Cytological events in tomato leaves inoculated with conidia of <Emphasis Type="Italic">Blumeria graminis</Emphasis> f. sp. <Emphasis Type="Italic">hordei</Emphasis> and <Emphasis Type="Italic">Oidium neolycopersici</Emphasis> KTP-01

Leaves of tomato and barley were inoculated with conidia of Blumeria graminis f. sp. hordei race 1 (R1) or Oidium neolycopersici (KTP-01) to observe cytological responses in search of resistance to powdery mildew. Both conidia formed appressoria at similar rates on tomato or barley leaves, indicating that no resistance was expressed during the prepenetration stage of these fungi. On R1-inoculated tomato leaves, appressoria penetrated the papillae, but subsequent haustorium formation was inhibited by hypersensitive necrosis in the invaded epidermal cells. On the other hand, KTP-01 (pathogenic to tomato leaves) successfully developed functional haustoria in epidermal cells to elongate secondary hyphae, although the hyphal elongation from some conidia was later suppressed by delayed hypersensitive necrosis in some haustorium-harboring epidermal cells. Thus, the present study indicated that the resistance of tomato to powdery mildew fungi was associated with a hypersensitive response in invaded epidermal cells but not the prevention of fungal penetration through host papilla.     

A natural fungicide for the control of <Emphasis Type="Italic">Erysiphe betae</Emphasis> and <Emphasis Type="Italic">Erysiphe cichoracearum</Emphasis>

This study examines the effects of a vegetable fungicide on sugar beet powdery mildew (Erysiphe betae) and cucumber powdery mildew (Erysiphe cichoracearum). The formulations consisting of a dispersion of Brassicaceae meal in vegetable or mineral oils on infected leaves of sugar beet, reared in the greenhouse, and of musk melons cultivated under plastic tunnels, were tested in comparison to each oil taken separately. Both formulations containing Brassicaceae meals, caused 94% of conidia to be distorted while for the untreated group only 2% were distorted. Furthermore, the leaf area infected by E. betae was 56% for untreated plants and 2.7 and 9.9% respectively, for plants treated with meal containing mineral and vegetable oil. Vegetable oil considered separately or with Brassicaceae meals showed no phytotoxicity, while the formulations based on mineral oil showed a significantly lower fresh and dry weight on tomato plants. The low level or absence of phytotoxicity of plants treated with vegetable oil formulations suggests that to improve the efficacy of powdery mildew control, they could be used mixed with sulphur. The efficiency of the vegetable formulations in the powdery mildew control observed during these trials encourages further investigation on other parasitic fungi and foliar pathogens.     

Morphological and molecular characterization of <Emphasis Type="Italic">Oidium</Emphasis> subgenus <Emphasis Type="Italic">Reticuloidium</Emphasis> (powdery mildew) newly occurred on cucumber in Japan

In 2002, a powdery mildew with catenate conidia lacking fibrosin bodies was found on cucumber in a greenhouse in Kanagawa Prefecture, Japan. Morphological observation revealed that the fungus belongs to Oidium subgenus Reticuloidium, anamorph of the genus Golovinomyces. Molecular phylogenetic analyses of the nucleotide sequences of the rDNA ITS regions and D1/D2 domains of the 28S rDNA indicated that the fungus belongs to the clade of G. orontii with other Golovinomyces fungi from a wide range of host plants, suggesting that the fungus was newly transported from abroad. Because there has been no prior report of cucumber powdery mildew caused by Reticuloidium, further research on the physiology, epidemiology, control and resistant cucumber varieties is required.     

Effects of light quality on conidiophore formation of the melon powdery mildew pathogen <Emphasis Type="Italic">Podosphaera xanthii</Emphasis>

The lengths of conidiophores in fungal colonies of the melon powdery mildew pathogen Podosphaera xanthii Pollacci KMP-6 N cultured under greenhouse (natural) conditions differed markedly from those cultured in a growth chamber. We hypothesized that light wavelength was responsible for the differences in conidiophore length. In this study, we examined the effects of light-emitting diode (LED) irradiation (purple, blue, green, orange, and red light) and white light on colony development and conidiophore formation in KMP-6 N using a stereomicroscope and a high-fidelity digital microscope. Colonies on leaves were flat under greenhouse conditions and under red LED light irradiation but were stacked under growth chamber conditions and under purple, blue, green, and orange LED light irradiation. In addition, KMP-6 N formed catenated conidia comprising six conidia per conidiophore under greenhouse conditions and red light but more than seven conidia per conidiophore under growth chamber conditions and purple, blue, green, and orange light. Furthermore, almost none of the conidia on top of the conidiophores grown under blue light were fully constricted. Therefore, these fungi could not scatter their conidia and spread infection. This is the first report of the effects of LED lights on conidiophore formation in the melon powdery mildew fungus P. xanthii. The results provide insight into the mechanisms underlying the responses of conidiophores to light of specific wavelengths and conidial scatter from conidiophores of melon powdery mildew fungi.     

Resistance levels of cucumber to <Emphasis Type="Italic">Podosphaera xanthii</Emphasis> in a growth chamber are related to haustorial formation and hyphal branching frequency

To clarify relationships between powdery mildew resistance of cucumber and hyphal developmental state of the pathogen, haustorial formation and the hyphal branching frequency were compared among cucumber varieties that differ in resistance levels to powdery mildew pathogen Podosphaera xanthii. Cotyledons of four cultivars were inoculated with P. xanthii. By 2 days after inoculation, secondary haustoria had developed from the first hyphal cell that formed beside the conidium in susceptible cultivars. The fungus on susceptible cultivars also tended to have hyphal branches just after the hyphal cell producing haustoria.     

Discrimination of <Emphasis Type="Italic">Xanthomonas campestris</Emphasis> pv. <Emphasis Type="Italic">campestris</Emphasis> races among strains from northwestern Spain by <Emphasis Type="Italic">Brassica</Emphasis> spp. genotypes and rep-PCR

Black rot, caused by Xanthomonas campestris pv. campestris (Xcc) is a severe seedborne disease of Brassica crops around the world. Nine races are recognized, being races 1 and 4 the most aggressive and widespread. The identification of Xcc races affecting Brassica crops in a target area is necessary to establish adequate control measures and breeding strategies. The objectives of this study were to isolate and identify Xcc strains from northwestern Spain by using semi-selective medium and pathogenicity tests, determine the existing races of Xcc in this area by differential series of Brassica spp., and evaluate the use of repetitive DNA polymerase chain reaction-based fingerprinting (rep-PCR) to differentiate among the nine existing Xcc races. Seventy five isolates recovered from infected fields were identified as Xcc. Race-typing tests determined the presence of the following seven pathogen races: 1, 4, 5, 6, 7, 8 and 9. Race 4 was the most frequent in Brassica oleracea and race 6 in Brassica rapa crops, therefore breeding should be focussed in obtaining resistant varieties to both races. Cluster analysis derived from the combined fingerprints showed four groups, but no clear relationship to race, crop or geographical origin was found. Rep-PCR analysis was found not to be a reliable method to discriminate among Xcc races, therefore race typing of Xcc isolates should be done by using the differential series of Brassica spp. genotypes or another alternative approach.     

Impact of <Emphasis Type="Italic">Erysiphe alphitoides</Emphasis> on transpiration and photosynthesis in <Emphasis Type="Italic">Quercus robur</Emphasis> leaves

Oak powdery mildew, (Erysiphe alphitoides) causes one of the most common diseases of oaks. We assessed the impact of this pathogen on photosynthesis and water relations of infected leaves using greenhouse-grown oak seedlings. Transpiration of seedlings infected by oak powdery mildew was also investigated. Altogether, E. alphitoides had a low impact on host gas exchange whether at the leaf or whole plant scale. Maximal stomatal conductance of infected leaves was reduced by 20–30% compared to healthy controls. Severely infected seedlings did not experience any detectable change of whole plant transpiration. The reduction in net CO₂ assimilation, A_n, was less than proportional to the fraction of leaf area infected. Powdery mildew reduced both the maximal light-driven electron flux (J_max) and the apparent maximal carboxylation velocity (Vc_max) although Vc_max was slightly more impacted than J_max. No compensation for the infection occurred in healthy leaves of partly infected seedlings as the reduced photosynthesis in the infected leaves was not paralleled by increased A_n levels in the healthy leaves of the seedlings. However, E. alphitoides had a strong impact on the leaf life-span of infected leaves. It is concluded that the moderate effect of E. alphitoides on oak might be related to the small impact on net CO₂ assimilation rates and on tree transpiration; nevertheless, the severe reduction in leaf life-span of heavily infected leaves may lead to decreased carbon uptake over the growth season.     

Screening of cultivated and wild adzuki bean for resistance to race 3 of <Emphasis Type="Italic">Cadophora gregata</Emphasis> f. sp. <Emphasis Type="Italic">adzukicola</Emphasis>, cause of brown stem rot

Two diseases of adzuki bean, brown stem rot (BSR, caused by Cadophora gregata f. sp. adzukicola) and adzuki bean Fusarium wilt (AFW, caused by Fusarium oxysporum f. sp. adzukicola), are serious problems in Hokkaido and have been controlled using cultivars with multiple resistance. However, because a new race of BSR, designated race 3, was identified, sources of parental adzuki bean for resistance to race 3 were needed. Therefore, we examined 67 cultivars and lines of cultivated and wild adzuki bean maintained at the Tokachi Agricultural Experiment Station using a root-dip inoculation method. Consequently, nine adzuki bean cultivars, one wild adzuki bean accession and 30 lines (including two lines resistant to all the three races of BSR and AFW) were confirmed to be resistant or tolerant to race 3 of BSR, and we found a cultivar Akamame as well as a wild adzuki bean Acc2515 to be a new source for a resistance gene to the race 3. This cultivar also holds promise as a source of resistance against other races of BSR and AFW.     

The first occurrence of leaf mold of tomato caused by races 4.9 and 4.9.11 of <Emphasis Type="Italic">Passalora fulva</Emphasis> (syn. <Emphasis Type="Italic">Fulvia fulva</Emphasis>) in Japan

Tomato leaf mold caused by Passalora fulva was found on two tomato varieties carrying the Cf-9 gene in Japan, in 2007. The isolates obtained from Chiba and Fukushima were identified as race 4.9.11, and those from Gunma were races 4.9 or 4.9.11. This is the first report in Japan of tomato leaf mold caused by P. fulva strains that can overcome the Cf-9 gene.     

Relation between pathogenicity and genetic variation within <Emphasis Type="Italic">Plasmodiophora brassicae</Emphasis>

The relation between diversity of pathogenicity on clubroot-resistant (CR) cultivars of Chinese cabbage (Brassica rapa subsp. pekinensis) bred in Japan and DNA polymorphisms in 17 populations of Plasmodiophora brassicae from cruciferous plants was examined by inoculation tests and random amplified polymorphic DNA (RAPD) analysis using 18 arbitrary primers. Four pathotypes (A–D) were identified after inoculation of six CR cultivars of Chinese cabbage in the 17 populations from cruciferous crops. A relatively high level of genetic diversity was also detected among these populations in the RAPD analysis. Although the four pathotypes could not be clearly differentiated using the RAPD data, most populations of three pathotypes had a consistent location on the dendrogram. All pathotype B (virulent on five cultivars except Utage 70) and D (avirulent on all cultivars) populations, which were common in incompatible interactions with cv. Utage 70, were located in a single subcluster. All five pathotype C populations (virulent only on cv. Utage 70) except for one population grouped in another single subcluster. Because four pathotype A populations (virulent on all six cultivars, races 4 and 9) fell in different subclusters, the populations may be genetically polyphyletic. Populations from cruciferous weed Cardamine flexuosa differed remarkably from those from cruciferous crops in pathogenicity on common cultivars of Chinese cabbage and turnip and C. flexuosa, but they grouped in a single cluster with all race 9 populations from crops. Race 9 populations from crops may thus be closely related to populations from the weed rather than to races 1 and 4 from crops.     

Infectivity of a Japanese isolate of <Emphasis Type="Italic">Oidium neolycopersici</Emphasis> KTP-01 to a European tomato cultivar resistant to <Emphasis Type="Italic">O. lycopersici</Emphasis>

The infectivity of a Japanese isolate of tomato powdery mildew, Oidium neolycopersici KTP-01, to tomato cultivars was examined using a resistant cultivar Grace bred in The Netherlands to O. lycopersici, which was recently proposed to be renamed O. neolycopersici. Grace was severely infected with KTP-01, and its susceptibility was similar to that on susceptible tomato cultivars Moneymaker and Ponderosa, suggesting that KTP-01 differs in pathogenicity on tomatoes from those of European and American isolates.     

First report of spinach downy mildew caused by race Pfs:8 of <Emphasis Type="Italic">Peronospora farinosa</Emphasis> f. sp. <Emphasis Type="Italic">spinaciae</Emphasis> in Japan

The race of field isolates of Peronospora farinosa f. sp. spinaciae (Pfs), causal agent of spinach downy mildew, were identified using race-differential cultivars. One isolate was similar to race Pfs:6. Three isolates were identified as race Pfs:8, the first time the race has been reported in Japan as far as we know. The differential reaction caused by the other two isolates did not match any known to be caused by races Pfs:1 through Pfs:11; thus, this strain appears to a new pathogenic strain in Japan.     

Host range expansion in a powdery mildew fungus (<Emphasis Type="Italic">Golovinomyces</Emphasis> sp.) infecting <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>: <Emphasis Type="Italic">Torenia fournieri</Emphasis> as a new host

Since 2003, Torenia fournieri plants grown for experimental purposes were repeatedly infected by powdery mildew in a laboratory in Hungary. Based on morphological characteristics, the pathogen belonged to the mitosporic genus Oidium subgen. Reticuloidium, the anamorph stage of Golovinomyces. The rDNA ITS sequence was identical to that of two other powdery mildew fungi, infecting Arabidopsis and Veronica, respectively, in different parts of the world. According to a previous phylogenetic analysis of ITS and 28S rDNA sequences, those two powdery mildews belong to a recently evolved group of Golovinomyces characterized by multiple host range expansions during their evolution. Both the ITS sequence and the morphological data indicate that the powdery mildew anamorph infecting Torenia also belongs to this group. It is likely that the powdery mildew infections of the experimental T. fournieri plants, native to south-east Asia, were the result of a very recent host range expansion of a polyphagous Golovinomyces because (i) T. fournieri is absent from our region, except as an experimental plant grown in the laboratory, (ii) the powdery mildew fungus infecting this exotic plant belongs to a group of Golovinomyces where host range expansion is a frequent evolutionary scenario, (iii) cross-inoculation tests showed that this pathogen is also able to infect other plant species, notably A. thaliana and tobacco, and (iv) no Golovinomyces species are known to infect T. fournieri anywhere in the world. Although host range expansion has often been proposed as a common evolutionary process in the Erysiphales, and also in other biotrophic plant pathogens, this has not been clearly demonstrated in any case studies so far. To our knowledge, this is the first convincing case of a host range expansion event in the Erysiphales.     

Differentiation of two powdery mildews of sunflower (<Emphasis Type="Italic">Helianthus annuus</Emphasis>) by a PCR-mediated method based on ITS sequences

Powdery mildew can be found in most sunflower fields during the winter season in Taiwan and causes severe yellowing on the blade, petiole, stem, and calyx, as well as serious defoliation. Two types of powdery mildew fungi isolated from sunflower leaves showed variable status for fibrosin bodies. But only the cleistothecium of Podosphaera xanthii, one of the pathogens causing this disease, was observed on samples from Chungpu County at the beginning of 2005. With a species-specific primer pair, PN23/PN34, no specific PCR product was amplified from the pathogen’s genomic DNA. Based upon the ITS sequence of rDNA, three PCR primer sets (S1/S2, G1/G2, and L1/L2) specific to P. xanthii, Golovinomyces cichoracearum and Leveillula taurica, respectively, were designed to detect and differentiate pathogens causing powdery mildews on sunflower. Only the primer pairs S1/S2 and G1/G2 could amplify PCR products, with product sizes of 454 and 391 bp, respectively. Four samples of fungal DNA were subjected to a multiplex PCR amplification with primer pairs S1/S2 and G1/G2; P. xanthii and G. cichoracearum were successfully detected. These results suggest that the multiplex PCR method is a rapid, simple, and effective technique to detect and differentiate powdery mildews, for example P. xanthii and G. cichoracearum, found on sunflower. With morphologic characteristics, ITS sequence analysis and pathogenicity testing, P. xanthii and G. cichoracearum, the first case, are two powdery mildews on sunflower in Taiwan.     

Characterization of <Emphasis Type="Italic">Phytophthora clandestina</Emphasis> races on <Emphasis Type="Italic">Trifolium subterraneum</Emphasis> in Western Australia

Phytophthora clandestina is a causal agent of root rot disease of subterranean clover in Western Australia (W.A). As a significant number of isolates of P. clandestina from W.A. could not previously be designated using existing differentials, a comprehensive set of subterranean clover (Trifolium subterraneum) cultivars was used as differentials to delineate a broader range of races of the pathogen. One hundred and one isolates of the pathogen collected from W.A. were screened on nine subterranean clover cultivars, of which seven were found to be useful as host differentials. A total of 10 races (in contrast to the five recognized previously) were defined and differentiated using octal nomenclature, presenting a clearer picture of the racial distribution of P. clandestina among W.A. isolates. Differences were found in the race populations between Australian states and are therefore important to the selection/breeding of cultivars for specific regions of Australia to counter the predominant race populations and for enforcing quarantine measures in relation to seed movements within and outside Australia. The octal nomenclature used provides a sound basis for follow-up studies and future race designations. Races 173 and 177 in this study were widely distributed and were the most common races in W.A., and together constitute 80% of the isolates characterized. While six of the seven host differentials were resistant to isolates belonging to race 001 and all were resistant to 000, it is of concern that only one differential was resistant to 157 and 173 and that none of the host differentials were resistant to 177. Our approach to P. clandestina race delineation is clearly conservative and is different from previous studies. The octal nomenclature we applied in this study is not only scientifically sound but also will facilitate rapid recognition and characterization of the races.     

Insecticidal activity of crude seed extracts of<Emphasis Type="Italic">Annona</Emphasis> spp.,<Emphasis Type="Italic">Lansium domesticum</Emphasis> and<Emphasis Type="Italic">Sandoricum koetjape</Emphasis> against lepidopteran larvae

Crude ethanolic seed extracts ofAnnona muricata, A. squamosa (Annonaceae),Lansium domesticum andSandoricum koetjape (Meliaceae) collected from different locations and years in Maluku, Indonesia, were screened for inhibition of larval growth against the polyphagous lepidopteranSpodoptera litura (Noctuidae). Extracts ofA. squamosa were significantly more active (20-fold) than those ofA. muricata. A. squamosa collected from Namlea yielded the extracts with the greatest inhibitory activity. There were significant differences among locations for bothA. squamosa andA. muricata but not forL. domesticum andS. koetjape. Extracts ofA. squamosa, collected from Namlea, inhibited larval growth in a dose-dependent manner, with a dietary EC₅₀ (effective concentration to inhibit growth by 50% relative to controls) of 191.7 ppm fresh weight. Extracts ofA. squamosa collected from individual trees in Namlea also varied in growth inhibitory effect againstS. litura andTrichoplusia ni larvae. This species is a candidate for development of a botanical insecticide for local use in Indonesia. http://www.phytoparasitica.org posting Dec. 1, 2003.     

Quantification of camalexin in several accessions of<Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> following inductions with<Emphasis Type="Italic">Peronospora parasitica</Emphasis> and UV-B irradiation

Five accessions ofArabidopsis thaliana, Ws-3, Nd-1, Ler, Col-5 and Oy-0, were inoculated withPeronospora parasitica isolate Emoy2 and the accumulation of camalexin within infected tissues was measured. The variations in camalexin accumulation in various accessions ofA. thaliana induced by a specific (P. parasitica isolate Emoy2) and a non-specific elicitor (UV-B irradiation) were investigated. Phenotypic examination of Emoy2/Oy-0 revealed that susceptibility was characterized by extensive asexual sporulation of the pathogen, whereas early restriction of the pathogen in infected plant tissues, accompanied by chlorosis and necrosis — which are associated with the hypersensitive response — was observed in Nd-1, Ws-3 and Ler. Partial resistance detected in Col-0 was characterized by low to medium sporulation of the pathogen. Camalexin was monitored by high pressure liquid chromatography (HPLC) and found to accumulate during both compatible and incompatible interactions and also following treatment with the abiotic inducer, UV-B. Among the accessions tested, Ws-3 yielded significantly more camalexin than the other accessions, regardless of which inducers (biotic or abiotic) were used. There was no significant correlation between resistance and camalexin accumulation in theA. thaliana/P. parasitica interaction. The results suggest that there is genetic variation in camalexin biosynthesis among accessions ofA. thaliana, rather than variation in types of induction. http://www.phytoparasitica.org posting Dec. 16, 2002.     

Unsaturated fatty acids from zoospores of <Emphasis Type="Italic">Sclerospora graminicola</Emphasis> induce resistance in pearl millet

Downy mildew of pearl millet, caused by Sclerospora graminicola, is a devastating disease, resulting in high economic losses in the semi-arid regions of the world. Recently, induction of host plant resistance using biotic and abiotic inducers are included among disease management practices as an eco-friendly approach. Unsaturated fatty acids are considered as a new generation of plant disease resistance inducers. In the present study, six unsaturated fatty acids, viz. docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), arachidonic acid (AA), linolenic acid, linoleic acid and oleic acid, all originally detected in the zoospores of S. graminicola,were applied to seeds of susceptible cultivars of pearl millet to examine their ability to protect against downy mildew under greenhouse and field conditions. In greenhouse experiments, EPA and AA induced a maximum of 78.6% and 76.5% protection, whereas linoleic acid, DHA and linolenic acid provided up to 66.3%, 61.2% and 24.5% protection, respectively. Oleic acid was not effective in protecting pearl millet (only 5.1% protection). A time interval of four days between treatment of seeds and challenge inoculation was required to obtain optimum protection. Plants raised from treated seeds and challenge inoculated at the tillering and inflorescence stages showed enhanced resistance, resulting in higher grain yield compared to untreated plants of the same cultivar. Chitinase activity was found to be higher in susceptible seedlings of pearl millet after treatment with the fatty acids and pathogen inoculation than in seedlings only inoculated with the pathogen. This indicates that host defence responses are activated and thus that induced resistance is involved in the protection observed. The role of unsaturated fatty acids as activators of resistance against downy mildew in pearl millet is discussed.     

Synergistic interaction between BABA and mancozeb in controlling<Emphasis Type="Italic">Phytophthora infestans</Emphasis> in potato and tomato and<Emphasis Type="Italic">Pseudoperonospora cubensis</Emphasis> in cucumber

Spray mixtures consisting of the plant activator BABA (DL-3-aminobutyric acid) and the protectant fungicide mancozeb were significantly more effective than BABA or mancozeb alone in controlling late blight (Phytophthora infestans) in potato and tomato and downy mildew (Pseudoperonospora cubensis) in cucumber. A mixture composed of 5 parts BABA and 1 part mancozeb (w/w, a.i.) exhibited a higher synergy factor than the 1+1 or the 1+5 (BABA + mancozeb) mixtures. No synergistic interaction was detected between BABA plus mancozeb in controlling sporangial or cystospore germination, nor mycelial growth ofP. infestans in vitro. The results showed enhanced effect of mancozeb in BABA-induced plants, suggesting, therefore, that lower dosages of this fungicide may be sufficient to control late blight or downy mildew under field conditions. http://www.phytoparasitica.org posting July 15, 2003.     

Note: Tachinid parasitoids of<Emphasis Type="Italic">Ancyrosoma leucogrammes</Emphasis> and notes on parasitization rates of<Emphasis Type="Italic">Clytiomya dupuisi</Emphasis>

The study deals with the tachinid parasitoids ofAncyrosoma leucogrammes (Gmelin) (Heteroptera: Pentatomidae):Clytiomya dupuisi Kugler,Clytiomya sola (Rondani) andGymnosoma clavatum (Rohdendorf) (Diptera: Tachinidae). All species are recorded for the first time as parasitoids ofA. leucogrammes, andC. dupuisi was reared for the first time from a host. The parasitization rates ofC. dupuisi on adults ofA. leucogrammes varied from 7% to 9% between 1994 and 1999.C. sola andG. clavatum were reared in only small numbers. http://www.phytoparasitica.org posting July 10, 2003.