Diversity of defence mechanisms in plant–oomycete interactions: a case study of <Emphasis Type="Italic">Lactuca</Emphasis> spp. and <Emphasis Type="Italic">Bremia lactucae</Emphasis>

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.     

Phenotypic and histological expression of different genetic backgrounds in interactions between lettuce, wild Lactuca spp., L. sativa × L. serriola hybrids and Bremia lactucae

Phenotypic and histological responses of cultivated lettuce (Lactuca sativa) and wild relatives L. saligna, L.␣virosa as well as interspecific crosses derived from L. sativa × L. serriola to two races of Bremia lactucae (CS2, CS9) were investigated. With the exception of L. sativa genotypes, all accessions and hybrids expressed incomplete or complete resistance to both pathogen races, with slight differences at seedling and adult plant stages, respectively. Histological features of the interactions (development of pathogen infection structures and host hypersensitive response to attempted infection) were studied on leaf discs 48 h after inoculation. Interactions with similar phenotypic expression of resistance were characterized by significant variation in rate of development of pathogen infection structures and hypersensitive reactions. Differences found within eight Lactuca spp. accessions and hybrids challenged by two distinct pathogen races are interpreted and discussed.     

Sexual compatibility types of Bremia lactucae isolates originating from Lactuca serriola

Results are given on the occurrence of sexual compatibility types of seven isolates ofBremia lactucae originating fromLactuca serriola (prickly lettuce). It is concluded that the isolates studied are heterothallic. Both compatibility types (B1 en B2) were determined, but type B2 was prevalent. Sexual recombination ofB. lactucae isolates originating from wild and cultivated lettuce may occur.Samenvatting Zeven isolaten vanBremia lactucae, afkomstig vanLactuca serriola in Tsjechoslowakije, zijn onderzocht op hun sexuele compatibiliteitstype door ze te combineren met Nederlandse fysio's vanB. lactucae, afkomstig van cultuursla (L. sativa), waarvan het compatibiliteitstype (B1 of B2) bekend is. Alle isolaten vanL. serriola bleken heterothallisch te zijn, waarbij type B2 meer werd aangetroffen dan type B1. Sexuele recombinatie vanBremia-isolaten van wildeLactuca-soorten en cultuursla blijkt goed mogelijk te zijn.     

ROS and NO production in compatible and incompatible tomato-<Emphasis Type="Italic">Meloidogyne incognita</Emphasis> interactions

Nitric oxide (NO) has been shown to be an essential regulatory molecule in plant response to pathogen infection in synergy with reactive oxygen species (ROS). At the present, nothing is known about the role of NO in disease resistance to nematode infection. We used a resistant tomato cultivar with different sensitivity to avirulent and virulent populations of the root-knot nematode Meloidogyne incognita to investigate the key components involved in oxidative and nitrosative metabolism. We analyzed the superoxide radical production, hydrogen peroxide content, and nitric oxide synthase (NOS)-like and nitrate reductase activities, as potential sources of NO. A rapid NO accumulation and ROS production were found at 12 h after infection in compatible and incompatible tomato-nematode interactions, whereas the amount of NO and ROS gave different results 24 and 48 h after infection amongst compatible and incompatible interactions. NOS-like arginine-dependent enzyme rather than nitrate reductase was the main source of NO production, and NOS-like activity increased substantially in the incompatible interaction. We can envisage a functional overlap of both NO and ROS in tomato defence response to nematode invasion, NO and H₂O₂ cooperating in triggering hypersensitive cell death. Therefore, NO and ROS are key molecules which may help to orchestrate events following nematode challenge, and which may influence the host cellular metabolism.     

Taxonomy,distribution and biology of lettuce powdery mildew (Golovinomyces cichoracearum sensu stricto)

This paper reviews the taxonomy, biology, importance, host–pathogen interactions and control of lettuce powdery mildew. The main causal agent of this disease, Golovinomyces cichoracearum s.s., is an important powdery mildew pathogen of many members of the family Asteraceae. The pathogen is distributed worldwide and occurs on Lactuca sativa as well as wild Lactuca spp. and related taxa (e.g. Cichorium spp.). Powdery mildew of lettuce can be a major problem in production areas with favourable environmental conditions for disease development (dry, hot weather). The fungus grows ectophytically and appears as white, powdery growth on both the upper and lower sides of leaves. There is rather limited information on the geographic distribution of powdery mildew on wild Lactuca spp. Most L. sativa cultivars have been found to be susceptible. Large variability in virulence was confirmed and existence of different races is supposed. Resistance in L. sativa and some related wild Lactuca spp. is characterized by race‐specificity, but the genetic background of resistance is poorly understood. Sources of resistance are known in L. saligna and L. virosa. Lettuce powdery mildew can be effectively controlled by common fungicides (e.g. sulphur, myclobutanil, quinoline, strobilurins, etc.) and protective compounds (e.g. extract of neem oil, Reynoutria sachaliensis extracts). However, fungicide resistance may arise. Non‐fungicidal activators of plant systemic acquired resistance (SAR) had no direct effect on the causal agent. Future issues regarding lettuce powdery mildew research are summarized.     

Wild Lactuca species,their genetic diversity,resistance to diseases and pests,and exploitation in lettuce breeding

Current knowledge of wild Lactuca L. species, their taxonomy, biogeography, gene-pools, germplasm collection quality and quantity, and accession availability is reviewed in this paper. Genetic diversity of Lactuca spp. is characterized at the level of phenotypic and phenological variation, variation in karyology and DNA content, biochemical traits, and protein and molecular polymorphism. The reported variation in reaction to pathogens and pests of wild Lactuca spp. is summarized, including the viral pathogens (Lettuce mosaic virus-LMV, Mirafiori lettuce virus/Lettuce big vein virus-LBV, Beet western yellows virus-BWYV, Tomato spotted wilt virus-TSWV, Cucumber mosaic virus-CMV, Lettuce necrotic stunt virus-LNSV), bacterial pathogens (corky root-Rhizomonas suberifaciens, bacterial leaf spot-Xanthomonas campestris pv. vitians), fungal pathogens (downy mildew-Bremia lactucae, powdery mildew-Golovinomyces cichoracearum, anthracnose-Microdochium panattoniana, stemphylium leaf spot-Stemphylium spp., sclerotinia drop-Sclerotinia spp., verticillium wilt-Verticillium dahliae, fusarium wilt-Fusarium spp., pythium wilt-Pythium tracheiphylum, P. uncinulatum), nematodes (potato cyst nematode-Globodera rostochiensis, root-knot nematode-Meloidogyne spp., incognita, hapla, javanica, enterolobii), insects and mites (the green lettuce aphid-Nasonovia ribisnigri, the green peach aphid-Myzus persicae, the potato aphid-Macrosiphum euphorbiae, leafminer-Liriomyza spp., L. langei). The approaches used to exploit wild Lactuca spp. in lettuce breeding (interspecific hybridization, cell and tissue culture, transformation) are dicussed, and known examples of lettuce cultivars with traits derived from wild Lactuca spp. are described.     

Structure and variation in the wild-plant pathosystem: <Emphasis Type="Italic">Lactuca serriola–Bremia lactucae</Emphasis>

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.     

Virulence patterns of<Emphasis Type="Italic">Bremia lactucae</Emphasis> 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. http://www.phytoparasitica.org posting Dec. 21, 2006.     

Classical and molecular genetics of <Emphasis Type="Italic">Bremia lactucae</Emphasis>, 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.     

Phylogenetic investigations in the downy mildew genus <Emphasis Type="Italic">Bremia</Emphasis> reveal several distinct lineages and a species with a presumably exceptional wide host range

Bremia lactucae is one of the most devastating and widespread pathogens in lettuce production worldwide. Despite its economical importance, uncertainty prevails about the species delimitation in the genus Bremia. Commonly, Bremia is considered to be monotypic, containing only Bremia lactucae, while taxonomists have described additional species, and molecular phylogenetic studies have shown significant sequence divergence between accessions from different hosts. Here, we report that several previously described species are genetically highly distinct from Bremia lactucae parasitic to Lactuca sativa. These include Bremia lapsanae, Bremia sonchicola, and Bremia taraxaci. In addition to these host-specific species, a plurivorous species is revealed, which infects hosts from three different tribes in the Asteraceae subfamilies Asteroideae and Carduoideae. The broad host range of clade 1 is exceptional for downy mildews and only paralleled by Pseudoperonospora cubensis, which infects a broad range of Cucurbitaceae. The taxonomic status of Bremia cirsii and of Bremia centaureae remains unresolved, as the accessions from Cirsium and Centaurea, respectively, did not form a monophylum but were partly contained in the plurivorous clade 1. Bremia lactucae was found to be restricted to Lactuca sativa and Lactuca serriola. Thus, it can be assumed that Bremia infections on weeds apart from Lactuca species do not pose a significant risk for lettuce production. However, it is unlikely that breeding resistance genes from Lactuca serriola into Lactuca sativa will result in durable resistance of lettuce to downy mildew disease, because the current study provides additional evidence that Bremia accessions from both hosts form a population continuum.     

First record of the Q biotype of the sweetpotato whitefly, <Emphasis Type="Italic">Bemisia tabaci</Emphasis>, in Guatemala

Bemisia tabaci (Gennadius) adults and immatures were collected from poinsettia plants at two commercial production greenhouses in Guatemala during an invited tour to observe IPM practices within the facilities. Despite extensive scouting, only low numbers of insects were collected from vegetable, weed and wild ornamentals species located close to these facilities. Prior to molecular and biochemical analyses, whitefly immatures were initially identified as B. tabaci using morphological characters of the pupae to distinguish them from the greenhouse whitefly. The biotype status of adults and immatures was then established using esterase isozyme patterns and MTCO1 sequencing. The Q biotype was the only biotype found on commercially grown poinsettia plants. The previously recorded B biotype was observed outside the greenhouse facilities on Lactuca spp., Hibiscus spp. and Euphorbia spp. (wild poinsettia). The New World biotype was observed on wild poinsettia and field-grown beans (Phaseolus spp.). This is the first report of the Q biotype in Guatemala, and serves notice of the need for greater vigilance in the management of whiteflies on poinsettia mother stock used as a source of cuttings for export to the USA.     

Nitric oxide is involved in chitosan‐induced systemic resistance in pearl millet against downy mildew disease

BACKGROUND: The nature and durability of resistance offered by chitosan and the involvement of nitric oxide (NO) in chitosan‐induced defence reactions in pearl millet against downy mildew disease were investigated. RESULTS: It had previously been reported that chitosan seed priming protected pearl millet plants against downy mildew disease. Further elucidation of the mechanism of resistance showed that chitosan seed priming protects the plants systemically. A minimum 4 day time gap is required between the chitosan treatment and pathogen inoculation for maximum resistance development, and it was found to be durable. Chitosan seed priming elevated NO accumulation in pearl millet seedlings, beginning from 2 h post‐inoculation, and it was found to be involved in the activation of early defence reactions such as hypersensitive reaction, callose deposition and PR‐1 protein expression. Pretreatment with NO scavenger C‐PTIO and nitric oxide synthase (NOS) inhibitor L‐NAME before pathogen inoculation reduced the disease‐protecting ability of chitosan, and defence reactions were also downregulated, which indicated a possible role for NO in chitosan‐induced resistance. CONCLUSION: Protection offered by chitosan against pearl millet downy mildew disease is systemic in nature and durable. Chitosan‐induced resistance is activated via NO signalling, as defence reactions induced by chitosan were downregulated under NO deficient conditions. Copyright © 2009 Society of Chemical Industry     

Post infection application of DL-3-amino-butyric acid (BABA) induces multiple forms of resistance against <Emphasis Type="Italic">Bremia lactucae</Emphasis> in lettuce

DL-3-amino-butyric acid (BABA) induces local and systemic resistance against disease in numerous plant species. In a recent study we showed that preventive application of BABA to lettuce (Lactuca sativa) plants induced resistance against downy mildew caused by the oomycete Bremia lactucae by callose encasement of the primary infection structures of the pathogen. Now we show that post-infection application of BABA to the foliage or the roots, even at progressive stages of disease development, is highly protective against B. lactucae. Resistance induced by BABA is manifested in multiple microscopic forms, depending on the time of its application. When applied at 1 day post inoculation (dpi) BABA induced HR in penetrated epidermal cells; at 2 dpi it caused massive encasement with callose of the primary haustoria; and, at 3 or 4 dpi it enhanced the accumulation of H₂O₂ in the developing mycelia runners and altered their colour to red. The pronounced change in the colour of the mycelium was visually apparent to the naked eye. In all cases the pathogen failed to sporulate on the treated plants. This is the first indication that an immunizing compound may be protective at advanced stages of disease development.     

A new race of Fusarium oxysporum f. sp. lactucae of lettuce

Fusarium oxysporum f. sp. lactucae, the causal agent of fusarium wilt of lettuce (Lactuca sativa), occurs in most countries in which lettuce is grown and causes serious economic losses. Three races (1, 2 and 3) of the pathogen have previously been identified on the basis of their ability to cause disease on differential lettuce cultivars, as well as by means of molecular tools developed to characterize different races of this pathogen. Only race 1 has been detected in Europe so far. In this study, two isolates of F. oxysporum, obtained from lettuce plants grown in the Netherlands showing symptoms of wilt, have been characterized by combining the study of pathogenicity with differential cultivars of lettuce and molecular assays to determine whether the isolates are different from the known races of F. oxysporum f. sp. lactucae. This study reports the presence of F. oxysporum f. sp. lactucae for the first time in the Netherlands. The causal pathogen has been identified, using the IRAP‐SCAR technique, as a new race of F. oxysporum f. sp. lactucae. Specific primers have been designed to identify this new race.     

Inhibition on brown rot disease and induction of defence response in harvested peach fruit by nitric oxide solution

Nitric oxide (NO) is an important signal molecule involved in numerous plant responses to biotic and abiotic stresses. The effect of nitric oxide (NO) solution on pathogen infection and defence response of peach (Prunus persica (L.) Batsch) fruit against brown rot disease caused by Monilinia fructicola was investigated. The results showed that 15 μmol l^?1 NO solution did not significantly inhibit spore germination, germ tube length or pathogenicity of M. fructicola, but significantly reduced disease incidence and lesion areas in the fruit. Although 100 μmol l^?1 NO solution effectively inhibited the spore germination, germ tube elongation and pathogenicity of M. fructicola, the high concentration of NO solution caused damage to the fruit. Moreover, 15 μmol l^?1 NO enhanced the activities of chitinase (CHI) and β-1,3-glucanase (GNS) in the fruit. RT-PCR analysis showed that the expression of four genes, CHI, GNS, pathogenesis-related protein 1 and 10 genes (PR-1, PR-10) all increased after NO treatment. Conversely, pretreatment with 100 μmol l^?1 NO scavenger, 2-4-carboxyphenyl-4,4,5,5- tetramethylimidazoline-1-oxyl-3-oxide (cPTIO), rendered the fruit relatively susceptible to pathogen infection and inhibited the defence response of the fruit. These results suggest that NO solution treatment can protect peach fruit from pathogen infection by inducing the activities of the defence enzymes and the expression of PR genes.     

Signals induced by exogenous nitric oxide and their role in controlling brown rot disease caused by <Emphasis Type="Italic">Monilinia fructicola</Emphasis> in postharvest peach fruit

Recent evidence suggests that nitric oxide (NO) signaling plays an important role in plant–pathogen interactions and that aconitase is a major target of NO. In the present study on the signaling role of NO in the elicitation of defense responses in peach fruit against Monilinia fructicola and subsequent effect on brown rot disease, 15 μM NO solution induced disease resistance in harvested peaches. As a potentiated elicitor, NO induced high levels of endogenous NO and superoxide (O₂ ^?), hydrogen peroxide (H₂O₂), and NADPH oxidase and Ca²⁺-ATPase activity in the fruit. Aconitase activity in peach fruit was inhibited by NO. Activity of partially purified aconitase was inhibited in vitro by sodium nitroprusside (SNP) and H₂O₂; however, the inhibition could be relieved by carboxy-2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (cPTIO) or catalase (CAT), indicating that the defense response and signals induced by NO transduction depend on aconitase and conditions leading to elevated levels of NO; otherwise, H₂O₂ would inactivate aconitase directly in fruit. Treatment with NO resulted in salicylic acid (SA) accumulating during storage. Higher levels of jasmonic acid (JA) were detected in NO-treated fruit 48 h after the treatment. But after NO was removed, the level of SA and JA were lower than in the control. The results suggest that exogenous NO enhances resistance of harvested peach fruit against the fungus by inducing signals such as endogenous NO, reactive oxygen species (ROS), SA and JA and by inhibiting aconitase activity.     

Colonization of lettuce cultivars and rotation crops by Fusarium oxysporum f. sp. lactucae,the cause of fusarium wilt of lettuce

The severity of fusarium wilt is affected by inoculum density in soil, which is expected to decline during intervals when a non‐susceptible crop is grown. However, the anticipated benefits of crop rotation may not be realized if the pathogen can colonize and produce inoculum on a resistant cultivar or rotation crop. The present study documented colonization of roots of broccoli, cauliflower and spinach by Fusarium oxysporum f. sp. lactucae, the cause of fusarium wilt of lettuce. The frequency of infection was significantly lower on all three rotation crops than on a susceptible lettuce cultivar, and the pathogen was restricted to the cortex of roots of broccoli. However, F. oxysporum f. sp. lactucae was isolated from the root vascular stele of 7·4% of cauliflower plants and 50% of spinach plants that were sampled, indicating a greater potential for colonization and production of inoculum on these crops. The pathogen was also recovered from the root vascular stele of five fusarium wilt‐resistant lettuce cultivars. Thus, disease‐resistant plants may support growth of the pathogen and thereby contribute to an increase in soil inoculum density. Cultivars that were indistinguishable based on above‐ground symptoms, differed significantly in the extent to which they were colonized by F. oxysporum f. sp. lactucae. Less extensively colonized cultivars may prove to be superior sources of resistance to fusarium wilt for use in breeding programmes.     

Effect of different organic amendments on lettuce fusarium wilt and on selected soilborne microorganisms

This study evaluated the effect of different organic amendments on lettuce fusarium wilt caused by Fusarium oxysporum f. sp. lactucae in pots under controlled conditions. Their effects on the density of the pathogen, on the total fungi and on fluorescent Pseudomonas spp. were also evaluated after two subsequent lettuce crops. A significant reduction in the severity of the symptoms of F. oxysporum f. sp. lactucae was found after the use of Brassica carinata pellets (52–79% reduction) and compost (49–67% reduction), while Brassica green manure and cattle and chicken manure only provided partial control of fusarium wilt. However, variations in effectiveness were observed for the same treatment in repeated trials. In general, an increase was observed in Pseudomonas and a decrease in fungal populations in the growing medium, which was obtained by mixing a blonde sphagnum peat and a sandy loam soil with B. carinata pellets and compost after two consecutive cropping cycles. Prolonging the Brassica and compost treatments from 30 to 60 days did not significantly affect disease severity, plant growth or the microbial population of the total fungi or Pseudomonas. The largest lettuce biomass was obtained in the non‐inoculated growing medium amended with brassica flour, chicken manure, B. carinata pellets and compost, as a consequence of fertilization. The treatment with B. juncea green manure, B. carinata (pellets and flour) and compost applied 30 days before planting led to promising results and merits further investigation for use under field conditions.     

Involvement of nitric oxide generation in hypersensitive cell death induced by elicitin in tobacco cell suspension culture

Recent studies suggest that nitric oxide (NO), an important signaling and defense molecule in mammals, plays a key role in activating disease resistance in plants. We characterized NO production by tobacco Bright Yellow-2 cells pharmacologically after treatment with INF1, the major elicitin secreted by the late blight pathogen Phytophthora infestans, prepared from Escherichia coli. NO production rapidly occurred within 1h and reached a maximum level 3–6h after the addition of INF1. Carboxy-PTIO, a NO-specific scavenger, abolished INF1-induced NO production in a dose-dependent manner. Pretreatment of protein synthesis inhibitor cycloheximide and protein kinase inhibitor K252a blocked NO production 3–12h after INF1 treatment, indicating that NO production requires de novo protein synthesis and protein phosphorylation. In an investigation of the relations between NO generation and several defense responses induced by INF1, carboxy-PTIO completely suppressed activation of a 41-kDa protein kinase and cell death by INF1. Carboxy-PTIO also suppressed the induction of hypersensitive-related (hsr) genes HSR515 and HSR203J, the expression of which is strongly correlated with the hypersensitive response in plants. The results suggest that NO plays a crucial role in the induction of hypersensitive cell death.     

Resistance mechanisms of wild tomato germplasm to infection of Oidium neolycopersici

Tomato powdery mildew (Oidium neolycopersici) is one of the most devastating diseases of cultivated tomatoes worldwide. Although the first epidemics were recorded more than 25 years ago many aspects of this host-pathogen interaction are still not well understood. Detailed morphological and molecular studies of the anamorphs confirmed that O. neolycopersici is phylogeneticaly close to Erysiphe aquilegiae var. ranunculi. Host range is rather broad, apart from Solanaceae hosts were found in the families Apocynaceae, Campanulaceae, Crassulaceae, Cistaceae, Cucurbitaceae, Linaceae, Malvaceae, Papaveraceae, Pedialiaceae, Scrophulariaceae, Valerianaceae a Violaceae. Non-host resistance within these families is not based on inhibition of formation of primary haustorium, however, on post-haustorial hypersensitive reponse and another type of non-hypersensitive resistance. Screening of wild Solanum species (previous Lycopersicon spp.) germplasm revealed valuable sources of resistance (S. habrochaites, S. pennellii, S. cheesmaniae, S. chilense, S. peruvianum). The main resistance mechanism was found to be a hypersensitive response (HR), in some cases followed by limited development of the pathogen. However, there is a broad variation in resistance response on the histological and cytological level. Interaction between many wild Solanum spp. and O. neolycopersici is race-specific, at least three races were differentiated. In some interspecific crosses (S. lycopersicum × S. habrochaites) adult plant resistance was observed. Biochemical studies focusing on production of reactive oxygen species (ROS) and peroxidase activity during infection of O. neolycopersici showed that production of ROS and activity of corresponding enzymes is related to activation of defence responses in genotypes of wild Solanum sect. Lycopersicon. The significance of nitric oxide (NO) in O. neolycopersici pathogenesis was supported by experiments with NO donors and scavengers. In moderately resistant genotype S. chmielewskii, treatment by heat stress caused slight deceleration of pathogen development, increased production of jasmonic acid (JA) and abscisic acid (ABA) and increased peroxidase activity in infected plants. The different degree of tomato resistance/susceptibility did not markedly change the rate and extent of photosynthetic response to O. neolycopersici; only minimal impairment of photosynthesis was found in both susceptible and moderately resistant genotypes during the first 9 days after inoculation. The accumulated evidence confirm a crucial role of localised increased production of ROS and reactive nitrogen species (RNS) in response to pathogen penetration into plant tissue and its involvement in the plant resistance responses including the initiation and progression of plant cell death in host wild Solanum species. Crucial points of further research are discussed.