasis Type="Italic">carotovorum</Emphasis> causing soft-rot disease on <Emphasis Type="Italic">Pinellia ternata</Emphasis> in China

Pinellia ternata is a traditional Chinese herb which has been used in China for over 1,000 years. A soft-rot disease characterized by water-soaked lesions and soft-rot symptoms with a stinking odour was commonly observed in cultivated fields of this plant, and Pectobacterium-like bacteria were consistently isolated from the infected tissues. Two typical strains (SXR1 and ZJR1), isolated from Shanxi and Zhejiang, respectively, were identified. Pathogenicity tests revealed that these strains were virulent to P. ternata and induced the same symptoms as observed in the field. Characterization involving fatty acid profile, metabolic and physiological properties, 16S rDNA sequence and PCR-RFLP identified both isolates as P. carotovorum subsp. carotovorum (Pcc). The 16S rDNA of both isolates shared 97–99% sequence similarity with that of Pcc strains. The phylogenetic trees showed that both isolates were clustered in the group of Pcc and P. carotovorum subsp. odorifera and both PCR-RFLP profiles were consistent with the pattern E produced by the minority of Pcc strains. Thus, isolates SXR1 and ZJR1 were characterized as Pcc in spite of some differences. This is the first report that Pcc has been proven as a causal agent of soft-rot disease on P. ternata.     

Monitoring of <Emphasis Type="Italic">Phytophthora</Emphasis> species on fruit trees in Bulgaria

Disease on fruit trees in Bulgaria caused by Phytopthora cactorum and P. citrophthora was found in the period 1998–1999. Leaves of some trees become reddish during July, and later in the season fall off. Infected trees die during the same season, or the next season. Observations on symptom development and spread of Phytophthora root and crown rot of fruit trees was undertaken from 1999 to 2009. Disease incidence is between 2% and 14% in some gardens and nurseries. The disease was registered in the regions of Plovdiv, Kjustendil, Sliven, Yambol, Karnobat, Bourgas and Svishtov. Samples from infected plant tissues were taken and isolations were done on selective PARP media, or by applying a baiting bioassay. Based on morphological and cultural characteristics and temperature requirements the following Phytophthora species have been identified: Phytophthora cactorum, P. citrophthora, P. drechsleri, P. cryptogea, hybrid and Pythium. Pathogenicity of the isolates was tested on green apple fruits or one-year-old apple rootstocks. Laboratory studies of the effect of temperature on mycelia growth showed that most isolates can grow from 5° up to 30°C, with an optimum from 18° to 25°C. Only three strains grew at 35–36°C, two developed slowly, one grew well. The optimal pH for mycelia development was tested. Aiming at control of disease, in vivo pot trials have been carried out for studying resistance of rootstocks to P. cactorum. At the end of the growing season a good level of resistance has been shown in the rootstocks M29C, Gizela 6, and MAXMA 14.     

Growth and sporulation of <Emphasis Type="Italic">Stemphylium vesicarium</Emphasis>, the causal agent of brown spot of pear,on herb plants of orchard lawns

The inoculum sources of ascospores of Pleospora allii and of conidia of its anamorph Stemphylium vesicarium were investigated in relation to the brown spot disease epidemiology on pear. Dead and living leaves of three pear varieties (Abate Fétel, Conference and William), seven grasses (Poa pratensis, Festuca rubra, Festuca ovina, Lolium perenne, Digitaria sanguinalis and Setaria glauca) and Trifolium repens, which are used in pear orchard lawns, were inoculated with conidia of Stemphylium vesicarium virulent on pear and incubated under controlled-environment. Stemphylium vesicarium was always re-isolated from dead leaves of the considered plants, but not from symptomless green or yellowish living leaves. The fungus was occasionally re-isolated from leaf segments showing unspecific necrosis. Inoculation of pear leaves with isolates from grasses demonstrated that the fungus did not lose pathogenicity. Pseudothecia, ascospores and conidia were produced on all the dead inoculated leaves; differences between specimens were found for phenology of pseudothecia, their density and size, and for the number of conidia produced. Pseudothecia were produced faster in the lawn species than in pear leaves, and their density was higher, especially for S. glauca, L. perenne and P. pratensis. Ascospore maturation and ejection was more concentrated for the pseudothecia developed on pear leaves than for those on F. ovina and S. glauca. All the lawn species produced more conidia than pear leaves.     

Multigene phylogenetic analysis of inter- and intraspecific relationships in <Emphasis Type="Italic">Venturia nashicola</Emphasis> and <Emphasis Type="Italic">V. pirina</Emphasis>

Isolates of Venturia species isolated from pear in Japan, China, Taiwan and Israel were used in this study to analyze their molecular phylogenetic relationship. The nucleotides of rDNA-ITS, partial β-tubulin and elongation factor 1α genes were sequenced directly after PCR. Based on these sequence data two phylogenetic groups could be distinguished. Isolates collected from Asian pears such as Japanese and Chinese pears formed a distinct evolutionary lineage from those derived from European and Syrian pears. This result corroborated the early taxonomic separation of V. nashicola from V. pirina. In addition, trees from single-locus data sets and the combined data set showed that all isolates of V. nashicola were included in a monophyletic group and representative isolates of five pathological races originating from different locations and cultivars formed a single lineage. In contrast, two distinct evolutionary lineages were revealed in V. pirina and isolates of five races were scattered in two lineages. Israeli isolates of race 2 as well as two Japanese isolates of V. pirina formed a distinct lineage from other isolates of this species, while other Israeli isolates belonging to races 1, 3, 4, and 5 were closely related to each other and formed another lineage. It was indicated that the evolution of pathological races in V. nashicola might have occurred relatively recently as compared with V. pirina.     

Characterization of three Colletotrichum acutatum isolates from Capsicum spp.

Colletotrichum acutatum causes anthracnose on peppers (Capsicum spp.), resulting in severe yield losses in Taiwan. Fungal isolates Coll-153, Coll-365 and Coll-524 collected from diseased peppers were found to differ in pathogenicity. Pathogenicity assays on various index plants revealed that Coll-524 was highly virulent and Coll-153 was moderately virulent to three commercially available pepper cultivars. Both isolates induced anthracnose lesions and produced abundant conidia. Coll-365 was only weakly virulent on pepper fruit, where it caused small lesions and hardly produced conidia on pepper fruit. However, Coll-365 was highly pathogenic to tomato fruit and mango leaves, where it caused anthracnose lesions and formed acervuli and conidia. All three isolates showed similar abilities in the attachment and germination of conidia, formation of highly branched hyphae and appressoria, penetration of cuticles, and infection of epidermal cells on chili peppers. Coll-365 accumulated less turgor pressure in appressoria but produced higher levels of cutinase and protease activity than Coll-153 and Coll-524 did. All three isolates invaded the neighbouring cells through plasmodesmata in chili peppers and showed similar pectinase or cellulase activities in culture. However, the most virulent strain Coll-524 expressed stronger laccase activity and was more resistant to capsaicin compared to Coll-153 and Coll-365. The three isolates are different in numbers and sizes of double-stranded RNAs. Depending on the cultivar genotypes, cellular resistance of chili pepper to C. acutatum might rely on the ability to restrict penetration, colonization, or conidiation of the pathogen. We conclude that the differences in pathogenicity among the three C. acutatum isolates of pepper are attributed to their ability to colonize the host plant.     

DNA sequence analysis of ribosomal ITS region 1 of Phytophthora vignae f. sp. adzukicola, the pathogen that causes stem rot of adzuki bean

Sequences of the internal transcribed spacer (ITS) region 1 were used to examine the phylogenetic relationships among races of 19 isolates of Phytophthora vignae f. sp. adzukicola and between this forma specialis and three isolates of the closely related P. vignae f. sp. vignae. The ITS 1 sequences were highly conserved (> 98.7% similarity) among representatives of both formae speciales groups. The results of this study indicate that P. vignae is a monophyletic group. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession nos. AB120062–AB120080 and AB120122     

Molecular characterization of <Emphasis Type="Italic">Phoma tracheiphila</Emphasis>, causal agent of Mal secco disease of citrus,in Israel

Mal secco disease of citrus caused by Phoma tracheiphila is a devastating disease in the Mediterranean basin. Susceptible citrus species include lemon, citron, lime and others. Trees attacked by the fungus show characteristic symptoms; the smallest twigs die first, followed by the larger branches. Eventually, the whole tree is killed. The symptoms are clear in the orchards but by the time they are visible the disease is already well established. The need for a sensitive, reliable and rapid diagnostic method for the early identification of the fungus in trees and fruit exists. We have developed a PCR-based method for the identification of P. tracheiphila from plant tissues including fruit. Any such method must take into account the genetic variability in the pathogen population. Molecular methods were used to compare different isolates of P. tracheiphila. This study found no significant differences between different isolates from different citrus species from different parts of Israel.     

Fruit spot of sweet lime (<Emphasis Type="Italic">Citrus limetta</Emphasis>) caused by <Emphasis Type="Italic">Septoria</Emphasis> sp. in Peru

In 2002, a severe fruit spot of sweet lime (Citrus limetta) was observed in Piura and Lambayeque provinces in northern Peru. Affected fruits showed large oval and sunken lesions, often surrounded by chlorotic haloes. Septoria sp. was isolated from affected fruits. Sweet lime isolates showed larger pycnidia and pycnidiospores than those of Septoria spp. previously described on citrus. In addition, phylogenetic analysis of the ITS sequences clearly separated the sweet lime isolates from S. citri and S. citricola. Isolates were pathogenic to detached sweet lime fruits and the fungus was isolated from lesions on inoculated fruits.     

Pathogenicity,colony morphology and diversity of isolates of <Emphasis Type="Italic">Guignardia citricarpa</Emphasis> and <Emphasis Type="Italic">G. mangiferae</Emphasis> isolated from <Emphasis Type="Italic">Citrus</Emphasis> spp.

In the present study, the pathogenicity of 36 isolates of Guignardia species isolated from asymptomatic ‘Tahiti’ acid lime fruit peels and leaves, ‘Pêra-Rio’ sweet orange leaves and fruit peel lesions, and a banana leaf were characterized. For pathogenicity testing, discs of citrus leaves colonized by Phyllosticta citricarpa under controlled laboratory conditions were kept in contact with the peels of fruit that were in susceptible states. In addition, pathogenicity was related to morphological characteristics of colonies on oatmeal (OA) and potato dextrose agar (PDA). This allowed the morphological differentiation between G. citricarpa and G. mangiferae. Polymerase chain reactions (PCRs) were also used to identify non-pathogenic isolates based on primers specific to G. citricarpa. A total of 14 pathogenic isolates were detected during pathogenicity tests. Five of these were obtained from leaf and fruit tissues of the ‘Tahiti’, which until this time had been considered resistant to the pathogen. Given that the G. citricarpa obtained from this host was pathogenic, it would be more appropriate to use the term insensitive rather than resistant to categorize G. citricarpa. A non-pathogenic isolate was obtained from lesions characteristic of citrus black spot (CBS), indicating that isolation of Guignardia spp. under these conditions does not necessarily imply isolation of pathogenic strains. This also applied to Guignardia spp. isolates from asymptomatic citrus tissues. Using fluorescent amplified fragment length polymorphism (fAFLP) markers, typically pathogenic isolates were shown to be more closely related to one another than to the non-pathogenic forms, indicating that the non-pathogenic isolates display higher levels of genetic diversity.     

Differences in virulence of <Emphasis Type="Italic">Phytophthora capsici</Emphasis> isolates from a worldwide collection on host fruits

Phytophthora capsici causes root, crown, and fruit rot of vegetable and tropical hosts. Cucumber, zucchini, tomato, and pepper fruits were inoculated using 6-mm-diameter agar plugs of P. capsici, incubated in clear plastic boxes at room temperature (25 ± 2°C and 100% relative humidity), and virulence was estimated by measuring the lesion diameter, pathogen growth diameter, and pathogen sporulation density three (cucumber, zucchini) or four (tomato, pepper) days later. When isolates were grouped by genetic cluster, significant differences in virulence were observed on cucumber and zucchini, with isolates belonging to genetic cluster five causing larger lesions than isolates from genetic cluster six. On tomato, no significant differences were observed for isolates grouped by genetic cluster, but isolates from vegetable crops were generally more virulent than isolates from tropical hosts. Isolates from fabaceous hosts sporulated better on cucumber fruits than isolates from solanaceous hosts. Isolates from vegetable hosts sporulated better on zucchini than isolates from tropical hosts. No significant differences in lesion diameter were noted on pepper when isolates were grouped by host family of origin or genetic cluster, but differences in pathogen sporulation were apparent by host family. Our findings suggest that isolate characteristics such as host family of origin and genetic cluster membership may be used to guide initial isolate selection for cucurbit fruit resistance screening. Final isolate selection should incorporate the phenotypic and genetic diversity of P. capsici, including isolates with differing virulence to the host organ of interest.     

Host specificity,but not high-temperature tolerance,is associated with recent outbreaks of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> in chrysanthemum in the Netherlands

Two hypotheses which might explain a recent increase in the incidence of verticillium wilt of chrysanthemums in glasshouses in the Netherlands were investigated, viz whether selection for increased resistance to elevated temperatures has occurred due to frequent steaming of soils in the glasshouses, or whether the strains of Verticillium dahliae occurring in chrysanthemum glasshouses are particularly virulent towards this host. Following artificial inoculation, five isolates of V. dahliae from chrysanthemum were pathogenic on chrysanthemum but five isolates from potato were non-pathogenic for this host. When inoculated onto potato plants, all isolates caused early senescence with no significant difference between the two groups of isolates. In amplified fragment length polymorphism analysis, the potato isolates formed a cluster distinct from all other isolates. As a group the chrysanthemum isolates were no more diverse than the potato isolates but did not form a cluster distinct from 12 other isolates tested. This suggests that high pathogenicity to chrysanthemum has developed on several occasions but that the group of potato isolates were possibly monophyletic. Microsclerotia produced in vitro from the chrysanthemum isolates had significantly lower average lethal temperature tolerance than those from the five potato isolates suggesting that being able to resist the effects of soil sterilisation by steam is not a factor in wilt of chrysanthemums in the Netherlands.     

Occurence of sclerotinia stem rot of fenugreek caused by <Emphasis Type="Italic">Sclerotinia trifoliorum</Emphasis> and <Emphasis Type="Italic">S. sclerotiorum</Emphasis> in Tunisia

Fenugreek is an annual leguminous crop grown for hay and grains in Tunisia. It is also considered a valuable rotation crop with cereals. Sclerotinia rot was observed in production fields since 2010. The survey conducted in 2013 revealed that the incidence of diseased plants varied between 5 and 20%. The identification of isolates of Sclerotinia obtained from fenugreek plants with symptoms of stem rot was determined using morphological and molecular criteria. The size, shape and abundance of sclerotia in potato dextrose agar (PDA) cultures were used to classify isolates as S. sclerotiorum or S. trifoliorum. A comparison of colony diameter on PDA after 24, 48 and 72 h at 25 °C, showed that one isolate grew faster (36 mm/day) than the other 10 isolates (14.8 mm/day). There was a significant difference in sclerotial size between the fast and the slow growing isolates, but there was no significant difference in the number of sclerotia produced after 3 weeks on PDA. Two of the slow growing isolates exhibited ascospore dimorphism, whereas the fast growing isolate did not. PCR amplification with the primer pair ITS5/ITS4 produced a fragment of 560 base pairs from the fast growing isolate and 1000 base pairs from all of the slow growing isolates. The ITS sequences of the fast growing isolate had 100% homology with S. sclerotiorum, whereas those of the slow growing isolates had 100% homology with S. trifoliorum. Isolates of both species were pathogenic on fenugreek seedlings in the greenhouse assay and there was no significant difference in the percentage of dead plants two weeks after inoculation between the two species.     

<Emphasis Type="Italic">In vitro</Emphasis> screening for sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis>L.) resistant calli to <Emphasis Type="Italic">Diaporthe helianthi</Emphasis> fungal culture filtrate

Diaporthe helianthi the causal agent of sunflower (Helianthus annuus) stem canker, causes significant reductions in yield and oil content in most sunflower-growing areas. With the aim of enhancing host resistance, we selected in vitro sunflower calli against culture filtrates of two pathogen isolates (7/96 and 101/96). This technique may be an effective and rapid tool to discriminate the most virulent D. helianthi isolate and to screen for host resistance in the early stage of a breeding programme. Further investigation on the mechanisms involved in defence pathways showed no induction of salicylic acid and pathogenesis-related proteins in calli, indicating that the host resistance is not associated with Systemic Acquired Resistance but probably other biochemical mechanisms.     

Characterisation of <Emphasis Type="Italic">Phoma tracheiphila</Emphasis> by RAPD-PCR,microsatellite-primed PCR and ITS rDNA sequencing and development of specific primers for <Emphasis Type="Italic">in planta</Emphasis> PCR detection

Thirty six isolates of Phoma tracheiphila from Italy, the causal agent of the mal secco disease on Citrus species, were characterised by different molecular tools in comparison with representative isolates of other phytopathogenic Phoma species. These included analysis of the distribution of RAPD and microsatellite markers and sequencing of the internal transcribed spacer (ITS) region of the nuclear rRNA genes. The results obtained with 12 RAPD primers (92 markers) and 7 microsatellite primers (56 markers) suggest that Italian isolates of P. tracheiphila are genetically homogeneous, leading to identical patterns upon amplification with all the tested primers. Accordingly, ITSI-5.8S-ITS2 sequences were highly conserved (98–100% identity along a 544-characters alignment) among all the isolates of P. tracheiphila. A neighbor-joining analysis of ITS sequences of P. tracheiphila in comparison with those of other Phoma species, as well as with alignable sequences from anamorphic and teleomorphic taxa retrieved in BLAST searches, revealed a close relationship between P. tracheiphila and Leptosphaeria congesta. A pair of P. tracheiphila-specific primers was designed on the consensus sequence (555 residues) obtained from the alignment of the newly generated P. tracheiphila ITS sequences. A PCR-based specific assay coupled to electrophoretic separation of amplicons made it possible to detect P. tracheiphila in naturally infected Citrus wood tissue collected from both symptomatic and symptomless plants. The limit of detection was 10 pg of genomic DNA and 5 fg of the ITS target sequence.     

广东果树上17种拟茎点霉的RAPD分析

 自320个随机引物中筛选出适于拟茎点霉属真菌种间亲缘关系分析的15个随机引物,并优化了RAPD分析的扩增体系,在此基础上,对广东果树上17种拟茎点霉进行了RAPD分析。各菌株间的Nei相似系数UPGMA法聚类结果表明:来源于不同地区的2个Phomopsis mangiferae Ahmad菌株和2个P.macadami Z.D.Jiang et P.K.Chi菌株都分别以0.636和0.589的相似系数两两首先聚在一起,而不同的种则只在小于0.54的相似系数范围内聚类,体现了种间及种内的亲缘关系差异程度;聚类群与寄主植物不具相关性,同种植物上的不同拟茎点霉,即使是分自相同寄生部位也不能聚在一类;支持形态学上将生于柑桔枝和黄皮茎、沙梨叶和果、杨梅叶和枝以及同是生于龙眼叶的共8个拟茎点霉分别鉴定为不同的种,而不支持将P. cytosporella Penz.et Sacc.与P. mangiferae合并为一个种的观点;RAPD技术可作为拟茎点霉属真菌种间的亲缘关系分析的重要手段。     

New Media for Increasing Sporulation and Germination of Phaeoisariopsis Griseola Conidia

The slow growth rate of the fungus Phaeoisariopsis griseola and the availability of a homogeneous highly concentrated inoculum is an important constraint for pathogenicity or virulence studies, where plant inoculations are needed. Therefore, the purpose of this paper was to evaluate the effect of supplementing culture media with Amaranthus cruentus seed meal on fungal growth and sporulation of isolates of P. griseola belonging to the Mesoamerican and Andean groups. The amendment of PDA or V8 media with A. cruentus seed meal resulted in a considerable increase in the number of conidia and also in their capacity to germinate; this depended mostly on the stage of maturity of conidia. Mesoamerican and Andean isolates produced a different number of conidia when cultured in vitro. Furthermore, while in Mesoamerican isolates a second degree polynomial represented the relationship between number of conidia and amount of A. cruentus supplementation, in Andean isolates the relationship was linear. It seems that either one or several of the nutritional factors provided by A. cruentus contributed to the increased production of conidia and their development, resulting in faster development of the disease and an earlier appearance of symptoms. Therefore, for cultural studies, especially for inoculum production and for pathogenicity evaluations, supplementation of the media with A. cruentus seed meal proved to be a good alternative.     

Identification of W-type and R-type isolates of Pseudocercosporella herpotrichoides

W-type isolates of Pseudocercosporella herpotrichoides grown on a maize-based agar and exposed to near- ultra-violet radiation at c . 13°C produced a greenish black colour, whilst R-type isolates produced a pink or pale brown colour in the agar medium. More colonies from directly plated lesions or from spore suspensions could be recognized as P. herpotrichoides and could be more easily differentiated as W-type or R-type or as mixtures of both by colour production on maize agar (MA) than by colony morphology on potato dextrose agar (PDA), despite the presence of other fungi. Isolates with intermediate morphology on PDA were positively identified as W-type or R-type on MA; their pathogenicities to wheat and rye seedlings were usually similar to those of W-type or R-type isolates with typical colony morphology, confirming their identification on MA. Drops of mixed suspensions of W-type and R-type spores on PDA formed fast-growing colonies with smooth margins which sometimes had slow-growing sectors with feathery margins. Drops of the same mixtures on MA formed greenish black colonies which sometimes had pink or pale brown sectors. However, when these mixtures were spread onto MA, W-type and R-type colonies could easily be differentiated by colour.     

Testing variability in pathogenicity ofPhytophthora cactorum, P. citrophthora andP. syringae to apple, pear, peach, cherry and plum rootstocks

The relative virulence ofPhytophthora cactorum andP. syringae originating from almond trees, and ofP. citrophthora originating from citrus, to apple, pear, peach, cherry and plum rootstocks, was studiedin vivo andin vitro. Results of the different experiments were in good agreement. All testedPhytophthora isolates showed little virulence to pear rootstocks-causing only minor crown rot symptoms - and no virulence at all to apple rootstocks. In contrast, they were highly virulent to stone fruit rootstocks, causing crown rot disease. The non-pathogenicity of these isolates to pome rootstocks could be interpreted as strict host specificity.