Lettuce Downy Mildew Research Articles

MALDI‐based intact spore mass spectrometry of downy and powdery mildews

Fast and easy identification of fungal phytopathogens is of great importance in agriculture. In this context, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has emerged as a powerful tool for analyzing microorganisms. This study deals with a methodology for MALDI-TOF MS-based identification of downy and powdery mildews representing obligate biotrophic parasites of crop plants. Experimental approaches for the MS analyses were optimized using Bremia lactucae, cause of lettuce downy mildew, and Oidium neolycopersici, cause of tomato powdery mildew. This involved determining a suitable concentration of spores in the sample, selection of a proper MALDI matrix, looking for the optimal solvent composition, and evaluation of different sample preparation methods. Furthermore, using different MALDI target materials and surfaces (stainless steel vs polymer-based) and applying various conditions for sample exposure to the acidic MALDI matrix system were investigated. The dried droplet method involving solvent evaporation at room temperature was found to be the most suitable for the deposition of spores and MALDI matrix on the target and the subsequent crystallization. The concentration of spore suspension was optimal between 2 and 5 × 10(9) spores per ml. The best peptide/protein profiles (in terms of signal-to-noise ratio and number of peaks) were obtained by combining ferulic and sinapinic acids as a mixed MALDI matrix. A pretreatment of the spore cell wall with hydrolases was successfully introduced prior to MS measurements to obtain more pronounced signals. Finally, a novel procedure was developed for direct mass spectra acquisition from infected plant leaves.

Effector identification in the lettuce downy mildew Bremia lactucae by massively parallel transcriptome sequencing.

Lettuce downy mildew (Bremia lactucae) is a rapidly adapting oomycete pathogen affecting commercial lettuce cultivation. Oomycetes are known to use a diverse arsenal of secreted proteins (effectors) to manipulate their hosts. Two classes of effector are known to be translocated by the host: the RXLRs and Crinklers. To gain insight into the repertoire of effectors used by B. lactucae to manipulate its host, we performed massively parallel sequencing of cDNA derived from B. lactucae spores and infected lettuce (Lactuca sativa) seedlings. From over 2.3 million 454 GS FLX reads, 59 618 contigs were assembled representing both plant and pathogen transcripts. Of these, 19 663 contigs were determined to be of B. lactucae origin as they matched pathogen genome sequences (SOLiD) that were obtained from >270 million reads of spore-derived genomic DNA. After correction of cDNA sequencing errors with SOLiD data, translation into protein models and filtering, 16 372 protein models remained, 1023 of which were predicted to be secreted. This secretome included elicitins, necrosis and ethylene-inducing peptide 1-like proteins, glucanase inhibitors and lectins, and was enriched in cysteine-rich proteins. Candidate host-translocated effectors included 78 protein models with RXLR effector features. In addition, we found indications for an unknown number of Crinkler-like sequences. Similarity clustering of secreted proteins revealed additional effector candidates. We provide a first look at the transcriptome of B. lactucae and its encoded effector arsenal.

Silicon and increased electrical conductivity reduce downy mildew of soilless grown lettuce

Downy mildew of lettuce, caused by Bremia lactucae, is difficult to control in soilless systems by using conventional methods of disease management because few chemicals are registered, while resistant cultivars face the problem of resistance break down; therefore other methods for disease control need to be investigated. The effect of silicon salt as well as increased electrical conductivities against downy mildew was evaluated in four experiments carried out in hydroponically systems, using the cultivar of lettuce “Cobham Green”, known for its susceptibility to the pathogen. Silicon, as potassium silicate, was added at 100 mg l−1 of nutrient solution at three levels of electrical conductivity: 1.5–1.6 mS cm−1 (EC1), 3.0–3.5 mScm−1 (EC2, 0.70 g l−1 NaCl) and 4.0–4.5 mS cm−1 (EC3, 0.95 g l−1 NaCl) respectively. Lettuce plants, grown for 14–20 (trials 1 and 2) and 36–45 (trials 3 and 4) days in the different nutrient solutions tested, were inoculated with B. lactucae conidia with a maximum of two inoculations before final disease assessment carried out 14–21 days after the inoculation able to give symptoms. EC and potassium silicate significantly influenced downy mildew incidence and severity, while their interaction was not a significant factor. The addition to the standard nutrient solution (EC1) of potassium silicate resulted in a significant reduction of downy mildew severity in trials 1 and 2 where plants were artificially inoculated 15 and 20 days after transplanting. This efficacy was slight on plants grown for 36 and 45 days before inoculation in a soil drenched with EC1 amended with potassium silicate. EC2 gave a significantly similar downy mildew reduction than EC2 added with potassium silicate in trial 3. Plants grown for 36 and 45 days at the highest electrical conductivity (EC3) showed a significant reduction in severity of downy mildew compared with that observed at EC2 level. The best results, in terms of disease control, were given by the addition of potassium silicate to the EC3 solution. This combination also led to a significantly increased plant biomass. The possibility and benefits of applying potassium silicate and increased EC amendments in practice is discussed.

Novas raças de Bremia lactucae, agente causador do míldio da alface, identificadas no estado de São Paulo

O objetivo deste trabalho foi relatar o surgimento de raças de Bremia lactucae, agente causal do míldio nas principais regiões produtoras de alface do estado de São Paulo. O estudo foi realizado no Laboratório de Melhoramento Genético de Hortaliças do Departamento de Produção Vegetal da UNESP, Campus de Jaboticabal. No período de 2006 e 2007, foram coletados 36 isolados de B. lactucae de diferentes regiões produtoras de alface no estado. Para identificação das raças foram utilizadas as cultivares diferenciadoras conforme o código "Sextet". Foram identificadas três novas raças, SPBl:02, SPBl:03 e SPBl:04 com os referidos comportamentos do fungo: (63/31/19/00), (63/63/19/00) e (63/63/03/00). Os genes Dm-14, Dm-17, Dm-18, Dm-37 e Dm-38 conferem resistência a essas novas raças identificadas.

New races of Bremia lactucae on lettuce in Australia

Three distinct patterns of virulence in Bremia lactucae were identified in samples of lettuce downy mildew collected between 2005 and 2010 in Australia. These are unique to Australia and it is proposed they be named AUS4, AUS5 and AUS6.

Influence of nitric oxide and reactive oxygen species on development of lettuce downy mildew in Lactuca spp.

The role of nitric oxide and reactive oxygen species, molecules indispensable for plant-pathogen signalling, was studied in the Lactuca spp.-Bremia lactucae pathosystem. Using a leaf disc model the translaminar effect of various compounds affecting their metabolism was studied by light microscopy. Time course studies revealed a slowdown in the development of B. lactucae (race BL16) infection structures by rutin (scavenger of reactive nitrogen and oxygen species) and SNP (NO donor) within 48 h post inoculation, followed by a retardation of sporulation. Application of the specific NO scavenger, PTIO, accelerated penetration of B. lactucae but had no further effects on the plant-pathogen interaction. Inhibitors of NO synthase (L-NAME) and nitrate reductase (sodium tungstate) did not influence pathogen development. Our results suggest that drastic change in the NO: hydrogen peroxide ratio seems to determine the pathogen’s fate. NO synthase-like activity significantly increased early after B. lactucae challenge in resistant L. virosa. Confocal laser scanning microscopy revealed the accumulation of nitric oxide in the penetrated cells, pointing to a role in the initiation of the hypersensitive reaction. The tips of germ tubes and appressoria of B. lactucae also accumulated NO, suggesting an essential role for this molecule in penetration of the biotrophic pathogen. Additionally, temporal changes in endogenous levels of rutin and quercetin in extracts from Lactuca spp. leaves will be discussed in connection to their role as part of the antioxidative machinery that influences the plants’ susceptibility/resistance to lettuce downy mildew.

New Honorary Member of BSPP

In almost 40 years of research and teaching John Mansfield has made several outstanding contributions to our understanding of plant-pathogen interactions and mechanisms of plant innate immunity. He has had an important impact as an inspirational teacher and supervisor, passing on his enthusiasm for the all-embracing subject of plant pathology. His interest in pathology was kindled as an undergraduate at University College of North Wales, Bangor (1965–68) following lectures and project supervision by Dave Shaw and Bob Whitbread. He was fortunate to move to Imperial College London for his PhD (1968–72) with Brian Deverall, during a period of focus on the role of newly identified phytoalexins in disease resistance. His challenge was to examine the role of antifungal plant metabolites in the differential pathogenicity of Botrytis species to Vicia faba. The concept of a balance operating between phytoalexin production by the plant and degradation by the adapted pathogen emerged from his early work. Research on phytoalexins and host specificity with Botrytis spp. including the epidemiology of narcissus smoulder, was continued as a lecturer in plant pathology at the University of Stirling (1972–80). Working with excellent postgraduate students and collaborating colleagues led to the identification of novel phytoalexins in bean (nine in total) and daffodil. The book, Phytoalexins, published in 1982, edited with John Bailey a contemporary at Imperial College, remains the definitive text on the antimicrobial secondary metabolites. At Stirling, John developed an interest in the use of electron microscopy to define the structural framework within which pathogens and plant cells interact, and began experiments on gene-for- gene interactions and the hypersensitive response (HR). Moving to Wye College in 1981 allowed interests in the HR to be expanded, using lettuce downy mildew and bean halo-blight diseases as model interactions. The emerging power of molecular genetics was harnessed to clone avirulence (avr) genes from pathovars of Pseudomonas syringae that controlled both varietal and non-host resistance. A successful and ongoing collaboration with colleagues at UWE, Bristol and HRI, Wellesbourne led to the identification of several avr genes, and more significantly the first cloning of an effector through its virulence function (virPphA). Two landmark papers demonstrating the suppression of the HR by effector proteins VirPphA and AvrPphF have provided the framework for much of the current research on effector functions. Working with colleagues from Helsinki, ably assisted by the technical expertise of the electron microscopist Ian Brown, John was also able to visualize the delivery of effectors through the Hrp pilus, a component of the type III secretion system. He has also used microscopy to demonstrate the importance of localized cell wall interactions in the basal defences of plants against non-pathogenic bacteria. The two strands of research, cloning effectors and analysis of basal defences have come together in his group’s demonstration of the suppression of wall alterations by effector proteins, notably AvrPtoB which is now known to target plant receptor-like kinases. During 28 years at Wye, John supervised a succession of postgraduate students from the UK and overseas, working on plant –pathogen interactions and more applied projects ranging from cashew powdery mildew in Tanzania to cauliflower diseases in Nepal. He was promoted to Reader in 1986 and offered a Research Professorship in Plant Pathology from the University of London in 1991. John’s academic career came full circle when he returned to Imperial College London after the closure of the Wye campus. He has acted as head of the group of plant scientists transferred to South Kensington. He is currently on a part-time appointment in the Division of Biology at Imperial. John continues to be active in the laboratory, with ongoing projects on the mobility of effector genes, the metabolomics of disease and plant-aphid interactions. As a founder member of the BSPP he has had significant roles within the Society, the highpoint being his Presidency of BSPP in 2000. The presidential meeting held at Wye focused on how research on disease has impacted on our fundamental understanding of microbial and plant biology. His tenure of the Presidency coincided with the launch of Molecular Plant Pathology and he has been on the editorial board of both MPP and Plant Pathology. An enthusiastic conference delegate, John has often acted as a member of organizing committees, notably for the successful ICPP1998 and ICPPB 2006 meetings, both held in Edinburgh. He is currently helping to organize the 8th Symposium on Pseudomonas syringae and related pathogens, to be held in Oxford 2010. It is with great pleasure that the Society confers Honorary Membership on John Mansfield.

Race of Bremia lactucae (Downy mildew of lettuce) in Hyogo Prefecture

Race of Bremia lactucae (Downy mildew of lettuce) in Hyogo Prefecture

Fungal pathogens in Norwegian lettuce fields

Norwegian field production of lettuce has increased considerably since the early 1990s. Disease problems rarely required fungicide applications before 1996, when lettuce downy mildew (Bremia lactucae) caused severe losses. From 2002 to 2004, surveys were conducted to identify fungal diseases in Buskerud, Vestfold and Østfold counties in the south-east and Rogaland County in the south-west, representing the main lettuce production regions of Norway. The distribution and incidence of B. lactucae was highly variable, but this pathogen was the most important due to the destructive nature of uncontrolled epidemics. Septoria lactucae caused severe damage, but was found in only one field. Sclerotinia sclerotiorum was the most widespread pathogen, found in 32% of the fields, but usually affecting less than 10% of the plants. Pythium tracheiphilum was reported from 33% of the fields in south-east Norway, but was not found in the south-west. Disease incidence was usually less than 5%, and a disease incidence of more than 10% was reported in one field only. Other pathogens of potential economic importance in Norwegian lettuce fields are Alternaria spp., Botrytis cinerea and Rhizoctonia solani, although they were sporadically distributed in relatively few fields in this survey.

Activity of carboxylic acid amide (CAA) fungicides against Bremia lactucae

Four carboxylic acid amide (CAA) fungicides, mandipropamid (MPD), dimethomorph (DMM), benthiavalicarb (BENT) and iprovalicarb (IPRO) were examined for their effects on various developmental stages of Bremia lactucae, the causal agent of downy mildew in lettuce, in vitro and in planta. Spore germination in vitro or on leaf surfaces was inhibited by all CAA fungicides (technical or formulated). MPD was more effective in suppressing germination than DMM or BENT, whereas IPRO was least effective. CAA induced no disruption of F-actin microfilament organisation in germinating spores of B. lactucae. CAA applied to germinating spores in vitro prevented further extension of the germ tubes. When applied to germinated spores on the leaf surface they prevented penetration. Preventive application of CAA to intact plants inhibited infection. MPD was more effective in suppressing infection than DMM or BENT, whereas IPRO was least effective. Curative application was effective at ≤3 h post-inoculation (hpi) but not at ≥18 hpi. CAA (except IPRO) applied to upper leaf surfaces inhibited spore germination on the lower surface and hence reduced infection. CAA suppressed sporulation of B. lactucae on floating leaf discs and when sprayed onto infected plants two days before onset of sporulation. BENT and DMM were more effective in suppressing sporulation than MPD or IPRO. Epidemics of downy mildew in shade-house grown lettuce were suppressed by CAA. A single spray applied to five-leaf plants before transplanting controlled the disease for 50 days. The results suggest that CAA are effective inhibitors of spore germination and therefore should be used as preventive agents against downy mildew of lettuce caused by B. lactucae.

Structure and variation in the wild-plant pathosystem: Lactuca serriola–Bremia lactucae

Over the past decade, extensive research on the wild-plant pathosystem, Lactuca serriola (prickly lettuce)–Bremia lactucae (lettuce downy mildew), has been conducted in the Czech Republic. Studies focused on pathogen occurrence and distribution, host range, variation in symptom expression and disease severity, interactions of B. lactucae with another fungal species (Golovinomyces cichoracearum) on L. serriola, variation in resistance within natural populations of L. serriola, the structure and dynamics of virulence within populations of B. lactucae, sexual reproduction of B. lactucae, and a comparison of virulence structure and changes in B. lactucae populations occurring in wild (L. serriola) and crop (L. sativa) pathosystems. The incidence of B. lactucae on naturally growing L. serriola and other Asteraceae was recorded. Lactuca serriola was the most commonly occurring host species, followed by Sonchus oleraceus. Over the duration of these studies, the incidence of B. lactucae in L. serriola populations varied between 45–87%. Disease incidence and disease prevalence were partly related to the size, density and different habitats of L. serriola populations. In addition to B. lactucae infection, infection by the lettuce powdery mildew fungus (Golovinomyces cichoracearum) was quite common, including co-infection. Variation in resistance to B. lactucae was studied by using ten isolates (NL and BL races with known virulence patterns) at a metapopulation level, i.e. 250 L. serriola samples representing 16 populations from the Czech Republic (CZ). Our comparisons revealed broad variation in host resistance among host populations and also intrapopulation variability. In the CZ populations, 45 resistance phenotypes were recorded, the most frequent were race-specific reaction patterns. Structural and temporal changes in virulence variation of B. lactucae populations on L. serriola were studied during 1998–2005. Altogether, 313 isolates of B. lactucae originating from the Czech Republic were examined for the presence of 32 virulence factors (v-factors), and 93 different virulence phenotypes (v-phenotypes) were recorded. A study of v-factor frequency showed that common v-factors in B. lactucae populations match some of the race-specific resistance genes/factors (Dm genes or R-factors) originating from L. serriola. The highest frequency was recorded by v-factors v7, v11, v15–17, and v24–30. In contrast, v-factors (e.g. v1–4, 6, and 10) matching Dm genes originating from L. sativa were very rare. This demonstrates the close adaptation of B. lactucae virulence to the host (L. serriola) genetic background. Temporal changes in virulence frequencies over the period were recorded. In many v-factors (v11, v14, v16, and v25–28), fluctuations were observed, some (v14 and v17) shifting to higher frequencies, and others (v5/8 and v23) decreasing. The occurrence of mating types was studied (1997–1999) in a set of 59 B. lactucae isolates. Both compatibility types (B1 and B2) were recorded; however the majority of the isolates (96%) were type B2. A comparative study of B. lactucae virulence variation between the wild (L. serriola) and crop (L. sativa) pathosystems showed major differences. Migration and gene flow between both pathosystems and the potential danger of wild B. lactucae populations for cultivated lettuce are discussed. This paper summarizes comprehensive and unique research on an oomycete pathogen (B. lactucae) that is shared between a crop (lettuce, L. sativa) and its close wild relative (prickly lettuce, L. serriola). The data demonstrate clear evidence about race-specific interactions, variation and changes in virulence, and coevolutionary relationships in the wild pathosystem L. serriola–B. lactucae. Conclusions contribute to the broadening and better understanding of gene-for-gene systems in natural host–pathogen populations and their relationships to crop pathosystems.

Diversity of defence mechanisms in plant–oomycete interactions: a case study of Lactuca spp. and Bremia lactucae

Plant pathogenic oomycetes, including biotrophic downy mildews and hemibiotrophs/necrotrophs such as Phytophthora and Pythium, cause enormous economic losses on cultivated crops. Lettuce breeders and growers face the threat of Bremia lactucae, the causal agent of lettuce downy mildew. This pathogen damages leaf tissues and lettuce heads and is also frequent on wild Asteraceae plants. The interactions of Lactuca spp. with B. lactucae (abbr. lettuce–Bremia) display extreme variability, due to a long co-evolutionary history. For this reason, during the last 30 years, the lettuce–Bremia pathosystem has been used as a model for many studies at the population, individual, organ, tissue, cellular, physiological and molecular levels, as well as on genetic variability and the genetics of host–parasite interactions. The first part of this review summarizes recent data on host–parasite specificity, host variability, resistance mechanisms and genetics of lettuce–Bremia interactions. The second part focuses on the development infection structures. Phenotypic expression of infection, behaviour of B. lactucae on leaf surfaces, the process of penetration, development of primary infection structures, hyphae and haustoria are discussed in relation to different resistance mechanisms. In the third part, the components of host resistance and the variability of defence responses are analysed. The role of reactive oxygen species (ROS), antioxidant enzymes, nitric oxide (NO), phenolic compounds, reorganization of cytoskeleton, electrolyte leakage, membrane damage, cell wall disruption, hypersensitive reaction and plant energetics are discussed in relation to defence responses. In general, the extreme variability of interactions between lettuce and Bremia, and their phenotypic expression, results from diversity of the genetic background. Different mechanisms of resistance are conditioned by an orchestra of defence responses at the tissue, cell, and molecular levels. The various events responsible for defence involve a complex interaction of the processes and reactions mentioned above. This review also provides an overview on the timing of pathogen development, host pathological anatomy, cytology and physiology of lettuce–Bremia associations. The significance of these factors on the expression of different resistance mechanisms (non-host and host resistance, race-specific and race non-specific resistance, field resistance) is discussed.

Basic Compatibility of Albugo candida in Arabidopsis thaliana and Brassica juncea Causes Broad-Spectrum Suppression of Innate Immunity

A biotrophic parasite often depends on an intrinsic ability to suppress host defenses in a manner that will enable it to infect and successfully colonize a susceptible host. If the suppressed defenses otherwise would have been effective against alternative pathogens, it follows that primary infection by the "suppressive" biotroph potentially could enhance susceptibility of the host to secondary infection by avirulent pathogens. This phenomenon previously has been attributed to true fungi such as rust (basidiomycete) and powdery mildew (ascomycete) pathogens. In our study, we observed broad-spectrum suppression of host defense by the oomycete Albugo candida (white blister rust) in the wild crucifer Arabidopsis thaliana and a domesticated relative, Brassica juncea. A. candida subsp. arabidopsis suppressed the "runaway cell death" phenotype of the lesion mimic mutant lsd1 in Arabidopsis thaliana in a sustained manner even after subsequent inoculation with avirulent Hyaloperonospora arabidopsis (Arabidopsis thaliana downy mildew). In sequential inoculation experiments, we show that preinfection by virulent Albugo candida can suppress disease resistance in cotyledons to several downy mildew pathogens, including contrasting examples of genotype resistance to H. arabidopsis in Arabidopsis thaliana that differ in the R protein and modes of defense signaling used to confer the resistance; genotype specific resistance in B. juncea to H. parasitica (Brassica downy mildew; isolates derived from B. juncea); species level (nonhost) resistance in both crucifers to Bremia lactucae (lettuce downy mildew) and an isolate of the H. parasitica race derived from Brassica oleracea; and nonhost resistance in B. juncea to H. arabidopsis. Broad-spectrum powdery mildew resistance conferred by RPW8 also was suppressed in Arabidopsis thaliana to two morphotypes of Erysiphe spp. following pre-infection with A. candida subsp. arabidopsis.

Classical and molecular genetics of Bremia lactucae, cause of lettuce downy mildew

Lettuce downy mildew caused by Bremia lactucae has long been a model for understanding biotrophic oomycete–plant interactions. Initial research involved physiological and cytological studies that have been reviewed earlier. This review provides an overview of the genetic and molecular analyses that have occurred in the past 25 years as well as perspectives on future directions. The interaction between B. lactucae and lettuce (Lactuca sativa) is determined by an extensively characterized gene-for-gene relationship. Resistance genes have been cloned from L. sativa that encode proteins similar to resistance proteins isolated from other plant species. Avirulence genes have yet to be cloned from B. lactucae, although candidate sequences have been identified on the basis of motifs present in secreted avirulence proteins characterized from other oomycetes. Bremia lactucae has a minimum of 7 or 8 chromosome pairs ranging in size from 3 to at least 8 Mb and a set of linear polymorphic molecules that range in size between 0.3 and 1.6 Mb and are inherited in a non-Mendelian manner. Several methods indicated the genome size of B. lactucae to be ca. 50 Mb, although this is probably an underestimate, comprising approximately equal fractions of highly repeated sequences, intermediate repeats, and low-copy sequences. The genome of B. lactucae still awaits sequencing. To date, several EST libraries have been sequenced to provide an incomplete view of the gene space. Bremia lactucae has yet to be transformed, but regulatory sequences from it form components of transformation vectors used for other oomycetes. Molecular technology has now advanced to the point where rapid progress is likely in determining the molecular basis of specificity, mating type, and fungicide insensitivity.

Podosphaera fusca에 의한 상추 흰가루병의 발생원인과 방제

Podosphaera fusca에 의한 상추 흰가루병은 상추 발생 병해중에서 가장 중요한 병해중 하나이다. 흰가루병 발생으로 상추재배가 피해로 인하여 재배자와 소비자를 위하여 좋은 품질의 상추를 생산하기 위한 방제법 개발이 필요하다. 상추 흰가루병 발생이 증가되는 원인은 계속적인 연중재배, 병든 식물체의 미제거, 흰가루병을 노균병으로 오인에 의한 부적절한 약제 처리 등이다. 경기도 수원시 근교에 상추 시설 재배 포장에서 미생물농약을 포함한 5가지 살균제에 대한 상추 흰가루병 방제 실험을 실시하였다. 상추 흰가루병이 발생 초기에 10일 간격 3회 분무 처리하여 최종 약제 처리 8일 후에 흰가루병 발생을 조사하였다. kresoxim-mothy SC, azoxystrobin SC, Ampelomyces quisqualis AQ94013 WP, Paenibacillus polymyxa AC-1 SC와 Bacillus Y 1336 WP를 처리한 흰가루병의 발생은 0.7%, 0.7%, 26.0%, 36.7% 각각 발생하였으며, 무처리는 55.3% 발병하였다. 상추유묘에 대하여 이들 약제의 기준량과 배량에서 약해는 없었다. Powdery mildew caused by Podosphaera fusca is one of the most important diseases in leafy lettuce (Lactuca sativa L.). Since the disease has been a threat to safe cultivation of leafy lettuce, its control methods have to develop to produce good quality of lettuce for farmer and consumer. Occurrence of lettuce powdery mildew is increasing more and more due to continuous cultivation of lettuce all through the year, non-removal of diseased plant parts of lettuce, spray of inadequate fungicides by mistaken acknowledge of lettuce powdery mildew for lettuce downy mildew, etc. The control effect of five fungicides against lettuce powdery mildew was examined in a plastic greenhouse located in Suwon. When fungicides were sprayed three times at 10 days-intervals in the early stage of occurrence of powdery mildew, the incidence of powdery mildew in the plants treated with kresoxim-methy SC, azoxystrobin SC, Ampelomyces quisqualis AQ94013 WP, Paenibacillus polymyxa AC-1 SC and Bacillus subtilis Y 1336 WP was 0.7%, 0.7%, 26.0%, 36.7% and 42.0%, respectively, whereas the incidence of non-treated control was 55.3% on eight days after final application. Phytotoxicity of five fungicides tested was not observed in lettuce seedling plants.

Impact of Diurnal Periodicity, Temperature, and Light on Sporulation of Bremia lactucae

ABSTRACT We evaluated direct and interactive effects of light quality and intensity, temperature and light, diurnal rhythms, and timing of high relative humidity during long day lengths on sporulation of Bremia lactucae, the causal agent of lettuce downy mildew, using inoculated lettuce seedlings and detached cotyledons. Suppression of sporulation by light was strongly dependent upon temperature and there was little suppression at </=10 degrees C. The most suppressive waveband was in the range from 400 to 450 nm, although a lesser effect of wavebands from 450 to 500 and 500 to 550 nm could be detected. At 15 degrees C, near the lower threshold for suppression of sporulation by light, a clear diurnal pattern of sporulation was observed independent of light and darkness. This diurnal rhythm potentially could interact with light and temperature to confound the results of controlled environment studies, and may be the controlling factor in timing of sporulation at low temperatures. Forecasting models that currently use sunrise and sunset to delimit periods conducive to sporulation can be adapted to short nights and extended twilight conditions by incorporating the effects reported herein. Additionally, models of sporulation could be adapted to better reflect a decrease or absence of the suppressive effect of light at <15 degrees C.

Downy Mildew Caused by Bremia lactucae on Strawflower (Helichrysum bracteatum) in California.

In 2006, field-planted strawflower (Helichrysum bracteatum (Venten.) Andr.) grown for cutflowers in coastal San Mateo County was severely infected with a downy mildew disease. Initial symptoms consisted of irregularly shaped chlorotic lesions. Lesions were typically vein delimited, and in particular, did not cross the central longitudinal vein of the lanceolate leaves. Lesions were large and could exceed 6 cm long. Profuse white sporulation developed on the abaxial sides of the lesions. As disease progressed, lesions became gray brown and necrotic. Cutflower stems having symptomatic foliage were unmarketable. Hyaline conidiophores emerged from stomata, branched dichotomously, and had branch tips ending in swollen vesicles bearing sterigmata. Conidia were hyaline and ovoid to globose in shape. The pathogen was identified as Bremia lactucae Regel. To establish pathogenicity, strawflower plants were spray inoculated with a conidial suspension of 1 × 104 conidia/ml, incubated for 24 h in a dew chamber (18 to 20°C), and then maintained in a greenhouse (22 to 24°C). After 14 days, symptoms and signs of downy mildew developed on inoculated plants and the pathogen was confirmed to be B. lactucae. Untreated control plants did not develop downy mildew. Because lettuce (Lactuca sativa L.) is grown extensively in this coastal region, we investigated the pathogenicity of B. lactucae from strawflower on lettuce by inoculating a lettuce downy mildew differential series that includes 20 lettuce lines, four wild L. serriola lines, and strawflower. A conidial suspension of 1 × 106 conidia/ml was applied to all plants; plants were then incubated in a growth chamber with irradiation at 15°C. After 14 days, only the strawflower plants showed downy mildew lesions and sporulation. Collected from diseased field-grown strawflower, 100 downy mildew conidia were measured and had dimensions of 24.4 × 17.4 μm. These conidia were significantly longer than the B. lactuca conidia collected from lettuce (21.8 × 18.1 μm) grown in a nearby area (LSD [P = 0.05] = 2.2), though dimensions were within the reported length range for B. lactucae (12 to 31 μm) (2). To our knowledge, this is the first report of downy mildew caused by B. lactucae on strawflower in California. Inoculation studies indicate that these strawflower and lettuce pathogens from California likely belong to distinct formae speciales (2), though a B. lactucae isolate in Italy was able to infect both hosts (1). The pathogen has been reported on strawflower in Florida, Egypt, Italy, and the United Kingdom (1).

Extreme size and sequence variation in the ITS rDNA of Bremia lactucae.

Bremia lactucae Regel (Chromista, Peronosporaceae) is an economically destructive pathogen, which causes downy mildew disease on lettuce (Lactuca sativa L.) worldwide. The ribosomal internal transcribed spacer (ITS) of Bremia lactucae isolates was analyzed for the first time. The ITS region of lettuce downy mildew was observed to have a size of 2458 bp; thereby, having one of the longest ITS sizes recorded to date. The majority of the extremely large sized ITS2 length of 2086 was attributed to the additional presences of nine repetitive elements with lengths of 179-194 bp, which between them shared the low homology of 48-69%. Comparison of the ITS2 sequences with the B. lactucae isolates from other host plants showed that isolates present on Lactuca sativa were distinct from those on L. indica var. laciniata, as well as Hemistepta and Youngia. We suggest the high degree of sequence heterogeneity exhibited in the ITS2 region of B. lactucae may warrant the specific detection and diagnosis of this destructive pathogen or its division into several distinct species.

Virulence patterns ofBremia lactucae in Israel

The variation and distribution of virulent phenotypes ofBremia lactucae Regel, the causal agent of lettuce downy mildew, were studied during 2002–2003 in lettuce fields (Lactuca sativa) in Israel. A total of 21 isolates ofB. lactucae were collected from nine locations in three regions of Israel: Galilee, the Coastal Plain, and the Shefela. The isolates were examined for the presence of 21 virulence factors (v-factors) and their combinations with differential sets of lettuce lines/varieties. There were clear differences in v-factors, and a broad diversity of v-phenotypes among the isolates was found. Although 17 different v-phenotypes and 20 v-factors were detected, a composite of similar v-phenotypes generally occurred between isolates within the three regions. They differed mostly in the presence or absence of only a few v-factors. The Coastal Plain region averaged the highest virulence complexity (0.63), significantly different from that of the Shefela (0.45) and of Galilee (0.4). Comparison of the IsraeliB. lactucae isolates that were tested in this study with data of other countries showed that factor v18, which did not occur in the Israeli populations, was detected only in Czech and German pathogen populations.

UV SUPPLEMENTATION FOR GROWTH REGULATION AND DISEASE CONTROL

The benefits of supplementary ultraviolet radiation (UVB: 280-320nm) in protected lettuce were investigated in controlled environment studies. Exposure to UV-B prior to inoculation with a conidial suspension of Bremia lactucae (lettuce downy mildew) led to significant reduction in the subsequent sporulation of the pathogen. In addition to disease control, UV treatments increased leaf thickness, reduced leaf area and increased leaf pigmentation in a red-leaved cultivar. Such changes in leaf area and thickness are important quality factors that would be expected to improve plant performance after transplantation into the field. The aim of further studies will be to test such findings in a commercial glasshouse environment.