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.     

Efficacy of a formulated product containing <Emphasis Type="Italic">Quillaja saponaria</Emphasis> plant extracts for the control of root-knot nematodes

The nematicidal effect of a formulated product containing extract from Quillaja saponaria was evaluated against the root-knot nematodes. The product QL Agri® 35 (QL) was tested to record the effect on second stage juveniles motility, egg hatch and also against field populations in greenhouse experiments contacted in three different locations of Greece. Convulsive movement of second stage juveniles of Meloidogyne incognita was recorded after exposure for 8 days at a series of doses, while the most paralyzed juveniles were counted at the dose of 8 mg l^?1. There was also a gradual decrease in the number of juveniles emerging from egg masses of the same nematode species when the dose of Q. saponaria was increased from 0 to 8 mg l^?1. In greenhouse experiments, the use of Q. saponaria could control root-knot nematodes and prevent nematodes increase in soil. The present study demonstrates that the use of Q. saponaria extract has the ability to control root-knot nematodes. Control given by Q. saponaria in field populations infecting cucumber was similar to that of cadusafos (Rugby®) and oxamyl (Vydate®) under the tested dosages and the specific conditions of the experiments.     

Toxicity of selected insecticides to <Emphasis Type="Italic">Trichogramma chilonis:</Emphasis> Assessing their safety in the rice ecosystem

Nine insecticides, namely, imidacloprid, thiamethoxam, chlorantraniliprole, clothianidin, pymetrozine, ethofenprox, BPMC, endosulfan, acephate, and the product Virtako® (Syngenta; chlorantraniliprole 20%?+?thiamethoxam 20%) were tested to determine their toxicity to the parasitoid Trichogramma chilonis using an insecticide-coated vial (scintillation) residue bioassay. All the insecticides tested showed different degrees of toxicity to the parasitoid. Thiamethoxam showed the highest toxicity to T. chilonis with an LC₅₀ of 0.0014 mg a.i. l ^?1, followed by imidacloprid (0.0027 mg a.i. l ^?1). The LC₅₀ values of acephate and endosulfan were 4.4703 and 1.8501 mg a.i. l ^?1, exhibiting low toxicity when compared with other insecticides tested. Thiamethoxam was found to be 3,195, 1,395 and 1,322 times more toxic than acephate, chlorantraniliprole and endosulfan, respectively, as revealed by the LC₅₀ values to T. chilonis. Based on risk quotient, which is the ratio between the field-recommended doses and the LC₅₀ of the beneficial, only chlorantraniliprole was found to be harmless to T. chilonis. The insecticides thiamethoxam, imidacloprid, Virtako®, ethofenprox and BPMC were found to be dangerous to the parasitoid. Since T. chilonis is an important egg parasitoid of leaf folders, reported to reduce the pest population considerably and often released augmentatively in rice IPM programs, the above noted dangerous chemicals should be avoided in the rice ecosystem.     

Multi-resistance to thiophanate-methyl,diethofencarb, and procymidone among <Emphasis Type="Italic">Alternaria alternata</Emphasis> populations from tobacco plants,and the management of tobacco brown spot with azoxystrobin

In 2014 and 2015, a total of 151 tobacco brown spot (Alternaria alternata) isolates were collected from Guizhou Province in China to evaluate their resistance to the benzimidazole thiophanate-methyl, the carbamate diethofencarb, and the dicarboximide procymidone. Resistance to thiophanate-methyl and diethofencarb was observed in all isolates. Resistance to all the three fungicides, thiophanate-methyl, diethofencarb, and procymidone was detected at a frequency of 6.0%. The F167Y single mutation in the β-tubulin gene was found to be associated with resistance to thiophanate-methyl,but no mutation was found in the coiled-coil region of the histidine kinase-encoding gene OS1, a fungal gene for dicarboximide resistance. Procymidone applied at the rate of 20 mg l^?1 inhibited spot lesion formation on tobacco leaves with an efficacy of 51.7% for the low resistance (LR) isolates and 74.2% for the procymidone-sensitive isolates. Thiophanate-methyl applied at 100 mg l^?1, however, slightly promoted the expansion of disease lesions with an efficacy of ?7.7%. Azoxystrobin applied at 10 and 20 mg l^?1 provided efficacies of 91.1 and 100%, respectively, regardless of whether the isolates were thiophanate-methyl resistant or procymidone-LR. Further studies suggested that azoxystrobin exhibited excellent protective activity and good curative activity against A. alternata in plants. The baseline sensitivity to azoxystrobin was then determined. In the presence SHAM, the mean EC₅₀ values for conidial germination inhibition were 0.49?±?0.22 (Mean?±?SD) mg l^?1. Interestingly, no resistance was recovered through UV irradiation or Agrobacterium tumefaciens-mediated mutagenesis. This research indicated widespread resistance to thiophanate-methyl and diethofencarb, low frequency of (6.0%) resistance to procymidone in A. alternata populations from tobacco, and suggested that azoxystrobin could potentially constitute a good alternative for the management of tobacco brown spot disease.     

Fungicidal activity of Thai medicinal plant extracts against <Emphasis Type="Italic">Alternaria brassicicola</Emphasis> causing black spot of Chinese kale

Crude ethanol extracts and six organic solvent fractions of 10 Thai medicinal plants were evaluated for their antifungal activity against Alternaria brassicicola in laboratory and under greenhouse conditions. The results showed that the ethanol extracts of Coscinium fenestratum, Piper betle, Syzygium aromaticus and Zingiber cassumunar displayed complete mycelial growth inhibition of A. brassicicola at a concentration of 0.1%. Meanwhile, the crude ethanol extract and methanol fraction obtained from the stems of C. fenestratum revealed the greatest inhibition against A. brassicicola at 10%, forming inhibition zones 2.55–2.58 cm in diameter. In the greenhouse experiments, crude ethanol extracts of C. fenestratum and P. betle at 1% significantly (P?<?0.05) reduced the disease incidence at up to 67%, indicating promising preventive and curative activities against A. brassicicola. This activity is similar to that of iprodione, a widely used commercial fungicide. Interestingly, Illicium verum extract showed a greater curative effect (58% disease reduction) than protective effect (47% disease reduction). Because the C. fenestratum extract showed the highest activity against the black spot pathogen both in vitro and under greenhouse conditions, its methanol fraction was further analyzed by spectroscopic techniques. We found that berberine is a key active substance inhibiting mycelial growth of A. brassicicola. The results of this study showed the potential of Thai medicinal plants as alternatives to the use of synthetic fungicides for controlling black spot in Chinese kale caused by A. brassicicola.     

Natural occurrence of <Emphasis Type="Italic">Fusarium</Emphasis> species and fumonisin on maize grains in Ethiopia

Fusarium species causing maize kernel rot are major threats to maize production, due to reduction in yield as well as contamination of kernels by mycotoxins that poses a health risk to humans and animals. Two-hundred maize kernel samples, collected from 20 major maize growing areas in Ethiopia were analyzed for the identity, species composition and prevalence of Fusarium species and fumonisin contamination. On average, 38 % (range: 16 to 68 %) of maize kernels were found to be contaminated by different fungal species. Total of eleven Fusarium spp. were identified based on morphological characteristics and by sequencing the partial region of translation elongation factor 1-alpha (EF-1α) gene. Fusarium verticillioides was the dominant species associated with maize kernels (42 %), followed by F. graminearum species complex (22.5 %) and F. pseudoanthophilium (13.4 %). The species composition and prevalence of Fusarium species differed among the areas investigated. Fusarium species composition was as many as eight and as few as four in some growing area. The majority of the maize samples (77 %) were found positive for fumonisin, with concentrations ranging from 25 μg kg^?1 to 4500 μg kg^?1 (mean: 348 μg kg^?1 and median: 258 μg kg^?1). Slight variation in fumonisin concentration was also observed among areas. Overall results indicate widespread occurrence of several Fusarium species and contamination by fumonisin mycotoxins. These findings are useful for intervention measures to reduce the impact of the main fungal species and their associated mycotoxins, by creating awareness and implementation of good agricultural practices.     

Molecular and experimental evidence of multi-resistance of <Emphasis Type="Italic">Cercospora beticola</Emphasis> field populations to MBC,DMI and QoI fungicides

Binucleate Rhizoctonia (BNR) spp. isolates were collected from taro (Colocasia esculenta (L.) Schott) and ginger (Zingiber officinale (Willd.) Roscoe) (Yunnanxiaojiang cv.) in Yunnan province. These Yunnan (YN) isolates did not anastomose with any of the tester isolates of the known AGs of binucleate Rhizoctonia spp. The growth of YN cultures on PDA was appressed, mealy and matlike after 4 days of incubation, then turned white brown, producing brown to dark brown, irregularly shaped sclerotia were embedded in the PDA medium after 14 days. All attempts to induce basidiospore production were unsuccessful, but the length and sequence of the internal transcribed spacer (ITS1 + 5.8S rDNA + ITS2) regions of 5.8S rDNA from the YN isolates were identical in length and sequence to isolates of all the other AGs of binucleate Rhizoctonia /Ceratobasidium spp. The sequences of 5.8S rDNA-ITS from the YN isolates were unique among AGs of BNR. The YN isolates had sequence similarities of 94% with isolates of AG Fb and P, 93% with AG E, 91% with AG R, 79–94% with AG S, and 74–87% with AG A, Ba, Bb, Bo, C, DI, DII, DIII, Fa, G, H, I, K, L, O, and Q. Four isolates of AG YN caused minor virulence (lesions ≦1mm²⁾ to ginger or taro in growth chamber studies. It was concluded that the YN isolates belong to a new anastomosis group AG-V of the Ceratobasidium spp..     

Challenging the larvae of <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> and assessing the immune responses to nematode-bacterium complex

Helicoverpa armigera is a strong insecticidal resistance developed insect pest. The understanding of its innate immune responses to emerging biocontrol agent entomopathogenic nematode-bacterial complex can provide an opportunity to control this insect in an environmentally benign manner. Study was focused on role of hemocytes changes and PO activity in Steinernema abbasi-Xenorhabdus indica challenged larvae of H. armigera over the time. Total cell count changed effectively from 10.2?±?1.81?×?10⁵ to 15.5?±?3.3?×?10⁵ cells/mm³ upto 9 h and reduced distinctly up to 8.0?±?2.49?×?10⁵ cells/ mm³ in 24 h. PO activity inclined significantly and was recorded highest at 9 h (24.67?±?1.08?×?10² units) and lowest at 24 h (14.34?±?0.74?×?10² units) in total hemolymph with a similar pattern in plasma and the cellular fraction. Phenoloxidase activity in total and cellular component of hemolymph was positively correlated with prohemocytes, granulocytes and oenocytoids. Study showed the hemocytes and PO accounted as active immune responses against nematode infection. The results provide the first insight to understand the hemolytic activity, quick immunosuppression responses of S. abbasi-X. indica and vulnerability of H. armigera.     

Quantitative PCR shows propagation of <Emphasis Type="Italic">Plasmodiophora brassicae</Emphasis> in Swedish long term field trials

Clubroot (Plasmodiophora brassicae) is a serious soil-borne disease in brassica crops world-wide. We report on a time series of soil samples from Swedish long-term fertility trials started in 1957, 1963 and 1966, which were analyzed for the amount of P. brassicae DNA. The crop rotations included a brassica crop every 4 or 6 years. All experimental sites with a 4-year rotation of oilseed rape, except one with calcium carbonate in the soil profile, showed high (>1000 fg DNA g^?1 soil) levels of P. brassicae DNA after 9, 11 and 12 rotations. In contrast, detectable levels (>5 fg DNA g^?1 soil) of P. brassicae were found only at one of five sites with a 6-year rotation of spring oilseed rape. In years with high levels of P. brassicae DNA, low yield was reported and a subsequent decline in P. brassicae DNA in soil was observed. Different NPK (nitrogen/phosphorus/potassium) fertiliser regimes resulted in similar P. brassicae DNA levels. The robustness and reliability of the method applied was verified by analyses of soil from individual plots compared with a mixture of plots and by repeated analyses of selected samples, which showed that P. brassicae DNA remained stable during dry storage.     

Improved assessment of mycelial growth stimulation by low doses of mefenoxam in plant pathogenic <Emphasis Type="Italic">Globisporangium</Emphasis> species

Globisporangium Uzuhashi, Tojo & Kakish. (syn. Pythium Pringsheim) species cause many plant diseases, including Pythium damping-off, leaf and fruit blights, and root rots. Fungicide resistant isolates are selected by repeated use of a single active ingredient on infected crops without rotation. Previous studies demonstrated increased pathogenicity and radial growth in a mefenoxam resistant isolate of Pythium aphanidermatum when exposed to sub-lethal doses of fungicides and ethanol. In those studies, reproducibility of in vitro assays was difficult to achieve due to large variations among trials. This study aimed to examine two protocols for improved reproducibility during the assessment of biphasic dose-responses in mefenoxam-resistant isolates of Globisporangium ultimum and G. irregulare. Two different growth related endpoints, total growth area and total dry mass weight, were assessed. Assays were conducted using ten concentrations of mefenoxam ranging from 0.01 to 1,000 μg/ml. Statistically-significant stimulatory effects were observed in the two Globisporangium species using the two growth related endpoints. Because of its better reproducibility, mycelial growth area is recommended as an endpoint for future studies of chemical hormesis on growth of Globisporangium spp.     

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.     

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.     

Influence of abiotic factors on behaviour and adult emergence pattern of coconut white grub, <Emphasis Type="Italic">Leucopholis coneophora</Emphasis> Burmeister (Coleoptera: Scarabaeidae)

Leucopholis coneophora Burmeister is a subterranean pest associated with coconut based cropping systems in south India. Feeding damage causes yellowing of fronds and yield reduction. To develop appropriate IPM strategy a basic knowledge on insect behaviour is essential. Four years studies indicated that, adult emergence of L. coneophora was commenced with summer shower in April in Kerala. Delay in summer shower delayed the emergence. After a pause in May, the emergence resumed with the setting of south west monsoon in June. The beetles did not emerge during dry spells in between the rainy days, when the soil temperature (at 10 cm depth) was ≥34.5 °C. Emergence of the beetles started at an illuminance of 124.37?±?75.5 l in evening and remained active till 2?±?0.4 l with a maximum swarming at 32.6?±?15.1 l. Female emergence and mating occurred at 12.04?±?8.1 l. Female based sex pheromone mediated communication is evident. Strong competition among the males for mating with emerging female, which was evident by a wider operational sex ratio in the initial period (1:10.11) that narrowed down to 1:4.33 in later days. The beetles neither congregate on any host plant nor exhibit phototaxis. Number of beetles entrapped in light traps varied from 1.5–16.5% and hand picking is highly significant over light trapping. Hence hand picking of beetles daily in the evening for 2 weeks commencing from the onset of south west monsoon in Kerala, in Indian subcontinent is suggested as a tool in IPM.     

Long-term dynamics of causative agents of stem base diseases in winter wheat and reaction of Czech <Emphasis Type="Italic">Oculimacula</Emphasis> spp. and <Emphasis Type="Italic">Microdochium</Emphasis> spp<Emphasis Type="Italic">.</Emphasis> populations to prochloraz

Trichoderma aggressivum is an aggressive contaminant mould in the cultivation of Agaricus bisporus leading to severe reductions in mushroom yields. Production of fully colonised A. bisporus substrate in Europe is commonly carried out in large tunnels (Phase III), after which the substrate undergoes several bulk handling (mixing) operations before ending up on shelves in mushroom growing facilities. The work presented here studied the effect of Trichoderma aggressivum inoculum, substrate mixing and supplementation on Agaricus bisporus yields and evaluated four methods to detect T. aggressivum in bulk handled substrate. Inoculum dilution level was shown to correlate well with mushroom yield (P < 0.0001) with reductions of 2–6 % at the most dilute level (10^?4) and 60–100 % at the most concentrated level (10^?1), depending on the experiment. Supplementation, with or without T. aggressivum, had no significant effect on mushroom yield (P ≥ 0.85) but a high degree of substrate mixing was shown to significantly increase (P < 0.0001) T. aggressivum-associated crop losses. Four T. aggressivum detection methods were evaluated and a quantitative polymerase chain reaction (qPCR) method gave the most consistent and least variable results. Cycle threshold (CT) values ranged from 24 to 40, depending on the experiment and the inoculum dilution level, and false negatives (CT = 40) were reported on one occasion with the most dilute samples. The results indicate that Phase III mushroom substrate is vulnerable to infection by T. aggressivum when the fully colonised substrate is broken up and mixed during bulk handling operations, identifying a previously unidentified risk for Phase III substrate producers.     

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%).     

Development of qPCR assays to monitor the ability of <Emphasis Type="Italic">Gliocladium catenulatum</Emphasis> J1446 to reduce the cereal pathogen <Emphasis Type="Italic">Fusarium graminearum</Emphasis> inoculum in soils

A qPCR approach was developed to specifically monitor in soils Fusarium graminearum, the main agent responsible for Fusarium Head Blight, and the biocontrol agent Gliocladium catenulatum J1446 (Prestop®). For both fungi, the amplification efficacy of standard curves obtained by mixing pure fungal DNA and soil background DNA was high (qPCR efficacy>96% with R²?>?0.97) with a linear range from 10^?3 ng to 10 ng/μL. Our qPCR method allowed quantifying down to 1 μg of F. graminearum and G. catenulatum J1446 mycelium per g of soil. The strong correlation observed between fungal biomass and quantified DNA (R²?=?0.9927 and 0.9356 for F. graminearum and G. catenulatum J1446, respectively) supported the use of the primers to monitor both fungi in soils. Under our experimental conditions, the ability of Prestop® to reduce F. graminearum growth was significantly higher in autoclaved soil compared to living soils, suggesting that there is an antagonistic effect of the soil microbial communities. In contrast, G. catenulatum J1446 growth was mostly not affected by the presence of F. graminearum and was able to persist in both autoclaved and living soils after 15 days of incubation. These results indicate that our qPCR approach may be used to assess the success of soil colonization by a biocontrol agent and its control efficacy by monitoring the dynamics of the BCA and the targeted pathogen in soil.     

In vitro and field efficacy of three fungicides against <Emphasis Type="Italic">Fusarium</Emphasis> bulb rot of garlic

Fusarium proliferatum has been identified as the main causal agent of bulb rot of garlic (Allium sativum L.). This disease occurs after the drying process and can rot almost 30 % of the bulbs. Few studies are available regarding the effectiveness of chemical treatments to reduce F. proliferatum incidence in garlic. The efficacy of three commercial fungicides of different chemical groups to reduce seven strains of F. proliferatum mycelial growth was tested in vitro. These three fungicides were also evaluated by foliar spreading of aqueous suspension in a field crop. Fluopyram 20 % + tebuconazole 20 % and tebuconazole 50 % + trifloxystrobin 50 % were highly effective at reducing mycelial growth in F. proliferatum with EC₅₀ values <2 ppm. In general, the effectiveness of the fungicides was enhanced with increasing dosage. Our results indicate that the fungicides evaluated in this study may lead to a risk of resistance appearing in F. proliferatum at low concentrations and this risk is maintained at higher doses for the fungicide dimethomorph 7.2 % + pyraclostrobin 4 %. Although several of the fungicides affected in vitro mycelial growth of F. proliferatum, as a part of an strategy to measure the efficacy of resistance management it is necessary to monitor the ongoing efficacy of fungicides under commercial conditions. All fungicidal treatments tested in field application failed to control garlic bulb rot during storage.     

Factors affecting <Emphasis Type="Italic">Neofuscicoccum ribis</Emphasis> infection and disease progression in blueberry

Botryosphaeria stem blight is an economically important disease of blueberry worldwide. In this study, factors affecting inoculum production, infection and disease progression of Neofusicoccum spp. in blueberries were investigated. Under laboratory conditions conidia of the main three Neofusicoccum species (N. australe, N. parvum and N. ribis) were released from pycnidia at 15–30 °C and under relative humidities (RHs) of 80–100%, with greatest numbers released by N. parvum. The greatest numbers of oozing pycnidia and conidial release occurred at higher temperatures (25–30 °C) and RHs (92–100%). Inoculation of green shoots with different N. parvum and N. ribis conidial concentrations (50 μL of 5 × 10⁴?5 × 10⁶ conidia/ mL) caused 100% incidence but lesion lengths increased with increasing concentrations. Wound age affected N. ribis lesion development, with lesions only observed for 0–7-day-old wounds in soft green shoots and 0–4-day-old wounds for both hard green shoots and trunks. Colonisation length decreased with increasing wound age. Lesions developed on wounded shoots when plants were exposed to 20 or 25 °C and 90 or 100% RH during the early infection processes; and in non-wounded shoots spot-like lesions were observed although N. ribis colonised the stem tissue. Seasons (summer, autumn and winter) had no effect on susceptibility of wounded plants to N. ribis. External lesions only developed in summer-inoculated plants and colonisation length was lower in winter-inoculated plants. Information on host and environmental factors that affect disease development determined by the study will be used to inform the development of control strategies.     

Genetic structure of <Emphasis Type="Italic">Pyrenophora teres</Emphasis> f. <Emphasis Type="Italic">teres</Emphasis> and <Emphasis Type="Italic">P. teres</Emphasis> f. <Emphasis Type="Italic">maculata</Emphasis> populations from western Canada

Infection by Pyrenophora teres f. teres (Ptt) or P. teres f. maculata (Ptm), the causal agents of the net and spot forms of net blotch of barley, respectively, can result in significant yield losses. The genetic structure of a collection of 128 Ptt and 92 Ptm isolates from the western Canadian provinces of Alberta (55 Ptt, 27 Ptm), Saskatchewan (58 Ptt, 46 Ptm) and Manitoba (15 Ptt, 19 Ptm) were analyzed by simple sequence repeat (SSR) marker analysis. Thirteen SSR loci were examined and found to be polymorphic within both Ptt and Ptm populations. In total, 110 distinct alleles were identified, with 19 of these shared between Ptt and Ptm, 75 specific to Ptt, and 16 specific to Ptm. Genotypic diversity was relatively high, with a clonal fraction of approximately 10 % within Ptt and Ptm populations. Significant genetic differentiation (PhiPT = 0.230, P = 0.001) was found among all populations; 77 % of genetic variation occurred within populations and 23 % between populations. Lower, but still significant genetic differentiation (PhiPT = 0.038, P = 0.001) was detected in Ptt, with 96 % of genetic variation occurring within populations. No significant genetic differentiation (PhiPT = 0.010, P = 0.177) was observed among Ptm populations. Isolates clustered in two distinct groups conforming to Ptt or Ptm, with no intermediate cluster. The high number of haplotypes observed, combined with an equal mating type ratio for both forms of the fungus, suggests that P. teres goes through regular cycles of sexual recombination in western Canada.     

Shift in the level of susceptibility and relative resistance of brinjal shoot and fruit borer <Emphasis Type="Italic">Leucinodes orbonalis</Emphasis> (Guen) to diamide insecticides

Brinjal shoot and fruit borer Leucinodes orbonalis Guen. is a major pest of brinjal in India. The field collected larvae of L.orbonalis were tested for their susceptibility to three diamide insecticides by fruit dip bioassay technique. Cyantraniliprole and chlorantraniliprole were 5.23 and 2.80 times more toxic to L. orbonalis as compared to flubendiamide. Large variation in the susceptibility of L. orbonalis to cyantraniliprole, chlorantraniliprole, flubendiamide was observed and the LC₅₀ values were 0.084, 0.157 and 0.439 mg a.i. L^?1, respectively. In span of two years there was a significant increase in the LC₅₀ values of cyantraniliprole (0.062 to 0.085 mg a.i. L^?1), chlorantraniliprole (0.097 to 0.157 mg a.i. L^?1), flubendiamide (0.284 to 0.439 mg a.i. L^?1) to population of L. orbonalis, which showed 1.35, 1.62 and 1.55 fold resistance, respectively indicating faster development of resistance to diamide insecticides.