Bacterial fruit blotch of melon: screens for disease tolerance and role of seed transmission in pathogenicity

Bacterial fruit blotch (BFB) of cucurbits, caused by Acidovorax avenae subsp. citrulli, is a serious threat to the watermelon and melon industries. To date, there are no commercial cultivars of cucurbit crops resistant to the disease. Here we assessed the level of tolerance to bacterial fruit blotch of various commercial cultivars as well as breeding and wild lines of melon, using seed-transmission assays and seedling-inoculation experiments. Selected cultivars were also tested in a greenhouse experiment with mature plants. All tested cultivars/lines were found to be susceptible to the pathogen, and most of them showed different responses (relative tolerance vs. susceptibility) in the different assays; however, some consistent trends were found: cv. ADIR339 was relatively tolerant in all tested assays, and cv. 6407 and wild lines BLB-B and EAD-B were relatively tolerant in seed-transmission assays. We also provide evidence supporting a strong correlation between the level of susceptibility of a cultivar/line and the ability of the pathogen to adhere to or penetrate the seed. To the best of our knowledge, this is the first attempt to assess melon cultivars/lines for bacterial fruit blotch response.     

The incubation period of beet yellowing viruses in sugar-beet under field conditions

In three years field trials, the incubation period, i.e. the time between infection and the appearance of symptoms, of beet yellows virus (BYV) and beet mild yellowing virus (BMYV) increased with later infection during the growing season. The incubation period of BYV, a closterovirus, increased from 3 weeks in young plants infected before canopy closure, to 9 weeks in old plants infected in August. The incubation period of BMYV, a luteovirus, increased from 4 to 5 weeks in young plants to 9 weeks in old plants. Symptoms were observed c. one week earlier on the inoculated leaves than on those infected systemically, except on young BYV-infected plants. On these plants, symptoms developed in 3 weeks on both leaf types.The incubation period decreased at increasing temperature, a fixed temperature sum being required for the development of symptoms on plants of a certain age. This temperature sum increased with plant age. Symptom development was related to leaf growth; the systemic symptoms appeared after the infected leaves attained their final size. Young, expanding leaves did not show symptoms. Thus the development of symptoms seems to be related to physiological conditions occurring only in full-grown leaves. A low rate of leaf expansion may constitute the underlying reason for the long incubation period of virus symptoms in old plants and at low temperatures.The incubation period was not substantially affected by: (1) the number ofMyzus persicae used to inoculate the plants, (2) the number of leaves inoculated, (3) the development stage of the inoculated leaf and (4) the source plant of BMYV, beet or shepherd's-purse,Capsella bursapastoris. The incubation period can be used to obtain rough estimates of the infection-date of individual plants, given the date on which symptoms appear.Samenvatting De incubatieperiode van het bietevergelingsvirus, BYV, en het zwakke-bietevergelings-virus, BMYV, nam toe naarmate suikerbieteplanten later in het seizoen geïnfecteerd werden. Jonge planten ontwikkelden BYV-symptomen na ongeveer 3 weken terwijl na gewassluiting de incubatieperiode geleidelijk toenam tot 9 weken. De incubatieperiode van BMYV nam toe van 4 à 5 weken na inoculatie in juni tot 9 weken na inoculatie in augustus. Geïnoculeerde bladeren ontwikkelden ongeveer een week eerder symptomen dan de systemisch geïnfecteerde bladeren, behalve bij jonge planten, geïnfecteerd met BYV, waar de symptomen zich op beide typen bladeren tegelijkertijd ontwikkelden.De incubatieperiode nam bij hogere temperatuur af en, afhankelijk van de leeftijd van de plant (aantal bladeren) was een bepaalde temperatuursom nodig voor de ontwikkeling van symptomen. Deze temperatuursom nam toe met de ouderdom van de plant. Van alle systemisch besmette bladeren, vertoonden de oudste, welke juist verschenen op het moment dat de plant werd geïnfecteerd, als eerste symptomen. Dit gebeurde zodra of kort nadat ze hun uiteindelijke grootte hadden bereikt. Groeiende bladeren vertoonden nooit vergelingssymptomen. De trage bladexpansie in oude planten en bij lage temperaturen is een mogelijke oorzaak van de lange incubatieperiode aan het einde van het seizoen.De incubatieperiode werd niet duidelijk beïnvloed door inoculatieomstandigheden, zoals (1) het aantal groene perzikluizen,Myzus persicae, dat werd gebruikt voor inoculatie, (2) het aantal geïnoculeerde bladeren, (3) de ouderdom van het geïnoculeerde blad, (4) de bronplant van BMYV, biet of herderstasje, of (5) de vector species. Omdat de incubatieperiode niet in belangrijke mate afhankelijk is van deze factoren kan bij kennis van de datum waarop symptomen verschenen de infectiedatum worden bepaald op basis van de incubatieperiode.     

Further characterization of melon necrotic spot virus causing severe disease in glasshouse cucumbers in the Netherlands and its control

A severe leaf necrosis, observed since 1978 in glasshouse cucumbers grown on rockwool and later also in crops on soil, is described. A virus could be isolated and the disease be reproduced in cucumber and melon. The virus could be transmitted by leaf inoculation with expressed sap and by pouring rockwool leakage water onto sterilized soil containing cucumber seedlings. Infectivity steeply declined in expressed sap between dilutions 10 and 100 (dilution endpoint ca 10⁶), at temperatures between 55 and 65°C (thermal inactivation point 75°C) and during storage between 1 and 1 1/2 month at room temperature.Out of 40 plant species tested only three species, viz. cucumber, melon and watermelon, were susceptible. All 21 cucumber cultivars and all 8 melon cultivars tested reacted severely with local lesions and some with systemic necrosis, but systemic infection and reaction were erratic under experimental conditions.Purified virus sedimented in sucrose and CsCl gradients and during analytical ultracentrifugation in a single peak. Thes ₂₀ was 134S and buoyant density in CsCl was 1.33 g.cm^–3. Virus particles in crude sap and purified suspensions were spherical and ca 30 nm in diameter. They contained one type of protein with a relative molecular mass of 46 000 and one RNA species. An antiserum with a titre of 1024 did not react with cucumber and tobacco necrosis viruses, nor did their antisera react with our cucumber virus. Serologically and in physicochemical properties the virus is similar to if not identical with the melon necrotic spot virus incompletely described in Japan.Disease control may be through improved hygiene, including steam sterilization of rockwool, soil disinfection by steam sterilization or with methyl bromide, and addition of a surfactant to nutrient solutions, and prevention may be by grafting cucumber ontoCucubita ficifolia rootstocks, immune to the virus.Samenvatting Sinds 1978 komt in de op steenwol en in grond geteelde kaskomkommer een ernstige bladnecrose voor, die vooral in het najaar tot afsterving van planten kan leiden en in wel 45% van de planten van een aangetast gewas is geconstateerd. Uit zieke planten kon een virus worden geïsoleerd dat gemakkelijk overging door sap-inoculatie en in lekvocht uit besmette steewol (waarschijnlijk door tussenkomst van eenOlpidium-soort), nadat dit werd gegoten op gesteriliseerde grond waarin komkommerzaailingen groeiden. Met dit virus konden de symptomen van de ziekte worden greproduceerd.Het infectivermogen van ruw platesap nam snel af bij verdunning tussen 10 en 100× (verdunningseindpunt ca 1 millioen), en bij warmtebehandelingen tussen 55 en 65°C (inactiveringstemperatuur 75°C) en bij bewaring bij kamertemperatuur tussen 1 en 1 1/2 maand.Slechts 3 van de 40 getoetste plantesoorten bleken vatbaar voor het virus, te weten komkommer, meloen en watermeloen. Alle 21 getoetste komkommercultivars en alle 8 getoetste meloenerassen reageerden hevig met lokale lesies en enkele, onder de heersende proeformastandigheden onvoorspelbaar, met systemische necrose. De wel als onderstam gebruikteCucurbita ficifolia is onvatbaar.Gezuiverd virus sedimenteerde in suiker- en CsCl-gradiënten en bij analytische ultracentrifugering in één piek. Des ₂₀ was 134S en de zweefdichtheid in CsCl 1.33 g.cm^–3. In ruw sap en gezuiverde suspensies deden de virusdeeltjes zich voor als bolletjes met een diameter van ongeveer 30 nm. Ze bevatten slechts éé soort eiwit met een relatieve moleculaire massa van 46 000 en één RNA-soort. Een antiserum met titer 1024 werd bereid. Het reageerde niet met komkommernecrosevirus en tabaksnecrosevirus. Wel reageerde het virus met een uit Japan ontvangen antiserum tegen het daar sinds 1966 bekende melon necrotic spot virus, terwijl het Japanse virus reageerde met het Nederlandse antiserum. Serologisch, zowel als in biologische en fysisch-chemische eigenschappen lijken de Nederlandse en Japanse isolaten identiek. Voor het virus wordt daarom de Nederlandse namm meloenenecrosevirus voorgesteld. Het verschilt van drie andere, onlangs min of meer gelijktijdig in Oost-Duitsland, op Kreta en in Libanese grond aangetroffen, via de bodem overgaande komkommervirussen, die evenals tabaks- en komkommernecrosevirus ook andere plantesoorten dan cucurbitaceeën kunnen infecteren.Waarschijnlijk is meloenencerosevirys al sinds 1967 bekend in Frankrijk als verwekker van criblure du melon. Het is ook nauw verwant aan de verwekker van een in een veredelingsprogramma van meloen in Californië opgedoken necrosevirus, waarvan echter wordt beweerd dat het overgaat met zaad van meloen en wordt overgebracht door bladkevertjes,Diabrotica-soorten. Het meloenenecrosevirus is in ons land voor het eerst geconstateerd als ziekteverwekker van kaskommer. Ook in England is het daarin onlangs aangetroffen. De ziekte kan op verschillende manieren bestreden, respectievelijk voorkomen worden. De grond dient gestoomd te worden of begast met methylbromide. Steenwolmatten kunnen bij hergebruik gestoomd worden, terwijl aan de voedingsoplossing uitloeier (Agral) toegediend kan worden. Zowel bij grond- als steenwolteelten is de ziekte te voorkomen door komkommerplante te enten op de onvatbare onderstamCucurbita ficifolia.     

Systemic toxicity of spinosad to the greenhouse whiteflyTrialeurodes vaporariorum and to the cotton leaf wormSpodoptera littoralis

When spinosad was administered at the stem base of tomato plants grown in rockwool, the main substrate used in greenhouses in northern Europe, it was taken up by the roots and transported to the leaves. Laboratory toxicity experiments showed that systemically applied spinosad, at doses as low as 2 mg active ingredient per plant, gave excellent control of nymphs of the greenhouse whiteflyTrialeurodes vaporariorum, but was less toxic to adults. The toxicity of spinosad on Homoptera has not been documented yet and a dose-response relationship was established. The persistence of toxicity was at least 22 days for whitefly nymphs. A dose-response relationship of systemically applied spinosad was also determined on third-instar larvae of the cotton leaf wormSpodoptera littoralis; a dose of 5 mg per plant consistently killed all larvae. The persistence of systemically applied spinosad reached up to 45 days after treatment. Systemically applied spinosad was harmless to the parasitic waspEncarsia formosa. http://www.phytoparasitica.org posting Nov. 29, 2005.     

Population Dynamics of <Emphasis Type="Italic">Monosporascus</Emphasis><Emphasis Type="Italic">cannonballus</Emphasis> Ascospores in Marsh Soils in Eastern Spain

The population dynamics of Monosporascus cannonballus ascospores in the soils of four muskmelon fields located in a marsh area in Castellón province (eastern Spain) was studied for a 3 year period. Two of these fields were cropped to muskmelon with fallow periods between muskmelon cropping, and the others were in fallow and had extensive flooding periods. Muskmelon cultivation resulted in a progressive increase of the number of ascospores in soil, reaching a maximum 7 months after muskmelon planting (2–4 months after plant death), and a subsequent decline during fallow periods between muskmelon crops. During muskmelon cropping, in-bed and between-bed ascospore numbers were compared and, in general, there were no statistical differences between them. In the fields which were in fallow and flooded, the dynamics found was a progressive decline of the population of ascospores. Soilborne inoculum was viable and capable of infecting muskmelon at the end of the 3 year period in all fields, demonstrating that ascospores of M.␣cannonballus are able to survive for this period of time in the absence of muskmelon cultivation and also that this fungus seems to be well adapted to survive in soils which maintain a high water table during the crop or under flooding conditions.     

Retrospective estimation of the data of infection with beet yellowing viruses in sugar-beet under field conditions

Sugar-beet plants, infected with beet yellows virus (BYV, closterovirus group) or beet mild yellowing virus (BMYV, luteovirus group) develop symptoms on the inoculated leaves on which aphids infected the plant. Symptoms develop also on the systemically-infected leaves to which virus has been transported via the phloem. Systemic infection occurs in the leaves which have just, or not yet appeared at the moment of infection of the plant. All other, older leaves remain uninfected. The infection-date can be estimated by assessing the date of appearance of the oldest systemicallyinfected leaf of a plant. This approach was tested in the field and gave good results.Samenvatting Suikerbieteplanten die besmet zijn met het bietevergelingsvirus, BYV, of met het zwakke vergelingsvirus, BMYV, ontwikkelen symptomen op de geïnoculeerde bladeren, waarop infectieuze bladluizen virus hebben overgedragen, én op de systemisch besmette bladeren waarheen het virus vanuit de geïnoculeerde bladeren is getransporteerd via het vaatsysteem. Bladeren die op het moment van infectie nog niet verschenen zijn of vlak ervóór zijn verschenen, worden systemisch besmet, terwijl oudere bladeren gezond blijven. De infectiedatum kan worden bepaald door aan de hand van temperatuursommen de verschijningsdatum van het oudste systemisch besmette blad te berekenen. Deze methode bleek bij toetsing in het veld goed te voldoen.     

Current status of the greenhouse whitefly, Trialeurodes vaporariorum, susceptibility to neonicotinoid and conventional insecticides on strawberries in southern California

Since 1998, the greenhouse whitefly, Trialeurodes vaporariorum Westwood (Homoptera: Aleyrodidae), has emerged as a major insect pest of many horticultural crops in coastal California. Control of this pest has been heavily dependent upon chemical insecticides. Objectives of this study were to determine the status of the greenhouse whitefly susceptibility to neonicotinoid and conventional insecticides on strawberries in Oxnard/Ventura, a year-round intensive horticultural production area of southern California. For bioassay tests, adult whiteflies were collected from commercial strawberry crops, and immatures were directly developed from eggs laid by these adults. LD(50) values of soil-applied imidacloprid, thiamethoxam and dinotefuran were respectively 8.7, 3.2 and 4.9 times higher for the adults, 1.8, 1.2 and 1.5 times higher for the first-instar nymphs and 89.4, 390 and 10.4 times higher for the third-instar nymphs than their top label rates. LC(50) values of foliar-applied imidacloprid, thiamethoxam and acetamiprid were respectively 6.1, 6.0 and 1.7 times higher for the adults and 3.8, 8.7 and 4.4 times higher for the second-instar nymphs than their top label rates. For the adults, LC(90) values of endosulfan, malathion, methomyl, bifenthrin and fenpropathrin were 2.2, 1.2, 1.9, 2.3 and 4.9 times lower than their respective top label rates. Chlorpyrifos was not very effective against the adults, as indicated by its LC(90) being 120% higher than its top label rate. The present results strongly emphasize the need to develop resistance management strategies in the region.     

Quantitative detection of <Emphasis Type="Italic">Monosporascus cannonballus</Emphasis> in infected melon roots using real-time PCR

A method was developed for the specific detection, identification and quantification of Monosporascus cannonballus in infected melon roots based on real-time PCR (SYBR® Green chemistry) targeting the ITS1 region of the rDNA conserved between different strains of the pathogen. The specificity of the reaction was assessed using a number of fungi taxonomically and ecologically related to M. cannonballus. The method was highly sensitive and M. cannonballus was first detected in the roots of a susceptible Piel de Sapo cultivar 2 days after inoculation, before symptom appearance. Although conventional PCR methods could also provide such a specific and sensitive detection, real-time PCR was also able to produce reliable quantitative data over a range of 4 orders of magnitude (from 5 ng to 0.3 pg). The method allowed the quantitative monitoring of fungal growth from the very first stages of infection, and was successfully employed in the early screening of resistance. The assessment of disease progress and severity obtained with real-time PCR was more accurate than that obtained with the visual scoring of root lesions or root biomass losses. Therefore, there exists a great potential for its implementation in those steps of breeding programmes where high accuracy is required.     

Incidence and characterisation of resistance to neonicotinoid insecticides and pymetrozine in the greenhouse whitefly,Trialeurodes vaporariorum Westwood (Hemiptera: Aleyrodidae)

BACKGROUND: Trialeurodes vaporariorum (Westwood), also known as the greenhouse whitefly, is a serious pest of protected vegetable and ornamental crops in most temperate regions of the world. Neonicotinoid insecticides are used widely to control this species, although resistance has been reported and may be becoming widespread. RESULTS: Mortality rates of UK and European strains of T. vaporariorum to a range of neonicotinoids and pymetrozine, a compound with a different mode of action, were calculated, and significant resistance was found in some of those strains. A strong association was found between neonicotinoids and pymetrozine, and reciprocal selection experiments confirmed this finding. Expression of resistance to the neonicotinoid imidacloprid and pymetrozine was age specific, and resistance in nymphs did not compromise recommended application rates. CONCLUSION: This study indicates strong parallels in the phenotypic characteristics of neonicotinoid resistance in T. vaporariorum and the tobacco whitefly Bemisia tabaci Gennadius, suggesting possible parallels in the underlying mechanisms. Copyright © 2010 Society of Chemical Industry     

Detection of Pseudomonas syringae pv. aptata in Irrigation Water Retention Basins by Immunofluorescence Colony-staining

Bacterial blight of cantaloupe (Cucumis melo) caused by Pseudomonas syringae pv. aptata was first observed in south-western France and has since spread to all cantaloupe-growing areas of this country. Use of pesticides registered for this disease has proved ineffective and no commercial cultivars of cantaloupe are resistant to this blight. To develop control strategies for this disease, the principal sources of inoculum were investigated. Among the different sources of inoculum studied, we report the isolation of P. syringae pv. aptata from irrigation water retention basins in south-western France using the immunofluorescence colony-staining (IFC) method. In this study, the pathogen was detected at a low concentration (12 and 70cful^–1) in two different retention basins. These results suggest that P. syringae pv. aptata can survive in water used to irrigate cantaloupe crops and could be a source of inoculum for epidemics of bacterial blight. To develop control strategies for this bacterial disease, the importance of water retention basins as sources of inoculum for bacterial blight of cantaloupe needs to be evaluated relative to other potential sources such as seeds, plants from nurseries and plant debris in the soil.     

浅黄恩蚜小蜂和丽蚜小蜂对温室白粉虱的寄生潜能分析

浅黄恩蚜小蜂Encarsia sophia GiraultDodd和丽蚜小蜂Encarsia formosa Gahan是防治粉虱类害虫的优势寄生蜂,通过生命表技术方法分析了2种寄生蜂对温室白粉虱Trialeurodes vaporariorum(Westwood)的防治潜能。结果表明,丽蚜小蜂在羽化第3天和第10天出现2次寄生高峰,占其总寄生量的13.7%和8.0%,在2次高峰之间逐日寄生粉虱数量比较平稳,单雌逐日平均产雌数保持在10.6~13.4头,10 d后寄生量呈明显的下降趋势;而浅黄恩蚜小蜂羽化10 d内逐日寄生粉虱量变化不大,单雌逐日产雌数稳定在4.2~5.4头,羽化14 d后寄生量呈明显下降趋势。丽蚜小蜂和浅黄恩蚜小蜂的R0、T、rm、λ值分别为171.5、18.0、0.2854、1.3303和61.6、16.2、0.2544、1.2897;粉虱若虫充足时,丽蚜小蜂平均单雌寄生若虫数是浅黄恩蚜小蜂的2.7倍,而后者平均单雌取食若虫数为60.6头,明显高于前者42.7头,总的来看,丽蚜小蜂通过寄生和取食杀死粉虱总量220.8头,明显高于浅黄恩蚜小蜂的127.9头。表明在应用寄生蜂防治温室白粉虱时,单独释放丽蚜小蜂比浅黄恩蚜小蜂显示出更好的防治潜能。     

Isolation and pathogenicity of Colletotrichum spp. causing anthracnose of strawberry in south west Spain

Anthracnose, caused by Colletotrichum spp., is a major disease of cultivated strawberry, Fragaria × ananassa. This study identifies the Colletotrichum spp. which causes strawberry anthracnose in the southwest of Spain. A survey of the region was carried out, and the strains isolated were identified as C. acutatum by using the polymerase chain reaction (PCR) with genus and species-specific primers, demonstrating that this species is currently the causal agent of strawberry anthracnose in the studied region. The pathogenicity of C. acutatum and C. gloeosporioides strains was evaluated on two principal strawberry cultivars (cvs Camarosa and Ventana) under field conditions, the latter being more pathogenic than the former.     

Monitoring physiological races of<Emphasis Type="Italic">Podosphaera xanthii</Emphasis> (syn.<Emphasis Type="Italic">Sphaerotheca fuliginea</Emphasis>), the causal agent of powdery mildew in cucurbits: Factors affecting race identification and the importance for research and commerce

Identification of the physiological races ofPodosphaera xanthii (syn.Sphaerotheca fuliginea), the causal agent of powdery mildew in cucurbits, is based upon the differing responses of various melon cultigens to the pathogen. Eight races of the pathogen have been identified to date in the USA, Africa, Europe and around the Mediterranean Sea, and four new races were reported from greenhouse melons in the major growing area of Japan. Plant responses to powdery mildew may be affected by environmental factors such as light intensity, temperature and humidity, as well as by age and nutritional status of the plants. The same factors affect the accuracy and reliability of race identification. In an attempt to overcome those obstacles, the genetic diversity ofP. xanthii was studied using molecular markers. Unfortunately, no correlation was found between DNA polymorphism and the race of the pathogen as identified by biological tests. The usefulness of race identification as a guide for the grower in selecting appropriate cultivars is limited because changes or shifts in the pathogen population are common. Such changes may be found among growing seasons, geographic regions and hosts, and also within a single greenhouse during a single season. On the other hand, race identification is important for basic research and is especially important for the commercial seed industry, which requires accuracy in declaring the type and level of resistance to powdery mildew in its products. http://www.phytoparasitica.org posting March 2, 2004. Contribution from the Agricultural Research Organization. No. 501/04.     

Colonisation and histological changes in muskmelon and autumn squash tissues infected by <Emphasis Type="Italic">Acremonium cucurbitacearum</Emphasis> or <Emphasis Type="Italic">Monosporascus cannonballus</Emphasis>

Muskmelon (Cucumis melo cv. Temprano Rochet) and autumn squash (Cucurbita maxima) seedlings were inoculated either with Acremonium cucurbitacearum or Monosporascus cannonballus, two of the soil-borne fungi implicated in ‘melon collapse’. Inoculation was achieved in two different ways: by growing the plants in pots containing infested soil to study the histological changes produced in the infected tissues using light microscopy and by growing seedlings in Petri dishes together with fungal colonies in order to observe the colonisation of the plant tissues using scanning electron microscopy. Both muskmelon and autumn squash roots infected with A. cucurbitacearum showed a suberised layer in the epidermis and the outermost layers of the parenchymatic cortex, but these symptoms developed earlier in the muskmelon plants. Muskmelon plants infected by this fungus also presented hypertrophy and hyperplasia, which led to a progressive separation of the vascular bundles in the lower stems of the affected plants. This response was not observed in autumn squash during the study. On the other hand, few histological changes were observed in tissues infected with M. cannonballus and only a slight increase in the size of cortical intercellular spaces was noted in the lower stems of muskmelon plants, and infected autumn squash tissues remained free of these symptoms throughout the study. The scanning electron microscope observations revealed that both fungi were able to colonise the tissues of the two host plants which were studied. A. cucurbitacearum colonised the epidermis and cortex of both muskmelon and autumn squash. The hyphae grew both inter- and intracellularly, and the density of the colonisation decreased within the endodermis. The same colonisation of host plants was observed as a result of M. cannonballus infection. The xylem vessel lumina of both muskmelon and autumn squash showed hyphae and tylose formation as a result of both fungal infections. However, non-fungal structures were detected in the hypocotyl vascular tissues. The present study demonstrates that both fungi are capable of infecting the tissues of a species which is resistant (autumn squash) and a species which is susceptible (muskmelon) to melon collapse.     

Viral dieback of carrot and other umbelliferae caused by the Anthriscus strain of parsnip yellow fleck virus,and its distinction from carrot motley dwarf

Viral dieback of carrot, chervil, coriander, dill and wild Umbelliferae is described. Disease incidence in carrot crops grown for seed is often high but low in ware carrot. There is no secondary spread in carrot crops.The causal virus was identified as theAnthriscus strain of parsnip yellow fleck virus (PYFV) transmitted byCavariella aegopodii from cow parsley(Anthriscus sylvestris). Nicotiana benthamiana was practically indespensable for isolation of PYFV by sap transmission from plants with viral dieback.No immunity was found in 12 carrot cultivars or in wild carrot. Disease control with a systemic insecticide had limited effect.Carrot red leaf virus and carrot mottle virus were commonly found in carrot, but they did not cause dieback symptoms. Cucumber mosaic virus, parsnip mosaic virus and a virus resembling that of carrot yellow leaf were occasionally isolated from carrot. Symptoms due to mycoplasma were also observed.Samenvatting Bij de zaadteelt van peen is in ons land reeds lang een schadelijke, vroeg in het seizoen optredende instervingsziekte bekend als voorjaarsziekte of het zwart. Planten vallen op door necrose van jonge spruiten (insterving). Soms gaat meer dan de helft van het gewas verloren. Voor consumptie geteelde peen wordt echter nauwelijks aangetast. De ziekte is nu ook gevonden bij dille, kervel, koriander en wilde schermbloemigen.Uit zieke planten en ook vaak uit symptoomloze fluitekruidplanten werd een virus geïsoleerd waarmee de insterving kon worden gereproduceerd. Het werd herkend als de fluitekruid-(ofAnthriscus-)stam van pastinakegeelvlekvirus (PYFV) op grond van waardplanten, symptomen, serologie en overdracht doorCavariella aegopodii met als onmisbare helper hetAnthriscus-vergelingsvirus (AYV), dat ook in fluitekruid voorkomt. Het gebruik vanNicotiana benthamiana als toetsplant maakte isolatie uit planten met virusinsterving mogelijk. Voor de ziekte wordt nu de naam virusinsterving van schermbloemigen voorgesteld.Peenroodbladigheid veroorzaakt door peenroodbladvirus, dat meestal samengaat met peenvlekkenvirus, bleek ook algemeen voor te komen. Deze twee virussen spelen geen rol bij het veroorzaken van virusinsterving, zoals wel werd aangenomen. Beide ziekten zijn geheel verschillend in symptomatologie en epidemiologie. Incidenteel werden komkommermozaïekvirus, pastinakemozaïekvirus en een virus gelijkend op peengeelbladvirus in aangetroffen. Ook werd eenmaal een aan een mycoplasma toe te schrijven ziekte geconstateerd.Virusinsterving bleek epidemiologisch te kunnen worden verklaard door de massale jaarlijkse migratie vanC. aegopodii in het voorjaar, waarbij PYFV van fluitekruid naar peen en andere schermbloemigen wordt verspreid. Door onvatbaarheid van peen voor het helpervirus (AYV) treedt in dit gewas geen secundaire verspreiding op.In geen van 12 peenrassen en wilde peen werd resistentie aangetroffen. Toepassing van een systemisch insekticide bleek in eerder onderzoek slechts een beperkt effect te hebben. Peenzaadteelt in gebieden met minder bladluizen, zoals het noorden des lands, lijkt aan te bevelen, maar verder lijkt de ziekte niet te bestrijden.Work in partial fulfillment of requirements for master's training at Agricultural University, Wageningen     

Fluorescent staining of resting spores of Polymyxa betae as a fungal vector of rhizomania disease of sugar beet in soil

When the resting spores of Polymyxa betae were pretreated with 2% sodium dodecyl sulfate (SDS) and then stained with various fluorochromes including 3,3′-dihexyloxacarbocyanine iodide [DiOC6(3)], calcofluor, and a fluorescein isothiocyanate (FITC)-conjugated lectin, such as wheat germ lectin (WGA) or caster bean lectin, most spores fluoresced brightly. FITC-WGA mainly stained the cell surface, while DiOC6(3) stained the cytoplasm. After pretreatment with SDS and addition of FITC-WGA or DiOC6(3) to a soil suspension containing resting spores, the resting spores were distinguishable from the soil particles. This staining method is easy to use for the detection of resting spores in the soil.     

Dissemination of rhizomania by soil,beet seeds and stable manure

Polymyxa betae, the vector of beet necrotic yellow vein virus (BNYVV), (the causal agent of rhizomania of sugar-beet), forms cystosores which are very persistent and might be dispersed by soil, beet seeds, plant material and stable manure. Research has been carried out into the risk of dissemination; relative importance was not determined. Inoculation with diseased soil in a field caused rhizomania in sugar-beets within one year. This implies that even small amounts of soil adhering to plant roots constitute a potential danger. Direct transmission of BNYVV by sugar-beet seeds could not be demonstrated, but, after processing and cleaning seed lots originating from infested fields, the seed waste proved to be contaminated. Cystosores ofP. betae and, to a lesser extent, BNYVV could pass through the intestine of sheep in fodder experiments carried out with heavily infested sugar-beet tails.Samenvatting De vector van het bieterhizomanievirus,Polymyxa betae, vormt zeer persistente ruststructuren (cystosoren) die verspreid worden met grond, bietezaad, plantmateriaal en stalmest.Onderzoek is uitgevoerd naar de risico's van verspreiding, maar door het ontbreken van kwantitatieve methoden kon de relatieve belangrijkheid niet worden vastgesteld.Wanneer een gezond perceel werd besmet met geïnfecteerde grond (20 ng dm^–3 op bouwvoor), werd binnen één jaar aantasting door rhizomanie waargenomen. Dit wijst erop dat geringe hoeveelheden grond, die meekomen met wortelmateriaal van plantgoed, een potentieel gevaar vormen.Directe overdracht van het bieterhizomanievirus door bietezaad kon niet worden aangetoond, maar na het schonen en bewerken van zaadpartijen afkomstig van zieke percelen bleek dat schoningsafval en in het bijzonder de grondfractie daarvan, wel was besmet.Cystosoren vanP. betae en in mindere mate het bieterhizomanievirus konden het maag-darmkanaal van schapen passeren, hetgeen werd aangetoond in enkele voederproeven, uitgevoerd met zwaar besmette suikerbietestaartjes.     

Ovicidal and nymphicidal effects of some fungicides on the greenhouse whitefly<Emphasis Type="Italic">Trialeurodes vaporariorum</Emphasis>

Eggs and L1 nymphs of the greenhouse whiteflyTrialeurodes vaporariorum (Westwood) (Hem.: Aleyrodidae) on bean leaves were directly sprayed with the fungicides maneb and mancozeb at recommended rates for practical use. Strong ovicidal and nymphicidal effects were observed. These fungicides may prove to have practical use for integration in IPM programs for whitefly control, as they also exert low or no toxicity to the whitefly parasitoidEncarsia formosa. http://www.phytoparasitica.org posting April, 30, 2004.     

The development of rhizomania in two areas of the Netherlands and its effect on sugar-beet growth and quality

Since detection of beet necrotic yellow vein virus (BNYVV) by serological techniques was not possible 10 years ago, there is no relaible information about the occurrence of the disease in the Netherlands prior to 1983. By evaluating historical information on the sugar production of fields, which have been proved to be infested by rhizomania recently, the time of occurrence of the first infections could be estimated roughly.Analysis of root samples from diseased fields and comparison with uninfected fields, revealed a correlation between sugar and sodium content and the occurrence of rhizomania. Therefore both quality parameters can be used as indicators for the disease under Dutch conditions.Differences between infected and uninfected fields were detected from mid-June onwards. Only a weak correlation between sugar and -amino N content could be established. At later stages of crop growth the -amino N level of diseased fields was always lower than that of healthy ones. However sugar content remained at a low level and did not increase substantially.Samenvatting Aangezien tien jaar geleden geen serologische technieken beschikbaar waren, kon het voorkomen van bieterhizomanie in Nederland niet worden aangetoond. Door het beoordelen van de suikeropbrengsten van percelen die met bieterhizomanie besmet bleken te zijn, over een reeks voorafgaande jaren, kon een periode waarin de infectie vermoedelijk had plaatsgevonden worden vastgesteld.Door analyse en vergelijking van bietmonsters van een reeks aangetaste proefplekken te een aantal gezonde referentiepercelen, kon worden aangetoond dat onder Nederlandse omstandigheden suiker- en Na-gehalten de meest betrouwbare indicatoren voor de aanwezigheid van bieterhizomanie zijn.Afwijkingen in de gehalten van deze kwaliteitsparameters werden in de zwaar aangetaste percelen vanaf half juni gevonden. Er werd een zwakke relatie tussen de gehalten aan suiker en -amino N aangetoond. Gedurende de tweede helft van de zomer was het gehalte aan -amino N in de zieke percelen lager dan dat van bieten in de gezonde. Het suikergehalte bleef op een laag niveau en vertoonde geen of een geringe toename.