Effect of azadirachtin on the control of <Emphasis Type="Italic">Anticarsia gemmatalis</Emphasis> and its impact on <Emphasis Type="Italic">Trichogramma pretiosum</Emphasis>

Anticarsia gemmatalis has great potential to reduce soybean productivity, and the egg parasitoid Trichogramma pretiosum is a major agent in the biological control of this pest in soybean fields. We show that azadirachtin is able to control velvetbean caterpillars in soybean plants and also that it has no effects on the parasitoid T. pretiosum. Soybean plants were sprayed with solutions containing control (water), 50 and 100 mg.l ⁻¹ of azadirachtin (AzaMax™ 12 g a.i.l ⁻¹). Third instar larvae of A. gemmatalis were confined to soybean plants after the spraying. Leaf consumption and larval mortality of A. gemmatalis were evaluated. Newly emerged females of T. pretiosum were placed for 24 h in tubes with a piece of cardboard containing the same doses of azadirachtin used against velvetbean caterpillars. After 24 h, cardboard with 20 eggs of A. gemmatalis was offered for parasitism during 24 h, and the emergence and sex ratio of progenies were determined. Azadirachtin at 50 or 100 mg.l ⁻¹ reduced leaf consumption and caused 100% mortality in A. gemmatalis larva. Azadirachtin did not negatively affect the parasitism, emergence or sex ratio of the progeny. This indicates that the product can be used with mass release of T. pretiosum to control A. gemmatalis.     

Susceptibility of eggs of <Emphasis Type="Italic">Tribolium confusum</Emphasis>, <Emphasis Type="Italic">Ephestia kuehniella</Emphasis> and <Emphasis Type="Italic">Plodia interpunctella</Emphasis> to four essential oil vapors

Susceptibility of eggs of Tribolium confusum du Val. (Coleoptera: Tenebrionidae), Ephestia kuehniella (Zell.) (Lepidoptera: Phycitidae) and Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae) to vapors of essential oil from garlic (Allium sativum L.), birch (Betula lenta L.), cinnamon (Cinnamonum zeylanicum (Blume)) and aniseed (Pimpinella anisum L.) was studied. Preliminary bioassay tests indicated that vapors of the essential oils had a significant effect on the eggs of tested insect species when exposed to a concentration of 20 μl l ⁻¹ air for 24 h. Generally, garlic and birch essential oils were more toxic to the eggs of tested insect species than cinnamon and aniseed essential oils (except for eggs of T. confusum). There was also a significant difference between susceptibility of eggs of T. confusum, E. kuehniella and P. interpunctella to tested essential oils. Toxicity data indicated that eggs of T. confusum were more susceptible to tested essential oils, with LC₉₀ values ranging from 3.11 to 33.49 μl l ⁻¹ air, than those of E. kuehniella and P. interpunctella; eggs of P. interpunctella were the most tolerant to the essential oils, with LC₉₀ values ranging from 22.02 to 72.42 μl l ⁻¹ air. Concentration × time (Ct) products of 0.29, 0.22, 0.13 and 1.37 mg h l ⁻¹ for garlic, birch, cinnamon and aniseed essential oil, respectively, were required to obtain 90% kill of T. confusum eggs. Although cinnamon essential oil had a much closer Ct product value to methyl bromide, garlic and birch essential oils were found to be the most promising ones since they had also high fumigant toxicity on eggs of both E. kuehniella and P. interpunctella.     

Biological traits and prey consumption of <Emphasis Type="Italic">Anthocoris minki</Emphasis> fed on <Emphasis Type="Italic">Agonoscena pistaciae</Emphasis> and <Emphasis Type="Italic">Brachycaudus (Thuleaphis) amygdalinus</Emphasis>

The predatory insect Anthocoris minki Dohrn (Heteroptera: Anthocoridae) is an indigenous Anthocoris species for the biological control of pests in pistachio orchards. The pistachio psylla Agonoscena pistaciae Burckhardt and Lauterer (Homoptera: Psyllidae) is an important insect pest in pistachio trees in Turkey. Similarly, Brachycaudus (Thuleaphis) amygdalinus (Schouteden) (Hemiptera: Aphididae) is a pest of almond trees that is considered as alternative prey for A. minki when pistachio psylla are not available in early spring on pistachio trees. The development time, survival percentage of immature stages, longevity, fecundity, prey consumption, and life table parameters of A. minki fed on A. pistaciae and B. amygdalinus nymphs were determined at 25 ± 1°C, 70 ± 5% r.h., and a 16 h:8 h (L:D) photoperiod under laboratory conditions. The nymphal survival rate was significantly higher when nymphs were fed on A. pistaciae (an average of 96.7%) than on B. amygdalinus (an average of 71.4%). The development time of A. minki was significantly shorter when nymphs were fed on B. amygdalinus (10.3 days) as opposed to A. pistaciae (11.0 days). No significant differences among prey species were found for longevity and fecundity. The total female longevity and fecundity of A. minki was 38.0 days and 247.2 eggs, respectively, when nymphs were fed on A. pistaciae; and 35.4 days and 233.0 eggs, respectively, when nymphs were fed on B. amygdalinus. On average, 104.4 A. pistaciae and 77.7 B. amygdalinus nymphs were consumed during the nymphal development time for A. minki. Adults of A. minki consumed significantly more psyllids than aphids throughout their life span. The greater difference did not significantly inpact the longevity and fecundity of A. minki. Females of A. minki consumed an average of 631.0 A. pistaciae and an average of 273.3 B. amygdalinus nymphs, while female predators consumed significantly more prey than males. The intrinsic rate of increase (r _m ) of A. minki fed on A. pistaciae (0.174) was significantly greater than those fed on B. amygdalinus (0.148). The successful development and reproduction of both A. pistaciae and B. amygdalinus indicates that they are suitable prey for A. minki.     

Visualisation of <Emphasis Type="Italic">hrp</Emphasis> gene expression in <Emphasis Type="Italic">Xanthomonas euvesicatoria</Emphasis> in the tomato phyllosphere

The plasmid pUFZ75 conferred constitutive GFP expression on the bacterial pathogen Xanthomonas euvesicatoria (syn. X. campestris pv. vesicatoria). Colonisation of the tomato phyllosphere and invasion of tomato leaves by X. euvesicatoria was examined using both fluorescence and confocal laser scanning microscopy. Xanthomonas euvesicatoria established a limited population on the tomato leaf surface, primarily occupying the depressions between epidermal cells and around the stomata, prior to invasion of the leaf via the stomata and subsequent growth within the substomatal chamber and the leaf apoplast. Additionally, hrp-gfp fusions were used to report on the temporal and spatial expression of hrp genes during epiphytic colonisation and invasion. Xanthomonas euvesicatoria cells carrying hrpG- and hrpX-gfp reporter constructs were not fluorescent in vitro on non-hrp-inducing LB agar but did exhibit a low level of fluorescence on the leaf surface within 24 h of inoculation, particularly in the vicinity of stomata. Cells carrying hrpG- and hrpX-gfp fusions exhibited high levels of fluorescence 72 h after inoculation in the substomatal chamber and the leaf apoplast. Cells carrying the hrpF-gfp fusion were slightly fluorescent on LB agar and showed no further increase in fluorescence on the leaf surface by 24 h after inoculation, but did show a significant increase in fluorescence 72 h after inoculation in the substomatal chamber and apoplast. The apparent low-level induction of the regulators hrpG and hrpX on the tomato leaf surface may suggest that some of the genes of the X. euvesicatoria HrpG/HrpX regulon are up- or down-regulated prior to invasion of the stomata while still on the leaf surface.     

Reduced fitness associated with spinosad resistance in <Emphasis Type="Italic">Helicoverpa armigera</Emphasis>

The fitness cost of spinosad resistance was investigated in the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). Laboratory experiments were conducted to compare relative fitness of H. armigera between the spinosad-susceptible and -resistant strains. During the experiments, the average development periods of the resistant strain were lengthened by 4–5 days, reflected in a prolongation of egg, larval and pupal periods. Furthermore, pupal survival, pupal weight, the mean life span of emerged adults, eggs laid and hatched decreased greatly in the resistant strain in comparison with the susceptible strain. Other life-cycle parameters such as larval survival, larval wet weights, prepupal periods, pupation ratio, and sex ratio did not change significantly. As a result, both net replacement rate (R ₀) and intrinsic rate of increase (r _m) were reduced for the resistant strain. Our results clearly indicated that relative fitness of resistant individuals was reduced in the absence of spinosad. Rational measures including pesticide rotations should be expected to delay development of resistance to spinosad in H. armigera field populations from China.     

Vascular blackening of wasabi rhizomes caused by <Emphasis Type="Italic">Pectobacterium carotovorum</Emphasis> subsp. <Emphasis Type="Italic">carotovorum</Emphasis>

Wasabi (Wasabia japonica) is grown for its highly-valued rhizome which is used as a condiment in Japanese food. Symptoms of vascular blackening in the rhizome were first observed in 2005 in plants grown in British Columbia, Canada. Microscopic observations and microbial isolation from infected tissues revealed that most of the xylem tracheid cells were blackened and bacteria were consistently associated with symptomatic plants. The bacterium most frequently recovered was identified as Pectobacterium carotovorum subsp. carotovorum (Pcc) using BioLog™ and sequencing of a specific ~510 bp IGS region. Pathogen-free plants obtained using meristem-tip micropropagation were inoculated with a wasabi isolate of Pcc. Vascular blackening symptoms developed in the rhizome after 8 weeks when the rhizome was first wounded by stabbing or cutting, or if the roots were pre-inoculated with Pythium species isolated from rhizome epidermal tissues, followed by inoculation with Pcc at 1 × 10⁸ cells ml⁻¹. Xylem tracheid cells were blackened and Pcc was reisolated from all diseased tissues. The highest frequency of rhizome vascular blackening occurred at 22°C and 27°C and these tissues occasionally succumbed to soft rot at higher temperatures, but not when inoculated tissues were incubated at 10°C. The rooting medium used by growers for vegetative propagation of wasabi was shown to contain Pcc but the pathogen was not recovered from the irrigation water. Entry of Pcc through wounds on wasabi rhizomes and the host tissue response result in symptoms of vascular blackening.     

<Emphasis Type="Italic">Trichoderma harzianum</Emphasis> elicits defence response genes in roots of potato plantlets challenged by <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

Biological control of Rhizoctonia solani with Trichoderma harzianum has been demonstrated in several studies. However, none have reported the dynamics of expression of defence response genes. Here we investigated the expression of these genes in potato roots challenged by R. solani in the presence/absence of T. harzianum Rifai MUCL 29707. Analysis of gene expression revealed an induction of PR1 at 168 h post-inoculation (hpi) and PAL at 96 hpi in the plants inoculated with T. harzianum Rifai MUCL 29707, an induction of PR1, PR2 and PAL at 48 hpi in the plants inoculated with R. solani and an induction of Lox at 24 hpi and PR1, PR2, PAL and GST1 at 72 hpi in the plants inoculated with both organisms. These results suggest that in the presence of T. harzianum Rifai MUCL 29707, the expression of Lox and GST1 genes are primed in potato plantlets infected with R. solani at an early stage of infection. Mycothèque de l’Université catholique de Louvain of S. Cranenbrouck's affiliation is part of the Belgian Coordinated Collections of Micro-organisms (BCCM).     

Effect of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> mixed with diatomaceous earth on mortality,mycosis and sporulation of <Emphasis Type="Italic">Rhyzopertha dominica</Emphasis> on stored wheat

A laboratory assay was designed to determine the insecticidal efficacy of Beauveria bassiana (Balsamo) Vuillemin (Hyphomycetes: Moniliales) and diatomaceous earth (Diafil 610) against Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae). The fungus B. bassiana was applied at 2.23 × 10⁷, 2.23 × 10⁸ and 2.23 × 10⁹ conidia kg⁻¹ of wheat individually as well as mixed with 200 and 400 ppm of Diafil 610. The conditions for the trials were 30 ± 2^oC with 55% r.h. and the counts for mortality were made after 8, 16 and 24 d. All the dead adults were removed after each count and the vials were kept for the next 60 d to assess the emergence of the F₁ generation. The findings from these studies proved that the extended exposure interval and the highest combined dose rate of the entomopathogenic fungus and the diatomaceous earth gave the maximum mortality of the beetles. The emergence of the progeny was also highly suppressed where the maximum dose rate of the synergized treatments was applied. The rate of mycosis and sporulation in the cadavers of R. dominica was maximum where the low dose rates of B. bassiana were applied.     

Bioactivity of pyrogallol against melon fruit fly, <Emphasis Type="Italic">Bactrocera cucurbitae</Emphasis>

The insecticidal effects of pyrogallol were studied by treating eggs and larvae of the melon fruit fly, Bactrocera cucurbitae (Coquillett) (Tephritidae: Diptera), with various concentrations (1, 5, 25, 125, 625 and 3125 ppm) of the phenolic compound. Although egg hatching decreased following treatment of 0–8-h old eggs with pyrogallol, the decrease was not significantly different from the control. Larval period and total development period declined significantly in 64–72-h-old and 88–96-h-old B. cucurbitae larvae fed on pyrogallol-treated diet. However, in the 44–48-h-old larvae, the larval period and total development period were not affected by pyrogallol treatment at any of the tested concentrations. None of them survived up to the pupal stage at the highest concentration. Number of pupae formed and adult emergence decreased significantly in all larval instars following feeding on pyrogallol-treated diet. The analysis of enzymes in 64–72-h-old larvae treated with LC₄₀ concentration (16.21 ppm) of pyrogallol at three time intervals, i.e., 24 h, 48 h and 72 h, showed significant induction in the activities of ascorbate peroxidase (APOX) and glutathione S-transferases (GSTs) at 24 h but a decrease was observed following prolonged treatment. On the other hand, superoxide dismutase (SOD) and peroxidases (POX) activity remained suppressed during the initial treatment interval but increased with prolonged treatment in 136–144-h-old larvae. The catalase (CAT) activity was suppressed at all treatment durations whereas glutathione reductase (GR) activity was not affected by pyrogallol treatment. An increase in the activities of ascorbate peroxidase, superoxide dismutase, peroxidases and glutathione S-transferases indicates an induction of defensive response of the melon fruit fly to the toxic effects produced by ingestion of pyrogallol. Although the effects of the compound on enzyme activity were tested on second instar, it would be interesting to see the effects on other instars too.     

Fumigant toxicity of<Emphasis Type="Italic">Eucalyptus blakelyi</Emphasis> and<Emphasis Type="Italic">Melaleuca fulgens</Emphasis> essential oils and 1,8-cineole against different development stages of the rice weevil<Emphasis Type="Italic">Sitophilus oryzae</Emphasis>

Essential oils extracted fromEucalyptus blakelyi (1,8-cineole, 77.5%),Melaleuca fulgens (1,8-cineole, 56.9%) and 1,8-cineole were shown to have fumigant toxicity against different development stages ofSitophilus oryzae. The eggs ofS. oryzae were the most tolerant, followed by pupae, larvae and adults in that order.M. fulgens oil,E. blakelyi oil and 1,8-cineole at 100 μl per liter of air gave, respectively, LT₅₀ values of 16.2, 17.4 and 9.1 h for adults, 31.1, 19.3 and 17.5 h for larvae, 55.6, 75.2 and 39.7 h for pupae, and required >7 days for eggs. Only 1,8-cineole (200 μl ⁻¹ air) gave a significant egg kill by 7 days and the LT₉₅ was 134.5 h. 1,8-Cineole could be a useful new fumigant. http://www.phytoparasitica.org posting Oct. 3, 2004.     

Suitability of different artificial diets for development and survival of stages of the predaceous ladybird beetle <Emphasis Type="Italic">Eriopis connexa</Emphasis>

The ladybug Eriopis connexa (Germar) (Coleoptera: Coccinellidae) is an important natural enemy of various pests. The potential of rearing it on 17 different diets was evaluated. The percentage of E. connexa adults was higher when its larvae received only eggs of Anagasta kuehniella (Zeller) (Lepidoptera: Pyralidae) after freezing for 1 day (92.5%) or combined in artificial diets with honey and water (82.5% to 100.0%). The viability from larvae to adult was 72.5% with eggs of A. kuehniella (after 1 day’s freezing) plus an artificial diet based on pet food. No adults of E. connexa were obtained with artificial diets as a stand-alone food source. The duration of the larval period to adult of this predator was longer, but with low viability, with only A. kuehniella eggs (after 6 months’ freezing) or with eggs + artificial diets. Eggs of A. kuehniella (after 1 day’s freezing) supplied separately or along with artificial diets were more appropriate to rear E. connexa and both diets can be used for mass rearing of this natural enemy.     

<Emphasis Type="Italic">Citrus exocortis viroid</Emphasis> transmission through commercially-distributed seeds of <Emphasis Type="Italic">Impatiens</Emphasis> and <Emphasis Type="Italic">Verbena</Emphasis> plants

Surveys of Impatiens and Verbena species in local nurseries in Fredericton, Canada and Verbena species in New Delhi, India showed widespread infection of Citrus exocortis viroid (CEVd) in vegetatively-propagated and seed-grown plants. To determine viroid seed transmission, samples of eight varieties of Impatiens and 11 varieties of Verbena were obtained from four commercial sources. All 19 samples collected contained viroid infection irrespective of variety. The presence of viroid in non-germinated seed was 21%, while the transmission rate in seedlings was 66% in Impatiens walleriana in 2006. Following 2 years of seed storage, the respective figures were 6% and 26%. Similarly, in Verbena x hybrida the presence of viroid in seed was 13% in 2006 with a seed-transmission rate in seedlings of 28%, while the respective figures after 2 years of storage were 5% and 45%.     

Comparison of the development <Emphasis Type="Italic">in planta</Emphasis> of a pyrrolnitrin-resistant mutant of <Emphasis Type="Italic">Botrytis cinerea</Emphasis> and its sensitive wild-type parent isolate

Botrytis cinerea is able to build-up resistance to pyrrolnitrin, an antibiotic produced by diverse biocontrol agents, possibly compromising the durability of this method of disease control. The development of two near-isogenic lines of B. cinerea differing in their level of resistance to pyrrolnitrin was compared in tomato plants and on PDA medium. In tomato plants, significant differences in the percentage of infected petioles 1 day after inoculation and in symptom progression on petioles and stems were observed between the resistant mutant and the sensitive wild-type parent, suggesting a difference in their level of aggressiveness. Cytohistological investigations revealed that conidia of both near-isogenic lines germinated 6 h after inoculation and mycelium developed within petiole tissues 12 h after inoculation. However, while the wild-type parent isolate spread throughout the petiole and rapidly invaded the stem tissues via the leaf-abscission zone 72 h after inoculation, the pyrrolnitrin-resistant mutant failed to extend beyond petiole tissues to invade the stem. Moreover, 72 h after inoculation, the mycelial development of the pyrrolnitrin-resistant mutant was accompanied by abnormal glycogen accumulation and chlamydospore-like cell formation. In contrast, wild-type parent mycelium was normally structured with intensive colonization of stem tissues. Additionally, on PDA medium the mycelium of the pyrrolnitrin-resistant mutant was less vigorous than the wild-type isolate. These results suggest that the acquisition of pyrrolnitrin-resistance in B. cinerea is accompanied by changes in mycelial structure and reduction in mycelial growth, leading to a noticeable loss of aggressiveness on tomato plants.     

Suppression of <Emphasis Type="Italic">Papaya ringspot virus</Emphasis> infection in <Emphasis Type="Italic">Carica papaya</Emphasis> with CAP-34, a systemic antiviral resistance inducing protein from <Emphasis Type="Italic">Clerodendrum aculeatum</Emphasis>

CAP-34, a protein from Clerodendrum aculeatum inducing systemic antiviral resistance was evaluated for control of Papaya ringspot virus (PRSV) infection in Carica papaya. In control plants (treated with CAP-34 extraction buffer) systemic mosaic became visible around 20 days that intensified up to 30 days in 56% plants. During this period, CAP-34-treated papaya did not show any symptoms. Between 30 and 60 days, 95% control plants exhibited symptoms ranging from mosaic to filiformy. In the treated set during the same period, symptoms appeared in only 10% plants, but were restricted to mild mosaic. Presence of PRSV was determined in induced-resistant papaya at the respective observation times by bioassay, plate ELISA, immunoblot and RT-PCR. Back-inoculation with sap from inoculated resistant plants onto Chenopodium quinoa did not show presence of virus. The difference between control and treated sets was also evident in plate-ELISA and immunoblot using antiserum raised against PRSV. PRSV RNA was not detectable in treated plants that did not show symptoms by RT-PCR. Control plants at the same time showed a high intensity band similar to the positive control. We therefore suggest that the absence/delayed appearance of symptoms in treated plants could be due to suppressed virus replication.     

Cloning and characteristics of <Emphasis Type="Italic">Brn1</Emphasis> gene in <Emphasis Type="Italic">Curvularia lunata</Emphasis> causing leaf spot in maize

The full length cDNA of the Brn1 was first cloned, and then expression of the Brn1 was analyzed and the function was identified by silencing technology. Results show that the full length cDNA of the C. lunata Brn1 gene contains 1001 base pairs and an 801 bp open reading frame encoding 267 amino acids. Semi-quantitative PCR analysis shows that the expression of Brn1 at 96 h is significantly higher than at 24 and 72 h (p < 0.05) in both the highly virulent isolate CX-3 and the weakly virulent isolate DD60. Brn1-silenced transformants were light brown in culture filtrate, and have significantly reduced toxin production relative to the wild-type. These results imply that Brn1 gene in C. lunata is not only involved in 1,8-dihydroxynaphthalene melanin synthesis, but is also relatively associated with toxin biosynthesis of the pathogen.     

Possible function of isoleucine in the methyl jasmonate response of <Emphasis Type="Italic">Arabidopsis</Emphasis> to <Emphasis Type="Italic">Phelipanche aegyptiaca</Emphasis>

Resistance of Arabidopsis thaliana to Phelipanche aegyptiaca (syn. Orobanche aegyptiaca) can be induced by exposure of the host to low concentrations of gaseous methyl jasmonate, in a concentration-dependent fashion. Application of methyl jasmonate at marginal concentrations, 10⁻⁸ M, for 12 h reduced infection by 50%. However, if the host plants were supplied with isoleucine at 10⁻³ M, and the isoleucine removed prior to exposure to methyl jasmonate, infection was reduced to less than 20%. The presence of isoleucine throughout the experiment—before, during and after exposure to methyl jasmonate—reduced infection of the host to almost zero. These results suggest that when Arabidopsis is exposed to methyl jasmonate, full activation of the defense mechanisms, including formation of the isoleucine jasmonic acid conjugate, might be limited by the availability of isoleucine in the host. These findings add another factor to the complex interaction between infecting parasite, and evocation of host defense mechanisms.     

Daily consumption and predation rate of different<Emphasis Type="Italic">Stethorus punctillum</Emphasis> instars feeding on<Emphasis Type="Italic">Tetranychus urticae</Emphasis>

Laboratory experiments were conducted to evaluate the prey stage preference and the daily consumption of each stage of the coccinellid predatorStethorus punctillum Weise (Coleoptera: Coccinellidae) feeding on the two-spotted spider miteTetranychus urticae (Koch) (Acari: Tetranychidae). Groups of different life stages of the prey were offered (eggs, larvae, nymphs and adults). The prey preference varied with the stage ofS. punctillum. First larval instars had no significant preference among theT. urticae stages offered. Second larval instars consumed significantly more spider mite larvae in comparison with nymphs. In contrast, third larval instars indicated a strong preference for mite eggs. Significantly fewerT. urticae larvae were consumed by the fourth larval instars ofS. punctillum, in comparison with the three other mite stages. Finally, adult predators consumed significantly more mite eggs than the other stages offered. This preferential trend was similar for all adults tested, whether during the pre-oviposition or the oviposition period. http://www.phytoparasitica.org posting Feb. 17, 2004.     

Conventional PCR and real time quantitative PCR detection of <Emphasis Type="Italic">Phytophthora cryptogea</Emphasis> on <Emphasis Type="Italic">Gerbera jamesonii</Emphasis>

A conventional PCR and a SYBR Green real-time PCR assays for the detection and quantification of Phytophthora cryptogea, an economically important pathogen, have been developed and tested. A conventional primer set (Cryp1 and Cryp2) was designed from the Ypt1 gene of P. cryptogea. A 369 bp product was amplified on DNA from 17 isolates of P. cryptogea. No product was amplified on DNA from 34 other Phytophthora spp., water moulds, true fungi and bacteria. In addition, Cryp1/Cryp2 primers were successfully adapted to real-time PCR. The conventional PCR and real-time PCR assays were compared. The PCR was able to detect the pathogen on naturally infected gerbera plants and on symptomatic artificially infected plants collected 21 days after pathogen inoculation. The detection limit was 5 × 10³ P. cryptogea zoospores and 16 fg of DNA. Real-time PCR showed a detection limit 100 times lower (50 zoospores, 160 ag of DNA) and the possibility of detecting the pathogen in symptomless artificially infected plants and in the re-circulating nutrient solution of closed soilless cultivation systems.     

Clutch size decisions of <Emphasis Type="Italic">Cotesia glomerata</Emphasis>, a gregarious parasitoid of <Emphasis Type="Italic">Pieris brassicae</Emphasis>

This work investigated the ability of the gregarious larval endoparasitoid Cotesia glomerata L. (Hymenoptera: Braconidae) to adjust the progeny sex ratio and clutch size with repeated oviposition experience and the effect of a honey-based diet on the clutch size. In the field-collected clusters many clusters were female-biased but some clusters (3.8%) produced only male wasps, suggesting that there is a low percentage of unmated females in the field. Superparasitism was common in the field, and females were believed to increase progeny sex ratio when attacking previously parasitized hosts. In the laboratory, the number of eggs laid in a day tended to decrease with increasing female age. For females that were offered two hosts per day and for those offered three hosts per day, this value became nearly the same at 9 days after the start of oviposition. Old females which attacked many hosts tended to lay fewer eggs in a day than young ones. However, the degree of this tendency was not the same for all the parasitoid females of all three groups because sperm remained viable throughout a female’s lifetime. The amount of sperm used in a single oviposition bout seemed fixed and was not dependent on the number of eggs laid. Over the 2 days of the clutch size response experiment, the number of hosts a female attacked per day was not affected by the presence or absence of honey.