Tight regulation of allene oxide synthase (AOS) and allene oxide cyclase-3 (AOC3) promote <Emphasis Type="Italic">Arabidopsis</Emphasis> susceptibility to the root-knot nematode <Emphasis Type="Italic">Meloidogyne javanica</Emphasis>

Biosynthesis of the oxylipin jasmonic acid (JA) in Arabidopsis thaliana is catalyzed by a single allene oxide synthase (AOS)-encoding gene and four genes encoding four functional allene oxide cyclase (AOC) polypeptides (AOC1, AOC2, AOC3, and AOC4). To elucidate the biological activities of the JA pathway in regulating the plant defense response to plant-parasitic nematodes, transgenic lines carrying the GUS reporter gene under the control of individual AOC or AOS promoters were examined. Upon penetration by second-stage juveniles (J2 s), promoter activities of AOC1, AOC3 and AOC4 appeared in the root tip and root-elongation zone, with AOC3 demonstrating highest induction. At 5 days AOC3 activity continued to be highly pronounced in the stele and root cortex, associated with nematode invasion throughout gall initiation and maturation. AOS expression appeared 3 days postinfection and accompanied all later infection stages. Mutant lines were analyzed: disruption in AOS rendered plants more resistant to nematode infection, as reflected by the decreased number of females produced on this line; loss-of-function of AOC3 rendered plants more susceptible to nematode infection. Oxylipins derived from the 9- and 13-lipoxygenase pathways were assayed for their direct inhibitory activity toward M. javanica J2 s. Clear nematicidal activity of the bioactive 9- and 13-hydroperoxides was observed. Oxylipins produced by divinyl ether synthase, colneleic acid, colnelenic acid and ω5(Z)-etherolenic acid demonstrated strong inhibitory activity. These data, along with those of other assayed oxylipins, suggest that temporal and spatial fine tuning of the JA route allowing nematodes parasitism on plant host.     

Antifungal activity of <Emphasis Type="Italic">Biscogniauxia</Emphasis> sp. culture filtrates against the rice pathogen <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

An ethyl acetate extract of a culture filtrate (ECF) from an unidentified fungal isolate O821 was evaluated for antifungal activity against the rice pathogen Magnaporthe oryzae. The O821-ECF significantly inhibited spore germination, appressorium formation, and mycelial growth of M. oryzae, and its antifungal activity was heat-stable. It also significantly suppressed the number and size of blast lesions. In an analysis of the ITS sequence of this isolate, it shared similarities with species of the fungus Biscogniauxia. These results suggest that isolate O821 of the genus Biscogniauxia produces a heat-stable antifungal compound(s) in its culture filtrate.     

Screening for Antifungal Potential of Plant Extracts of Fifteen Plant Species Against Four Pathogenic Fungi Species

The antifungal activity of fifteen wild plant species grown in the Al-Qassim region, Saudi Arabia, was investigated against the fungi species Fusarium solani, Botrytis cinerea, Alternaria alternata and Stemphylium botryosum at concentrations of 10, 20, 40 and 80?g/L. The inhibitory effect of test extracts varied among examined fungi, and Fusarium solani exhibited the least sensitivity compared to other fungi species. Antifungal activity of the tested extracts was proportional to the applied dose. Extracts of Lactuca virosa, Neurada procumbens, Ochradenus baccatus and Cyperus conglomerates showed relatively low inhibition effects on the fungi species. The most effective plants were Pulicaria undulata, Artemisia monosperma, Prosopis juliflora, Withania somnifera and Rumex vesicarius. At 80?g/L, extracts of these plant species reduced mycelial growth of Botrytis cinerea by 16.5–32.2%, of Fusarium solani by 11.1–27.9%, of Alternaria alternaae by 26.9–63.5% and of Stemphylium botryosum by 22–40%. The methanolic extract of the most effective plant species was further fractioned with hexane, methylene chloride and ethyl acetate. The obtained fractions varied in their effects on mycelial growth of the four tested fungi. Using the same fraction resulted in different inhibition effects on mycelial growth of all tested fungi. The antifungal activity of each crude extract tended to be distributed among its three fractions, probably because the bioactive components were also distributed among the fractions.     

Assessment of the antifungal activity of selected biocontrol agents and their secondary metabolites against <Emphasis Type="Italic">Fusarium graminearum</Emphasis>

Fusarium graminearum (teleomorph: Gibberella zeae) is the causal agent of several destructive diseases in cereal crops worldwide. In the present study we have evaluated the potential of two strains of Trichoderma sp. (T23, and T16), a strain of Paecilomyces sp. (PS1), and their secondary metabolites (SMs) in suppressing F. graminearum. Results from dual culture experiments show that in the presence of either Trichoderma sp., or Paecilomyces sp. mycelial growth of F. graminearum is considerably inhibited. Strain T23 causes the greatest inhibition (83.8%), followed by strain T16 (72.2%), and strain PS1 (61.9%). Likewise, mycelial growth of the pathogen is completely inhibited (≥ 98%) when grown under exposure to volatile metabolites excreted from Trichoderma cultures. Bioautographic analyses using culture filtrates revealed that several antifungal SMs are excreted. Among five metabolites tested, 6-pentyl-alpha-pyrone (6PAP) from strain T23, and PF3 from strain PS1 exhibit pronounced antifungal activity against F. graminearum. A new method for mass production of perithecia of F. graminearum which is simple and more effective than traditional methods was developed, which allows an increase in perithecial formation of more than 5-fold. Using this method, we found, that in the presence of SMs perithecial formation was negatively affected. Perithecial production was suppressed by 81.4% and 76.6% using 200 μg ml^?1 of either 6PAP or PF3, respectively. Moreover, ascospore discharge was significantly suppressed (67.0%) when perithecia were exposed to the metabolite F116 produced by T16. Including 6PAP or PF3 in conidial suspensions impeded germination of conidia completely. Similarly, both metabolites strongly inhibited ascospore germination (? 90%).     

Inhibitory effect of <Emphasis Type="Italic">Lycium europaeum</Emphasis> extracts on phytopathogenic soil-borne fungi and the reduction of late wilt in maize

The ability to control soil-borne pathogens in agriculture is highly conditioned by the restricted use of synthetic pesticides. Allelopathy, the antimicrobial activity of plant extracts, is a promising option against crop pathogens. Extracts from Lycium spp. such as L. barbarum, L. chinense and L. intricatum possess biological and therapeutic properties. Individual methanolic extracts from leaves and stems of the Mediterranean medicinal species L. europaeum collected in two locations of Tunisia were each evaluated in vitro against Verticillium dahliae (Vd), Sclerotinia sclerotiorum (Ss) and Harpophora maydis (Hm). The mycelial growth of the three fungi was significantly reduced by all the extracts at doses of 10 and 30 μl mL^?1 (equivalent to 1 and 3 mg plant tissue mL^?1). The sporulation of Hm was almost completely inhibited in all the amendments, but that of Vd was stimulated by one of the leaf extracts when 1 and 3 mg dried plant tissue mL^?1 were used. Sclerotia of Ss were formed in a smaller number, their total weight increasing at extract doses equivalent to 1 mg plant tissue mL^?1 and higher. In greenhouse, the pathogenicity of Hm was confirmed as early as 6 weeks after inoculation, since it caused significant decreases of weights in both roots and aboveground parts of maize. The detrimental effect of Hm on maize root weight in greenhouse was significantly counteracted by one of the leaf extracts added by watering. In total, 11 phenolic compounds were separated in the four extracts. The hydroxycinnamic acid family, including chlorogenic acid as a major compound, represented more than 50% of the total content in all the samples. Rutin was the most abundant flavonoid. The results of this work show the detrimental effect of L. europaeum extracts against the soil-borne pathogens Hm, Ss and Vd, and highlight their potential in crop protection if adequately developed into final products and used in combination with other tools.     

Mechanism of in vitro antagonism of phytopathogenic <Emphasis Type="Italic">Scelrotium rolfsii</Emphasis> by actinomycetes

Sclerotium rolfsii (Sr), a soil-borne fungal pathogen, causes disease in a wide range of crops. Recently, we identified five actinomycetes (Streptomyces globisporus subsp. globisporus, S. globisporus, S. flavotricini, S. pactum, and S. senoensis) showing significant inhibitory effects on plant pathogens. In this study, the effects of the five actinomycetes for the biocontrol of Sr were investigated using the plate culture method and microscopy examination. Two actinomycetes with higher inhibitory effect were subsequently examined for the inhibition of sclerotial germination of Sr in unsterile soil in vitro. The cell-free cultures of five actinomycetes mediated significant inhibition of hyphal growth and sclerotial formation and germination of Sr. All actinomycete strains exhibited the ability to produce extracellular cell wall degrading enzymes in the culture conditions. The crude enzyme suspensions of S. flavotricini and S. pactum hydrolyzed the cell wall of Sr. At a dose of 1 g per kilogram soil, the solid formulations of S. flavotricini and S. senoensis prevented germination of 24% and 68% of sclerotia, respectively. Our results provide evidence of effective strains for the biocontrol of Sr, in addition to a further understanding of the underlying mechanism.     

Influence of exogenous cholesterol in Pythiaceae resistance to inhibition by <Emphasis Type="Italic">Trichoderma</Emphasis> antibiosis

The purpose of this study was to determine if exogenous cholesterol availability influenced Pythiaceae resistance to antibiosis. Characterisation of an isolate of Phytophthora erythroseptica and Pythium ultimum for tolerance to antibacterial compounds found that 0.05 g.l^?1 chloramphenicol inhibited mycelial growth by 96.6 % and 23.5 % respectively. However, the addition of cholesterol (0.01 g l^?1) to potato dextrose agar (PDA) containing 0.05 g l^?1 chloramphenicol was found to increase mycelial growth of P. erythroseptica, indicating a role for cholesterol in tolerance to inhibitory antibacterial compounds. To determine if this property extended to suppressive effects of a potential biocontrol agent, P. erythroseptica and P. ultimum were then tested against a cell-free filtrate of diffusible metabolites produced by a suppressive Trichoderma harzianum isolate in the presence and absence of cholesterol in PDA. In the absence of cholesterol, diffusible metabolites of the T. harzianum isolate were found to inhibit mycelial growth of P. erythroseptica and P. ultimum on PDA by 98 % and 63.6 % respectively (P?<?0.0001). However, the inhibitory effect of the metabolites was mitigated when 0.005 g l^?1 of cholesterol was present in PDA, with mycelial growth of P. ultimum and P. erythroseptica reduced by only 60.4 % and 41.8 %, respectively (P?<?0.0001), much less inhibition than was observed in the absence of cholesterol. These results demonstrated that access to exogenous cholesterol can influence the sensitivity of Pythiaceae species to antibiosis by positively influencing mycelial growth.     

Control of infection of tomato fruits by <Emphasis Type="Italic">Alternaria</Emphasis> and mycotoxin production using plant extracts

Tomato fruits are susceptible to infection by Alternaria species. In addition, Alternaria species may contaminate the fruits with mycotoxins. There is thus interest in control systems to minimise pathogenicity and control toxin production. The objectives of this study were to examine the effect of plant extracts of Eucalyptus globulus and Calendula officinalis on the growth of strains of Alternaria alternata and A. arborescens, on pathogenicity of tomato fruits and mycotoxin production. The growth bioassays showed that the ethanolic and chloroformic fractions of E. globulus were the most effective in reducing growth of A. alternata (66–74 %) and A. arborescens (86–88 %), respectively at 2500 μg/g. The effects of plant extracts on mycotoxin biosynthesis were variable and strain dependent. The most effective fractions in decreasing mycotoxin accumulation were the ethanolic and chloroformic extracts of E. globulus, which reduced tenuazonic acid by 89 %, alternariol by 75–94 % and almost complete inhibition of alternariol monomethyl ether. All the tested fractions reduced percentage of infected tomato fruits when compared to the controls. The ethanolic and chloroformic fractions of E. globulus completely inhibited growth of A. alternata and A. arborescens on unwounded fruits and reduced the aggressiveness on wounded fruits of strains of both species significantly.     

Antifungal activity of a novel compound purified from the culture filtrate of <Emphasis Type="Italic">Biscogniauxia</Emphasis> sp. O821 against the rice blast fungus <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis>

Rice blast is a devastating fungal disease resulting in major losses to rice crops. Owing to continuous acquisition of resistance by the causal fungus, several fungicide chemicals are no longer effective. Therefore, there is a need to identify natural components and develop new agents to control fungal pathogens. We previously demonstrated that the culture filtrate of Biscogniauxia sp. O821 inhibited infection behavior of Magnaporthe oryzae and subsequent blast lesion formation. In the present study, we isolated a new compound, (3aS,4aR,8aS,9aR)-3a-hydroxy-8a-methyl-3,5-dimethylenedecahydronaphto[2,3-b]furan-2(3H)-one (HDFO), from the culture filtrate of Biscogniauxia sp. O821 and determined its molecular weight as 248. The HDFO structure was determined by electrospray ionization-mass spectrometry and nuclear magnetic resonance spectroscopy after purification with column chromatography and high-performance liquid chromatography. The structure of this antifungal compound was similar to that of alantolactone and isoalantolactone. The growth inhibition zone against M. oryzae in presence of HDFO was observed at Rf 0.5–0.6 on a thin layer chromatography plate. HDFO inhibited conidial germination of M. oryzae in a dose-dependent manner (1–200 ppm). Furthermore, blast lesion formation was significantly suppressed by HDFO at over 5 ppm. These results suggest that HDFO from the culture filtrate of Biscogniauxia sp. O821 can protect rice from rice blast disease caused by M. oryzae. This is the first report that HDFO produced by Biscogniauxia sp. can serve as an antifungal compound against M. oryzae.     

The potential of fludioxonil for anthracnose control on China chili fruit

Anthracnose, caused by Colletotrichum scovillei, is one of the most destructive disease which causes massive yield losses in chili. Our preliminary study has demonstrated that fludioxonil had high activity against mycelial growth and spore germination of C. scovillei but it has not been labeled on chili. The aim of this study was to investigate the impact of fludioxonil on C. scovillei infection. In greenhouse and field trials, fludioxonil was applied to chili plants to determine the duration of protection of fruit provided by this compound against chili anthracnose. Fludioxonil may have impact on the early stage of chili anthracnose infection by inhibiting the germ tube elongation and appressorium formation. Moreover, it inhibited the secondary infection of C. scovillei by inhibiting the sporulation, germination of spores and formation of appressoria. Fludioxonil provided high level of protective activity for up to 96 h, as well as excellent curative activity of within 24 h in vivo tests. In field trials, fludioxonil can reduce the incidence and severity of chili anthracnose while giving a higher chili yield. These results suggest that fludioxonil could be a promising fungicide for anthracnose control in chili production.     

Mapping of easy to screen SSR markers for selection of RFLP markers-bracketed downy mildew resistance QTLs in pearl millet

There is a growing necessity to replace chemical agents with ecofriendly materials, arising from the impact on the environment and/or human health, which calls for the design of new broad-spectrum fungicides. In this work, chitosan oligomers (COs), propolis (Ps) and silver nanoparticles (AgNPs) mixtures in solution were assessed to control the growth of different phytopathogenic fungi and oomycetes in vitro. Binary solutions of COs-Ps and COs-AgNPs evinced the highest antifungal effect against Fusarium circinatum and Diplodia pinea fungi, respectively, with a ca. 80% reduction in their mycelial growth. The COs solution by itself also proved to be greatly effective against Gremmeniella abietina, Cryphonectria parasitica and Heterobasidion annosum fungi, causing a reduction of 78%, 86% and 93% in their growth rate, respectively. Likewise, COs also attained a 100% growth inhibition on the oomycete Phytophthora cambivora. On the other hand, Ps inhibited totally the growth of Phytophthora ×alni and Phytophthora plurivora. The application of AgNPs reduced the mycelial growth of F. circinatum and D. pinea. However, the AgNPs in some binary and ternary mixtures had a counter-productive effect on the anti-fungal/oomycete activity. In spite of the fact that the anti-fungal/oomycete activity of the different treatments showed a dependence on the particular type of microorganism, these solutions based on natural compounds can be deemed as a promising tool for control of tree diseases.     

Effect of boron on mycelial growth,sporangiogenesis and zoosporogenesis of <Emphasis Type="Italic">Phytophthora nicotianae</Emphasis> and the possible inhibitory mechanisms

Phytophthora nicotianae is one of the most important soil-borne plant pathogens. Sporangia and zoospores of P. nicotianae are responsible for primary infection and disease dissemination. The disease caused by P. nicotianae was difficult to control by fungicide. Boron, an essential plant micronutrient, was found to have a direct effect on other pathogens. In this paper, the effects of B on the growth, antioxidant system and gene differential expression of P. nicotianae were tested. The results showed that 0.1 mM B could dramatically decrease the sporangiogenesis and zoosporogenesis of P. nicotianae. Mycelial growth of P. nicotianae was significantly inhibited when the concentration of B reached 8 mM. A high-quality differential expression sequence csn4 was obtained by gene differential expression analysis. Under the treated of B, csn4 expression was inhibited, activity of superoxide dismutase (SOD) and catalase (CAT) significantly decreased and the malondialdehyde (MDA) content notably increased compared to control. It is suggested that B could serve as a potential fungicide for the control of plant disease caused by P. nicotianae.     

Polyphenols and natural resistance of apple leaves against Venturia inaequalis

Phloridzin was oxidized via 3-hydroxyphloridzin to the correspondingo-quinone by a polyphenoloxidase present in homogenates and in acetone powders of apple leaves. Phloridzin could also be hydrolyzed by a β-glucosidase to phloretin, which was oxidized, although at a lower rate, to 3-hydroxyphloretin and its correspondingo-quinone. Phloretin and its oxidation products can couple with theo-quinone of 3-hydroxyphloridzin.Oxidation products of phloridzin inactivated pectinase. Moreover, the intermediate oxidation products showed fungicidal activity. Apparently, in this way apple leaves possess a defense mechanism against those organisms which are able to cause cell collapse resulting in phloridzin oxidation. Leaves of plants with a hypersensitive reaction to infection byVenturia inaequalis were also highly sensitive to toxic proteins secreted by this fungus in vitro.     

Biocontrol of <Emphasis Type="Italic">Fusarium</Emphasis> wilt of <Emphasis Type="Italic">Capsicum annuum</Emphasis> by rhizospheric bacteria isolated from turmeric endowed with plant growth promotion and disease suppression potential

In the present study, 129 rhizospheric bacteria isolated from Curcuma longa were screened for their antagonistic potential against six fungal phytopathogens. Among them, 32 isolates that showed significant antagonistic potential were screened for their in vitro plant growth promoting (PGP) traits. The identification of potential isolates was confirmed by 16S rRNA gene sequencing and results revealed Bacillus as the dominant genus followed by Staphylococcus, Pseudomonas, Sphingomonas and Achromobacter. Based on the antagonistic activity and PGP traits; two strains (BPSRB4 and BPSRB14), identified as Bacillus amyloliquefaciens, were further tested for their in vivo PGP and disease suppression potential on Capsicum annuum seedlings under greenhouse conditions. The results demonstrated that BPSRB4 and BPSR14 strains suppress fungal pathogen infection and promote plant growth. Further, the BPSRB4 strain was positive for the production of the phytohormone indole acetic acid (IAA) detected by thin layer chromatography (TLC). In addition, nitrogen fixation and plant growth promotion activity were also confirmed by amplification and sequencing of nitrogen fixation gene (nifH) and ACC (1-aminocyclopropane-1-carboxylate) deaminase (acdS) gene from strains BPSRB4 and BPSRB14. The present study demonstrated that the B. amyloliquefaciens strains BPSRB4 and BPSR14 possess antagonistic activity and PGP potential which could be explored for the development of biofertilizers and biocontrol agents for the growth of chilli seedlings.     

Postharvest evaluation of natural coatings and antifungal agents to control <Emphasis Type="Italic">Botrytis cinerea</Emphasis> in <Emphasis Type="Italic">Rosa</Emphasis> sp.

Botrytis cinerea is a fungal pathogen that limits rose production and commercialization worldwide. Therefore, we evaluated a novel postharvest treatment against Botrytis cinerea in roses (Rosa sp. cv Vendela) using coating bases and antifungal agents of natural origin. Aloe vera pulp, cassava starch and gelatin were used as coating bases. Oregano essential oil (Origanum vulgare), thyme essential oil (Thymus vulgaris) and chitosan were used as natural antifungal agents. The coating bases were evaluated in different concentrations to observe effects of toxicity and opening diameter in rose buds. Gelatin and cassava starch coatings inhibited rose opening and showed petal damage in all concentrations tested. However, Aloe vera pulp at 25% allowed normal buds’ opening and no damage was observed, indicating that Aloe vera could be an ideal coating base for rose postharvest treatments. During in vitro assays, natural antifungal agents efficiently inhibited Botrytis cinerea growth in the concentrations tested. Further, mixture treatments of Aloe vera pulp (25%) with oregano essential oil (1%), thyme essential oil (0.1%) and chitosan (0.1%) showed independently neither damage nor opening inhibition in rose buds. Selected combinations of Aloe vera pulp and natural antifungal agents were applied in roses infected with Botrytis cinerea to evaluate their control of this pathogen. Unfortunately, the selected combinations did not reduce pathogen growth during postharvest treatments since they were similar to untreated controls. Further research has to be performed to find ideal combinations with Aloe vera that could inhibit B. cinerea during postharvest treatments in roses.     

Efficacy of phosphonate in controlling white powdery rot of fig caused by <Emphasis Type="Italic">Phytophthora palmivora</Emphasis>

Pre- and post-infection activities of phosphonate and fungicides (azoxystrobin, chlorothalonil, cyazofamid, copper hydroxide, and copper sulfate) against white powdery rot on fig leaves caused by Phytophthora palmivora were determined. Phosphonate and fungicides were applied at 3, 7, or 14 days before inoculation and 16, 24, or 40 h after inoculation. Phosphonate and four fungicides (except for chlorothalonil) had protectant activity for up to 14 days before inoculation. Protectant activity between phosphonate and the four fungicides did not differ significantly. These fungicides did not have a curative activity, whereas only phosphonate had a curative activity when applied 16 h after inoculation. The pre-infection activity of phosphonate was determined in fig fields. Phosphonate provided excellent protectant activity, and its activity did not differ significantly from that of the copper fungicides. The activity of phosphonate in the life cycle of P. palmivora was compared with that of the five fungicides. Half-maximal effective concentration of the inhibition of mycelial growth was low for phosphonate, chlorothalonil, and the two copper fungicides. Mycelial growth was least affected by azoxystrobin and cyazofamid, but n-propyl gallate, an alternative oxidase inhibitor, in combination with these fungicides, significantly inhibited mycelial growth. Sporangium formation was sensitive to all compounds. Germination of encysted zoospores was sensitive to azoxystrobin, cyazofamid, and chlorothalonil and least sensitive to phosphonate and the two copper fungicides. The activity of phosphonate on mycelial growth and sporulation of P. palmivora suggests it has potential for controlling the disease.     

Phytotoxin produced by the netted scab pathogen, <Emphasis Type="Italic">Streptomyces turgidiscabies</Emphasis> strain 65, isolated in Sweden

Streptomyces spp. are a highly diverse group of bacteria most of which are soil-inhabiting saprophytes. A few are plant pathogens that produce a family of phytotoxins called thaxtomins and cause significant economic losses, e.g., by reducing the marketability of potato tubers (Solanum tuberosum). In northern Europe, S. scabies, S. turgidiscabies and S. europaeiscabiei are the most common plant pathogenic species. In this study, a Streptomyces strain isolated from a netted scab lesion on a tuber of potato cv. Bintje in northern Sweden was identified as S. turgidiscabies but was found to differ in the genomic region carrying genes required for thaxtomin biosynthesis. Our results showed that the strain did not produce thaxtomin but rather phytotoxin fridamycin E, which is an anthraquinone novel to plant pathogenic Streptomyces spp. Fridamycin E was shown to reduce or inhibit sprouting of potato microtubers in vitro. While fridamycin E is known to have antibiotic activity against Gram-positive bacteria, the inhibitory activity of fridamycin E on plant growth is a novel finding.     

Effects of triazole fungicides on sterol biosynthesis during spore germination of Botrytis cinerea,Venturia inaequalis and Puccinia graminis f. sp. tritici

With three plant pathogens,Botrytis cinerea, Venturia inaequalis and Puccinia graminis f. sp.tritici, the time course of sterol biosynthesis during spore germination was examined by labeling experiments along with the question whether this pathway could be inhibited by triazole fungicides. Conidia ofB. cinerea andV. inaequalis are able to synthesize sterols immediately after the beginning of the germination process when the germ tubes have not yet emerged. On the contrary uredospores ofP. graminis start sterol biosynthesis after 6 to 8 h germination time almost at the end of the germ tube phase, indicating that sterol reserves of the spores are likely to be used for the germ tube growth.The sterol C-14 demethylation appeared to be the rate limiting step within the sterol biosynthetic pathway: the half life of 24-methylenedihydrolanosterol was less than 1 h forB. cinerea. It was more than 1 h forV. inaequalis and 3 h forP. graminis. Independent of these differences in the time course of sterol biosynthesis and in the C-14 demethylation rate, the synthesis of sterols in germinating spores was strongly inhibited by triazole fungicides in all three pathogens examined. In contrast toP. graminis, this inhibition could be demonstrated withB. cinerea andV. inaequalis even in ungerminated conidia, indicating that the fungicides were rapidly taken up and reached their target within 1 or 2 h. These results are discussed along with the question whether spore germination can be used as a bioassay for the estimation of sensitivities of triazole fungicides.     

Comparative colonisation by virulent versus avirulent <Emphasis Type="Italic">Pyricularia oryzae</Emphasis> on wild <Emphasis Type="Italic">Oryza australiensis</Emphasis>

Pyricularia oryzae (rice blast) conidial development at pre-penetration stage determines success or otherwise of infection inside the rice host plants. Studies on conidial germination and growth on the leaf surface in commercial rice (Oryza sativa) report differently, dependent upon host type and level of blast resistance. Although wild rice (O. australiensis) is known to be an alternative host of blast, the interaction between P. oryzae conidia and wild O. australiensis on its leaf surface has not been previously studied. We found significant (P?<?0.001) differences in conidial development between two blast isolates with different virulence in terms of conidial germination, germ tube growth and appressoria formation on both wild and cultivated rice. Conidial germination at 6 h post-inoculation (hpi) for the virulent isolate was significantly (P?<?0.001) delayed. Germ tubes of the avirulent isolate conidia grew significantly (P?<?0.001) faster and with significantly (P?<?0.001) longer germ tubes than from virulent conidia. Appressoria development for the virulent isolate was significantly (P?<?0.001) faster at its later growth stages of 12 and 18 hpi when approximately 100% of germ tubes formed appressoria. In contrast, formation rate of appressoria for the avirulent isolate was significantly (P?<?0.001) slower and only reached 76% of germ tubes forming appressoria. Appressoria formation on O. australiensis was significantly (P?<?0.001) greater than the formation on O. sativa for both virulent and avirulent P. oryzae at 12 hpi, a clear indication that host type influences the extent of appressoria formation.     

Interaction between polygalacturonase-inhibiting protein and jasmonic acid during defense activation in tomato against <Emphasis Type="Italic">Botrytis cinerea</Emphasis>

Oligogalacturonic acids (OGAs) generated from in vitro interaction between fungal polygalacturonase (PG) and bean PG-inhibiting protein (PGIP) were shown to activate phytoalexin biosynthesis in soybean. Based on this observation, it was hypothesized that PGIP-dependent generation of OGAs activates plant defence responses in vivo. We tested the hypothesis that PGIP activates jasmonic acid-dependent responses to pathogens. For this purpose, a population of tomato plants segregating for a mutation in the jasmonate receptor CORONATINE INSENSITIVE1 (coi1) and for overexpression of pear PGIP (pPGIP) was challenged with Botrytis cinerea. The coi1 mutant was hypersensitive to B. cinerea, but overexpression of pPGIP in the coi1 mutant background reduced pathogen susceptibility, suggesting that these two genes independently alter defence responses. In addition, pPGIP overexpression suppressed pathogen induction of salicylic acid in the coi1 mutant and activated expression of acidic ß-1,3-glucanase independently of the coi1 mutation. However, expression of proteinase inhibitor II (PIN II) in pPGIP overexpressing tomato plants was dependent on COI1. Effects of pPGIP overexpression on defence are therefore complex and only in the case of PIN II pPGIP acts through COI1.