A myosin passenger protein gene (<Emphasis Type="Italic">FaSmy1</Emphasis>) is an essential regulator of cell development,pathogenicity, DON biosynthesis,and resistance to the fungicide phenamacril in <Emphasis Type="Italic">Fusarium asiaticum</Emphasis>

Wheat streak mosaic virus (WSMV) and Triticum mosaic virus (TriMV) are important viruses of wheat (Triticum aestivum L.) in the Great Plains of United States. In addition to agronomic practices to prevent damage from these viruses, temperature sensitive resistance genes Wsm1, Wsm2 and Wsm3, have been identified. However, threshold temperatures for Wsm1 and Wsm3 have not been clearly defined. To better understand these two resistance genes, wheat lines C.I.15092 (Wsm1), KS96HW10–3 (Wsm1), and KS12WGGRC59 (Wsm3) were evaluated for WSMV resistance at 27, 30, 33 and 35 °C and for TriMV resistance at 18, 21, 24, 27, 30, 33 and 35 °C. The results showed that only C.I.15092 remained resistant at 30 °C for both viruses. This line also tolerated TriMV at 33 and 35 °C with less sever symptom and lower infection rates. Wheat lines KS96HW10–3 and KS12WGGRC59 hold resistance to TriMV up to 21 °C. Molecular marker results suggested that the resistance in C.I.15092 is most probably conditioned by the resistance gene Wsm1 and additional gene(s) other than Wsm2 and Wsm3.     

Thermal effects on the development of <Emphasis Type="Italic">Bactrocera dorsalis</Emphasis> (Hendel) (Diptera: Tephritidae) and model validation in Taiwan

The oriental fruit fly, Bactrocera dorsalis (Hendel), is a major polyphagous insect pest with a worldwide distribution. The effects of temperature on stage-specific development in B. dorsalis were investigated at eight constant temperatures (13.0 °C, 14.4 °C, 16.2 °C, 19.5 °C, 23.8 °C, 27.7 °C, 31.8 °C and 34.8 °C). B. dorsalis developed successfully from the egg stage to the adult stage at all the tested temperatures, except at the lowest temperatures (13.0 °C and 14.4 °C). Stage-specific lower developmental thresholds and thermal constants were determined using linear regression. The lower and higher temperature threshold (TL and TH, respectively) were estimated using the Sharpe-Schoolfield-Ikemoto model. The lower developmental threshold and thermal constant from egg to adult emergence were 9.8 °C and 325.5 degree-days, respectively. The intrinsic optimum temperatures of the egg, larval, pupal and egg to pupal stage were 20.7 °C, 21.8 °C, 21.1 °C, and 22.4 °C, respectively. The temperature range of the B. dorsalis total immature stage from TL to TH was 20.4 °C (13.8 °C - 34.2 °C). The stage-specific developmental completion of B. dorsalis was determined using a two-parameter Weibull function. The daily adult emergence frequency of B. dorsalis was estimated in relation to adult age and temperature using non-linear developmental rate functions and the Weibull function. The date of cumulative 50% adult emergence estimated using non-linear functions was approximately one day earlier than the experimentally observed date. Thermal performance was compared among B. dorsalis populations from different locations.     

Locomotory behaviour of susceptible and resistant <Emphasis Type="Italic">Plutella xylostella (Linnaeus)</Emphasis> (Lepidoptera: Plutellidae)

Locomotory behaviour of insecticide-resistant and susceptible populations of diamondback moth, Plutella xylostella (Linnaeus), a challenging insect-pest of cruciferous vegetable crops round the globe, was studied with the help of Ethovision. Larvae from the susceptible population of P. xylostella travelled more distance on insecticide-treated or untreated surface and consequently their speed was also more as compared to those taken from the insecticide-resistant population.Whereas, the turn angle of larvae from insecticide-resistant population was significantly higher as compared to those from susceptible population. The resistant larvae travelled significantly less distance (260.68 cm/5 min) on treated-surface (with LC₅₀ value of endosulfan, quinalphos, fenvalerate and spinosad) with more value of turn angle (231.16°) than the susceptible ones which moved faster (517.23 cm/5 min) with less value of turn angle (100.63°). Influence of varying temperatures on locomotory behaviour of larvae from the susceptible and resistant population of P. xylostella was also studied and observed that susceptible P. xylostella larvae travelled significantly greater distance as compared to the larvae from insecticide-resistant population, at different temperatures. Locomotory behaviour of larvae of P. xylostella also changes with food conditions. Resistant larvae starved for 24 h travelled significantly lesser distance (147.29 cm) as compared to unstarved ones (332.50 cm). Similar trend in behaviour was also recorded with respect to speed of larvae of P. xylostella. Larvae turn more frequently when kept without food (starved) than those fed normally; the turn angle was significantly higher (290.07°) for resistant larvae and for susceptible larvae it was lesser i.e. 151.55°, when kept at starved conditions. Hence, certain pronounced behavioural differences were registered in locomotion of insecticide-resistant P. xylostella as compared to the susceptible one and this knowledge would help to find effective management solutions to P. xylostella.     

Toxicity of butene-fipronil,in comparison with seven other insecticides,in <Emphasis Type="Italic">Leptinotarsa decemlineata</Emphasis> and <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The speed of toxic action of an insecticide is an indicator for control efficacy and has considerable practical importance. For agricultural pest control, fast-acting is an important feature for an insecticide to consistently reduce the amount of feeding damage. Butene-fipronil is a novel compound obtained via the structural modification of fipronil. However, information about the toxicity and speed of toxic action is still limited. In the present paper, we compared the toxic feature of butene-fipronil with seven other insecticides, of which imidacloprid and abamectin are slow-acting insecticides, and acephate, endosulfan, methomyl, α-cypermethrin and spinosad are fast-acting insecticides. We found that the contact and stomach toxicities of butene-fipronil were among the highest ever estimated to Leptinotarsa decemlineata and Drosophila melanogaster. The speed of toxic action of butene-fipronil was determined using median lethal time (LT₅₀) at a dose (concentration) equivalent to LD₈₀ values. For L. decemlineata, the values for butene-fipronil, imidacloprid, abamectin, acephate, endosulfan, methomyl, cypermethrin and spinosad were calculated to be 39.9, 36.5, 37.5, 20.2, 22.4, 23.8, 16.4 and 23.1 h, respectively. Those for D. melanogaster were 29.8, 31.5, 29.4, 14.0, 20.3, 18.1, 13.5, and 20.1 h, respectively. ANOVA analysis showed that butene-fipronil, imidacloprid, abamectin had similar LT₅₀ values, whereas acephate, endosulfan, methomyl, spinosad and cypermethrin had comparable LT₅₀ values. Thus, butene-fipronil belongs to slow-acting insecticides. Our results provide more empirical information for butene-fipronil potential application.     

Temperature and plant age drive downy mildew disease epidemics on oilseed <Emphasis Type="Italic">Brassica napus</Emphasis> and <Emphasis Type="Italic">B. juncea</Emphasis>

Studies were undertaken on the effects of temperature (14/10 °C and 22/17 °C day/night) and plant age (15, 23, 31 and 40 day-old-plants) on the severity of downy mildew (Hyaloperonospora parasitica) on oilseed Brassica cultivars (temperature: Brassica juncea Montara, B. napus Atomic, ATR-Hyden, Hyola 432, Hyola 450 TT, Thunder TT; plant age: B. juncea Dune, B. napus Surpass 402 and Hyola 450 TT). For temperature studies, there were significant (P?<?0.001) effects of temperature, cultivar, and cultivar x temperature interaction. On cotyledons of susceptible cultivars (B. napus Hyola 450 TT and Thunder TT), plants were symptomatic at 22/17 °C by 48 h post inoculation (hpi) and with abundant sporulation evident by 72 hpi, and with all cotyledons of B. napus Thunder TT collapsed by 7 days post inoculation (dpi). However, at 14/10 °C, there were no symptoms on the same cultivars until 5 dpi, and sporulation only observed at 7 dpi. Percent disease index values (DI%) at 22/17 °C of B. juncea Montara and B. napus ATR-Hyden, Hyola 432, Atomic, Hyola 450 TT and Thunder TT were 4.5, 49.0, 51.4, 65.8, 86.3 and 96.0, respectively, with all except B. juncea Montara having significantly lower (P?<?0.001) disease at 14/10 °C with DI% values of 2.8, 30.4, 27.9, 31.1, 44.4 and 76.4, respectively. For plant age studies, there were significant (P?<?0.001) effects of plant age, cultivar, and cultivar x plant age interaction. DI% was significantly higher at 15 compared to 40 day-old-plants (dop) across all cultivars. B. juncea Dune showed greatest resistance, particularly on 40 dop, with DI% values of 25.8, 24.6, 22.9 and 7.5, for 15, 23, 31 and 40 dop, respectively. B. napus Surpass 402 showed high susceptibility on cotyledons of 15 dop but moderate resistance on leaves of other ages, with DI% values of 59.0, 31.2, 27.1 and 26.2 for 15, 23, 31 and 40 dop, respectively. B. napus Hyola 450 TT showed very high susceptibility at the cotyledon stage on 15 dop, but some resistance on 23 dop and more so on 31 and 40 dop, with DI% values of 84.0, 41.2, 35.4 and 32.9 for 15, 23, 31 and 40 dop, respectively. Together, these findings explain for the first time why development of downy mildew epidemics on susceptible cultivars occurs early in the growing season when warmer seasonal temperatures in autumn coincide with presence of seedlings; in contrast to later in the growing season on less susceptible older plants coinciding with cooler and less favourable winter temperatures. Increasing maximum and minimum temperatures associated with climate change have likely fostered the increased severity of downy mildew over the past 15 years.     

Effect of controlled fluctuating low temperatures on survival of <Emphasis Type="Italic">Puccinia striiformis</Emphasis> f. sp. <Emphasis Type="Italic">tritici</Emphasis>

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is an important disease of wheat worldwide. Understanding the survival of Pst during the winter is critical for predicting Pst epidemics in the spring. We used a real-time quantitative PCR (qPCR) method to quantify Pst CYR32 biomass in infected wheat seedlings under several fluctuating temperature regimes (three average temperatures 0, ?5 and ?10 °C, each with two daily fluctuating amplitudes 8 and 13 °C). The survival of Pst CYR32 increased with increasing average temperature but also varied greatly with the amplitude – larger amplitude led to lower survival, particularly at 0 and ?5 °C. Nevertheless the survival at both amplitudes was still significantly greater than under the corresponding constant temperatures. There were small, albeit statistically significant, differences between the two cultivars (Xiaoyan 22, low winter-hardiness; Lantian 15, high winter-hardiness) in Pst CYR32 survival. This study indicated potential errors that could result from using daily average temperatures to predict Pst survival during the winter.     

Unusual behavior of oviposition and development of <Emphasis Type="Italic">Sitophilus zeamais</Emphasis> (Coleoptera: Curculionidae) in peach and apple fruits

The maize weevil Sitophilus zeamais Motschulsky, 1855 (Coleoptera: Curculionidae) is a worldwide pest on stored products, as maize, wheat and rice grains. However, in South Brazil, these weevils have been found attacking fruits in field conditions. It was reported that adults attack fruits to feeding, but there was no information of oviposition and larval development on these hosts. Furthermore, due to monitoring apple orchards, was observed that some fruits showed distinct kind of punctures, suggesting a oviposition by these weevils. The present study reports first record an unusual behavior of S. zeamais by oviposition and larval development in peach and apple fruits. The study was conducted in laboratory conditions (temperature: 25?±?1 °C, RH: 70?±?10% and photophase of 14 h). Peach and apple fruits were offered to S. zeamais females. The oviposition and duration of immature stages in peach were measured in fruits and in corn grains, used as control observation. The oviposition and larval development occurred in peach and apple fruits, with a development until to adult. The average of egg-to-adult was 47.4 d in peach and and 45.1 d apple fruits, respectively, while in corn grains, its natural host, the duration was 38.2 d. These results show that the damage caused by S. zeamais in peach and apple fruits may be due to oviposition and the development of immature stages. So, the novelty is in demonstrating that the maize weevil does not just feed as an adult opportunistically on the fruits, but can complete a full life cycle using fresh fruits.     

Infection criteria,inoculum sources and splash dispersal pattern of <Emphasis Type="Italic">Colletotrichum acutatum</Emphasis> causing bitter rot of apple in New Zealand

Infection of Malus x domestica cv. Royal Gala fruit by Colletotrichum acutatum causing bitter rot was studied in the temperate climate of New Zealand. Temperatures above 15 °C were required for lesions to develop on detached apple wound-inoculated or inoculated without wounding with C. acutatum spores, regardless of maturity. A wetness period of 72 h was required for infection of mature detached apple fruit without wounding. On wound-inoculated detached apple fruits, sporulation was related to temperature and followed a similar pattern. In the field, a mean temperature above 15 °C for 72 h after wound-inoculation was required for lesions to develop. Buds were a more important source of inoculum than twigs, and it was shown that C. acutatum could be isolated more frequently from outer bud scales than from inner scales. Asymptomatic infection of vegetative and reproductive buds was detected. C. acutatum was detected on asymptomic surface-sterilised petals and fruit, more commonly during summer than spring. Symptomless sterilised leaves generally yielded C. acutatum throughout the season, but isolations were more frequent in summer. Recovery of inoculum using a splash meter to detect vertical dispersal showed that in summer inoculum was primarily splashed up from the ground. In spring, inoculum was recovered in similar quantities from all heights up to a metre, suggesting that splash dispersal occurs from the canopy as well as from the ground. A disease cycle for C. acutatum infecting apples and causing bitter rot in New Zealand is suggested.     

Characterization and pathogenicity of <Emphasis Type="Italic">Bipolaris peregianensis</Emphasis>: the causal organism for leaf spot of hybrid bermudagrass in China

Several golf courses established with hybrid bermudagrass (Cynodon dactylon × C. transvaalensis) were surveyed from April 2011 through April 2015 in Hainan Province, China. The hybrid bermudagrass in these golf courses showed a new leaf spot disease, and a filamentous fungus was consistently recovered from the infected leaves. Based on the morphological characteristics of colony color and appearance, shapes of conidiophores and conidia as well as sequences of internal transcribed spacer regions (ITS) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH), the fungus was identified as Bipolaris peregianensis. The pathogenicity test conducted on healthy hybrid bermudagrass produced leaf spot symptoms one week post inoculation. B. peregianensis mycelia grew in a temperature range of 5 to 35 °C with the optimum temperature being 28 °C.     

Environmental effects on growth and sporulation of <Emphasis Type="Italic">Fusarium</Emphasis> spp. causing internal fruit rot in bell pepper

Internal fruit rot in bell pepper (Capsicum annuum L.) is mainly caused by members of the Fusarium lactis species complex (FLASC) and to a lesser extent by Fusarium oxysporum and Fusarium proliferatum. Despite the importance of the disease, there is hardly no information about growth, sporulation and germination dynamics of FLASC. In order to understand the dominance of FLASC as main pathogen of internal fruit rot, the effects of temperature (5 °C – 35 °C), water activity (a_w 0.76–0.96), pH (pH 3 - pH 9) and oxygen concentration (2.5% - 20%) on growth and sporulation of all three Fusarium species were compared. In addition, germination kinetics were also investigated. FLASC showed optimal mycelium growth and sporulation in the narrow range of 25 °C, while both other strains were also tolerant for higher temperatures to 30 °C. FLASC was also characterized by a broad pH optimum from pH 3–7 while F. oxysporum (pH 4–7) and F. proliferatum (pH 5–8) were more demanding concerning pH. In addition, optimal sporulation occurred in the acid region for FLASC (pH 3) whilst neutral and alkaline pH were more favourable for the other species. Germination kinetics revealed that FLASC did not benefit from an earlier and/or faster germination process. A thorough understanding of the growth characteristics and dominance of FLASC as main pathogen for internal fruit rot is inevitable to develop sustainable control measures for the disease.     

Life-stage-dependent side effects of selected insecticides on <Emphasis Type="Italic">Trichogramma cacoeciae</Emphasis> (Marchal) (hymenoptera: Trichogrammatidae) under laboratory conditions

Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is classified as one of the most harmful pest of tomato crops. Many species of predators and parasitoids including Trichogramma cacoeciae (Marchal) (Hymenoptera: Trichogrammatidae) are noted as potential candidates used for biological control of this pest. Therefore, the use of selective insecticides is critical to conserve and protect natural enemies in the field. This study assessed the side effects of insecticides on different development stages of T. cacoeciae under laboratory conditions. For this, eleven pesticides such as: Indoxacarb, spiromesifen, cyromazin, chlorfenapyr, cypermethrin, diafenthiuron, chlorantraniliprole, spinosad, azadirachtin, Bacillus thuringiensis (Bt) and virus HaNPV were tested. This study shows that indoxacarb, spiromesifen, chlorfenapyr, cypermethrin, diafenthiuron and spinosad had a negative effect on immature stages of Trichogramma. All insecticides residues on tomato leaves were found to be toxic to Trichogramma adults except azadirachtin, Bt and virus HaNPV. Therefore, the use of the tested natural products (azadirachtin, Bt and HaNPV) at the recommended doses is viable, having no negative impact on T. cacoeciae in tomato crops.     

Chitinolytic <Emphasis Type="Italic">Streptomyces griseorubens</Emphasis> E44G enhances the biocontrol efficacy against <Emphasis Type="Italic">Fusarium</Emphasis> wilt disease of tomato

Streptomyces griseorubens E44G is a chitinolytic bacterium isolated from cultivated soil in Saudi Arabia (a hot, arid climatic region). In vitro, antifungal potential of S. griseorubens E44G was assessed against the phytopathogenic fungus, Fusarium oxysporum f. sp. lycopersici (the causative agent of the Fusarium wilt disease of tomato). An inhibition zone of 24 mm was recorded. The chitinolytic activity of S. griseorubens E44G was proved when the colloidal chitin agar plate method was used. A thermostable chitinase enzyme of 45 kDa molecular weight was purified using gel filtration chromatography. The optimum activity was obtained at 60 °C and pH 5.5. The purified enzyme has shown a very pronounced activity against the phytopathogenic fungus, F. oxysporum. The molecular characterization of the chitinase gene indicated that it consists of 1218 bp encoding 407 amino acids. The phylogentic analysis based on the nucleotide DNA sequence and the deduced amino acids sequence showed high similarity percentages with other chitinases isolated from different Streptomyces species. In the field evaluation, application of both S. griseorubens E44G treatments significantly increased all tested growth and yield parameters and decreased the disease severity compared with the infected-untreated tomato plants suggesting potential as a biocontrol agent.     

Environmental and inoculum effects on epidemiology of bacterial spot disease of stone fruits and development of a disease forecasting system

Bacterial spot disease of stone fruits, caused by Xanthomonas arboricola pv. pruni, is of high economic importance in the major stone-fruit-producing areas worldwide. A better understanding of disease epidemiology can be valuable in developing disease management strategies. The effects of weather variables (temperature and wet/dry period) on epiphytic growth of X. arboricola pv. pruni on Prunus leaves were analyzed, and the relationship between inoculum density and temperature on disease development was determined and modeled. The information generated in this study, performed under controlled environmental conditions, will be useful to develop a forecasting system for X. arboricola pv. pruni. Optimal temperature for growth of epiphytic populations ranged from 20 to 30 °C under leaf wetness. In contrast, multiplication of epiphytic populations was not only interrupted under low relative humidity (RH) (< 40%) at 25 °C, but also resulted in cell inactivation, with only 0.001% initial cells recovered after 72 h incubation. A significant effect of inoculum density on disease severity was observed and 10⁶ CFU/ml was determined as the minimal infective dose for X. arboricola pv. pruni on Prunus. Infections occurred at temperatures from 15 to 35 °C, but incubation at 25 and 30 °C gave the shortest incubation periods (7.7 and 5.9 days respectively). A model for predicting disease symptom development was generated and successfully evaluated, based on the relationship between disease severity and the accumulated heat expressed in cumulative degree day (CDD). Incubation periods of 150, 175 and 280 CDD were required for 5, 10 and 50% of disease severity, respectively.     

Efficacy of <Emphasis Type="Italic">Beauveria Bassiana</Emphasis> for the Management of Economically Important Wireworm Species (Coleoptera: Elateridae) in Organic Farming

Wireworms, the larvae of click beetle species Agriotes lineatus, A. obscurus and A. sputator are serious pests for several field crops. They are considered severe pests of potato tubers and the damages caused by them can be resulted a significant loss especially in organic crop production. Since synthetic insecticides are prohibited in Organic Farming; biological control methods have to be used in organic crop production. In the current study naturalis Beauveria bassianaa strain ATCC 74040 was used both under laboratory and field conditions using wheat and potato crops as a food source respectively. Fungus showed a significant mortality in high number of wireworms boxes (50%) compared to low number of wireworms (17%) and untreated boxes (13%) respectively. However, seed germination showed no effect in all three categories. Field data have shown mixed results when B. bassiana has been applied in a variety of application methods. Maximum infestation (3.99%) was recorded in untreated plots; while significantly lower damage (1.11%) was recorded in whole surface treated plots. During first year all the treatments were significantly different from each other; however, no significant differences were noted between furrows and whole surface applications but were different from control treatment during second year of experimentation. The results showed that the biological control of wireworms by using entomopathogenic fungi, such as B. bassiana is a promising target specific option without disturbing the other communities in the soil.     

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.     

Accumulation of toxigenic <Emphasis Type="Italic">Fusarium</Emphasis> species and <Emphasis Type="Italic">Stenocarpella maydis</Emphasis> in maize grain grown under different cropping systems

Entomopathogenic nematodes in the genus Steinernema are associated with Xenorhabdus spp. bacteria. When steinernematid colonise an insect host the nematode-bacterium association overcomes the insect immune system and kills the host within 48 h. Xenorhabdus spp. produce secondary metabolites that are antifungal to protect nematode-infected cadavers from fungal colonization. The concentrated, or cell-free metabolites of X. szentirmaii exhibit high toxicity against various fungal plant pathogens and show potential as natural bio-fungicides. In the current study, we determined 1) thermo-stability, 2) dose-response, and 3) shelf-life of antifungal metabolites of X. szentirmaii against Monilinia fructicola (cause of brown rot of peach and other stone fruit) and Glomerella cingulata (cause of antharacnose). Thermo-stability was determined by autoclaving bacterial culture broths (121 °C and 15 psi for 15 min) and measuring fungal growth on in potato dextrose agar (PDA) containing 10% of the supernatants. Autoclaving had no impact on the antifungal activity of the secondary metabolites. Over a test period of 9 months, the activity of both extract types did not decline when stored at 4 or 20 °C. A dose-response study (10, 20, 40, 60, 80 and 100% supernatant-containing metabolite) using both phytopathogens demonstrated that a greater dose of supernatant increased antifungal activity. The antifungal-metabolite containing supernatant of X. szentirmaii has potential as a bio-fungicide. These results demonstrate the metabolite(s) are thermo-stable, they have a long shelf-life and require no stabilizing formulation, even at room temperature.     

Bacterial canker of cherry trees, <Emphasis Type="Italic">Prunus avium,</Emphasis> in South Africa

Laboratory and nursery experiments were conducted to identify the causal agent of a needle blight of Pinus wallichiana, a species native to the Western Himalayas. The pathogen was identified as Myrothecium verrucaria, on the basis of morphological, cultural and molecular characterization. BLAST analysis of ITS sequences of the pathogen revealed maximum sequence identity of 99% with M. verrucaria. The sequence is the first of this fungus from P. wallichiana. Phylogenetic analysis grouped all M. verrucaria isolates in a single clade; M. roridum and M. inundatum clustered in separate clades. The pathogen grew optimally at 25 ± 1 °C on oat meal agar, pH 5.5. Inoculation experiments with M. verrucaria demonstrated pathogenicity on Pinus halepensis, Cedrus deodara and Cryptomeria japonica, in addition to Pinus wallichiana.     

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.     

Short-term toxicity of insecticides residues to key predators and parasitoids for pest management in cotton

The cotton ecosystem comprises various arthropod pest and natural enemies with simultaneous occurrence irrespective of growing region. The use of insecticides with reduced impact on natural enemies is a major goal to conserve them and, therefore, to reduce populations of arthropod pests. The survival of twelve key natural enemies for cotton pest management exposed to dried residues using the highest and lowest recommended rates representing old and new insecticides recommended to control cotton pests (chlorantraniliprole, chlofernapyr, spinosad, lambda-cyhalotrin, methidathion, pymetrozine, and thiamethoxam) was determined. The study included parasitoids [Aphelinus gossypii Timberlake, Bracon vulgaris Ashmead, Lysiphlebus testaceipes (Cresson), Telenomus podisi (Ashmead), Trichogramma pretiosum (Riley)] and predators [Hippodamia convergens Guérin-Méneville, Euborellia annulipes (Lucas), Podisus nigrispinus (Dallas), Solenopsis invicta Buren), Orius insidiosus (Say), Chrysoperla externa Hagen and Eriopis connexa (Germar)], with two different cohorts for these last two species. All natural enemies exposed to methidathion exhibited 100% mortality. Thiamethoxam, lambda-cyhalothrin and chlorfenapyr also caused high mortality of P. nigrispinus, S. invicta, H. convergens, O. insidiosus and all tested parasitoids. Among the natural enemies, E. annulipes exhibited high survival when exposed to all tested insecticides, except methidathion. Chlorantraniliprole and pymetrozine caused overall lower impact on the natural enemies tested followed by spinosad; hence, they are options for cotton pest management. Furthermore, the outcomes highlight the implication of knowing the background susceptibility of the species tested when addressing the impact of insecticides on natural enemies.     

Visual detection of <Emphasis Type="Italic">Didymella bryoniae</Emphasis> in cucurbit seeds using a loop-mediated isothermal amplification assay

A method was developed using a Loop-mediated isothermal amplification assay (LAMP) for detecting Didymella bryoniae in cucurbit seeds. The LAMP primers were designed based on the DNA-dependent RNA polymerase II RPB140 gene (RPB2) from D. bryoniae. Calcein was used as an indicator for the endpoint visual detection of DNA amplification. The LAMP assay was conducted in isothermal (65 °C) conditions within 1 h. The detection threshold of the LAMP assay was 10 pg of genomic DNA and D. bryoniae was detected in 100 % of artificially infested seedlots with 0.05 % infestation or greater. With the LAMP assay, 16 of 60 watermelon and muskmelon seedlots collected from Xinjang province were determined to be positive for D. bryoniae. In contrast, a real-time PCR assay determined that 11 of the 60 seedlots from Xinjiang province were positive for the pathogen. These results showed that the LAMP technique was simple, rapid and well suited for detecting D. bryoniae DNA, especially in seed health testing.