The effects of different temperatures and relative humidity on the development,mortality and nymphal predation of <Emphasis Type="BoldItalic">Anthocoris minki</Emphasis>

Mass rearing techniques are a necessity in developing a suitable and economic biological control method. Anthocoris minki Dohrn (Heteroptera: Anthocoridae), which is successfully reared for use in biological control, is a promising indigenous Anthocoris species for biological control of Agonoscena pistaciae Burckhardt and Lauterer (Homoptera: Psyllidae) in pistachio orchards. Development time of nymphal instars, mortality percentage and prey consumption of A. minki fed on Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs were studied at three constant temperatures (20, 25 and 30 ± 1°C) and two relative humidity levels (40 and 65 ± 5%) under laboratory conditions. Temperature and r.h. significantly affected development time and nymphal prey consumption of A. minki. Duration of incubation and total nymphal development time decreased significantly with each increase in temperature. Total nymphal development time of A. minki at 40% and 65% r.h. was 18.6 and 18.6 days at 20°C; 13.7 and 14.6 days at 25°C; and 10.8 and 11.8 days at 30°C, respectively. The shortest nymphal development time and highest nymphal mortality percentage were recorded at 30°C and 40% r.h. The lowest nymphal mortality was 30.1% and 32.1% at 40% and 65% r.h., respectively, at 25°C; the highest nymphal mortality was 51.5% and 52.1% at 20° and 30°C, respectively, and 40% r.h. The highest average number of E. kuehniella eggs consumed was 123.1 at 20°C and 65% r.h., and the lowest was 86.4 at 30°C and 40% r.h. during nymphal development time of A. minki.     

Prey consumption by the mealybug predator <Emphasis Type="Italic">Spalgis epius</Emphasis> on pink hibiscus mealybug (<Emphasis Type="Italic">Maconellicoccus hirsutus</Emphasis>)

The pink hibiscus mealybug Maconellicoccus hirsutus (Green) (Hemiptera: Pseudococcidae) is a major pest of economically important crops. The apefly Spalgis epius (Westwood) (Lepidoptera: Lycaenidae) is a potential predator of various species of mealybugs. Studies of its preying potential and preference for prey stages on M. hirsutus are incomplete. An investigation was undertaken to determine the daily prey consumption and preference for prey stages by different larval instars of S. epius reared on M. hirsutus in the laboratory. During the 8-day larval development period with four larval instars of S. epius, the daily prey consumption increased from the first to the seventh day and decreased on the eighth day prior to the prepupal stage. Generally, there was a significant difference in the prey consumption on different days. The 1st to 4th instar larvae of S. epius consumed, respectively, a mean of 181.3, 679.1, 1770.4 and 4333.0 eggs or 19.1, 67.7, 153.0 and 639.0 nymphs or 2.72, 6.26, 13.8 and 32.1 adults of M. hirsutus. When an S. epius larva was fed on M. hirsutus eggs, nymphs and adults separately, it consumed a mean of 6952.6 eggs, 878.8 nymphs or 53.9 adults during its entire development. A single larva of S. epius consumed 2358.3 eggs, 151.2 nymphs and 34.3 adults of M. hirsutus during its entire development when the prey stages were offered all together. The study revealed that S. epius is a voracious predator of M. hirsutus and thus could be utilized as a potential biological control agent.     

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.     

Prey density-dependent feeding activity and life history of <Emphasis Type="Italic">Scymnus subvillosus</Emphasis>

The functional response, development time, survival and reproduction of the lady beetle Scymnus subvillosus (Goeze) (Coleoptera: Coccinellidae) were evaluated at different densities of the mealy plum aphid Hyalopterus pruni (Geoffroy) (Homoptera: Aphididae). Treatments were carried out at 25 ± 1°C, 60 ± 10% r.h. and 16L:8D photoperiod in a controlled temperature room. The larvae and adults of S. subvillosus were fed with different densities (5, 10, 20, 40 and 80) of H. pruni in petri dishes. The shape of the functional response was determined by logistic regression, and the values of the coefficient of attack rates (α) and handling times (T _h) were estimated by using nonlinear least-squares regression. Behavior of each larval stage and adults matched Holling’s type II functional response. Estimates of α for all stages of S. subvillosus tested were similar, but estimates of T _h varied; it was the shortest for adult females and the longest for third instar larva. Larvae were able to complete their development at each of the five prey densities, but increased prey densities reduced development time and mortality rate. Increased prey consumption did not change longevity, but resulted in a higher intrinsic rate of increase (r), the finite rate of increase (λ), net reproduction rate (R ₀ ), gross reproductive rates (GRR), and shorter mean generation time (T) and doubling time (DT). This research was funded by the Yuzuncu Yil University/foundation of scientific research projects (No. 2002-ZF 044)     

Pathogenicity of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> and <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> against <Emphasis Type="Italic">Galleria mellonella</Emphasis>

The greater wax moth Galleria mellonella L. (Lepidoptera: Pyralidae) is occasionally found in beehives and is a major pest of stored wax. Entomopathogenic fungi have recently received attention as possible biocontrol elements for certain insect pests. In this study, 90 isolates of Beauveria bassiana and 15 isolates of Metarhizium anisopliae were screened for proteases and lipases production. The results showed significant variations in the enzymatic action between the isolates. In the bioassay, the selected isolates evinced high virulence against the 4th instar of the G. mellonella larvae. The isolates BbaAUMC3076, BbaAUMC3263 and ManAUMC3085 realized 100% mortality at concentrations of 5.5 × 10⁶ conidia ml⁻¹, 5.86 × 10⁵ conidia ml⁻¹, and 4.8 × 10⁶ conidia ml⁻¹, respectively. Strong enzymatic activities in vitro did not necessarily indicate high virulence against the tested insect pest. The cuticle of the infected larvae became dark and black-spotted, indicating direct attack of fungus on the defense system of the insects. The LC₅₀ values were 1.43 × 10³, 1.04 × 10⁵ and 5.06 × 10⁴ for Bba3263AUMC, Bba3076AUMC and Man3085AUMC, respectively, and their slopes were determined by computerized probit analysis program as 0.738 ± 0.008, 0.635 ± 0.007 and 1.120 ± 0.024, respectively.     

Effects of calcium salts in controlling <Emphasis Type="Italic">Phytophthora pistaciae,</Emphasis> the causal agent of pistachio gummosis

The ability of calcium chloride and calcium sulfate to control pistachio gummosis caused by Phytophthora pistaciae was evaluated both in vitro, on solid and liquid media, and in vivo, on broad bean, a potential annual host of the pathogen, and seedlings of three pistachio cultivars. In vitro, all concentrations of the salts (1.5, 4, 8 and 15 mM) reduced mycelial growth, dry weight of mycelial mats, and sporangial production of the pathogen. Calcium chloride reduced the size and volume of sporangia, antheridia and oogonia. Calcium sulfate also reduced the size and volume of sporangia, but it had less impact on the volume of oogonia and increased the size of antheridia and the volume of oospores. Moreover, concentrations of both calcium salts higher than 1.5 mM reduced zoospore discharge and induced cyst germination. In vivo, application of calcium salts before and after inoculation of sterilized soil with P. pistaciae reduced the mortality of broad bean plants and different cultivars of pistachio trees compared to the respective controls. Analyses using atomic absorption showed calcium concentrations to increase in the foliage of Ca-treated broad bean and pistachio plants. Based on the results, the application of calcium chloride to P. pistaciae infested soils could be more advantageous than calcium sulfate to control P. pistaciae gummosis in commercial pistachio crops.     

Pathogenicity of morphologically different isolates of <Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis> with <Emphasis Type="Italic">Brassica napus</Emphasis> and <Emphasis Type="Italic">B. juncea</Emphasis> genotypes

Sclerotinia stem rot caused by Sclerotinia sclerotiorum is a serious threat to oilseed production in Australia. Eight isolates of S. sclerotiorum were collected from Mount Barker and Walkway regions of Western Australia in 2004. Comparisons of colony characteristics on potato dextrose agar (PDA) as well as pathogenicity studies of these isolates were conducted on selected genotypes of Brassica napus and B. juncea. Three darkly-pigmented isolates (WW-1, WW-2 and WW-4) were identified and this is the first report of the occurrence of such isolates in Australia. There was, however, no correlation between pigmentation or colony diameter on PDA with the pathogenicity of different isolates of this pathogen as measured by diameter of cotyledon lesion on the host genotypes. Significant differences were observed between different isolates (P ≤ 0.001) in two separate experiments in relation to pathogenicity. Differences were also observed between the different Brassica genotypes (P ≤ 0.001) in their responses to different isolates of S. sclerotiorum and there was also a significant host × pathogen interaction (P ≤ 0.001) in both experiments. Responses between the two experiments were significantly correlated in relation to diameter of cotyledon lesions caused by selected isolates (r = 0.79; P < 0.001, n = 48). Responses of some genotypes (e.g., cv. Charlton) were relatively consistent irrespective of the isolates of the pathogen tested, whereas highly variable responses were observed in some other genotypes (e.g., Zhongyou-ang No. 4, Purler) against the same isolates. Results indicate that, ideally, more than one S. sclerotiorum isolate should be included in any screening programme to identify host resistance. Unique genotypes which show relatively consistent resistant reactions (e.g., cv. Charlton) across different isolates are the best for commercial exploitation of this resistance in oilseed Brassica breeding programmes.     

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.     

Resistance to <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> expresses a hypersensitive-like response in <Emphasis Type="Italic">Coffea arabica</Emphasis>

Root-knot nematodes (RKN) are obligate parasite species of the genus Meloidogyne that cause great losses in Arabica coffee (Coffea arabica L.) plantations. Identification of resistant genotypes would facilitate the improvement of coffee varieties aiming at an environmental friendly and costless nematode control. In this work, the C. arabica genotype ‘UFV 408-28’ was found to be resistant to the most destructive RKN species M. incognita. Pathogenicity assays indicated that the highly aggressive populations of M. incognita races 1, 2 and 3 were not able to successfully reproduce on ‘UFV 408-28’ roots and displayed a low gall index (GI = 2). An average reduction of 87% reduction of the M. incognita population was observed on ‘UFV 408-28’ when compared to the susceptible cultivar ‘IAC 15’. By contrast, ‘UFV 408-28’ was susceptible to the related species M. exigua and M. paranaensis (GI = 5 and 4, respectively). Histological observations performed on sections of UFV408-28 roots infected with M. incognita race 1 showed that nematode infection could be blocked right after penetration or during migration and establishment stages, at 6 days, 7 days and 8 days after infection (DAI). Fluorescence and bright field microscopy observations showed that root cells surrounding the nematodes exhibited HR-like features such as accumulation of phenolic compounds and a necrotic cell aspect. In the susceptible ‘IAC 15’ roots, 6 DAI, feeding sites contained giant cells with a dense cytoplasm. Necrotic cells were never observed throughout the entire infection cycle. The HR-like phenotype observed in the ‘UFV 408-28’—M. incognita interaction suggests that the coffee resistance may be mediated by a R-gene based immunity system and may therefore provide new insights for understanding the molecular basis of RKN resistance in perennial crops.     

<Emphasis Type="Italic">Odontoglossum ringspot virus</Emphasis> causing flower crinkle in <Emphasis Type="Italic">Phalaenopsis</Emphasis> hybrids

A new disorder exhibiting flower crinkle on Phalaenopsis orchids bearing white flowers has been observed in Taiwan, China and Japan for several years. This disorder decreased the flower longevity and was considered as a physiological syndrome. The objective of this study was to identify and characterize the real causal agent of this new Phalaenopsis disorder. Five plants of Phalaenopsis hybrids “V3” (Phal. Yukimai × Phal. Taisuco Kochdian) with flower crinkle symptoms were collected and tested by enzyme-linked immunosorbent assay with antisera against 18 viruses. The extract of leaves and flowers from one diseased plant (96-Ph-16) reacted positively only to antiserum against Odontoglossum ringspot virus (ORSV), while those from the other four plants (96-Ph-7, 96-Ph-17, 96-Ph-18 and 96-Ph-19) reacted positively to the antisera against ORSV and Cymbidium mosaic virus (CymMV). Five ORSV isolates, one each from flowers of those five diseased Phalaenopsis orchids, were established in Chenopodium quinoa. A CymMV culture was isolated from the flowers of one of the ORSV/CymMV mix-infected Phalaenopsis orchids (96-Ph-19). To determine the causal agent of the flower crinkle disease, healthy Phalaenopsis seedlings were singly or doubly inoculated with the isolated ORSV and/or CymMV. Results of back inoculation indicated that ORSV is the sole causal agent of the crinkle symptom on petals of Phalaenopsis orchid. The CP gene of the ORSV isolates from this study shared 97.3–100% nucleotide identity and 96.2–100% amino acid identity with those of 41 ORSV isolates available in GenBank. This is the first report demonstrating ORSV as the sole virus causing flower crinkle disease on Phalaenopsis orchids.     

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.     

Effects of photoperiod on the development and growth of <Emphasis Type="Italic">Podisus nigrispinus</Emphasis>, a predator of cotton leafworm

The effects of photoperiod on nymphal development, growth and adult size in Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) were studied. Predators were collected in cotton fields in Patos (7°S, 37°W), Paraíba State, Brazil. A randomized block experimental design was used, with treatments consisting of photoperiods of 10L:14D, 11L:13D, 12L:12D, 13L:11D, 14L:10D, 15L:9D and 16L:8D (LD, in h), at a constant temperature of 28 ± 1°C and relative air humidity of 70 ± 10%. Treatments were distributed in four replications, with each experimental unit composed of 40 nymphs. The development period for each instar of P. nigrispinus varied according to the photoperiod exposure. Regardless of the photophase (PhP), the 5th instar nymphs exhibited the longest development period, except for the 15-h PhP, in which the development period of 2nd instar nymphs (4.13 days) was as long as that of the 5th instar nymphs (4.23 days). In the 1st, 3rd and 5th instars of P. nigrispinus, the development period was inversely proportional to the increase in light period in which the nymphs developed, for the PhP intervals of 10–14 h, 12–14 h, and 12–15 h, respectively. Predators exposed to a 14-h PhP developed a wider pronotum than those exposed to extreme PhP’s (of 10 h, 11 h and 16 h). Conditions from 14 h to 15 h of light resulted in higher daily growth rates in P. nigrispinus than those obtained with the other PhP’s tested. P. nigrispinus females exhibited faster daily growth rates than did males.     

Feeding potential of <Emphasis Type="Italic">Cryptolaemus montrouzieri</Emphasis> against the mealybug <Emphasis Type="Italic">Phenacoccus solenopsis</Emphasis>

Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) is an exotic species native to the USA, damaging cotton and other plant families. The feeding potential of different development stages of Cryptolaemus montrouzieri Mulsant, a biological control agent against mealybugs, was investigated on different development stages of P. solenopsis. Fourth instar grubs and adults of C. montrouzieri were the most voracious feeders on different instars of mealybug. The number of 1^st instar nymphs of mealybug consumed by 1^st, 2^nd, 3^rd and 4^th instar larvae and adult beetles of C. montrouzieri was 15.56, 41.01, 125.38, 162.69 and 1613.81, respectively. The respective numbers of 2^nd and 3^rd instar nymphs of mealybug consumed were 11.15 and 1.80, 26.35 and 6.36, 73.66 and 13.32, 76.04 and 21.16, 787.95 and 114.66. The corresponding figures for adult female mealybugs were 0.94, 3.23, 8.47, 12.71 and 73.40, respectively. The results indicate that C. montrouzieri has the potential to be exploited as a biocontrol agent in North India; inoculative releases of 4^th instar larvae and adults may provide instant control of P. solenopsis. Field experiments should be conducted to determine the efficiency of the ladybird on this mealybug.     

<Emphasis Type="Italic">Paecilomyces lilacinus</Emphasis>, a potential biocontrol agent on apple rust mite <Emphasis Type="Italic">Aculus,schlechtendali</Emphasis> and interactions with some fungicides <Emphasis Type="Italic">in vitro</Emphasis>

The apple rust mite Aculus schlechtendali (Nal.) (Acari: Eriophyidae), is a main pest in apple-growing areas in Ankara, Turkey, and chemical control applications have some limitations. Entomopathogenic fungi have a potential for biological control of mites. In this study, an entomopathogenic fungus, Paecilomyces lilacinus (Thom) Samson (Deuteromycota: Hyphomycetes), was first isolated from the mite cadavers on Japanese crab apple leaves and pathogenicity of the fungus was observed in different inoculum densities and relative humidities. The pathogen caused up to 98.22% mortality of the mite population. The effects of some fungicides on the entomopathogenic fungus were determined in in vitro studies. Carbendazim, penconazole and tebuconazole were the most effective fungicides on mycelial growth of P. lilacinus, with EC₅₀ values under 3 μg ml⁻¹. In spore germination tests, captan, mancozeb, propineb were the most effective fungicides, followed by tebuconazole, penconazole, nuarimol and chlorothalonil. Sulphur could not inhibit the conidia germination totally at 5,000 μg ml⁻¹. Copper oxychloride and fosetyl-al prevented conidia formation at concentrations above 1,000 μg ml⁻¹.     

Effect of azadirachtin applied systemically through roots of plants on the mortality,development and fecundity of the cabbage aphid (<Emphasis Type="Italic">Brevicoryne brassicae</Emphasis>)

The effect of low concentrations of azadirachtin applied systemically through root tissues of rape plants (Brassica napus subsp.napus) on the mortality, period of development, longevity and fecundity of the cabbage aphid (Brevicoryne brassicae L.) (Sternorrhyncha: Aphididae) was studied. The concentrations of azadirachtin A tested were: 0.25, 0.05, 0.005, 0.0005 and 0.00005 mg ml⁻¹. The mortality of cabbage aphid nymphs, particularly during the period of ecdysis, increased significantly with increasing concentrations. Azadirachtin had no effect on the length of the development period of the respective immature stages of cabbage aphids. The average longevity of the cabbage aphids decreased with increasing azadirachtin concentration and time spent feeding. The fecundity of aphids decreased after the application of azadirachtin. http://www.phytoparasitica.org posting May 6, 2004.     

Functional response of the coccinellid predator <Emphasis Type="Italic">Adalia fasciatopunctata revelierei</Emphasis> to walnut aphid (<Emphasis Type="Italic">Callaphis juglandis</Emphasis>)

The functional response types and parameters of 3rd and 4th instar larvae, and adult females and males of a coccinellid predator, Adalia fasciatopunctata revelierei (Mulsant) (Col.: Coccinellidae), were evaluated at five different densities of Callaphis juglandis (Goeze) (Hemiptera: Aphididae) in order to understand their role for the aphid’s biological control. Experiments were carried out in petri dishes at 25 ± 1°C, 60 ± 10% r.h. and 16L:8D photoperiod in a controlled temperature room. All tested stages exhibited a Type II response determined by a logistic regression model. The attack rate (α) and handling time (T _h ) coefficients of a Type II response were estimated by fitting a “random-predator” equation to the data. Although the estimates of α for all stages of A. fasciatopunctata revelierei tested were similar, the longest T _h was obtained for 3rd instar larva because of the lower consumption rate at densities above 40 prey/day. Results indicated that the adult female has the highest predation of C. juglandis followed by 4th instar larvae, adult males and 3rd instar larvae. However, further field-based studies are needed to draw firm conclusions.     

Pathogenicity and reproductive fitness of <Emphasis Type="Italic">Pratylenchus coffeae</Emphasis> and <Emphasis Type="Italic">Radopholus arabocoffeae</Emphasis> on Arabica coffee seedlings (<Emphasis Type="Italic">Coffea arabica</Emphasis> cv. Catimor) in Vietnam

The pathogenicity and reproductive fitness of Pratylenchus coffeae and Radopholus arabocoffeae from Vietnam on coffee (Coffea arabica) seedlings cv. Catimor were evaluated in greenhouse experiments. The effect of initial population densities (Pi = 0, 1, 2, 4, 8, 16, 32, 64, 128, and 256 nematodes per cm³ soil) was studied for both species at different days after inoculation (dai). The data were adjusted to the Seinhorst damage model Y = m + (1-m).z^Pi-T. Tolerance limit (T) for P. coffeae was zero for the height and the diameter of the coffee plants. For the diameter, the T-value for R. arabocoffeae was 25.6 for 30 and 60 dai and 12.8 for 90 and 120 dai. After 4 months T was zero. The low tolerance limits indicate that Arabica coffee is highly intolerant to both nematode species. At the end of the experiment (180 dai), all plants were infected and most were dead when inoculated with R. arabocoffeae at initial densities of 32, 64, 128 and 256 nematodes/cm³ soil. For P. coffeae plant death was already observed at the lowest inoculation densities. Growth of coffee was reduced at all inoculation levels for both species. Pratylenchus coffeae and R. arabocoffeae caused intense darkening of the roots, leaf chlorosis and a strong reduction of root and shoot growth. It was observed that P. coffeae mainly destroyed lateral roots rather than tap roots, whereas R. arabocoffeae reduced tap root length rather than the lateral roots. At the lowest inoculum densities, the reproduction factor of P. coffeae was 2.38 and 2.01 for R. arabocoffeae, indicating that arabica coffee is a host for both species. Plant growth as expressed by shoot height and shoot and root weight measured 60 dai was negatively correlated with nematode (both species) density as expressed by the geometric mean of nematode numbers at 30 and 60 dai.     

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.     

Selection,characterization and genetic analysis of laboratory mutants of <Emphasis Type="Italic">Botryotinia fuckeliana</Emphasis> (<Emphasis Type="Italic">Botrytis cinerea</Emphasis>) resistant to the fungicide boscalid

Resistance to the fungicide boscalid in laboratory mutants of Botryotinia fuckeliana (Botrytis cinerea) was investigated. The baseline sensitivity to boscalid was evaluated in terms of colony growth (EC₅₀ = 0.3–3 μg ml⁻¹; MIC = 10–30 μg ml⁻¹) and conidial germination (EC₅₀ = 0.03–0.1 μg ml⁻¹; MIC = 1–3 μg ml⁻¹) tests. Mutants were selected in vitro from wild-type strains of the fungus on a fungicide-amended medium containing acetate as a carbon source. Mutants showed two different levels of resistance to boscalid, distinguishable through the conidial germination tests: low (EC₅₀ ∼ 0.3 μg ml⁻¹, ranging from 0.03 to 1 μg ml⁻¹; MIC > 100 μg ml⁻¹) and high (EC₅₀ > 100 μg ml⁻¹) resistance. Analysis of meiotic progeny from crosses between resistant mutants and sensitive reference strains showed that resistant phenotypes were due to mutations in single major gene(s) inherited in a Mendelian fashion, and linked with both the Daf1 and Mbc1 genes, responsible for resistance to dicarboximide and benzimidazole fungicides, respectively. Gene sequence analysis of the four sub-units of the boscalid-target protein, the succinate dehydrogenase enzyme, revealed that single or double point mutations in the highly conserved regions of the iron-sulphur protein (Ip) gene were associated with resistance. Mutations resulted in proline to leucine or phenylalanine replacements at position 225 (P225L or P225F) in high resistant mutants, and in a histidine to tyrosine replacement at position 272 (H272Y) in low resistant mutants. Sequences of the flavoprotein and the two transmembrane sub-units of succinate dehydrogenase were never affected.     

Survival of <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> in infected tissues of <Emphasis Type="Italic">Capsicum annuum</Emphasis> and in soils of the state of Pernambuco,Brazil

The survival of Ralstonia solanacearum A1-9^Rif race 1 phylotype I was studied in ten different soil types in the absence of the host plant as well as in infected tissues of the stem and root of bell peppers buried in the soil at 0, 5, and 15 cm. The survival time of R. solanacearum A1-9^Rif in the ten soil types ranged from 42 up to 77 days. Among the chemical and physical characteristics of the soil, clay content, residual moisture, and available water were positively correlated, and pH was negatively correlated, with survival time, population size at 42 days, and area under the population curve. The pathogen survival differed significantly in relation to the plant tissues, but not with respect to the incorporation depth of the infected tissues. The root tissue of bell pepper supported a larger bacterial population at 7 and 21 days (5 × 10⁴ and 3.1 × 10⁴ CFU g⁻¹ tissue, respectively) compared with the stem tissue (0.35 × 10⁴ and 0.48 × 10⁴ CFU g⁻¹ tissue, respectively) and also had a larger area under the population curve. On the other hand, the stem tissues presented a greater decomposition rate and pH compared with the roots. In conclusion, the different types of studied soils as well as the infected bell pepper tissues were considered potential primary sources of R. solanacearum inocula, but only for a short period.