Characterization of the pathogenicity of strains of Pseudomonas syringae towards cherry and plum

Bacterial canker is a major disease of Prunus avium (cherry), Prunus domestica (plum) and other stone fruits. It is caused by pathovars within the Pseudomonas syringae species complex including P. syringae pv. morsprunorum (Psm) race 1 (R1), Psm race 2 (R2) and P. syringae pv. syringae (Pss). Psm R1 and Psm R2 were originally designated as the same pathovar; however, phylogenetic analysis revealed them to be distantly related, falling into phylogroups 3 and 1, respectively. This study characterized the pathogenicity of 18 newly genome‐sequenced P. syringae strains on cherry and plum, in the field and laboratory. The field experiment confirmed that the cherry cultivar Merton Glory exhibited a broad resistance to all clades. Psm R1 contained strains with differential specificity on cherry and plum. The ability of tractable laboratory‐based assays to reproduce assessments on whole trees was examined. Good correlations were achieved with assays using cut shoots or leaves, although only the cut shoot assay was able to reliably discriminate cultivar differences seen in the field. Measuring bacterial multiplication in detached leaves differentiated pathogens from nonpathogens and was therefore suitable for routine testing. In cherry leaves, symptom appearance discriminated Psm races from nonpathogens, which triggered a hypersensitive reaction. Pathogenic strains of Pss rapidly induced disease lesions in all tissues and exhibited a more necrotrophic lifestyle than hemibiotrophic Psm. This in‐depth study of pathogenic interactions, identification of host resistance and optimization of laboratory assays provides a framework for future genetic dissection of host–pathogen interactions in the canker disease.     

Field evaluation of treatments for the control of the bacterial apical necrosis of mango (Mangifera indica) caused by Pseudomonas syringae pv. syringae

Bacterial apical necrosis is a critical disease in the main production area of mango in Europe. It is caused by Pseudomonas syringae pv. syringae, and produces necrotic lesions on mango buds and leaves, causing severe yield losses due to a decrease of flowering and fruit set. A field study to evaluate control treatments against bacterial apical necrosis was carried out during three seasons on mango trees cv. Tommy Atkins in Huelva (Spain). Experimental treatments included Bordeaux mixture, fosetyl-Al, acibenzolar-s-methyl, gibberelic acid, silicon gel, a mixture between acibenzolar-S-methyl and Bordeaux mixture, and combined applications of fosetyl-Al with Bordeaux mixture or silicon gel. The treatments which caused a consistent reduction in bacterial apical necrosis symptoms at similar levels to the conventional treatment with Bordeaux mixture, were the plant resistance activator acibenzolar-S-methyl and the phosphonate derivative fosetyl-Al applied singly or in combination with other compounds, which could be alternative treatments. These treatments showed a significant decrease in the necrotic buds and/or leaves numbers; however, minor differences in P. syringae-like population levels were observed. The analysis of the inhibitory and bactericidal concentrations of cupric compounds against P. syringae strains isolated from mango tissues suggests that the commercial copper-based treatments with Bordeaux mixture used in the management of mango crops do not work in a bactericidal mode of action.     

Pseudomonas syringae pv. syringae from cool climate Australian grapevine vineyards: new phylogroup PG02f associated with bacterial inflorescence rot

During the period 2006–2011, Pseudomonas syringae pv. syringae caused a bacterial inflorescence rot (BIR) epidemic in an Australian cool climate viticultural region. Molecular multilocus sequence typing of ‘housekeeping’ genes (MLST), biochemical testing and analysis of molecular variance (AMOVA) were used to characterize the genotypes and phenotypes of P. syringae pv. syringae grapevine isolates. Comparison of the MLST data with exemplars of phylogroups available at PAMDB demonstrated that the BIR isolates formed a new clade within P. syringae pv. syringae phylogroup 2 (PG02): putatively designated PG02f. Analysis of the MLST and phenotypic data by AMOVA demonstrated some genetic differences between the BIR isolates and the general vineyard P. syringae pv. syringae population. Isolates positive for syringopeptin, syringomycin and tyrosinase, tobacco leaf hypersensitivity reaction (HR), ampicillin resistance and grapevine leaf pathogenicity were genetically distinct from those negative for these factors. This study has shown that, generally, the core genome of P. syringae pv. syringae is only weakly associated with the virulence-associated traits. As the new phylogroup PG02f consists of the epidemic BIR isolates and nonpathogenic grapevine isolates, these genetically similar isolates differ greatly in pathogenicity and most of the other tested phenotypic traits. However, within the PG02f group, tobacco leaf HR and presence of sylC (the gene for phytotoxin syringolin A) are associated with the BIR and bacterial leaf spot (BLS) isolates, and negative for the nonpathogens, indicating that these two virulence factors may be associated with vineyard pathogenicity within the new Australian phylogroup.     

Susceptibility of European pear cultivars to Pseudomonas syringae pv. syringae using immature fruit and detached leaf assays

The susceptibility of thirty-three pear cultivars and two pear rootstocks to four virulent strains of Pseudomonas syringae pv. syringae was evaluated by inoculating detached immature fruits and young leaves. The four strains were similarly virulent and did not show cultivar specificity although they were isolated from different pear cultivars and exhibited different biochemical profiles. The most frequently planted pear cultivars, Conference, Abate Fetel, General Leclerc, Williams, D. Comice, El Dorado, Alexandrine, B. Anjou, Passe Crassane and the rootstock OHxF 333 were susceptible to P. syringae pv. syringae. Maximal severity values were obtained on 'Preguystar' leaves (about 90%). The rootstock Winter Nelis was less susceptible. Results with immature fruit and detached leaf assays agreed with field observations on cultivar susceptibility to bacterial blast. However, the detached leaf test gave a more accurate prediction and has the advantages that symptoms develop quickly (48 h), and leaves are available for a longer period of time than fruits. This method is proposed as a rapid and reproducible screening system of cultivar susceptibility to bacterial blast of pear.     

Phylogenetic,genetic, and phenotypic diversity of Pseudomonas syringae pv. syringae strains isolated from citrus blast and black pit in Tunisia

Citrus blast and black pit caused by Pseudomonas syringae pv. syringae (Pss) is the only bacterial disease reported in Tunisian Citrus orchards. The phylogenetic relationship between Pss strains was studied based on multilocus sequence analysis (MLSA), using partial sequences of housekeeping genes rpoD, rpoB, gyrB, cts, and pfk for 14 representative Pss Citrus strains, including the reference strain LMG5496. The MLSA revealed that the studied Tunisian Citrus strains are closely related to LMG5496 and cluster in phylogroup 02. Based on the cts gene, the majority of Citrus strains clustered in clades “a” and “b”. However, five strains were placed in a newly defined clade “g”. We describe the presence of six different type III secreted effectors (T3SEs). These were found with frequencies of 100% for the effector hopAN1 and the helper hrpK1, 65% for hopT1-2, and 14% for hopN1, hopR1, and hopQ1-2. Investigation of copper resistance showed that 67% of our Pss Citrus strains from Tunisia are resistant to copper sulphate in vitro, and the copper resistance genes copABCDR were detected in 23% of the strains. Our results present new data concerning the genetic diversity and phylogeny, presence of T3SEs, and copper resistance within the Pss populations that affect Citrus in Tunisia.     

Phenotypic,molecular and pathogenic characterization of Phlyctema vagabunda,causal agent of olive leprosy

Olive leprosy, caused by the fungus Phlyctema vagabunda, is a classic fruit rot disease widespread in the Mediterranean basin. From 2009 to 2013, new disease symptoms consisting of small circular necrotic leaf lesions, coin branch canker and shoot dieback were observed in Spanish and Portuguese olive orchards showing intense defoliation. Phlyctema‐like anamorphs were consistently isolated from leaves and shoots with symptoms. Representative isolates from affected leaves, shoots and fruits were characterized based on morphology of colonies and conidia, optimum growth temperature and comparison of DNA sequence data from four regions: ITS, tub2, MIT and rpb2. In addition, pathogenicity tests were performed on apple and olive fruits, and on branches and leaves of olive trees. Maximum mycelial growth rate ranged between 0.54 and 0.73 mm per day. Conidia produced on inoculated apple fruits showed slight differences in morphology among the representative fungal isolates evaluated. Phylogenetic analysis clustered all of the Phlyctema‐like isolates in the same clade, identifying them as Phlyctema vagabunda. On fruits, influence of wounding, ripening and cultivar resistance was studied, with cv. Blanqueta being the most susceptible cultivar. On branches, a mycelial‐plug inoculation method reproduced olive leprosy symptoms and caused shoot dieback. On leaves, Koch's postulates were fulfilled and the pathogen caused characteristic necrotic spots and plant defoliation. This is the first time that the pathogenicity of P. vagabunda in olive leaves has been demonstrated.     

Pathogenesis and control of bacterial speck,Pseudomonas syringae pv. tomato,on tomato*

Bacterial speck of tomato, caused by Pseudomonas syringae pv. tomato, was observed to cause severe symptoms, especially on protected tomato crops, during the winter season in the coastal area of Lebanon. This study was conducted to investigate the aetiology and pathogenesis of the bacterium involved and the efficacy of different chemicals for the control of the disease. Biochemical, physiological and pathological tests verified the identity of the bacterium involved as P. s. tomato. Periodic histological sectioning of inoculated tomato leaves showed that bacterial cells resided and multiplied in depressions and around trichome bases for 24 h before penetration through stomata and trichome basal cells. The bacteria invaded intercellular spaces and caused cell lignification, collapse and shrinkage, 48 h after inoculation. Necrotic lesions filled with bacterial masses and collapsed lignified cells were readily observable at and after 72 h. No detectable histological changes were observed in the yellow halo region surrounding the necrotic leaf specks. A thermostable toxin was produced by the pathogen and is involved in chlorotic symptom expression. An antibiotic mixture of streptomycin + oxytetracy‐cline was most effective in controlling infections followed by copper oxychloride + mancozeb, tribasic copper sulphate + sulphur, copper oxychloride and copper oxychloride + zineb.     

Environmentally friendly treatment alternatives to Bordeaux mixture for controlling bacterial apical necrosis (BAN) of mango

Bacterial apical necrosis (BAN), caused by the bacterium Pseudomonas syringae pv. syringae (Pss), is currently the most limiting disease affecting the mango crop in the Mediterranean area. The copper‐based compound Bordeaux mixture (BM) is considered to be the conventional treatment against BAN, but it does not act as a bactericide. Alternative experimental treatments to BM that are compatible with organic farming were tested for their ability to control BAN disease. Field trials were conducted over six seasons in different mango orchards with natural infestation of Pss. The experimental treatments included applications of Silicon gel (a commercial formulation of aqueous potassium silicate), dicalcium phosphate, Kaolinite, and Ulmasud B^® (bentonite, powder); BM was applied as the conventional treatment. During the first two seasons (small‐scale trial, 2002–2004), all these experimental compounds were applied in order to select those treatments providing the greatest reduction of BAN symptoms. In the next three seasons (2005–2008), a semi‐commercial scale trial was carried out with the best‐performing treatments, resulting in the selection of Silicon gel. Finally, Silicon gel was tested in a commercial scale trial during the last season (2008–2009). Trees treated with Silicon gel showed significantly fewer necrotic buds and leaves, reaching disease levels very similar to those using the conventional treatment with BM. However, minor differences in P. syringae‐like population levels were observed, as has been described in previous studies. The possible mode of action of the Silicon gel is discussed. Currently, the Silicon gel compound is undergoing the registration process for its commercial use in mango management in Spain.     

Effect of aerated compost tea on grapevine powdery mildew, botrytis bunch rot and microbial abundance on leaves

Aerated compost tea (ACT), prepared from immature compost, was applied to foliage and fruit of grapevines (Vitis vinifera) to assess its potential for suppressing two important diseases: botrytis bunch rot, caused by Botrytis cinerea, and powdery mildew, caused by Erysiphe necator. An ACT applied to leaves of Cabernet Sauvignon vines in pots 7 days before inoculation with E. necator conidia reduced mean powdery mildew severity on the three youngest expanded leaves (at inoculation) to less than 1 %; mean severity on non-treated, inoculated leaves was 15 %. Multiple applications of ACTs at two vineyards in different growing seasons suppressed powdery mildew to <1 % mean severity on Chardonnay leaves (non-treated 79 % severity) and bunches (non-treated 77 % severity), and on Riesling leaves (non-treated 24 % severity). The same treatments reduced the incidence of Chardonnay bunches with latent B. cinerea and Riesling bunches with sporulating B. cinerea, although the level of botrytis bunch rot in both experiments was not economically damaging. The numbers of culturable bacteria, fungi and yeasts on Chardonnay leaves were higher than pre-treatment levels 10 days after ACT application, as were fungal numbers on Riesling leaves 21 days after treatment. Suppression by ACTs of two fruit and foliar pathogens of grapevine with different biology and epidemiology indicated potential for their use as a tactic in integrated disease management. Further testing of ACTs in a range of viticultural environments and application regimes will contribute to a better understanding of the impact of this tactic on disease, grape and wine quality.     

Bacterial brown spot on Avena storigosa Schereb. caused by Pseudomonas syringae pv. alisalensis

Avena storigosa Schereb. (bristle oat) is used as a green manure in crop rotations and as an antagonist of nematodes in Nagano Prefecture, Japan. In 2011, necrotic, brown, water-soaked lesions were observed on young bristle oat plants. A pathogenic bacterium was isolated from symptomatic leaves of infected plants and produced the same symptoms after inoculation. Bacteriological properties of the bacterial isolates from bristle oat matched those of Pseudomonas syringae pathovars. The host range of the bristle oat isolates was identical to that of P. syringae pv. alisalensis. This is the first report of bristle oat disease caused by P. syringae pv. alisalensis.     

Genetic diversity and pathogenicity of Pseudomonas syringae pv. aptata isolated from sugar beet

Pseudomonas syringae pv. aptata is the causal agent of bacterial leaf spot disease of sugar beet (Beta vulgaris). During 2013, 250 samples were collected from leaf lesions with typical symptoms of bacterial leaf spot in commercial fields of sugar beet in Serbia, and 104 isolates of P. syringae pv. aptata were obtained. Identification and characterization was performed using biochemical, molecular and pathogenicity tests. Identification included LOPAT tests and positive reactions using primers Papt2F and Papt1R specific for P. syringae pv. aptata. Repetitive (rep) sequence‐based PCR typing with ERIC, REP and BOX primers revealed high genetic variability among isolates and distinguished 25 groups of different fingerprinting profiles. Pulse‐field gel electrophoresis (PFGE) and multilocus sequence analysis (MLSA) of representative isolates showed higher genetic variability than in rep‐PCR analysis and distinguished three and four major genetic clusters, respectively. A pathogenicity test performed with 25 representative isolates on four cultivars of sugar beet confirmed the occurrence of leaf spot disease and showed correlation between the most aggressive isolates and the genetic clusters obtained in MLSA. All these findings point to the existence of several lines of P. syringae pv. aptata infection in Serbia that are genetically and pathologically different.     

Induced resistance against Fusarium diseases of <Emphasis Type="Italic">Cymbidium</Emphasis> species by weakly virulent strain HPF-1 (<Emphasis Type="Italic">Fusarium</Emphasis> sp.)

The mechanism by which Fusarium diseases of cymbidium plants are suppressed by a weakly virulent strain HPF-1 of Fusarium sp. was studied. Strain HPF-1 produced microscopic, necrotic local lesions on cymbidium leaves, causing minor damage to palisade tissues at the infection sites. This weakly virulent strain remained near the site of infection and did not develop further. It systemically and nonselectively suppressed some diseases of cymbidium such as yellow spot of leaves caused by Fusarium proliferatum and F. fractiflexum, bulb and root rot caused by F. oxysporum, and dry rot of bulbs and roots caused by F. solani. Because endogenous salicylic acid levels increased in cymbidium leaves inoculated with strain HPF-1, the mechanism of disease suppression is thought to be systemic acquired resistance.     

Epidemiology of Grey Mould in Annual Waiting-bed Production of Strawberry

The epidemiology of Botrytis cinerea was studied in five annual strawberry crops using waiting-bed transplants, a system widely adopted in the Netherlands. On dead leaves of transplants the incidence of B. cinerea varied from 26.7% to 52.6%, but the leaf area with potential sporulation was low (3.5–15.6%). During each crop cycle, the availability of necrotic leaf substrate for spore production of B. cinerea was generally low and varied between seasons and with the quality of transplants. B. cinerea sporulated on a maximum of 15.5 cm² of leaf area per plant, measured as potential sporulation. The aerial concentration of B. cinerea conidia in untreated plots did not differ from the concentration in plots where all dead leaves had been removed, nor from the concentration at 25–50 m distance from the strawberry plots. B. cinerea incidence on flowers ranged from 5% to 96%, but no correlation was found with the potential spore production on necrotic leaves. Grey mould at harvest varied from 1.4% to 11.3% and was correlated with the average precipitation during the harvesting period but not with B. cinerea incidence on flowers. Post-harvest grey mould ranged from 2.1% to 32.6% and was correlated with petal colonisation by B. cinerea. The results suggest that in the annual cropping system with waiting-bed transplants, necrotic leaves are not a significant source of B. cinerea inoculum, unlike in other strawberry production systems. Therefore, control measures of grey mould in this annual system should focus on protection of flowers and young developing fruits, and not on the reduction of inoculum production on leaf debris.     

A strain ofPseudomonas syringae which does not belong to pathovarphaseolicola produces phaseolotoxin

A bacterial strain, CFBP 3388, isolated from Vetch (Vicia sativa, L.) was identified asP. s. pv.syringae on the basis of nutritional and biochemical patterns which were obtained with classical tests and the Biolog system. It caused necrotic symptoms typical ofP. s. pv.syringae on bean leaves and pods after artificial inoculation. However, the isolate caused a citrulline-reversible inhibition ofE. coli in phaseolotoxin bioassay. Furthermore, with CFBP 3388 DNA as template a 1900 bp DNA fragment, specific for the phaseolotoxin DNA cluster ofP. s. pv.phaseolicola, was amplified by PCR. This is the first demonstration that an isolate ofP. syringae that is not pv.phaseolicola can produce phaseolotoxinAbbreviations bp base pair - kb kilobase - OCT Ornithine Carbamoyl Transferase     

Role of phaseolotoxin production by Pseudomonas syringae pv. actinidiae in the formation of halo lesions of kiwifruit canker disease

Pseudomonas syringae pv. actinidiae, the causal bacterium of kiwifruit canker, induces the formation of chlorotic halo lesions on infected leaves and inhibits the growth of Escherichia coli. The inhibition ofE. coli growth was shown to be reversed by L -arginine or L -citrulline, but not by L -glutamine, suggesting that the pathogen produces a toxin similar to phaseolotoxin, which inhibits ornithine carbamoyltransferase. The toxin was purified from culture broth of P. syringae pv. actinidiae strain Kw11, and was shown by nuclear magnetic resonance to be identical to phaseolotoxin. Assays based on inhibition of E. coli growth and on amplification of a phaseolotoxin fatty acid desaturase gene (ptx) fragment revealed that, among the plant pathogenic bacteria examined, the production of phaseolotoxin is restricted to strains of P. syringae pv. phaseolicola and pv.actinidiae . A non-toxigenic mutant of strain Kw11 generated by disruption of the ptx gene induced the formation of necrotic lesions on kiwifruit leaves; however, the lesions were not surrounded by a chlorotic halo as were those induced by the parent strain. The growth rate of the non-toxigenic mutant in leaf tissue was similar to that of the parent strain. These results suggest that phaseolotoxin production contributes to the formation of chlorotic halo lesions in kiwifruit canker but is not required for multiplication of the pathogenic bacterium during infection.     

Basal glume rot (Pseudomonas syringae pv. atrofaciens) on wheat and barley in FRG and resistance screening of wheat1

A survey in 1987 and 1988 revealed that basal glume rot, caused by Pseudomonas syringae pv. atrofaciens, occurred nearly everywhere in FRG. The symptoms of the disease usually consisted of water-soaked dark green to brown lesions on unripe wheat heads, mainly at the basal end of the glumes, which later became dark brown. Forty-six isolates of P.s. atrofaciens were obtained from glumes, seeds and leaves of wheat and barley. For a fast identification of the isolated bacteria, a bio-assay was developed. Four to five-day-old wheat seedlings, grown on wet filter paper in Petri dishes, were pricked at two-three sites with a dissecting needle contaminated with bacteria. After 2–3 days, pathogenic isolates induced brown to black spots. The bacterial isolates from wheat inhibited the growth of several fungi grown on potato dextrose agar. In contrast, an authentic isolate of P.s. syringae obtained from wheat showed no inhibitory effect. During screening for resistance, several cultivars of spring and winter wheat were tested in the greenhouse and/or field tests. The results revealed marked differences in the susceptibility of different cultivars.     

Current situation and characterization of Pseudomonas syringae pv. actinidiae on kiwifruit in Galicia (northwest Spain)

Bacterial canker of kiwifruit, caused by Pseudomonas syringae pv. actinidiae (Psa), is a disease that is spreading rapidly in several kiwifruit‐producing countries, causing significant economic losses. In 2011, it was detected for the first time in Spain, in the south of Galicia (northwest Spain). Kiwifruit orchards were therefore inspected and sampled in 2011 and 2012 to determine the pathogen distribution, and the isolates obtained were characterized by morphology, fatty acids profile, biochemical tests and molecular techniques. Isolates were obtained from Actinidia deliciosa ‘Hayward’ (from leaves, canes, flower buds, fruits and roots), from A. deliciosa ‘Summer’, from Actinidia chinensis ‘Jin Tao’ (from canes and leaves) and from A. chinensis pollinator ‘Belén’ (from canes). Results of the analysis of the cfl gene (phytotoxin production‐related), the tox–argK gene cluster and phylogenetic analysis of the cts gene demonstrated that all Psa isolates from northwest Spain correspond to the Psa3 population, which includes strains of haplotype 2. This is the first record of Psa3 and haplotype 2 in Spain.     

Appearance of a new leaf rot disease on common ice plant

An unknown disease abruptly appeared on hydroponic cultures of common ice plant (Mesembryanthemum crystallinum) in the greenhouse, causing catastrophic damage. Although the symptoms of the plant were unlike typical Botrytis lesions on leaves and stems of other plants, fungi isolated from the necrotic lesions on the plant were similar to genus Botrytis in terms of conidial shape, colony color and nature. A representative isolate, Ice-2, caused similar symptoms on the host plants after inoculation with conidia, and the same fungus was isolated from the lesions. The conidia and conidiophores of Ice-2 were morphologically similar to those of B. cinerea but not to those of B. allii, B. fabae or B. squamosa. The tested Botrytis fungi grew at temperatures between 5 and 30°C. Ice-2 grew faster than the others at the lower end of the temperature range. Ice-2 was also more virulent than B. cinerea (Bay-1) in artificial inoculations, especially on common ice plant leaves. The glyceraldehyde-3-phosphate dehydrogenase gene (G3PDH) sequence of Ice-2 was determined and compared with those from four Botrytis species. The gene sequence of Ice-2 appeared to be identical to that of B. cinerea. In leaf tests on common ice plant and kidney bean, the diseases caused by Ice-2 and Bay-1 were controlled equally well by the primary Botrytis fungicides. Based on the results of the present studies, Ice-2 is thought to be Botrytis cinerea Person: Fries.     

Identification,potential inoculum sources and pathogenicity of botryosphaeriaceous species associated with grapevine dieback disease in New Zealand

This study investigated the prevalence and identity of botryosphaeriaceous dieback pathogens in necrotic grapevines tissues in New Zealand vineyards, and other woody hosts growing nearby. The presumptive identities of the isolates by conidial and cultural morphology were confirmed with ITS sequence data as Neofusicoccum australe, N. luteum, N. parvum and Diplodia seriata. They were isolated predominantly from necrotic stems of grapevine and other hosts, but also from leaves, flowers and wood debris of grapevines. Inoculation with conidia and mycelium of multiple isolates of each species onto excised and attached green shoots and trunks of five grapevine varieties, Cabernet sauvignon, Chardonnay, Pinot noir, Riesling, and Sauvignon blanc, showed that all varieties became infected to a similar extent. All species except D. seriata were pathogenic, irrespective of the host source, with N. luteum being the most and D. mutila the least pathogenic (P < 0.05). On trunks, N. parvum caused cankers and the other pathogenic species caused die-back when the inoculated vines became winter-dormant. Conidia were produced from green shoot lesions and die-back wood, which indicates potential inoculum sources for vineyard infection.     

Translocation of benomyl,prochloraz and procymidone in relation to control of Botrytis cinerea in strawberries

Benomyl, prochloraz or procymidone, applied as an overall plant spray at the openflower stage, effectively suppressed Botrytis cinerea fruit rot, whereas no control was achieved by foliar application only. Fruit rot was prevented using procymidone applied to the soil 12 days before inoculation of the flowers, whereas benomyl or prochloraz gave little or no control, respectively when applied in the same manner. Bioassays, using Penicillium expansum on leaf and flower extracts of strawberry plants growing in soil treated with procymidone, showed the presence of an inhibitory compound with the same R_F value on thin-layer chromatography as that of procymidone. Analysis by gas chromatography and identification by gas chromatography—mass spectrometry established that the fungicide procymidone was translocated from the root system of strawberry plants to the leaves and flowers.