Simple diagnosis using ethanol immersion of strawberry plants with latent infection by Colletotrichum acutatum, Dendrophoma obscurans, and Fusarium oxysporum f. sp. fragariae

Simple diagnosis by ethanol immersion (SDEI) to detect Glomerella cingulata was used to detect three other fungi that also cause latent infection of strawberry plants. Signs on strawberry leaves with asymptomatic latent infection by Colletotrichum acutatum became visible using SDEI. Salmon-pink conidial masses were produced in the acervuli on the treated leaves 5 days after incubation at 28°C. In the case of Dendrophoma obscurans, pycnidia with amber conidial masses formed 5 days after incubation at 28°C. The pycnidia were observed mainly on the ribs, and conidial masses exuded from the ostiole. These macroscopic conidial masses were similar to those of G. cingulata and C. acutatum. When water was dripped onto a lesion caused by D. obscurans, the pycnidia exuded white filamentous conidial masses, making the distinction of D. obscurans from G. cingulata or C. acutatum. On petioles with latent infection by Fusarium oxysporum f. sp. fragariae, white aerial hyphae grew out from the vascular tissues on the cut surface 3 days after incubation at 28°C and were easily observed by eye or with a loupe. Thus, SDEI was also useful for diagnosing latent infection of strawberry plants by C. acutatum, D. obscurans, and F. oxysporum f. sp. fragariae.     

Correlation between field disease and latent infection by Glomerella cingulata of strawberry plants as revealed by a simple diagnostic method using ethanol immersion

Leaves of strawberry plants growing in fields were collected and assayed by ethanol immersion treatment (SDEI) to detect latent infection by Glomerella cingulata and Dendrophoma obscurans. SDEI revealed that 83% of the plants in growers fields were latently infected by G. cingulata and 58% by D. obscurans. In such fields, only 0.8%–2.5% of the plants later became wilted or died because of G. cingulata. When the latently infected plants from naturally infested fields were kept under optimal conditions for disease development for 5 weeks, 70.0%–83.3% of them wilted or died from G. cingulata. However, only 5.6%–6.7% of the plants diagnosed by SDEI as being without latent infection wilted or died. The results indicate the importance of selecting healthy plants, suggesting that SDEI is an effective method for detecting latent infection and reducing losses from G. cingulata.     

Effects of inoculum density, leaf age, moisture, temperature, and wetness duration on black streak of edible burdock

Inoculum density, temperature, leaf age, and wetness duration were evaluated for their effects on the development of black streak (Itersonilia perplexans) on edible burdock (Arctium lappa L.) in a controlled environment. The effect of relative humidity (RH) on ballistospores production by I. perplexans was also evaluated. Symptoms of black streak on leaves increased in a linear fashion as the inoculum density of I. perplexans increased from 10² to 10⁶ ballistospores/ml. Rugose symptoms on young leaves were observed at densities of ≥10⁴ ballistospores/ml. Disease severity of I. perplexans in relation to leaf age followed a degradation curve when the leaves were inoculated with ballistospores. Disease severity was high in newly emerged leaves up to 5 days old, declined as leaf age increased to 29 days, and was zero when leaf age increased from 30 to 33 days. Disease development of edible burdock plants exposed to ballistospores of I. perplexans was evaluated at various combinations of temperature (10°, 15°, 20°, 25°C) and duration of leaf wetness (12, 24, 36, 48, and 72 h). Disease was most severe when plants were in contact with the ballistospore sources at 15° or 20°C. The least amount of disease occurred at 25°C regardless of wetness duration. Ballistospores required 24–36 h of continuous leaf wetness to cause visible symptoms by infection on edible burdock. Ballistospores production in infected lesions required at least 95.5% RH.     

Biological control ofBotrytis cinerea on tomato stem wounds withTrichoderma harzianum

The effectiveness ofTrichoderma harzianum in suppression of tomato stem rot caused byBotrytis cinerea was examined on tomato stem pieces and on whole plants. Ten days after simultanous inoculation withB. cinerea andT. harzianum, the incidence of infected stem pieces was reduced by 62–84%, the severity of infection by 68–71% and the intensity of sporulation by 87%. Seventeen days after inoculation of wounds on whole plants, the incidence of stem rot was reduced by 50 and 33% at 15 and 26 °C, respectively, and the incidence of rot at leaf scar sites on the main stem was reduced by 60 and 50%, respectively. Simultanous inoculation and pre-inoculation withT. harzianum gave good control ofB. cinerea (50 and 90% disease reduction, 10 days after inoculation). The rate of rotting was not reduced by the biocontrol agent once infection was established. However, sporulation byB. cinerea was specifically reduced on these rotting stem pieces. Temperature had a greater effect than vapour pressure deficit (VPD) on the efficacy of biocontrol. Suppression ofB. cinerea incidence byT. harzianum on stem pieces was significant at 10 °C and higher temperatures up to 26 °C. Control of infection was significantly lower at a VPD of 1.3 kPa (60% reduction), than at VPD<1.06 kPa (90–100% control). Reductions in the severity of stem rotting and the sporulation intensity of grey mould were generally not affected by VPD in the range 0.59–1.06 kPa. Survival ofT. harzianum on stems was affected by both temperature and VPD and was greatest at 10 °C at a low VPD and at 26 ° C at a high VPD.     

Relationship between Concentrations of Botrytis Cinerea Conidia in Air, Environmental Conditions, and the Incidence of Grey Mould in Strawberry Flowers and Fruits

Atmospheric concentrations of Botrytis cinerea conidia were monitored for two seasons in a strawberry crop in Moguer (Huelva, southwestern Spain). Concentrations of conidia were estimated using a Burkard volumetric spore sampler. A diurnal pattern of conidial release was observed. Airborne conidial concentration was significantly and positively correlated with the average solar radiation and mean temperature, and negatively with rainfall and relative humidity. Among the weather variables considered, solar radiation showed the most consistent results in the regression analysis, explaining over 40% of airborne conidial concentration variability. Correlation between Botrytis fruit rot incidence and accumulated number of conidia over seven days was significant and positive. Two regression models containing three variables explained over 62 and 52% of the fruit rot incidence variability. A positive but non-significant correlation was established between B. cinerea incidence in flowers and airborne conidial concentration. It was not possible to fit a consistent regression model to relate flower infection incidence to conidial concentration or weather variables.     

Line pattern in Limonium latifolium caused by tobacco rattle virus

Tobacco rattle virus (TRV) was isolated from plants ofLimonium latifolium showing bright yellow or red line patterns and ringspots on the leaves. It was proved that this virus, designated TRV-Lim, was the causal agent of the disease. In its reactions onNicotiana clevelandii it resembled a yellow strain of TRV from Oregon (USA), but the symptoms inN. glutinosa, N. megalosiphon, N. tabacum andPetunia hybrida were more comparable to those caused by socalled unstable variants of TRV. Dilution end-point was 10^–6–10^–7, thermal inactivation at 76–80°C, and ageing in vitro 55–60 days. The purified virus suspension contained particles of three normal lengths, 70, 102, and 194 nm. The virus sedimented as three components with average sedimentation coefficients of 129, 161 and 206 S, respectively. In purified suspensions TRV-Lim had two different buoyant densities. A serological relationship was found with TRV isolated from Europe and Brazil.Samenvatting Tabaksratelvirus (TRV) werd geïsoleerd uitLimonium latifolium planten die heldergeel of rood figuurbont op de bladeren vertoonden. Er werd aangetoond dat dit virus, aangeduid als TRV-Lim, de ziekteverwekker was. De reacties van dit virus opNicotiana clevelandii deden denken aan die van een gele stam van TRV afkomstig uit Oregon (VS), maar de symptomen opN. glutinosa, N. megalosiphon, N. tabacum enPetunia hybrida vertoonden meer gelijkenis met die welke veroorzaakt worden door de zogenaamde onstabiele varianten van TRV. De verdunningsgrens was 10^–6–10^–7, de inactiveringstemperatuur 75–80°C en de houdbaarheid in vitro 55–60 dagen. De gezuiverde virussuspensie bevatte deeltjes met drie normale lengtes, nl. 70, 102 en 194 nm. Het virus sedimenteerde, als drie componenten met gemiddelde sedimentatiecoëfficiënten van respectievelijk 129, 161 en 206 S. In gezuiverde suspensie vertoonde TRV-Lim twee verschillende zweefdichtheden. Het virus was serologisch verwant aan TRV-isolaten uit Europa en Brazilië.     

Germination of <Emphasis Type="Italic">Gibberella zeae</Emphasis> ascospores as affected by age of spores after discharge and environmental factors

The effects of age of ascospores (0–18 days after discharge), photon flux density (0–494 mol m^–2 s^–1 PAR), temperature (4–30 °C), frost (–15 °C for 30 min), relative humidity (RH; 0–100%), pH (2.5–6.5) and dryness (0 and 53% RH for up to 40 min) on the germination of the ascospores of the mycotoxin-producing fungus Gibberella zeae (anamorph Fusarium graminearum) were studied. Freshly discharged ascospores germinated within 4 h at 20 °C and 100% RH. The rate of germination and the percentage of viable ascospores decreased over time after the spores were discharged from perithecia. The time course of ascospore germination was not significantly affected by photon flux density. The period of time required to obtain 50% germinated ascospores at 100% RH was 26.90 h at 4 °C, 10.40 h at 14 °C, 3.44 h at 20 °C and 3.31 h at 30 °C. There was no significant effect of frost on the percentage of viable ascospores. A small percentage (6.6 ± 3.8%) of the ascospores germinated at 53% RH. At RH 84% and 20 °C almost 100% of the freshly discharged ascospores germinated. The time course of ascospore germination was affected by pH. The maximum rate of ascospore germination was estimated to be at pH 3.76. Ascospores lost their ability to germinate following exposure to 0% RH almost instantaneously. No germinating spores were detected after an incubation period of 1 min at 0% RH. Incubating the ascospores at 53% RH decreased the percentage of viable spores from 93 to 6% within 10 min. The data demonstrate that age of spores, relative humidity, temperature and pH, but not photon flux density, are key factors in germination of G. zeae ascospores.     

疏水表面诱导橡胶树炭疽菌侵染结构的发育分化过程

为了解橡胶树2种炭疽病菌的侵染结构发育分化过程,采用平板菌落生长速率法测定了3株胶孢炭疽菌Colletotrichum gloeosporioides和3株尖孢炭疽菌C.acutatum的菌丝生长速率,测量其分生孢子大小,显微观察2种炭疽菌在疏水表面诱导下侵染结构的发育分化过程。结果表明,胶孢炭疽菌菌丝生长速率为0.96~1.36 cm/d,显著高于尖孢炭疽菌的菌丝生长速率0.72~0.89 cm/d,但二者分生孢子大小无显著差异。在疏水表面诱导下,2种炭疽菌分生孢子在接种2~6 h后开始萌发,12 h孢子萌发率为71.70%~88.05%,13~16 h开始分化附着胞,24 h附着胞形成率为48.99%~70.74%,36 h菌丝诱发形成大量附着枝,48 h后分生孢子产生的次生菌丝也可诱发形成附着枝,附着枝呈圆形、姜瓣形、梨形或不规则形。分生孢子极易产生,可在菌丝顶端成簇或菌丝侧面排列产生,也可由分生孢子形成的芽管产生,或在芽管分化附着胞过程分枝形成分生孢子;附着胞多着生于芽管顶端,少数附着胞顶端可继续萌发类似短芽管结构,再次分化形成可黑色化的次级附着胞。表明橡胶树2种炭疽菌不同菌株间分生孢子萌发时间、孢子萌发率、附着胞形成时间和形成率有一定差异,但种间无明显差异;橡胶树炭疽菌分生孢子极易形成,在疏水表面容易分化形成附着胞和附着枝,说明具有极强的适生性。     

Multidrug-resistance plasmid of <Emphasis Type="Italic">Enterobacter cloacae</Emphasis>: transfer to <Emphasis Type="Italic">Erwinia herbicola</Emphasis> on the phylloplane of mulberry and weeds

A mulberry epiphytic Enterobacter cloacae MUL1 harbors plasmid pMUL1 encoding five drug-resistance genes. This plasmid was examined upon its conjugal transfer into epiphytic Erwinia herbicola on the phylloplane of mulberry and 12 species of weeds. The plasmid was transferred into Er. herbicola at a frequency of 10^–5–10^–3/recipient in mulberry and Lolium multiflorum LAM. 1–8 days after wound inoculation with 10⁶–10⁸/ml suspensions. In Chenopodium album L. and C. album L. var. centrorubrum, however, it was transferred only after wound inoculation with a 10⁸/ml suspension, but not with 10⁷/ml or 10⁶/ml suspensions, owing to the weak epiphytic fitness of Ent. cloacae on these weeds. Transconjugants were also obtained for seven other species of weeds in the case of inoculation with a 10⁸/ml suspension. In contrast, when bacterial suspensions were sprayed on mulberry leaves with or without fresh wounds, transconjugants were obtained only in wounded leaves, which were considered suitable for bacterial conjugation. These findings suggest that epiphytic bacteria, including Ent. cloacae and Er. herbicola, may be carriers of drug-resistance genes distributed among plant pathogenic bacteria in nature.     

Life-table analysis of faba bean rust

In controlled near-optimum conditions (18 °C), monocyclic sporulation capacity and spore infection efficiency were assessed for faba bean rust on the first and second leaves of field bean. After a latency period of 8–10 days, lesions sporulated duringc. 50 days. Spore production on the second leaf,c. 9×10⁴ spores per lesion, was two times as high as spore production on the first leaf. Infection efficiency was similar for both leaf layers, with a mean value of 0.11 lesion per inoculated spore. Infection efficiency decreased strongly when spores originated from mother lesions older than 20 days. Three life-table statistics (the net reproduction numberR _o , the mean generation timeT _g , and the maximum relative growth rater _max ) were calculated.R _o was larger andT _g was longer for the second than for the first leaf, butr _max was nearly the same for both leaf layers (0.31–0.33 day^–1).r _max was compared with the exponential growth rater measured in a field experiment. From the difference between the two rates, the fraction of inoculum lost in field conditions was estimated at 0.54–0.94. The life-table statistics were also compared to those of other legume rusts, and implications of life-table analysis for comparative epidemiology were discussed.     

Destruxin B produced by Alternaria brassicae does not induce accessibility of host plants to fungal invasion

It has been reported that Alternaria brassicae, the causal agent of gray leaf spot in Brassica plants, produces a host-specific or host-selective toxin (HSTs) identified as destruxin B. In this study, the role of destruxin B in infection of the pathogen was investigated. Destruxin B purified from culture filtrates (CFs) of A. brassicae induced chlorosis on host leaves at 50–100 μg ml⁻¹, and chlorosis or necrosis on non-host leaves at 250–500 μg ml⁻¹. Destruxin B was detected in spore germination fluids (SGFs) on host and non-host leaves, but not in a sufficient amount to exert toxicity to host plants. When spores of non-pathogenic A. alternata were combined with destruxin B at 100 μg ml⁻¹ and inoculated on the leaves, destruxin B did not affect the infection behavior of the spores. Interestingly, SGF on host leaves allowed non-pathogenic spores to colonize host leaves. Moreover, a high molecular weight fraction (>5 kDa) without destruxin B obtained by ultrafiltration of SGF had host-specific toxin activity and infection-inducing activity. From these results, we conclude that destruxin B is not a HST and does not induce the accessibility of the host plant which is essential for colonization of the pathogen. In addition, the results with SGF imply that a high molecular weight HST(s) is involved in the host–pathogen interaction.     

The Effect of Inoculation Treatment and Long-term Application of Moisture on Fusarium Head Blight Symptoms and Deoxynivalenol Contamination in Wheat Grains

Fusarium head blight (FHB) is an important disease of wheat, which can result in the contamination of grains with mycotoxins such as deoxynivalenol (DON). Artificial inoculation of flowering ears with conidial suspensions is widely used to study FHB diseases. Our goal was to compare four inoculation treatments in which a conidial suspension was sprayed on flowering ears and to study the effect of the application of moisture during kernel setting and filling with a mist-irrigation system. Ten wheat genotypes were inoculated with a DON-producing Fusarium culmorum strain. Inoculation treatments varied in time of application of the inoculum (morning or evening) and in the method of controlling humidity during inoculation (bagging or mist irrigation). A wet season was simulated with a mist-irrigation system, keeping the crop canopy wet for at least 26 days after flowering. The severity of FHB symptoms (area under disease progress curve (AUDPC)), yield loss and DON contamination in the grains were determined. AUDPC data obtained with the different inoculation treatments were highly correlated (r=0.85–0.95). Mist irrigation after inoculation resulted in a higher mean disease severity, but in a overall lower toxin contamination as compared to the non-irrigated treatments. Genotypic differences in DON accumulation were present: for one wheat line toxin contamination significantly increased when irrigated, while two genotypes accumulated significantly less toxin. The closest relationships (r=0.73–0.89) between the visual symptoms and the DON content were obtained under moderate mean infection pressure. This relation between visual symptoms and the DON content deteriorated at higher infection levels.     

Forecasting Infections of the Leaf Curl Disease on Peaches Caused by Taphrina deformans

An Israeli model forecasting leaf curl disease on peaches caused by Taphrina deformans was validated in the Emilia-Romagna region of northern Italy, during a three-year period (1996–1998), in 13 cases (year × location × cultivar). When the peach trees are susceptible to infection, the model uses mathematical functions to calculate the risk of infection on the basis of weather conditions (daily rainfall greater than 10mm, and maximum air temperature greater than 5°C), and it forecasts periods of possible symptom appearance based on the length of incubation. Peach trees became susceptible to infection between the end of January and mid March, when the first leaf buds attained phenological stage C, i.e. appearance of leaf apex. The trees remained susceptible for at least 9 weeks: the last infection occurred in mid-May.Since most of the leaf curl onsets observed in the orchards fell within the range of model forecasts, the model proved to be accurate in signalling both the first seasonal infection and repeated infections during the primary inoculum season. Few errors occurred, caused either by conditions of rainfall and temperature lower than the thresholds fixed in the model, or by discrepancies between forecast and actual length of incubation. Infection occurred also at 3.1–3.5°C, and with 9.6mm rainfall. Thus, thresholds should not be accepted too rigorously, and perhaps temperature should not be considered as a limiting factor for infection under the conditions of the present work. The length of incubation showed high variability: it was 23 days long on average, with a 95% confidence interval ranging from 20 to 27 days, and extreme values of 9 and 33 days.     

Phyllosphere yeasts antagonize penetration from appressoria and subsequent infection of maize leaves by Colletotrichum graminicola

In growth cabinet experiments, the common phyllosphere yeastsSporobolomyces roseus andCryptococcus laurentii var.flavescens were sprayed as a mixture (11) onto the fourth leaves of maize plants (Zea mays) two-three days prior to inoculation withColletotrichum graminicola. In four experiments the average yeast population of the treated leaves at the time of pathogen inoculation varied between 5× 10⁴ and 8× 10⁵ cells cm^–2 leaf, whereas on the untreated leaves the yeast population varied from <10³ to 10⁴ cells cm^–2 leaf. The yeasts reduced lesion density and necrosis fromC. graminicola infection by approximately 50%. Contrary to findings with other necrotrophic pathogens, conidial germination, superficial mycelial growth and appressorium formation were not affected. Instead, the reduction of infection could only be explained by a reduced number of penetrations from the normally formed appressoria, a site of interaction not previously recorded.Samenvatting In klimaatkastexperimenten werden maisbladeren (4e blad) twee-drie dagen voor inoculatie metColletotrichum graminicola bespoten met een mengsel (11) van de algemeen voorkomende fyllosfeergistenSporobolomyces roseus enCryptococcus laurentii var.flavescens. In vier experimenten varieerde de gemiddelde gistpopulatie op de behandelde bladeren, op het moment van inoculatie met het pathogen, van 5× 10⁴ tot 8× 10⁵ cellen cm^–2 blad, op de onbehandelde bladeren van <10³ tot 10⁴ cellen cm^–2 blad. De gisten reduceerden de lesiedichtheid en het necrotisch bladoppervlak tengevolge van deC. graminicola infectie voor ongeveer 50%. De stadia in de ontwikkeling van andere necrotrofe pathogenen, die gewoonlijk gevoelig zijn voor antagonisme door gisten, zoals sporekieming, oppervlakkige myceliumgroei en vorming van appressoria, werden bijC. graminicola niet beïnvloed. De waargenomen reductie van infectie kon alleen verklaard worden door een remming van de penetratie vanuit normaal gevormde appressoria. Interactie in dit stadium van het infectieproces is nog niet eerder waargenomen.     

Relationship between the incidence of latent infections caused by <Emphasis Type="Italic">Monilinia</Emphasis> spp<Emphasis Type="Italic">.</Emphasis> and the incidence of brown rot of peach fruit: factors affecting latent infection

Five field experiments were performed in commercial orchards located in Lleida (Spain) over three growing seasons, 2000–2002, in order to estimate the relationship between the incidence of latent infection caused by Monilinia spp. in peaches and the incidence of post-harvest brown rot. No latent infection was recorded at popcorn and the maximum incidence occurred pre-harvest; in some orchards a second peak was detected during the pit hardening period. Monilinia laxa is the most prevalent species isolated from peaches with brown rot. There was a positive correlation between the incidence of latent infection and that of post-harvest brown rot. The average incidence of latent infection during the crop season explained 55% of the total variation in the incidence of post-harvest brown rot. The effect of temperature (T) and duration of wetness (W) on the incidence of latent infection in peach and nectarine orchards was analysed using multiple regression. The regression analysis indicated that T and W jointly explained 83% of the total variation in the incidence of latent infection. The model predicts no latent infections when T < 8°C, and >22 h wetness are required when T = 8°C but only 5 h at 25°C are necessary for latent infection to occur. The incidence of brown rot and latent infection of peaches caused by M. laxa under controlled experimental conditions were also affected by T and W, as well as by fruit maturity and inoculum concentration. Latent infections were produced in fruit when T was not suitable for the development of brown rot symptoms. In these experiments more than 4–5 h of daily wetness were required after embryo growth in fruit sprayed to run-off with an inoculum concentration higher than 10⁴ conidia ml⁻¹ of M. laxa for brown rot and latent infections to develop. The fitted model obtained from the field data was able to predict the observed data obtained under controlled environmental conditions.     

Infection of wheat spikes by <Emphasis Type="Italic">Fusarium avenaceum</Emphasis> and alterations of cell wall components in the infected tissue

The infection process of Fusarium avenaceum on wheat spikes and the alteration of cell wall components in the infected host tissue were examined by means of electron microscopy and cytochemical labelling techniques following spray inoculation at growth stage (GS) 65 (mid-flowering). Macroconidia of the pathogen germinated with one to several germ-tubes 6–12 h after inoculation (hai) on host surfaces. The germ-tubes did not penetrate host tissues immediately, but extended and branched on the host surfaces. Hyphal growth on abaxial surfaces of the glume, lemma and palea was scanty 3–4 days after inoculation (dai) and no direct penetration of the outer surfaces of the spikelet was observed. Dense mycelial networks formed on the inner surfaces of the glume, lemma, palea and ovary 36–48 hai. Penetration of the host tissue occurred 36 hai by infection hyphae only on the adaxial surfaces of the glume, lemma, palea and upper part of ovary. The fungus penetrated the cuticle and hyphae extended subcuticularly or between the epidermal wall layers. The subcuticular growth phase was followed by penetration of the epidermal wall, and hyphae spread rapidly inter- and intracellularly in the glume, lemma, palea and ovary. During this necrotrophic colonization phase of the wheat spike, a series of alterations occurred in the host tissues, such as degeneration of cytoplasm and cell organelles, collapse of host cells and disintegration of host cell walls. Immunogold labelling techniques showed that cell walls of spike tissues contained reduced amounts of cellulose, xylan and pectin near intercellular hyphae or infection pegs compared to walls of healthy host tissues. These studies suggest that cell wall degrading enzymes produced by F. avenaceum facilitated rapid colonization of wheat spikes. The different penetration properties of abaxial and adaxial surfaces of the spikelet tissues as well as the two distinct colonization strategies of host tissues by F. avenaceum are discussed. The penetration and colonization behaviour of F. avenaceum in wheat spikelets resembled that of F. culmorum and F. graminearum, although mycotoxins produced by F. avenaceum differed from those of the latter two Fusarium species.     

Lonicera latent virus,a new carlavirus serologically related to poplar mosaic virus: some properties and inactivation in vivo by heat treatment

A virus with elongate particles (656 nm) was isolated from severalLonicera species. This virus, apparently belonging to the carlavirus group, is serologically distantly related to shallot latent virus and closely related to poplar mosaic virus. The inability to infect poplar and two other hosts of poplar mosaic virus characterizes the virus fromLonicera as a new virus which was namedLonicera latent virus.The virus was easily sap-transmissible but was not transmitted byMyzus persicae.Dilution end-point was about 10^–3, thermal inactivation between 65°C and 80°C and ageing in vitro 1–6 days.Heat treatment, combined with tip-rooting appeared to be a good method to eliminate the virus from severalLonicera species and cultivars.Samenvatting In verschillende soorten en cultivars van het geslachtLonicera (kamperfoelie) blijkt een virus voor te komen dat gemakkelijk door sapinoculatie kan worden overgebracht op kruidachtige planten.Een tegen gezuiverd virus bereid antiserum had een titer van ca. 4096. Er kon mee worden aangetoond dat het virus van kamperfoelie serologisch nauw verwant is met populieremozaïekvirus (Tabel 1). Het virus van kamperfoelie is echter niet in staat om populier,Phaseolus vulgaris Bataaf enVigna sinensis te infecteren en wordt mede daarom als een afzonderlijk virus beschouwd. Het wordt aangeduid als latent kamperfoelievirus (Lonicera latent virus) en behoort evenals populieremozaïekvirus tot de carlavirusgroep (aardappelvirus-S-groep).Het virus blijkt vrij gemakkelijk te kunnen worden geëlimineerd door besmette kamperfoelieplanten gedurende ongeveer zes weken een warmtebehandeling (37°C) te geven en daarna de uiterste toppen (1 cm) te stekken. Van verschillende cultivars werd op deze wijze virusvrij uitgangsmateriaal verkregen.     

Induction of Systemic Acquired Resistance in Pepper Plants by Acibenzolar-S-methyl against Bacterial Spot Disease

The ability of acibenzolar-S-methyl to induce resistance in pepper plants against Xanthomonas campestris pv. vesicatoria was investigated in both growth chamber and open field conditions. Growth chamber experiments showed that acibenzolar-S-methyl (300M) treatment protects pepper plants systemically and locally against X. campestris pv. vesicatoria. Evidence for this was a reduction in the number and diameter of bacterial spots and bacterial growth in planta. Systemic protection was also exerted by the acibenzolar-S-methyl acid derivative, CGA 210007, which may be produced by hydrolysis in the plant. The efficacy of acibenzolar-S-methyl was also found in open field conditions, where both leaves and fruit were protected from the disease. The highest efficacy (about 67%) was obtained by spraying the plants 6–7 times every 8–12 days with a mixture of acibenzolar-S-methyl and copper hydroxide (2.5 + 40ghl^–1 active ingredient). Persistence and translocation data obtained from the growth chamber experiments revealed a persistence of acibenzolar-S-methyl lasting five days after treatment with rapid translocation and negligible levels of acid derivative formation. Since the protection exerted by acibenzolar-S-methyl against bacterial spot disease was observed when the inducer was completely degraded, it would appear to be due to SAR activation.     

Environmental factors influencing sporocarp formation in <Emphasis Type="Italic">Typhula ishikariensis</Emphasis>

Environmental factors influencing sporocarp formation in Typhula ishikariensis were studied under controlled conditions. Sporocarp formation in T. ishikariensis was divided into two stages: stipe elongation from the sclerotium and fertile head development at the tip of the stipe. Factors required for each stage differed. At the stipe elongation stage, low temperature (10°/5°C; day/night) and high humidity were important, but light was not required. In contrast, at the fertile head stage, light and moderate day length (8h/day) were essential. Fertile heads developed at 46µEm^–2s^–1; and high intensity (137µEm^–2s^–1) did not suppress development. Moreover, adding unsterilized soil to the sea sand medium accelerated sporocarp formation. These findings imply that the sclerotium of T. ishikariensis recognizes several physical factors for sporocarp formation. Sporocarps of T. ishikariensis developed within 4 weeks after incubation under optimal conditions. The sporocarp produced basidiospores, and differential mating incompatibility was confirmed among monokaryons derived from basidiospores produced under artificial conditions. This method should be useful for obtaining monokaryons for genetic studies of T. ishikariensis.