Identifying resistance in wild and ornamental cherry towards bacterial canker caused by Pseudomonas syringae

Bacterial canker is a major disease of stone fruits and is a critical limiting factor to sweet cherry (Prunus avium) production worldwide. One important strategy for disease control is the development of resistant varieties. Partial varietal resistance in sweet cherry is discernible using shoot or whole tree inoculations; however, these quantitative differences in resistance are not evident in detached leaf assays. To identify novel sources of resistance to canker, we used a rapid leaf pathogenicity test to screen a range of wild cherry, ornamental Prunus species and sweet cherry × ornamental cherry hybrids with the canker pathogens, Pseudomonas syringae pvs syringae, morsprunorum races 1 and 2, and avii. Several Prunus accessions exhibited limited symptom development following inoculation with each of the pathogens, and this resistance extended to 16 P. syringae strains pathogenic on sweet cherry and plum. Resistance was associated with reduced bacterial multiplication after inoculation, a phenotype similar to that of commercial sweet cherry towards nonhost strains of P. syringae. Progeny resulting from a cross of a resistant ornamental species Prunus incisa with susceptible sweet cherry (P. avium) exhibited resistance indicating it is an inherited trait. Identification of accessions with resistance to the major bacterial canker pathogens is the first step towards characterizing the underlying genetic mechanisms of resistance and introducing these traits into commercial germplasm.     

Paecilomyces farinosus destroys powdery mildew colonies in detached leaf cultures but not on whole plants

Since 2001, several isolates of Blumeria graminis, the causal agent of cereal powdery mildew, maintained on detached leaves at the John Innes Centre, Norwich, UK, have spontaneously become infected with an unknown filamentous fungus whose mycelia have quickly overgrown the powdery mildew colonies and destroyed them completely. A total of five isolates of the contaminant were obtained and identified as Paecilomyces farinosus based on morphological characteristics and rDNA ITS sequence data. To determine whether these P. farinosus isolates can be considered as biocontrol agents (BCAs) of powdery mildews, we studied the interactions between P. farinosus and the following four powdery mildew species: B. graminis f.sp. hordei infecting barley, Oidium neolycopersici infecting tomato, Golovinomyces orontii infecting tobacco and Podosphaera fusca infecting cucumber. The powdery mildew colonies of all these four powdery mildew species were quickly destroyed by P. farinosus in leaf cultures but neither conidial suspensions nor cell-free culture filtrates of P. farinosus isolates could suppress the spread of powdery mildew infections on diseased barley, tomato, tobacco or cucumber plants in the greenhouse. It is concluded that P. farinosus cannot be considered as a promising BCA of powdery mildew infections although it can destroy powdery mildew colonies in detached leaf cultures and can be a menace during the maintenance of such cultures of cereal, apple, cucurbit and tomato powdery mildew isolates.     

Overwintering of Ampelomyces mycoparasites on apple trees and other plants infected with powdery mildews

Apple shoots and aerial parts of 13 other plant species infected with powdery mildews during the previous season were collected in late winter and early spring between 1998 and 2003 at a total of 34 sample sites in Hungary. Samples were examined for the presence of overwintering structures of Ampelomyces, common mycoparasites of powdery mildews. Pycnidia and resting hyphae resembling those of Ampelomyces were found on six plant species, including apple. Their viability and subsequent mycoparasitic activity of the hyphae emerging from the overwintered fungal structures were studied in vitro to determine whether they can serve as sources of primary inocula of Ampelomyces in the spring. Overwintered pycnidia of Ampelomyces collected in the spring, and produced in both the ascomata and the conidiophores of powdery mildews during the previous season, initiated the life cycle of these mycoparasites when placed close to fresh powdery mildew colonies in vitro. Similarly, thick-walled resting hyphae, found in the dried powdery mildew mycelia which covered the overwintered aerial parts of the host plants, also germinated and gave rise to new intracellular pycnidia of Ampelomyces when powdery mildew colonies were inoculated with them in vitro. On apple trees, Ampelomyces mycoparasites overwintered as resting hyphae in the dried powdery mildew mycelia covering the shoots and in the parasitized ascomata of Podosphaera leucotricha on the bark and the scales of the buds. Approximately 31% of the field samples collected from apple trees in spring between 1998 and 2003 contained overwintered structures of Ampelomyces. Artificial bursting of apple buds in the laboratory showed that both P. leucotricha and Ampelomyces start their life cycle during or soon after bud burst, but Ampelomyces can only slowly follow the spread of its mycohost on infected leaves. Most probably, the mycoparasites did not overwinter in the dormant hyphae of P. leucotricha in the buds, but only on the bark and the bud scales, as their hyphae were not found in the young hyphae of apple powdery mildew that appeared on the leaf tissues during bud burst. This study demonstrated that Ampelomyces mycoparasites can survive the winter in the field as pycnidia and as resting hyphae in the dried mycelia of their mycohosts.     

Resistance levels of cucumber to <Emphasis Type="Italic">Podosphaera xanthii</Emphasis> in a growth chamber are related to haustorial formation and hyphal branching frequency

To clarify relationships between powdery mildew resistance of cucumber and hyphal developmental state of the pathogen, haustorial formation and the hyphal branching frequency were compared among cucumber varieties that differ in resistance levels to powdery mildew pathogen Podosphaera xanthii. Cotyledons of four cultivars were inoculated with P. xanthii. By 2 days after inoculation, secondary haustoria had developed from the first hyphal cell that formed beside the conidium in susceptible cultivars. The fungus on susceptible cultivars also tended to have hyphal branches just after the hyphal cell producing haustoria.     

Differentiation of two powdery mildews of sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis>) by a PCR-mediated method based on ITS sequences

Powdery mildew can be found in most sunflower fields during the winter season in Taiwan and causes severe yellowing on the blade, petiole, stem, and calyx, as well as serious defoliation. Two types of powdery mildew fungi isolated from sunflower leaves showed variable status for fibrosin bodies. But only the cleistothecium of Podosphaera xanthii, one of the pathogens causing this disease, was observed on samples from Chungpu County at the beginning of 2005. With a species-specific primer pair, PN23/PN34, no specific PCR product was amplified from the pathogen’s genomic DNA. Based upon the ITS sequence of rDNA, three PCR primer sets (S1/S2, G1/G2, and L1/L2) specific to P. xanthii, Golovinomyces cichoracearum and Leveillula taurica, respectively, were designed to detect and differentiate pathogens causing powdery mildews on sunflower. Only the primer pairs S1/S2 and G1/G2 could amplify PCR products, with product sizes of 454 and 391 bp, respectively. Four samples of fungal DNA were subjected to a multiplex PCR amplification with primer pairs S1/S2 and G1/G2; P. xanthii and G. cichoracearum were successfully detected. These results suggest that the multiplex PCR method is a rapid, simple, and effective technique to detect and differentiate powdery mildews, for example P. xanthii and G. cichoracearum, found on sunflower. With morphologic characteristics, ITS sequence analysis and pathogenicity testing, P. xanthii and G. cichoracearum, the first case, are two powdery mildews on sunflower in Taiwan.     

Identification and distribution of mating‐type idiomorphs in populations of Podosphaera macularis and development of chasmothecia of the fungus

Podosphaera macularis, the causal agent of hop powdery mildew, is known to produce chasmothecia (formerly cleistothecia) in eastern North America and Europe. Ascocarps have not yet been reported from the Pacific Northwestern region of North America. Reasons for the apparent absence of chasmothecia in the Pacific Northwest were unknown. This study established that P. macularis is heterothallic and ascocarp ontogeny, maturation, dehiscence and ascospore infection proceed similarly to other powdery mildew fungi. Genome sequencing of a MAT1‐1 isolate revealed the structure of the MAT1 locus and presence of MAT1‐1‐3, demonstrating further similarities to other powdery mildew fungi. PCR assays with primers designed from conserved domains of the MAT1 idiomorphs were developed to characterize the frequency of idiomorphs in populations of P. macularis. Amongst 317 samples of P. macularis collected during 2012 and 2013 from the Pacific Northwest only the MAT1‐1 idiomorph was found. In contrast, among 56 samples from the eastern United States and Europe, MAT1‐1 and MAT1‐2 idiomorphs were detected at equivalent frequencies. At temperatures representative of late season conditions in the Pacific Northwest, chasmothecia formed readily when a Pacific Northwest MAT1‐1 isolate was paired with a MAT1‐2 isolate collected from outside the region. Although these findings do not encompass all climatic, geographic or temporal barriers that could inhibit the formation of chasmothecia, the current absence of the ascigerious stage of P. macularis in the Pacific Northwest could be explained by the absence of the MAT1‐2 mating type idiomorph.     

Efficacy of biocontrol agents and natural compounds against powdery mildew of zucchini

The activity of different types of natural compounds and of two biofungicides based on Bacillus subtilis and Ampelomyces quisqualis alone and in combination with fungicides was tested against powdery mildew of zucchini. The efficacy was compared to the activity of fungicides used alone in four experimental trials carried out in the open field and under greenhouse conditions. The Podosphaera xanthii population used throughout the work was partially resistant to azoxystrobin, whereas it was susceptible to mychlobutanil. Sulphur plus terpenes and mustard oil consistently controlled powdery mildew, followed by mychlobutanil alone or in combination with A. quisqualis. B. subtilis and A. quisqualis when tested alone were partially effective. The combination of azoxystrobin and B. subtilis only delayed the spread of the pathogen.     

Plant age and ambient temperature: significant drivers for powdery mildew (Erysiphe cruciferarum) epidemics on oilseed rape (Brassica napus)

Powdery mildew (Erysiphe cruciferarum) is an important disease in oilseed rape crops worldwide, but of sporadic importance in most southern Australian crops. Six Brassica napus cultivars were exposed to E. cruciferarum simultaneously in four plant age cohorts. First symptoms of powdery mildew appeared 9 days after inoculation (dai) on the oldest plants [42 days after seeding (das)], but 44 dai in the youngest plants that were exposed to inoculum from sowing, although final disease severity did not differ with the plant age at exposure. The maximum level of pod peduncle infestation was unaffected by plant age (P = 0.37) or cultivar (P = 0.28). The effect of temperature was also investigated. The development of disease on plants was slower and final severity reduced at a day/night temperature 14/10 °C compared with 22/17 °C. In vitro, maximum growth of germ tubes from conidia of E. cruciferarum was at 15–20 °C and survival of conidia reduced by temperatures >30 °C. The results explain the sporadic nature of powdery mildew outbreaks in winter‐grown oilseed rape in Australia, where slow rates of infection occur when seasonal colder prevailing winter conditions coincide with the presence of younger plants, together curtailing rapid disease development until temperatures increase in late winter/early spring. These results explain why epidemics are most severe in the two warmer cropping regions, viz. the northern agricultural region of Western Australia and New South Wales. This study suggests that with increases in winter temperatures under future climate scenarios, earlier and more severe powdery mildew outbreaks in Australia will be favoured.     

Species spectra,distribution and host range of cucurbit powdery mildews in the Czech Republic,and in some other European and Middle Eastern countries

The occurrence and geographic distribution of powdery mildew on cucurbits was studied in the Czech Republic, Austria, France, Germany, Great Britain, Italy, Slovakia, Slovenia, Spain, Switzerland, the Netherlands, as well as in Turkey and Israel, during the period 1995–2000. In total, 599 leaf samples with powdery mildew symptoms were collected from cucurbits from 166 locations within the Czech Republic; an additional 69 samples were collected from 42 locations elsewhere. Two powdery mildews (Golovinomyces cichoracearum and Podosphaera xanthii) were identified. The host range included the cultivated cucurbits (Cucumis melo, C. sativus, Cucurbita pepo, C. maxima, and Citrullus lanatus) and several other species. P. xanthii was the only powdery mildew pathogen found in Spain, Israel, and Turkey. P. xanthii and G. cichoracearum were detected in the other surveyed countries, occurring in both mixed infections and separately. In the more northerly latitudes and higher elevations, G. cichoracearum is more often the single species. G. cichoracearum was the dominant powdery mildew species in the Czech Republic (detected in 98.8% of the locations there); P. xanthii was found as the lone species in 1.2% of locations. At 28.4% of locations, G. cichoracearum was found with P. xanthii as mixed infections. The hyperparasitic fungus Ampelomyces quisqualis was found in 30% of the samples from the Czech Republic and was also recorded in Austria, Italy, Slovenia, Switzerland, and Great Britain.     

Concepts and methods in biological control of diseases in apple orchards1

The project ‘Promotion of Biological Methods in Apple Growing’ which started in Switzerland in 1985 concentrates on the control of Venturia inaequalis (scab) and Podosphaera leucotricha (powdery mildew). The project considers the selection of suitable cultivars resistant or only slightly susceptible to fungal diseases and the assessment of tolerable economic loss. It also includes curative treatments with sterol-synthesis-inhibiting fungicides, according to scab infection periods, as well as applications of plant extracts (proposed by the Institute of Biological Husbandry) and suspensions of antagonists or their culture filtrates as plant-protecting agents. Screening experiments on apple seedlings showed that root extracts of Rumex obtusifolium significantly reduced powdery mildew infections under greenhouse conditions. In field experiments treatments with R. obtusifolium extracts at intervals of 7–10 days were less effective. Saponin-containing extracts of different plants gave satisfying control of scab on seedlings in the growth chamber but were not very effective in the field. Applications of spore suspensions and culture filtrates of the phyllosphere fungi Chaetomium spp. reduced scab and mildew infections on apple seedlings to some extent. It was found that in the last 2 years even primary infections of P. leucotricha were severely parasitized by Ampelomyces quisqualis. Preliminary studies showed that the lack of efficacy of this biocontrol system is due to the delayed spread of the hyperparasite.     

一体化智能孢子捕捉系统在黄瓜霜霉病和黄瓜白粉病预测上的应用

为探索国内研制的新型一体化智能孢子捕捉系统在黄瓜霜霉病和黄瓜白粉病预测预报上的应用,在田间自然发病情况下,通过对捕捉孢子的形态进行识别,优化一体化智能孢子捕捉系统主要工作参数如有/无空气切割头、空气采集口高度和空气采集时间;通过病害及孢子的动态监测分析大棚黄瓜霜霉病和黄瓜白粉病病情指数与孢子捕捉量的关系。结果表明,当不加装空气切割头、空气采集口高度为70 cm、孢子捕捉时间在10:00—10:30时段有利于孢子的捕捉。黄瓜霜霉病和黄瓜白粉病病情指数与连续7 d孢子捕捉总量具有强正相关性。连续多日监测到黄瓜霜霉病菌孢子囊且数量快速增加是黄瓜霜霉病发生或快速上升的一个预测指标。黄瓜白粉病发病之前没有监测到黄瓜白粉病菌分生孢子,且在病害盛发期分生孢子捕捉量仍较少。研究表明,一体化智能孢子捕捉系统适用于黄瓜霜霉病的预测,但在黄瓜白粉病的预测上尚存在一定问题。     

利用28S rDNA D1/D2区和ITS rDNA序列鉴定甜瓜白粉病病原菌

为了明确宁夏干旱带压砂甜瓜白粉病病原菌,从病原菌分生孢子中提取DNA,PCR扩增ITSrDNA和28S rDNA D1/D2区段,测序后进行BLAST比对.结果表明,病原菌的ITS rDNA和28SrDNA D1/D2序列与菜豆叉丝单囊壳白粉菌Podosphaera phaseoli、凤仙花又丝单囊壳白粉菌P.bal-saminae、菊科叉丝单囊壳白粉菌P.fwca、苍耳单囊壳白粉菌P.xanthii、瓜类单囊壳白粉菌P.fuligi-nea等叉丝单囊壳属Podosphaera的多个种的ITS rDNA和28S rDNA D1/D2序列之间相似度均大于99%,鉴定甜瓜白粉病病原菌为叉丝单囊壳属Podosphaera.     

Pseudomonas syringae pv. avii (pv. nov.), the Causal Agent of Bacterial Canker of Wild Cherries (Prunus avium) in France

Bacterial strains isolated from cankers of wild cherry trees (Prunus avium) in France were characterized using numerical taxonomy of biochemical tests, DNA–DNA hybridization, repeat sequence primed-PCR (rep-PCR) based on REP, ERIC and BOX sequences, heteroduplex mobility assay (HMA) of internal transcribed spacer (ITS) as well as pathogenicity on wild cherry trees and other species of Prunus. They were compared to reference strains of Pseudomonas syringae pathovars isolated from wild and sweet cherry and various host plants. Wild cherry strains were closely related to P. syringae (sensu lato) in LOPAT group Ia (+ - - - +). Wild cherry strains were pathogenic to wild cherry trees and produced symptoms similar to those observed in orchards. They were pathogenic also, but at a lesser extent, to sweet cherry trees (cv. Napoléon). The wild cherry strains were collected from five different areas in France and appeared to constitute a very homogeneous group. They showed an homogenous profile of a biochemical and physiological characteristics. They were closely related by DNA–DNA hybridization and belonged to genomospecies 3 `tomato'. Rep-PCR showed that wild cherry strains constitute a tight group distinct from P. s. pv. morsprunorum races 1 and 2 and from other P. syringae pathovars. HMA profiles indicated that the ITS of all wild cherry strains were identical but different from P. s. pv. persicae strains since the two heteroduplex bands with reduced mobility were generated by hybridization with the P. s. pv. persicae pathotype strain CFBP 1573. The 8 genomospecies of Gardan et al. (1999) have not been converted into formal species as they cannot be differentiated by biochemical tests. Therefore, the pathovar system within P. syringae was currently used. P. syringae pv. avii is proposed for this bacterium causing a wild cherry bacterial canker and strain CFBP 3846 (NCPPB 4290, ICMP 14479) is designated as the pathotype.     

小豆白粉病菌的鉴定与小豆品种抗白粉病评价

为明确小豆白粉病病原菌的种类以及小豆种质资源对白粉病的抗性,采用形态学和系统发育学方法对近年来在北京市发生的小豆白粉病病原菌种类进行鉴定,并采用室内苗期人工接种法评价小豆常见栽培品种（系）对白粉病的抗性。结果表明,从北京市采集的感白粉病小豆病样中培养获得病原菌BJ1,该菌能在小豆叶片和茎上产生明显的白色粉斑,分生孢子梗直立,不分枝,分生孢子单细胞,成链状着生于分生孢子梗上,呈椭圆形或卵圆形。通过rDNA-ITS序列系统发育分析,小豆白粉病菌BJ1被鉴定为白粉菌目白粉菌科的苍耳叉丝单囊壳Podosphaera xanthii。室内苗期人工接种条件下,19个供试小豆品种（系）接种小豆白粉病菌BJ1后均可发病,其中9个审定品种均表现为中度感病或高度感病,10个优良品系发病略轻。     

Crop damage caused by powdery mildew on hop and its relationship to late season management

Powdery mildew of hop (Podosphaera macularis) may cause economic loss due to reductions in cone yield and quality. Quantitative estimates of crop damage from powdery mildew remain poorly characterized, especially the effect of late season disease management on crop yield and quality. Field studies in Washington State evaluated cone yield, bittering acid content and quality factors when fungicide applications were ceased at different stages of cone development. The incidence of cones with powdery mildew was linearly correlated with yield of cones, bittering acids and accelerated cone maturation. In cultivar Galena, the cumulative effect of every 1% increase in cones powdery mildew incidence was to reduce alpha‐acid yield by 0·33%, which was due to direct effects on cone yield but also indirect effects mediated by dry matter. In the more susceptible cultivar Zeus, alpha‐acid yield was increased 20% by controlling powdery mildew through the transition of bloom to early cone development compared to ceasing fungicide applications at bloom: additional applications provided only modest improvements in alpha‐acid yield. In both cultivars, the impact of powdery mildew on aroma characteristics and bittering acid content were less substantial than cone yield. The damage caused by powdery mildew to cone colour and alpha‐acid yield, as well as the effectiveness of fungicide applications made to manage the disease, appears inseparably linked to dry matter content of cones at harvest. Realising achievable yield potential in these cultivars requires control of the disease through early stages of cone development and harvest before maturity exceeds c. 25% dry matter.     

Effect of temperature on infection and development of powdery mildew on cucumber

Podosphaera xanthii and Golovinomyces orontii are the causal agents of cucurbit powdery mildew. The effect of temperature on conidial germination, infection and sporulation was studied under controlled conditions. Conidia were inoculated on cucumber leaf discs, and incubated at six constant temperatures (from 10 to 35 °C in 5 °C steps) for 3 to 72 h to evaluate conidial germination and infection, and for 6–15 days to evaluate sporulation intensity. Germination took place at all tested temperatures, but was close to zero at 35 °C. The longest germ tubes measured in this experiment were 141.74 μm for the secondary germ tube of P. xanthii at 20 °C after 48 h of incubation, and 67.92 μm for G. orontii for the primary germ tube at 20 °C after 48 h of incubation. The optimal temperatures for conidial germination, infection and sporulation were 24.4, 25.7 and 22.3 °C, respectively, for P. xanthii, and 17.9, 17.3 and 14.9 °C, respectively, for G. orontii. Equations were developed to describe conidial germination with a coefficient of determination (R²) of 0.85 and 0.90 for P. xanthii and G. orontii, respectively. Infection equations resulted in R² of 0.94 and 0.93 for P. xanthii and G. orontii, respectively; and for sporulation, R² of 0.75 and 0.76 for P. xanthii and G. orontii respectively, as a function of temperature. These results can be used to develop models for the risk of cucurbit powdery mildew under field conditions.     

Characterization of <Emphasis Type="Italic">Phytophthora clandestina</Emphasis> races on <Emphasis Type="Italic">Trifolium subterraneum</Emphasis> in Western Australia

Phytophthora clandestina is a causal agent of root rot disease of subterranean clover in Western Australia (W.A). As a significant number of isolates of P. clandestina from W.A. could not previously be designated using existing differentials, a comprehensive set of subterranean clover (Trifolium subterraneum) cultivars was used as differentials to delineate a broader range of races of the pathogen. One hundred and one isolates of the pathogen collected from W.A. were screened on nine subterranean clover cultivars, of which seven were found to be useful as host differentials. A total of 10 races (in contrast to the five recognized previously) were defined and differentiated using octal nomenclature, presenting a clearer picture of the racial distribution of P. clandestina among W.A. isolates. Differences were found in the race populations between Australian states and are therefore important to the selection/breeding of cultivars for specific regions of Australia to counter the predominant race populations and for enforcing quarantine measures in relation to seed movements within and outside Australia. The octal nomenclature used provides a sound basis for follow-up studies and future race designations. Races 173 and 177 in this study were widely distributed and were the most common races in W.A., and together constitute 80% of the isolates characterized. While six of the seven host differentials were resistant to isolates belonging to race 001 and all were resistant to 000, it is of concern that only one differential was resistant to 157 and 173 and that none of the host differentials were resistant to 177. Our approach to P. clandestina race delineation is clearly conservative and is different from previous studies. The octal nomenclature we applied in this study is not only scientifically sound but also will facilitate rapid recognition and characterization of the races.     

Host range expansion in a powdery mildew fungus (<Emphasis Type="Italic">Golovinomyces</Emphasis> sp.) infecting <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>: <Emphasis Type="Italic">Torenia fournieri</Emphasis> as a new host

Since 2003, Torenia fournieri plants grown for experimental purposes were repeatedly infected by powdery mildew in a laboratory in Hungary. Based on morphological characteristics, the pathogen belonged to the mitosporic genus Oidium subgen. Reticuloidium, the anamorph stage of Golovinomyces. The rDNA ITS sequence was identical to that of two other powdery mildew fungi, infecting Arabidopsis and Veronica, respectively, in different parts of the world. According to a previous phylogenetic analysis of ITS and 28S rDNA sequences, those two powdery mildews belong to a recently evolved group of Golovinomyces characterized by multiple host range expansions during their evolution. Both the ITS sequence and the morphological data indicate that the powdery mildew anamorph infecting Torenia also belongs to this group. It is likely that the powdery mildew infections of the experimental T. fournieri plants, native to south-east Asia, were the result of a very recent host range expansion of a polyphagous Golovinomyces because (i) T. fournieri is absent from our region, except as an experimental plant grown in the laboratory, (ii) the powdery mildew fungus infecting this exotic plant belongs to a group of Golovinomyces where host range expansion is a frequent evolutionary scenario, (iii) cross-inoculation tests showed that this pathogen is also able to infect other plant species, notably A. thaliana and tobacco, and (iv) no Golovinomyces species are known to infect T. fournieri anywhere in the world. Although host range expansion has often been proposed as a common evolutionary process in the Erysiphales, and also in other biotrophic plant pathogens, this has not been clearly demonstrated in any case studies so far. To our knowledge, this is the first convincing case of a host range expansion event in the Erysiphales.     

Phytogeny and Cross-infectivity of Powdery Mildew Isolates (Podosphaera fuliginea s. lat.) on Cosmos and Cucumber

To clarify the relationship between the phylogeny and infectivity of isolates of Podosphaera fuliginea s. lat. (= Sphaerotheca fuliginea s. lat.) from cosmos and cucumber, more than 50 powdery mildew isolates from these two plants were subjected to nucleotide sequencing or PCR-RFLP analysis of the rDNA internal transcribed spacer (ITS) region and cross-inoculation tests. The isolates from both cosmos and cucumber are genetically monotypic, and there are six nucleotide substitutions in the rDNA ITS region between isolates from cosmos and cucumber. Cross-inoculation tests of these isolates revealed that isolates from cosmos are not pathogenic on cucumber. Although isolates from cucumber produced conidia on leaves of cosmos in the laboratory, the conidial density was much lower than that from isolates from cosmos. This result, as well as the fact that the cucumber strain was not isolated from cosmos in fields, suggests that isolates from cucumber do not infect cosmos in the field. Therefore, powdery mildews on cosmos and cucumber can be regarded to have become specialized for their hosts both genetically and pathogenically. The present study reconfirms the close relationship between phylogeny and infectivity of powdery mildew fungi. Host specialization may be a trigger that causes genetic divergence of powdery mildew fungi. Received 28 June 2000/ Accepted in revised form 4 September 2000     

Identification and Discrimination of Pseudomonas syringae Isolates from Wild Cherry in England

A survey of wild cherry (Prunus avium) woodland plantations and nurseries was carried out in 2000/01. Trees with symptoms of bacterial canker were found in 20 of the 24 plantations visited and in three of seven nurseries. Fifty-four Pseudomonas syringae isolates from wild cherry together with 22 representative isolates from sweet cherry and 13 isolates from other Prunus spp., pear and lilac were characterised by physiological, biochemical, serological and pathogenicity tests. Isolates from wild cherry were predominantly P. syringae pv. syringae (Pss), but P. syringae pv. morsprunorum (Psm) races 1 and 2 were also found. Physiological and biochemical tests discriminated Psm races 1 and 2 from other P. syringae isolates. Agglutination and indirect-enzyme-linked immunosorbent assay tests with three different antisera showed that Psm race 1 and race 2 were very uniform and indicated high variability amongst other P. syringae isolates. However, pathogenic Pss isolates could not be distinguished from non-pathogenic isolates of P. syringae on the basis of physiological, biochemical or serological tests. Pathogenicity tests on rooted lilac plants and on micropropagated plantlets of lilac and two wild cherry clones differentiated Pss and Psm isolates and demonstrated a range of aggressiveness amongst Pss isolates. Serological tests could be used as an alternative to the classical physiological and biochemical tests to increase the speed of detection and discrimination of isolates, but pathogenicity tests are still necessary to discriminate the pathogenic Pss isolates.