Characterization of fungi (Fusarium and Rhizoctonia) and oomycetes (Phytophthora and Pythium) associated with apple orchards in South Africa

Several species of fungi and oomycetes including Fusarium, Rhizoctonia, Phytophthora and Pythium have been reported as root pathogens of apple where they contribute to a phenomenon known as apple replant disease. In South Africa, little is known about specific species in these genera and their pathogenicity toward apple. Therefore, these aspects were investigated along with the development and optimization of qPCR tests for detection and quantification of the most virulent oomycete species. In eight investigated orchards, the oomycete Phythophthora cactorum was widely distributed, while nine Pythium species were differentially distributed among the orchards. Pythium irregulare was the most widely distributed and the most virulent species along with P. sylvaticum, P. vexans and Ph. cactorum. Seven binucleate Rhizoctonia anastomosis groups (AGs) were also differentially distributed among the orchards, with the majority appearing to be non-pathogenic while certain AG-I and AG-F isolates exhibited low virulence on apple. In the genus Fusarium, F. oxysporum was widely distributed, but isolates were non-pathogenic. Fusarium solani and F. avenaceum were less frequently encountered, with only some isolates having low virulence. qPCR data obtained from seedling roots inoculated with the most virulent Pythium species (P. irregulare, P. sylvaticum and P. vexans) and the genus Phytophthora were not always reproducible between trials, or isolates of the same species. In general, seedling growth inhibition was associated with the presence of a low amount of pathogen DNA (±40 fg μl⁻¹ to 2 pg μl⁻¹) in roots. Pythium irregulare, although having the lowest DNA concentrations in roots, was the only species for which a significant negative correlation was found between seedling weight and pathogen DNA concentration.     

Characterization of root rot disease of kiwifruit in the Black Sea region of Turkey

Forty-two kiwifruit orchards from Rize and Samsun provinces (Black Sea region, Turkey) exhibiting symptoms of root rot disease were examined between 2009 and 2010. Twenty-four fungal isolates recovered from affected roots were included in this study. Morphological characteristics of all isolates were recorded on potato dextrose agar, malt extract agar and Spezieller Nährstoffarmer agar. The effect of temperature on radial colony growth was also evaluated at 5, 10, 15, 20, 25, 30, and 35 °C. Histone H3 gene (HIS) was amplified with primers CYLH3F and CYLH3R and the amplified fragments were sequenced. The HIS phylogeny grouped all the isolates into six well-supported clades which were in agreement with phenotypical characteristics. Isolates were identified as “Cylindrocarpon” pauciseptatum, Cylindrocladiella parva, Ilyonectria liriodendri, I. torresensis, I. robusta and I. europaea, I. liriodendri being the most frequent species. All of them are reported for the first time on kiwifruit in Turkey, with the exception of I. liriodendri. Pathogenicity tests with selected isolates showed that 10 out of 11 isolates tested were able to induce typical root rot disease symptoms, affecting plant development and leading to the death of some plants. This study shows the high diversity of root rot pathogens found in kiwifruit trees in the Black Sea region of Turkey, being the first step towards a better understanding and management of the disease in local conditions.     

Detection of black-foot disease pathogens in the grapevine nursery propagation process in Spain

Two commercial nurseries located in Comunidad Valenciana region (central-eastern Spain) were sampled in 2010 to evaluate whether the grapevine nursery propagation process could be a source of black-foot disease pathogens. Samples were taken from four sources of the propagation process: pre-grafting hydration tanks, scissors used for cutting buds, omega-cut grafting machines, and peat used for callusing. DNA from these samples was extracted and multiplex nested-PCR using primers specific for “Cylindrocarpon” pauciseptatum, Ilyonectria liriodendri and I. macrodidyma-complex (composed of I. alcacerensis, I. estremocensis, I. macrodidyma, I. novozelandica, I. torresensis, and two undescribed species) was used to identify the species present. Ilyonectria liriodendri and I. macrodidyma-complex were detected in hydration tanks, scissors, grafting machines and peat, I. macrodidyma-complex being the most frequent. Additionally, ten grafted cuttings each from five grapevine scion/rootstock combinations were collected from each nursery immediately after callusing, and again after one growing season in a nursery field. Roots of these grafted cuttings and plants were sampled to isolate the fungal pathogens. Only I. torresensis was isolated after callusing, while I. liriodendri, I. novozelandica and I. torresensis were isolated after one growing season, showing the highest incidence at this latter sampling time. Moreover, DNA was extracted from roots and analyzed as described before. Ilyonectria liriodendri and I. macrodidyma-complex were also detected at both sampling times. The use of the multiplex nested-PCR technique improved the detection of I. liriodendri and I. macrodidyma-complex from grafted cuttings and plants in both nurseries. This work shows that the grapevine nursery propagation process should be considered as a potential infection source for black-foot disease pathogens, and confirms that infections caused by Ilyonectria spp. in grapevine planting material increase markedly after one growing season in nursery fields.     

High genotypic and virulence diversity in Ilyonectria liriodendri isolates associated with black foot disease in New Zealand vineyards

A total of 57 Ilyonectria liriodendri isolates were identified by a combination of species‐specific PCR and DNA sequencing from a collection of 174 Ilyonectria‐like isolates recovered from 101 diseased grapevine samples. These samples were representative of the national vineyard, comprising material contributed by 49 grape growers across seven grape growing areas. This species was predominant, representing 33% of the recovered isolates, and has been reported as a major pathogen of grapevines in other countries. The genetic diversity of the 57 New Zealand isolates was compared to that of isolates from Australia and South Africa using universally primed polymerase chain reaction (UP‐PCR). A total of 66 informative loci distinguished 52 genotypes, of which five contained up to four clonal isolates. Four main clades were identified in a neighbour‐joining (NJ) tree. The international isolates (Australia and South Africa) were placed in a clade that did not include New Zealand isolates. There was a high level of intra‐ and inter‐vineyard genetic variation indicating the free movement of isolates between regions. A subset of nine isolates from different branches of the NJ tree produced two vegetative compatibility groups and hyphal fusion was observed between non‐self pairings. Pathogenicity tests using isolates from different genetic groups inoculated onto either detached roots or 1‐year‐old potted vines showed variability in virulence; however, no correlations were detected.     

Auxin‐mediated relationships between apple plants and root inhabiting fungi: impact on root pathogens and potentialities of growth‐promoting populations

This study aimed to elucidate the relationship between plant hosts and root‐colonizing fungi recovered from apple orchard soils that had been replanted over multiple generations. Functional relationships of three groups of filamentous fungi (Ceratobasidium sp., Cylindrocarpon‐like group and Fusarium acuminatum) with apple rootstocks were evaluated in plant growth bioassays. The Cylindrocarpon‐like group and Ceratobasidium sp. showed a relationship with the host plant varying from pathogenic to commensal through to mutualistic for the latter group, while that of F. acuminatum tended to be mutualistic. Seven fungal isolates of each group, which induced the highest plant growth in bioassays, were evaluated for auxin (IAA) and gibberellin (GA₃ and GA₄) production in culture filtrate. All isolates of F. acuminatum as well as most of those of the Ceratobasidium sp. and Cylindrocarpon‐like groups produced IAA in culture filtrate. IAA production was evaluated for additional isolates of endophytic fungal species from fruit tree orchards and the functionality of IAA was confirmed by growing in vitro micropropagated plantlets of apple rootstock on MS medium supplemented with fungal culture filtrate. Findings from this study may explain the difficulty in defining the precise role of diverse root‐colonizing fungal populations in replant disease aetiology of fruit tree orchards. However, the results demonstrate the presence of a positive and widely available biotic component of the orchard soil biology that may be exploited for the benefit of tree growth and production.     

Pathogenicity studies in avocado with three nectriaceous fungi,Calonectria ilicicola,Gliocladiopsis sp. and Ilyonectria liriodendri

Calonectria ilicicola, Gliocladiopsis sp. and Ilyonectria liriodendri were isolated from diseased roots of young avocado trees. Pathogenicity studies with seedlings of three avocado cultivars, Velvick, Hass and Reed, demonstrated that Calonectria ilicicola is a severe root rot pathogen, reducing the biomass of healthy roots, and reducing plant height over time. Calonectria ilicicola was re‐isolated from diseased roots. Ilyonectria liriodendri and Gliocladiopsis sp. were not pathogenic and plant height was increased after Gliocladiopsis sp. amendment compared to all other treatments in trials with cvs Velvick and Hass.     

Selection and orchard testing of antagonists suppressing conidial production by the apple scab pathogen <Emphasis Type="Italic">Venturia inaequalis</Emphasis>

Apple scab caused by Venturia inaequalis is a major disease in apple production. Epidemics in spring are initiated by ascospores produced on overwintering leaves whereas epidemics during summer are driven by conidia produced on apple leaves by biotrophic mycelium. Fungal colonisers of sporulating colonies of V. inaequalis were isolated and their potential to reduce the production of conidia of V. inaequalis was evaluated on apple seedlings under controlled conditions. The four most effective isolates of the 63 screened isolates were tested subsequently under Dutch orchard conditions in 2006. Repeated applications of conidial suspensions of Cladosporium cladosporioides H39 resulted in an average reduction of conidial production by V. inaequalis of approximately 40%. In 2007, applications of conidial suspensions of C. cladosporioides H39 reduced conidial production by V. inaequalis by 69% on August 6 and by 51% on August 16, but no effect was found on August 20. However, viability of available conidia of C. cladosporioides H39 was low at the end of the experiment. Epiphytic and endophytic colonisation by Cladosporium spp. of leaves treated during the experiment with C. cladosporioides H39 was significantly higher than on control leaves sampled 6 weeks after the last application. It is concluded that C. cladosporioides H39 has promising potential as a biological control agent for apple scab control. More information is needed on the effect of C. cladosporioides H39 on apple scab epidemics as well as on mass production, formulation and shelf life of conidia of the antagonist.     

Pathogenicity of fungi associated with winter loss and injury in white lupin

Fungi isolated from diseased white lupin plants were tested for pathogenicity in standardized glasshouse tests to assess their potential as causes of plant death in winter in autumn-sown crops. Pleiochaeta setosa and, to a slightly lesser extent, Fusarium avenaceum caused lesions or plant death when inoculated onto hypocotyls of lupin seedlings. Disease was more severe when the hypocotyls were wounded before inoculation. F. avenaceum also caused injury when inoculated onto roots; wounding the roots had no effect. Some isolates of F. solani also caused injury, especially after root inoculation, but isolates of F. oxysporum and Cylindrocarpon destructans were only slightly or non-pathogenic. Isolates of Fusarium and C. destructans , but not of P. setosa , differed in pathogenicity. There were differences in susceptibility to disease among three lupin cultivars.     

Transformation of soil microbial community structure and rhizoctonia-suppressive potential in response to apple roots

ABSTRACT Changes in the composition of soil microbial communities and relative disease-suppressive ability of resident microflora in response to apple cultivation were assessed in orchard soils from a site possessing trees established for 1 to 5 years. The fungal community from roots of apple seedlings grown in noncultivated orchard soil was dominated by isolates from genera commonly considered saprophytic. Plant-pathogenic fungi in the genera Phytophthora, Pythium, and Rhizoctonia constituted an increasing proportion of the fungal community isolated from seedling roots with increasing orchard block age. Bacillus megaterium and Burkholderia cepacia dominated the bacterial communities recovered from noncultivated soil and the rhizosphere of apple seedlings grown in orchard soil, respectively. Populations of the two bacteria in their respective habitats declined dramatically with increasing orchard block age. Lesion nematode populations did not differ among soil and root samples from orchard blocks of different ages. Similar changes in microbial communities were observed in response to planting noncultivated orchard soil to five successive cycles of 'Gala' apple seedlings. Pasteurization of soil had no effect on apple growth in noncultivated soil but significantly enhanced apple growth in third-year orchard block soil. Seedlings grown in pasteurized soil from the third-year orchard block were equal in size to those grown in noncultivated soil, demonstrating that suppression of plant growth resulted from changes in the composition of the soil microbial community. Rhizoctonia solani anastomosis group 5 (AG 5) had no effect on growth of apple trees in noncultivated soil but significantly reduced the growth of apple trees in soil from third-year orchard soil. Changes in the ability of the resident soil microflora to suppress R. solani AG 5 were associated with reductions in the relative populations of Burkholderia cepacia and Pseudomonas putida in the rhizosphere of apple.     

Fungi as potential factors limiting natural regeneration of pedunculate oak (Quercus robur) in mixed-species forest stands in Poland

Dieback of young Quercus robur seedlings can limit natural regeneration in mixed-species forest stands in Poland. The aim of this study was to examine the role of fungi in the dieback of oak seedlings in central Europe. Fungi were isolated from the stems and roots of Q. robur, from both healthy seedlings and seedlings with symptoms, that were sampled from four stands in Poland. In total, 111 distinct taxa were identified. Ascomycota was dominant, representing 95.6% of the isolates. Among the taxa identified, Alternaria alternata, Colletotrichum fioriniae, Colletotrichum godetiae, Coniella quercicola, Diaporthe eres, Gnomoniopsis paraclavulata, Ilyonectria rufa, Mucor genevensis, Penicillium glandicola, Tubakia dryina, and Umbelopsis changbaiensis were most frequently isolated, and were consistently found in stems and roots of both healthy and diseased plants. The community compositions of fungi in healthy and diseased stems were similar, although Fusarium species, especially Fusarium sp. 1 (FTSC 5) and F. sporotrichioides, were found mainly in seedlings with symptoms. The pathogenicity of the most consistently isolated species from stems and roots of diseased seedlings was tested on Q. robur seedlings. F. sporotrichioides caused the largest lesions on inoculated seedlings. Six weeks after inoculation, D. eres, C. fioriniae, G. paraclavulata, T. dryina, and F. sporotrichioides killed 0%–18.8% of seedlings, while I. rufa and Ilyonectria pseudodestructans did not cause any lesions or other symptoms. This study is the first comprehensive report suggesting that massive fungal attack can lead to oak seedling dieback in mixed-species forest stands in central Europe.     

Deleterious effects of fungi isolated from paddy soils on seminal root of rice

Soil samples were collected from rice paddies at 22 locations in northeastern Honshu, Japan. In 20 of the samples, seedling growth of rice was improved by soil pasteurization (aerated steaming at 60°C for 30 min), although no typical disease symptoms were observed in the seedlings grown in the untreated soil samples. In most locations, rice seedlings grew better in a potting medium containing root material from plants grown in the pasteurized portion of a soil sample than in a medium with root material from plants grown in the unpasteurized portion of the same sample. The results suggest that microorganisms that restrained rice seedling growth may be common in the soils of rice paddies. Approximately 800 isolates were obtained from seedling roots grown in unpasteurized soils and grouped by cultural and microscopic morphologies. The deleterious effects of 79 isolates from 21 major groups, most of which were soil-dwelling taxa, were examined after direct inoculation of the seminal roots. Isolates of Curvularia sp., Cirrenalia sp., Eppicoccum nigrum, Fusarium graminearum, F. oxysporum, Gliocladium virens, Humicola sp., Penicillium sp., Rhizoctonia oryzae-sativae, Sclerotium hydrophilum, Trichoderma aureoviride, and T. harzianum inhibited root growth, suggesting that deleterious root-infecting fungi were more common in paddy soil than previously thought. These fungi may be involved in the restraint of rice seedling growth.     

A case study of infection of rice roots by deleterious fungi and minor pathogenic<Emphasis Type="Italic"> Pythium</Emphasis> species in a paddy field

Roots of rice plants grown in paddy fields in a transplant culture system were collected seven times between 8 and 22 weeks after transplanting, for two crop seasons. Rice seedlings grown in a potting medium amended with the collected roots were significantly shorter than those grown in the same medium either without the addition of the roots or amended with pasteurized, collected roots indicating that seedlings were inhibited by heat-labile microorganisms on the collected rice roots. Ninety-five and 172 pure cultures of Pythium spp. and fungi, respectively, were isolated from the rice roots collected 5 or 7.5 weeks after transplanting in the fields. Among these microorganisms, Pythium aristosporum inhibited seedling growth in greenhouse experiments, and Acremonium sp., Alternaria sp., Epicoccum nigrum, Fusarium sp., Massarina sp., Penicillium spp., Rhinocladiella sp., Stemphylium sp., Trichocladium sp., and several unidentified fungi inhibited seminal root growth in in vitro experiments. These microorganisms might be involved in the inhibition of seedling growth in soils amended with the rice roots collected from the paddy field. Thus, roots of rice plants at the middle stage of growth transplanted into paddy fields can harbor pathogenic or deleterious fungi or Pythium sp(p). The effects of these microorganisms on rice growth in paddy fields are discussed.     

Potential allelopathic effects of Mikania micrantha on the seed germination and seedling growth of Coix lacryma‐jobi

The allelopathic potential of Mikania micrantha H.B.K. to affect the seed germination and seedling growth of Coix lacryma‐jobi L. was investigated. Water‐soluble allelopathic substances were found in the water extracts of M. micrantha. The effect of the water extracts on the seed germination and seedling growth of C. lacryma‐jobi was concentration‐dependent. The water extracts from the different plant parts (leaf, stem, and root) of M. micrantha differed in their effect on the germination and seedling growth of C. lacryma‐jobi, with the effect of the leaf extract being the least inhibitory. The malondialdehyde (MDA) content in the C. lacryma‐jobi seedlings increased by 64%, 45%, and 52% of the control with increasing concentrations of the extracts of the root, stem, and leaf (80, 400, and 400 g L^?1, respectively). The extract from the M. micrantha roots significantly increased the catalase (CAT) activity of the C. lacryma‐jobi seedlings (48% and 54% of the control at the concentrations of 20 g L^?1 and 80 g L^?1, respectively). The extracts from the leaves and stems at low concentrations increased the CAT activity, but at high concentrations, the extracts decreased the CAT activity. The extracts from the roots, stems, and leaves at concentrations of 80, 400, and 400 g L^?1 also significantly decreased the peroxidase (POD) activity of the C. lacryma‐jobi seedlings to 27%, 52%, and 34% of the control, respectively. These results indicate that the water extracts of M. micrantha could inhibit the seed germination and seedling growth of C. lacryma‐jobi through the regulation of anti‐oxidase activity, such as POD and CAT in the cells. The growth inhibition of the C. lacryma‐jobi seedlings is probably related to injury after oxidization of the cell membranes with the increase of MDA content.     

Resistance Reaction of Conifer Species (European Larch, Norway Spruce, Scots Pine) to Infection by Selected Necrotrophic Damping-off Pathogens

Three conifer species (European larch, Norway spruce, Scots pine) were investigated for their resistance to five damping-off pathogens (Rhizoctonia solani, Fusarium solani, F. oxysporum, F. culmorum, F. avenaceum). Inoculation of the primary roots of seedlings with these pathogens caused host cell death which did not prevent the invasive growth of these fungi; seedlings that had formed secondary and tertiary roots could overcome the infection to a significant degree. Infections with R. solani caused significant mortality to all the conifer species. In contrast, the tree species expressed different levels of resistance when challenged with the Fusarium isolates, with Norway spruce being the most resistant compared to uninoculated controls. Some of the Fusarium isolates were more pathogenic to certain hosts than others; F. oxysporum for European larch, F. avenaceum for Scots pine, F. solani for European larch; only F. culmorum was significantly pathogenic to Norway spruce. No significant differences in disease severity were observed at different soil pH (4.3–7.5). Disease progression was delayed at lower (10–15°C) rather than higher temperatures (20–25°C).     

Detection and quantification of Ilyonectria spp. associated with black‐foot disease of grapevine in nursery soils using multiplex nested PCR and quantitative PCR

Three nursery fields and three rootstock mother fields from commercial nurseries located in Comunidad Valenciana region (central‐eastern Spain) were surveyed in July 2011 to detect the presence and to quantify Ilyonectria spp. in the soil. In each field, ten soil samples were taken randomly with a soil probe at a depth of 10–30 cm, and 10–20 cm from the base of the plant. Three replicate subsamples (10 g each) were taken from each soil sample. DNA was extracted and a multiplex nested PCR with species‐specific primer pairs (Mac1/MaPa2, Lir1/Lir2 and Pau1/MaPa2) was used to identify the species present. Among the 180 soil DNA samples analysed, Ilyonectria spp. were detected in 172 of them. Ilyonectria macrodidyma complex was the most frequently detected, being identified in 141 samples from all the fields evaluated. However, I. liriodendri was detected in only 16 samples, but was present in all open‐root field nurseries and in two rootstock mother fields. In addition, quantitative PCR (qPCR) assays were done to assess the levels of I. liriodendri and I. macrodidyma‐complex DNA in the soil samples. Detection of Ilyonectria spp. DNA using qPCR correlated with the fields found positive with the nested multiplex PCR. DNA concentrations of Ilyonectria spp. ranged from 0·004 to 1904·8 pg μL^?1. In general, samples from rootstock mother fields showed the highest DNA concentrations. The ability to detect and quantify Ilyonectria spp. genomic DNA in soil samples from nursery fields and rootstock mother fields confirms soils from both field types as important inoculum sources for black‐foot pathogens.     

Factors Influencing Development of Root Rot on Ginseng Caused by Cylindrocarpon destructans

ABSTRACT The fungus Cylindrocarpon destructans (Zins) Scholten is the cause of root rot (disappearing root rot) in many ginseng production areas in Canada. A total of 80 isolates of C. destructans were recovered from diseased roots in a survey of ginseng gardens in British Columbia from 2002-2004. Among these isolates, 49% were classified as highly virulent (causing lesions on unwounded mature roots) and 51% were weakly virulent (causing lesions only on previously wounded roots). Pectinase and polyphenoloxidase enzymes were produced in vitro by C. destructans isolates when they were grown on pectin and phenol as a substrate, respectively. However, highly virulent isolates produced significantly (P < 0.001) higher enzyme levels compared with weakly virulent isolates. Histopathological studies of ginseng roots inoculated with a highly virulent isolate revealed direct hyphal penetration through the epidermis, followed by intracellular hyphal growth in the cortex. Subsequent cell disintegration and accumulation of phenolic compounds was observed. Radial growth of highly and weakly virulent isolates on potato dextrose agar was highest at 18 and 21 degrees C, respectively and there was no growth at 35 degrees C. Mycelial mass production was significantly (P 相似文献   

Molecular diversity of binucleate <Emphasis Type="Italic">Rhizoctonia</Emphasis> AG-A in China

AG-A belongs to the binucleate Rhizoctonia (BNR) anastomosis group (AG) of the Ceratobasidium teleomorph, which parasitizes the roots of many plant species. Ninety nine isolate species of AG-A were obtained from Tibet, Sichuan, and Yunnan Province in China. All isolates were divided into three types based on their cultural characteristics. Type I: abundant aerial mycelia, dense hyphae, loose sclerotia; Type II: abundant aerial mycelia, no sclerotia. Type III: sparse aerial mycelium and no sclerotia. All of the isolates infected the seedlings of Chinese mustard and Chinese cabbage, causing the formation of lesions on the stem and a brown discoloration of the roots. Sequence analysis of the 5.8S rDNA-ITS showed a similarity of 98–100% among the isolates. Inter Simple Sequence Repeat (ISSR) was used to detect genetic variation in binucleate Rhizoctonia spp. Forty two AG-A isolates were amplified using 15 random primers. From a total of 164 bands, 144 bands (87.8%) were polymorphic in the 42 tested isolates. A dendrogram showing genetic relationships between the isolates was constructed using unweighted pair-group averages based on genetic distances. According to the dendrogram, the 42 tested isolates could be aligned into three clusters with a genetic similarity coefficient of 0.29, the first clusters including 27 isolates with III of culture characteristics on PDA; the second clusters included eight isolates with I of cultural characteristics on PDA; the third cluster included seven isolates with II of cultural characteristics on PDA. The results of ISSR analysis showed an association between the hosts of these isolates. Our results showed that ISSR analysis can reveal more molecular variation among isolates of AG-A than sequence analysis using the 5.8S rDNA-ITS.     

Comparison of pathogenicity of the Fusarium crown rot (FCR) complex (<Emphasis Type="Italic">F. culmorum</Emphasis>, <Emphasis Type="Italic">F. pseudograminearum</Emphasis> and <Emphasis Type="Italic">F. graminearum)</Emphasis> on hard red spring and durum wheat

Fusarium species involved in the Fusarium crown rot (FCR) complex affect wheat in every stage of development from seedling to grain fill. This study was designed to compare the aggressiveness of the FCR complex members including F. culmorum, F. pseudograminearum and F. graminearum in causing seedling blight, decreased plant vigour and crown rot. To assess their relative pathogenicity, two hard red spring wheat cultivars and two durum wheat cultivars were inoculated in the field with five isolates from each of the three species for two years. Significant differences in patterns of pathogenicity were identified. In particular, F. culmorum caused greater seedling blight while F. pseudograminearum and F. graminearum caused greater crown rot. Greatest yield reductions were caused by F. pseudograminearum. Cultivar differences were identified with respect to seedling disease and late season crown rot. No interactions were identified between cultivar performance and isolates or species with which they were challenged.     

Performance of three endophytic actinomycetes in relation to plant growth promotion and biological control of <Emphasis Type="Italic">Pythium aphanidermatum</Emphasis>, a pathogen of cucumber under commercial field production conditions in the United Arab Emirates

In the current study, the performance of three endophytic actinomycetes identified as Actinoplanes campanulatus, Micromonospora chalcea and Streptomyces spiralis previously shown to reduce seedling damping-off, and root and crown rots of mature cucumber (Cucumis sativus) caused by Pythium aphanidermatum in pots under greenhouse conditions were further evaluated to determine their potential as biological control agents and as plant growth promoters in the field under the conditions of commercial production of cucumbers in the United Arab Emirates (UAE). When applied individually or in combination to cucumber seedlings, the three isolates significantly promoted plant growth and yield and reduced seedling damping-off and root and crown rots of mature cucumber plants. Individually the performance level of S. spiralis was relatively the best followed by A. campanulatus and then by M. chalcea. The three isolates (which were not inhibitory to each other) performed better, both as biological control agents as well as plant growth promoters, when applied together than when they were inoculated individually. The ability of these three isolates to colonize the internal tissues of roots, stems and leaves under field conditions, and to persist up to 8 weeks after seedling inoculation, showed that they can easily adapt to an endophytic habit systemically within healthy cucumber plants. As the three endophytic actinomycete isolates also colonized the rhizosphere and showed outstanding rhizosphere competency it is clear that they are facultative and not obligate endophytes. The success with the three inoculants indicated that they could well be used in place of the fungicide metalaxyl which is currently recommended for the management of Pythium diseases in the UAE. This is the first successful field use of endophytic actinomycetes as promising plant growth promoters and biological control agents against Pythium diseases of cucumber.     

Persistent,symptomless, systemic,and seed-borne infection of lettuce by <Emphasis Type="Italic">Botrytis cinerea</Emphasis>

Experiments are presented which show that Botrytis cinerea, the cause of grey mould disease, is often present in symptomless lettuce plants as a systemic, endophytic, infection which may arise from seed. The fungus was isolated on selective media from surface-sterilised sections of roots, stem pieces and leaf discs from symptomless plants grown in a conventional glasshouse and in a spore-free air-flow provided by an isolation propagator. The presence of B. cinerea was confirmed by immuno-labelling the tissues with the Botrytis-specific monoclonal antibody BC-12.CA4. As plants grew, infection spread from the roots to stems and leaves. Surface-sterilisation of seeds reduced the number of infected symptomless plants. Artificial infection of seedlings with dry conidia increased the rate of infection in some experiments. Selected isolates were genetically finger-printed using microsatellite loci. This confirmed systemic spread of the inoculating isolates but showed that other isolates were also present and that single plants hosted multiple isolates. This shows that B. cinerea commonly grows in lettuce plants as an endophyte, as has already been shown for Primula. If true for other hosts, the endophytic phase may be as important a component of the species population as the aggressive necrotrophic phase.