Molecular identification and pathogenicity of Rhizoctonia solani and Pythium spp. associated with damping-off disease on baby leafy vegetables in Greece

In the last years, leafy vegetables cultivated as baby leaves have been established in the market and have attracted the interest of consumers throughout the world. During the growing seasons of 2019 and 2020, 97 isolates of Rhizoctonia solani and 112 isolates of Pythium spp. were obtained from baby leaf vegetables exhibited damping-off symptoms. Representative isolates of R. solani from each surveyed plant species were characterized using sequence analysis of the internal transcribed spacer (rDNA-ITS) region. Isolates were identified as belonging to four anastomosis groups (AGs): AG2-1, AG-IB, AG4-HGI and AG4-HGIII. AG4-HGI was the most prevalent group and phylogenetic analysis showed that the isolates were distinctly separated according to their AGs. Pathogenicity among the four AGs on 23 plant species varied considerably, from not susceptible to highly susceptible, while, in general, AGs did not exhibit host specificity. Furthermore, a total of 112 Pythium spp. isolates were obtained. The ITS region and the cytochrome oxidase II (coxII) gene were amplified, and three Pythium spp. were identified (P. ultimum, P. aphanidermatum and P. sylvaticum), which were used further for maximum-likelihood phylogenetic analysis. The pathogenicity of representative isolates was assessed in vitro and in vivo on 10 plant species. In general, all three tested Pythium spp. were virulent when used in vitro, while P. ultimum was the most virulent in vivo. This is the first comprehensive study aimed at determining the occurrence of specific R. solani AGs and Pythium spp. derived from baby leafy vegetables exhibiting damping-off symptoms in Greece.     

Suitability of Ten Plant Baits for the Rapid Detection of Pathogenic Pythium Species in Hydroponic Crops

Ten types of plant baits were tested in the laboratory to assess their capacity to detect pathogenic Pythium species. These were orange tree leaves, tomato leaves, pepper leaves, geranium leaves, Bermuda grass leaves, pine needles, immature carnation petals, hemp-seed cotyledons, pepper and cucumber fruits. The Pythium spp. tested were P. aphanidermatum, P. irregulare and Pythium group F from hydroponic market garden crops in the Poniente region of Almería (south-east Spain). The test consisted of observing the velocity at which five baits were colonized and the day of colonization of the first bait. Results indicated that the slowest baits to be infected were immature carnation petals and pine needles. These two, together with Bermuda grass leaves, were also the baits infected in lowest number, such that practically no further infection was produced in the baits after the fifth day of contact with the inoculated water. The other plant baits tested were equally suitable for detection of Pythium spp. over the first two days, although only orange leaves and hemp-seed cotyledons were infected on the first day.     

Pathogenicity and fungicide sensitivity of Pythium and Phytopythium spp. associated with soybean in the Huang-Huai region of China

Pythium and Phytopythium spp. cause seed decay, damping off, and root rot in soybean, wheat, and many other crops. However, their diversity and importance as pathogens, particularly in different crop rotation systems, are largely unknown. A survey was conducted in the Huang-Huai region, one of the main areas of soybean–wheat rotation farming in China. In 2016–2018, we collected 300 soybean seedlings and 150 field soil samples from several representative locations, and identified 26 Pythium and 6 Phytopythium spp. from 212 isolates, based on internal transcribed spacer 2 (ITS2) and cytochrome oxidase subunit 1 sequences. The pathogenicity of these isolates was evaluated by growing soybean and wheat seeds in dishes and pots containing oomycete cultures. We found that 12 Pythium spp. (but no Phytopythium spp.) showed high pathogenicity on soybean and/or wheat, and nine of them (75%) were highly pathogenic on both crops. Among the nine species, Pythium spinosum, Pythium ultimum, Pythium species 1 (tentatively designated as ‘Candidatus Pythium huanghuaiense’), Pythium aphanidermatum, and Pythium myriotylum were highly abundant among all isolates (15%, 10%, 9%, 8%, and 5%, respectively). Nine species were selected for testing of sensitivity to the fungicides metalaxyl and mefenoxam. The EC₅₀ values were all less than 10 μg/ml, indicating little resistance. Minimum inhibitory concentration values indicated isolates were about twice as sensitive to mefenoxam as to metalaxyl. These results provide a systematic understanding of Pythium and Phytopythium species associated with soybean in the Huang-Huai region, which is important for disease management and breeding programmes.     

Antagonismusin vitro vonBacillus spp. gegenüberRhizoctonia solani undPythium spp.

Es konnten 132Bacillus-Stämme isoliert und gegenüber7 Rhizoctonia-solani- und 6Pythium-Stämmen auf Antagonismusin vitro untersucht werden. Die gefundenen Antagonisten hemmten die Pilzisolate unterschiedlich stark, so daß Gemische von Antagonisten für die Praxis zu empfehlen sind.There were 132Bacillus-strains isolated and tested for antagonism against 7 strains ofRhizoctonia solani and 6 strains ofPythium spp. The isolated antagonists didn't show a uniform effect against the tested strains ofRhizoctonia solani andPythium spp. For an application in practice it will be better to use a mixture of antagonists.

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Mit 4 Abbildungen und einer Tabelle     

Effect of Pythium spp. and glyphosate on phytoalexin production and exudation by bean (Phaseolus vulgaris L.) roots grown in different media

Kievitone, phaseollinisoflavan and phaseollin were detected in roots of bean seedlings (Phaseolus vulgaris L.) grown in natural soil. Comparison of phytoalexin production by roots grown in different media indicated that these phytoalexins were probably induced by microorganisms in soil. The influence of common root rot pathogens of bean, Pythium spp., on phytoalexin production was determined. Pythium ultimum elicited kievitone, phaseollinisoflavan and phaseollin in roots grown in sterilized silica sand. P. sylvaticum induced only kievitone and phaseollin in the same growth medium. Glyphosate did not significantly affect the accumulation of phytoalexins within 3 days. However, by day 5, significantly more phaseollin was detected in the roots of Pythium inoculated plants treated with glyphosate than in Pythium inoculated plants not treated with glyphosate. In a hydroponic system, both Pythium spp. elicited accumulation of kievitone and phaseollin in root tissue, and both phytoalexins were exuded into the bathing solution. Glyphosate application did not significantly affect accumulation or exudation of phytoalexins by bean roots in the hydroponic system. The results from this study illustrate the nature and extent of phytoalexin production by bean roots in the absence and presence of microbes.     

Characterisation of fungal pathogens causing basal rot of lettuce in Belgian greenhouses

Basal rot is a common disease in lettuce greenhouses. A 3-year study on the diversity of pathogens associated with basal rot in Belgium was carried out. A total of 150 isolates were collected originating from 56 greenhouses. Four pathogens appeared to be involved. Rhizoctonia solani was found to be the causal agent at 23 locations, Sclerotinia spp. at 14, Botrytis cinerea at 17 and Pythium spp. at seven. The isolates of R. solani were further characterised to anastomosis groups and subgroups using morphological characteristics, pectic zymogram and PCR-RFLP. Five anastomosis groups could be distinguished: AG1-1B, AG4 HGI, AG10, AG2-1, AG2-1 Nt and AG3, with isolates of AG4 HGI and AG1-1B being the most prevalent and the most aggressive. Sclerotinia sclerotiorum was found at 13 locations, while S. minor was found at only one location. Based on ITS-sequencing Pythium isolates were assigned to three different species. At 20°C, isolates of all pathogens were able to cause lesions on detached lettuce leaves, except isolates of R. solani AG3 and AG2-1 Nt. A correlation could be found between the occurrence of the pathogens and the growing season. Botrytis cinerea was the most common pathogen in winter, whereas R. solani was most frequently isolated in summer. Sclerotinia spp. and Pythium spp. were isolated in spring, summer and autumn. The information obtained in this study will be most useful in the development of an alternative control strategy for causal agents of basal rot.     

Identity and pathogenicity of Rhizoctonia spp. associated with bare patch disease of cereals at a field site in Western Australia

Roots of seedlings of wheat and barley affected by bare patch disease at a field site in Western Australia were assessed for root damage and plated to isolate fungi. The patches were variable in shape and size and had the most severely affected plants in the centre. Of the 165 isolates ofRhizoctonia spp. obtained, 90% were multinucleate and 10% binucleate, the former being predominant in the plants at the centre of the patch. The relative frequency of binucleate isolates increased with proximity to the periphery. The increase in activity of avirulent binucleate isolates towards the periphery of the patch may be related to the sharp and abrupt edging of the patch. A variety of other species of fungi such asFusarium spp.,Mortierella spp.,Bipolaris sorokiniana, Pythium sp. andTrichoderma sp. were encountered within the patches. The multinucleate isolates belonging to anastomosis groups (Ag) 2–1, 2–2 and 8 (Thanatephorus cucumeris) were most pathogenic to wheat. The binucleate isolates of Ag C, D, E, and K (Ceratobasidium sp.) were less pathogenic. It is suggested that the bare patch disease is caused by a complex of root rot fungi composed of one or more anastomosis groups ofRhizoctonia spp. and other associated fungi.Samenvatting Van kiemplanten van tarwe en gerst, afkomstig van een met kale-plekkenziekte besmet perceel in West Australië werd de mate van wortelbeschadiging bepaald en werden schimmels uit de wortels geïsoleerd. De plekken waren verschillend van vorm en afmeting; de zwaarst aangetaste planten werden in het centrum ervan aangetroffen. Van de 165 verkregen isolaten vanRhizoctonia spp. was 90% meerkernig en 10% tweekernig. De meerkernige overheersten in de centra van de plekken. Relatief gezien nam het aantal tweekernige isolaten toe naarmate de herkomst dichter bij de periferie van de plekken was. De scherpe begrenzing van de ziekte aan de randen van de plekken zou in verband kunnen staan met het toenemen van de activiteit van de avirulente tweekernige isolaten in de nabijheid van de periferie van de plekken. Een aantal andere schimmels, zoalsFusarium spp.,Mortierella spp.,Bipolaris sorokiniana, Pythium sp. enTrichoderma sp. werd eveneens in de plekken aangetroffen. De meerkernige isolaten die tot de anastomosegroepen Ag 2–1, 2–2 en 8 (Thanatephorus cucumeris) behoren, waren voor tarwe het meest pathogeen. De tweekernige isolaten van de anastomosegroepen Ag C, D, E en K (Ceratobasidium sp.) waren minder pathogeen. Gesuggereerd wordt, dat de kale-plekkenziekte veroorzaakt wordt door een complex van verschillende wortelschimmels, die behoren tot een of meer anastomosegroepen vanR. solani en andere daarmee geassocieerde schimmels.     

利用环介导等温扩增(LAMP)技术快速检测辣椒疫霉菌

<正>辣椒疫霉(Phytophthora capsici)是一种重要植物病原菌,能造成植株坏死、果实腐烂,严重影响产量[1]。辣椒疫霉侵染植物的早期病症并不明显,易被忽视。因此,对辣椒疫霉早期快速、准确检测显得尤为重要。聚合酶链式反应(PCR)为动植物病原物检测的重要方法,但需要较昂贵的仪器、试剂与耗材,后期的电泳检测也费时费力,致使这一技术很难在生产一线普及推广。Notomi等2000年研发了环介     

三类植物卵菌的全基因组比较

为有效防治由不同类型卵菌引起的植物病害,该研究对数据库中的霜霉、疫霉和腐霉3类不同卵菌的全基因组数据进行序列特征分析、同源性分析和共线性分析以及同源差异基因富集通路分析。结果显示,霜霉、疫霉和腐霉来自独立的谱系,且疫霉与霜霉的亲缘关系较近;疫霉的致病相关基因数量最多,主要有RxLR、CRN、NPP、NF和PcF基因家族,霜霉的致病相关基因数量次之,腐霉的致病相关基因数量最少;3类卵菌共确定了13 392个同源基因,其中3类卵菌均共有的同源基因为3 786个,不同菌种的同源基因数量差异较大,整体表现为疫霉>霜霉>腐霉;同源差异基因富集程度最高的前20个通路主要是基础代谢的通路和中间代谢通路,其中抗坏血酸和藻酸盐代谢、ABC转运蛋白和果糖和甘露糖代谢相对富集程度较高,基因数目也较多。     

Pathogenicity ofPythium species on cucumber in peat-sand,rockwool and hydroponics

Pythium spp. that cause damping-off of seedlings also can cause root rot of older plants and lead to yield reductions. This can be especially severe in soilless cultures where the fungus can spread easily with the nutrient solution. 39Pythium isolates obtained from discolored roots were assayed for their ability to cause damping-off on cucumber seedlings in sand-peat and for their pathogenicity in soilless culture of cucumber in rockwool or hydroponic solution. Isolates ofPythium aphanidermatum, P. irregulare, P. sylvaticum andP. ultimum were highly pathogenic in sand-peat, but onlyP. aphanidermatum strains were pathogenic in soilless conditions and led to root decay, plant death in rockwool culture and growth reduction in hydroponic culture. One strain ofP. aphanidermatum significantly reduced the yield of cucumber grown in rockwool under conditions similar to those of commercial cultures.     

Identity and variability of Pythium species associated with yield decline in aerobic rice cultivation in the Philippines

The cultivation of aerobic rice in the tropics enables farmers to save water without lowering productivity. Unfortunately, this system suffers from declining yields due to a disease complex involving nematodes, pathogenic Pythium spp. and nutrient deficiencies. Assessing the impact of each underlying factor can contribute to efficient disease control measures. This study therefore investigated pathogenic and genotypic variability among Pythium species from affected aerobic rice fields in the Philippines using pathogenicity assays and sequence information from the internal transcribed spacer (ITS) region and β‐tubulin gene. Three closely related Pythium spp., P. arrhenomanes, P. graminicola and P. inflatum, were recovered from affected aerobic rice fields. All P. arrhenomanes isolates reduced rice seedling growth, whereas only a few P. graminicola isolates and no P. inflatum isolates were pathogenic, indicating that P. arrhenomanes is probably the most important species affecting rice. Both P. arrhenomanes and P. graminicola isolates showed little genetic variation, despite the observed pathogenic variation within P. graminicola. Intraspecific variation was higher among P. inflatum isolates, but again no correlation was observed with phenotype. When screening P. arrhenomanes isolates from other hosts such as sugarcane, maize and several grasses, a link between pathogenic and genetic variability was detected. However, rice and maize isolates seemed to lack host specificity, and therefore crop rotation with maize might be a risky strategy to manage yield decline in Philippine aerobic rice fields.     

Targets for pathology research in protected crops

Due to the need to diminish the amount of pesticides used, alternative ways of controlling fungal diseases have to be developed. Foliar diseases have been managed mainly by chemicals, but research has been started to develop integrated control programmes. Data from studies on epidemiology, in combination with nutritional and climatic management and use of partial resistant cultivars will lead to a reduced use of chemicals. Models used are: Botrytis cinerea in gerbera and roses, and powdery mildew in roses and cucumbers. Research has been intensified on biological control of these pathogens. In closed systems, with recirculation of the nutrient solution, soil-borne fungi can cause serious problems. Not only do diseases known from traditional cultural systems, like fusarium wilt of carnation, fusarium crown and root rot of tomato and Phytophthora and Pythium spp. in several crops occur, but also new problems like a new Phytophthora sp. and Gnomonia sp. in roses and a Cylindrocladium sp. in Spathiphyllum spp. To prevent introduction of pathogens in recirculation systems, emphasis is put on developing an integrated disease management programme by using disease-free planting material, disease-free irrigation water, strict hygienic measures, resistant cultivars, methods of disinfecting the nutrient solution and biological control. Research over the last 5–6 years has revealed good prospects for biological control, especially in closed systems with a limited amount of substrate. Wilt in carnation, caused by Fusarium oxysporum f.sp. dianthi can be prevented effectively by adding a non-pathogenic isolate of F. oxysporum and/or Pseudomonas spp. There are indications that non-pathogenic isolates of F. oxysporum are also effective against wilt disease in other crops. An isolate of Trichoderma harzianum appears to be very effective against fusarium crown and root rot of tomato. More applications of this, and other, biocontrol agents seem possible. However, translation of the results of research to practical application and registration is still very difficult.     

Root rot of hyacinths caused by species of Pythium

The root rot widely seen in hyacinth was found to be caused byPythium spp. instead of byFusarium culmorum. Of sixPythium species isolated, three, includedP. ultimum andP. violae, were investigated in glasshouse experiments and their pathogenicity demonstrated. In these experiments plants were successfully grown in containers in which an aqueous mist was maintained, or in water cultures. In experimental plots on infected soil, Dexon, a fungicide selective for Pythiaceae, distinctly reduced the number of dead plants and increased yield and bulb size, thus confirming the role ofPythium in causing root rot. Practical application of Dexon deserves further attention.     

Evaluation of plant growth-promoting rhizobacteria for biological control of pythium root rot of cucumbers grown in rockwool and effects on yield

Three strains ofPseudomonas fluorescens (63-49, 63-28, and 15), one strain ofPseudomonas corrugata (13) and one strain ofSerratia plymuthica (R1GC4) were tested on rockwool-grown cucumbers for their ability to reduce Pythium root-rot caused byPythium aphanidermatum. These strains were previously selected for biocontrol ability from collections of >4000 bacteria. Strains 63-49 and 63-28 were tested on cucumber plants grown in rockwool in two replicatedPythium-inoculated trials conducted in British Columbia (B.C). Another inoculated, replicated trial was conducted in Quebec with all five strains. Cucumber yields (fruit number and weight) were measured over a ten-week harvest period. Strain 63-49 caused an early promotion of plant growth and increased cucumber yields at early harvests. No measurable effect ofPythium inoculation on disease development was observed in the Quebec trial, due to unfavourable cool weather. However, 63-49 significantly increased the total number of cucumbers (12%) and cucumber weight (18%), compared to the non-treated control. Strains 13, 15 and R1GC4 slightly increased the cumulative cucumber yields, but strain 63-28 had no effect. In the B.C. trial, inoculation withP. aphanidermatum reduced the number and weight of cucumbers by 27%. Treatments ofPythium-inoculated cucumbers with 63-49 significantly increased fruit number and weight by 18%, compared to thePythium-inoculated control. Strain 63-28 increased the cumulative number of cucumbers over time, compared to thePythium-inoculated control, but the increase was less than with 63-49. The use ofPseudomonas spp. in rockwool-grown cucumbers can increase yields, both in the presence and absence of Pythium root rot, and with variable seasonal conditions and disease pressures.     

Production of Indole-3-acetic Acid and Tryptophol by Pythium ultimum and Pythium Group F: Possible Role in Pathogenesis

Pythium group F is a minor pathogen which induces symptomless infections that occur frequently and results in yield losses in tomato soilless cultures. To elucidate the mode of action of this microorganism, the influence of culture filtrates of Pythium group F on tomato growth was investigated and compared to that of the pathogen Pythium ultimum. Depending on metabolite production by the fungus, marked differences were observed in plant response. Pythium group F crude culture filtrates or low molecular weight fractions (< 500) caused swelling behind the root tip and reduced root growth; the cohesion and adherence of cells within the cortical area were also affected. These symptoms were similar to those observed on plants treated with indole-3-acetic acid. By contrast, P. ultimum filtrates caused a marked distortion of cell shape accompanied with folding of host cell walls, particularly in the cortical area. These symptoms were characteristic of the activity of toxic compound(s) on host cells. Chemical analysis of the filtrates demonstrated that indole-3-acetic acid and tryptophol were produced by Pythium group F and P. ultimum. However, Pythium group F took up and metabolized more indole-3-acetic acid precursors, especially tryptophan, a key amino acid in the pathways leading to indole-3-acetic acid synthesis. The fact that Pythium group F and P. ultimum transformed tryptamine and indole-3-acetaldehyde into indole-3-acetic acid and tryptophol confirms the existence of a tryptamine pathway within both fungi. These results support the hypothesis that auxins facilitate Pythium group F infections. On the other hand, toxin(s) and hydrolytic enzymes are likely involved in P. ultimum pathogenesis.     

Detection of Pythium aphanidermatum in tomato using loop-mediated isothermal amplification (LAMP) with species-specific primers

A loop-mediated isothermal amplification (LAMP) reaction with a primer set designed from the rDNA ITS sequence of P. aphanidermatum was developed. Results of a specificity test using 57 strains of Pythium spp. indicated that the LAMP assay gave no cross reactions in other 39 Pythium species, 11 strains of Phytophthora spp. and eight other soil borne pathogens. The detection limit was 10 fg of genomic DNA, which was ten times the sensitivity of the polymerase chain reaction. The LAMP assay was applied to hydroponic solution samples from tomato fields, and the results were compared to those of the conventional plating method. LAMP was observed to be effective for the specific detection of P. aphanidermatum. Furthermore, P. aphanidermatum was detected directly in tomato roots infected with P. aphanidermatum without DNA extraction. The LAMP method established in this study is a simple, sensitive and rapid tool for the detection of P. aphanidermatum.     

Pythium uncinulatum sp. nov. and P. tracheiphilum pathogenic to lettuce

From mature lettuce heads with bottom rot symptoms severalPythium species were isolated, two of which were studied further. Pathogenicity tests with both fungi have been carried out only with seedlings. P. uncinulatum sp. nov. was isolated frequently. Its relation to otherPythium spp. with spiny oogonia is discussed. It proved to be very pathogenic to seedlings of lettuce and somewhat less to those of cucumber and tomato. P. tracheiphilum, which was isolated far less frequently, proved to be highly pathogenic to seedlings of lettuce, cucumber and cauliflower, and somewhat less to tomato and pea.Samenvatting Van slaplanten met smetsymptomen werden behalveBotrytis cinerea, Rhizoctonia sp. enSclerotinia spp. dikwijlsPythium-soorten geïsoleerd. Twee hiervan zijn nader onderzocht. P. uncinulatum (Fig. 1, 3a, 3b), die dikwijls werd geïsoleerd, wordt beschreven als nieuwe soort en de relatie met anderePythium-soorten met gestekelde oögoniën wordt besproken. In proeven bleekP. uncinulatum zeer pathogeen te zijn voor zaailingen van sla en iets minder voor zaailingen van komkommer en tomaat (Fig. 4a, Tabel 2). Het planten van slazaailingen in metP. uncinulatum geïnoculeerde grond had meer effect dan het dopen van de wortels in een myceliumsuspensie van de desbetreffende schimmel vóór het planten. Het effect uitte zich in groeivertraging van de plant en beschadiging (necrose) van het gehele wortelstelsel. P. tracheiphilum (Fig. 2, 3c), die slechts een paar maal werd geïsoleerd, was zeer pathogeen voor zaailingen van sla, komkommer en bloemkool (Fig. 4a) en iets minder voor zaailingen van tomaat en erwt (Tabel 2). Groeiremming van de planten was sterker wanneer de wortels van de zaailingen vóór het planten in een myceliumsuspensie vanP. tracheiphilum gedoopt werden (Fig. 4b), dan wanneer de zaailingen in geïnoculeerde grond geplant werden. De hoofdwortels van de zaailingen werden ernstig beschadigd, maar zich later vormende zijwortels werden weinig door de schimmel beïnvloed.Tot nu toe zijn de pathogeniteitsproeven slechts met zaailingen uitgevoerd. Proeven met volwassen planten moeten volgen om te onderzoeken of er een verband kan bestaan tussen de aanwezigheid van de betreffendePythium-soorten in de grond en het optreden van smet in sla.     

Genetic diversity,anastomosis groups and virulence of <Emphasis Type="Italic">Rhizoctonia</Emphasis> spp. from strawberry

Virulent Rhizoctonia spp. isolated from strawberry in Israel belonged to anastomosis groups (AG) of: binucleate Rhizoctonia (BNR) AG-A, AG-G, AG-K and AG-F, and to multinucleate Rhizoctonia (MNR) AG 4 subgroup HG-I. In addition, a soil isolate of AG 4 subgroup HG-III was also found to be virulent on strawberry. None of the Israeli isolates obtained in the present study belonged to BNR AG-I, or other MNR AGs. In the cluster analysis of rDNA-ITS sequences, all of the isolate sequences consistently clustered according to their known AGs and subgroups. One AG-F cluster included sequences of 10 strawberry isolates, while another AG-F cluster included sequences of two isolates submitted to GenBank. Additional work is needed to determine whether the isolates of these two clusters may belong to different AG-F subgroups. The current virulence bioassay used for Rhizoctonia spp. isolates on strawberry is based on inoculation of stolon-derived daughter plants with the isolates and estimation of the reduction in plant biomass, rather than on specific distinct disease severity symptoms. The duration of this test is relatively long (ca. 5 weeks or more) and the availability of daughter plants from runners is naturally limited to a certain season. Among the possible alternative methods evaluated in the present study (inoculation of fruits or seedlings developed from germinated strawberry seeds), the method based on seedlings was best. This method has a potential to replace the currently used stolon-daughter plant inoculation bioassay for testing virulence of strawberry root pathogens. This is the first report indicating that Rhizoctonia spp. isolates that belong to AG-F, AG-K, AG 4 HG-I and AG 4 HG-III are virulent to strawberry.     

PCR‐based identification of Pythium spp. causing cavity spot in carrots and sensitive detection in soil samples

On the basis of ITS sequences PCR primers were designed for the identification of the five Pythium species found to be most important for the development of carrot cavity spot in Norway: P. intermedium, P. sulcatum, P. sylvaticum, P. violae and P. ‘vipa’. The P. ‘vipa’ isolates had a unique ITS sequence, differed morphologically from all other Pythium isolates, and thus probably represent a new species. The PCR primers were species‐specific with no cross‐reaction to other Pythium species or to fungal isolates from carrot tested. The detection limits varied for the different primer pairs. The two most sensitive assays allowed detection of as little as 5 fg DNA. All five Pythium species could be detected in lesions from diseased carrots. Weak positive signals were obtained from some carrot samples without symptoms. PCR assays allowed detection of pathogens in soil. In samples of soil known to produce cavity spots on cropped carrots, strong signals were obtained. In several soil samples more than one of the five Pythium species could be detected. The utilization of this diagnostic PCR assay in analysis of field soil and carrot tissue might in the future be exploited to reduce the incidence of this serious carrot disease.     

Biocontrol traits of plant growth suppressive arbuscular mycorrhizal fungi against root rot in tomato caused by <Emphasis Type="Italic">Pythium aphanidermatum</Emphasis>

Arbuscular mycorrhizal (AM) fungi known to cause plant growth depressions in tomato were examined for their biocontrol effects against root rot caused by Pythium aphanidermatum. The main hypothesis was that plant growth suppressive AM fungi would elicit a defence response in the host plant reducing Pythium root rot development. To test this hypothesis a fully factorial experiment was performed with AM fungi (Glomus intraradices, G. mosseae, G. claroideum or nonmycorrhizal), Pythium (± P. aphanidermatum) and harvest (7 and 14 days after pathogen inoculation (dapi)) as the main factors. Two weeks after AM fungi inoculation, roots were challenged with P. aphanidermatum. Variables evaluated at each harvest were root colonization levels of the interacting fungi, plant growth responses, and expression of a plant pathogenesis related protein gene (PR-1). All of the tested AM fungi caused marked growth suppressions, but did not affect PR-1 gene expression or the phosphorous concentration in the host plant. Plants singly inoculated with P. aphanidermatum had an increased PR-1 expression and phosphorous concentration. Among the AM fungi included in the study only G. intraradices reduced the pathogen root infection level, measured both in terms of Pythium ELISA and by recovery on selective media and only at the first harvest. Likewise, P. aphanidermatum root infection reduced colonization levels of G. intraradices, but not that of the two other AM fungi. In conclusion, plant growth suppressive AM fungi may offer plant beneficial traits in terms of biocontrol of root cortical pathogens.