Prey consumption during development as well as longevity and reproduction of Typhlodromus pyri Scheuten (Acari, Phytoseiidae) at higher temperatures in the laboratory

The predatory mite Typhlodromus pyri Scheuten (Acari, Phytoseiidae) has been reported as an important predator of the European red mite, Panonychus ulmi (Koch) (Acari, Tetranychidae) in apple culture and vineyards at below 25°C. However, sufficient biological data was lacking on its efficiency at temperatures above 25°C. Therefore, the purpose of the present laboratory work was to obtain experimental data on prey consumption during development as well as longevity and reproduction of T. pyri on apple leaf discs and in Plexiglas cells at constant temperatures of 25±2°C and 30±2°C with P. ulmi as prey.The results showed that mean daily and total prey consumption by both the nymphs and adults of T. pyri decreased significantly on both the arenas as the temperature was increased from 25°C to 30°C, whereby adult prey consumption, both mean daily and total, was higher than that of nymphs. Prey consumption by both the nymphs and adults was significantly higher in the Plexiglas cells than on the leaf discs at both temperatures. Mean total prey consumption during nymphal development was 16.1 () and 12.8 () at 25°C compared to 7.0 () and 5.8 () preys at 30°C on the apple leaf discs and 46.0 () and 38.5 () at 25°C compared to 25.2 () and 20.3 () preys at 30°C in the Plexiglas cells. Mean duration of nymphal development was similar for the two sexes at the same temperature, but it was longer at 25°C than at 30°C. It was 6.0 and 4.0days on the apple leaf discs while 7.0 and 6.0days in the Plexiglas cells at 25°C and 30°C, respectively. Mean daily and total prey consumption by both male and female adults also decreased with the increasing temperature, whereby the females consumed more than double the mean total number of prey than the males on both the arenas of observation and at both temperatures: 355.4 versus 149.7 preys at 25°C and 192.2 versus 85.6preys at 30°C on the leaf discs and 826.8 versus 374.5 preys at 25°C and 488.9 versus 187.9 preys at 30°C in the Plexiglas cells. Longevity of the females was longer than males on both arenas and at both temperatures and it was longer at 25°C than at 30°C. Mean total longevity on the apple leaf discs was 68.3 () and 50.8 () days at 25°C compared to 52.5 () and 36.8 () days at 30°C, while in the Plexiglas cells it was 91.0 () and 65.8 () days at 25°C compared to 75.3 () and 48.5 () days at 30°C. Reproduction in females also decreased significantly with increasing temperature. It decreased from 62.0 to 39.0 eggs/female on the leaf discs and 75.0 to 47.1 eggs/female in the Plexiglas cells. The females laid significantly higher numbers of eggs at both temperatures in the Plexiglas cells than on the leaf discs. Oviposition period in females was 30days at 25°C on both the arenas, while at 30°C it was 26days on the apple leaf discs and 27days in the Plexiglas cells.     

Influence of mixed biocide GCSC-BtA on the pupae and adult stages of Apanteles plutellae Kurd. (Hym., Braconidae) and its host. Plutella xylostella (L.) (Lep., Plutellidae)

This paper deals with the influence of the mixed biocide GCSC-BtA on the pupal and adult stages of Apanteles plutellae Kurd. (Hym., Braconidae) and its host, Plutella xylostella (L.) (Lep., Plutellidae). The results show that mortalities of the pupae of P. xylostella in the direct-dip bioassay were 84.67%, that of the adults in the residue bioassay at 1.2500mg/ml concentration of GCSC-BtA were 78.00% which were significantly higher than the mortality values for the pupae with 54.62% and adults with 48.13% of A. plutellae. In contrast, cypermethrin showed extremely high toxicity to the pupae with 94.58% and adults with 86.00% mortality values of A. plutellae as compared to the low mortality values of 42.14% for the pupae and 32.11% for the adults of P. xylostella, with the same concentrations and bioassay methods. The LC₅₀ values of GCSC-BtA were 0.3402, 0.5516 and 1.2405, 1.9480mg/ml for the pupae and adults of P. xylostella and A. plutellae, respectively, while the LC₅₀ values for cypermethrin were 1.5652, 2.3471 and 0.1096, 0.1152mg/ml, respectively. GCSC-BtA was found more toxic to the pupae and adults of P. xylostella and safer to the pupae and adults of A. plutellae than cypermethrin. The possibilty of using GCSC-BtA against P. xylostella under partial control by A. plutellae in vegetable fields is discussed.     

Biological characteristics and life tables of Neoseiulus umbraticusChant (Acari, Phytoseiidae) at three constant temperatures

The development time, survival and fecundity of the generalist predatory mite, Neoseiulus umbraticusChant, were determined at 20, 25, and 30°C and 65±10% RH. N. umbraticus females completed development in 9.7, 8.0 and 5.9 days, respectively, using a diet of all life stages of Tetranychus cinnabarinus Boisduval. Total developmental times of males were relatively shorter at 25 and 30°C than at 20°C. In general, preoviposition, oviposition, and postoviposition periods of N. umbraticus shortened as temperature increased. The longest survival rate of N. umbraticus of 80.5 days occurred at 20°C, followed by 67.0 and 57.6 days at 25 and 30°C, respectively. Mated females laid an average 0.9, 1.3 and 1.4 eggs per female per day and 33.1, 44.0 and 43.6 eggs over their entire lives at 20, 25 and 30°C, respectively. The sex ratios of this species were 0.57, 0.57 and 0.54 female (female+male) at 20, 25 and 30°C, respectively. The intrinsic rate of increase (r_m) became greater with rising temperatures from 0.123 at 20°C to 0.180 at 30°C. The net reproduction rate (R_o) was highest at 25°C (25.0 females/female) and lowest at 20°C (18.8 females/female), while T_o decreased with increasing temperatures, from 23.8 days at 20°C to 17.5 days at 30°C.     

Effects of soil solarization on nematodes in Croatia

For the first time soil solarization was investigated in Croatia both in the field and in the greenhouse in 1991, 1992, 1993 and 1994. For two months (July and August), the soil was mulched with transparent polyethylene (PE) sheets of 0.015 or 0.050mm thickness. Soil temperatures at depths of 5, 10 and 20cm were recorded daily. In order to assess nematode population densities, soil samples were analysed before mulching and at the end of the mulching treatment. The results of these experiments showed that soil solarization drastically reduced the population of plant-parasitic nematodes (Meloidogyne, Pratylenchus, Paratylenchus, Tylenchus, Tylenchorhynchus spp.) by about 97–100% at a depth of 10cm and 92–97% at a depth of 20cm in the field, while in the greenhouse, the population of plant-parasitic nematodes was reduced by about 89–100% at a depth of 10cm and 98–100% at a depth of 20cm.In the same experiments, the population of saprophytic nematodes in the field was reduced by about 86–90% at a depth of 10cm and 72–89% at a depth of 20cm. In the greenhouse, the population of saprophytic nematodes was reduced by about 87–97% at a depth of 10cm and 87–93% at a depth of 20cm. This data shows that soil solarization was less effective in the control of saprophytic nematodes, which is considered to be an advantage.     

Infection of Tomicus piniperda (Col., Scolytidae) with Canningia tomici sp. n. (Microsporidia, Unikaryonidae)

Canningia tomici sp. n. (Microsporidia, Unikaryonidae) infects the midgut epithelium, the gut muscules, Malpighian tubules, connective tissues, adipose tissues and the gonads of the pine shoot beetle, Tomicus piniperda (L.) (Coleoptera, Scolytidae). The infection is present in populations of Tomicus piniperda in Europe and in the United States. Uninucleate oval single spores occur in two sizes: 2.8±0.4× 1.4±0.4m and 3.8±0.3×2.0±0.2m. The polar filament of this microsporidium is fixed subapically in a flat anchoring disc. The thick posterior lamellae of the binary polaroplast are asymmetric due to the lateral fixation of the polar filament.     

Gamma radiation sensitivity of larvae and adults of the red flour beetle, Tribolium castaneum Herbst

In this study, the susceptibility of larvae and adults of Tribolium castaneum (Herbst) to gamma radiation was investigated in the laboratory in Turkey. Gamma radiation was applied at 6 dose levels between 20 and 200Gy to 13–15day-old adults and at 5 dose levels between 20 and 180Gy to 18–20day-old larvae of T. castaneum. All experiments were performed in growth chambers maintained at 27±1°C and 70±5% r.h. The larvae proved to be the more susceptible stage, survival to the adult stage being prevented by exposure of the eggs to 100Gy; the adult stage was less susceptible. LD₅₀ and LD₉₉ values were determined as 19,75 and 42,97Gy for larvae and 33,21 and 64,50Gy for adult stage, respectively. It was concluded that 100Gy is the effective dose for both larval and adult stages.     

Investigations on side-effects of the mixed biocide GCSC-BtA on different predators of Plutella xylostella (L.) (Lep., Plutellidae) in southeastern China

The present paper deals with laboratory studies on side-effects of the mixed biocide GCSC-BtA on Plutella xylostella (L.) (Lep., Plutellidae) and its predators, e.g. Amblyseius longispinosus (Evans) (Acari, Phytoseiidae), Erigonidium graminicola (Sundvall) (Araneae, Linyphiidae), Orius similis Zheng (Het., Anthocoridae) and Coccinella septempunctata L.(Col., Coccinellidae), in comparison to the commercial insecticides, e.g. Abamectin, Tebufenozide, Dichlorvos, Cartap and Lambda-cyhalothrin.The results showed that GCSC-BtA was highly toxic to the 3^rd instar of P. xylostella with 91.18% mortality, followed by Cartap with 84.38%, Abamectin with 78.00%, Tebufenozide with 75.57%, Lambda-cyhalothrin with 63.75% and Dichlorvos with 50.86% mortality. On the other hand, GCSC-BtA was found to be comparatively less toxic to the predators, causing 31.11%, 13.33%, 11.54% and 6.00% mortalities in A. longispinosus, E. graminicola, O. similis and C. septempunctata, respectively. For comparison, the mortalities recorded for Abamectin, Tebufenozide, Dichlorvos, Cartap, Lambda-cyhalothrin were 72.94%, 55.55%, 70.00%, 53.26% and 98.85% in A. longispinosus, 46.51%, 55.10%, 60.00%, 46.00% and 73.68% in E. graminicola, 22.00%, 16.00%, 35.71%, 26.78% and 81.03% in O. similis, 15.55%, 19.64%, 28.00%, 16.66% and 41.79% in C. septempunctata, respectively.Cluster analysis was introduced to group the mortalities caused by the treatments into three toxicity groups with the distance of D=4.1. The 1^st group consisted of GCSC-BtA characterized with low toxicity to all the predators tested with 15.49% mortality on average (highest 31.11% and lowest 6.00%). The 2^nd group consisted of Abamectin, Tebufenozide, Dichlorvos and Cartap with moderate toxicity to the predators with 39.96% mortality on average. The 3^rd group included Lambda-cyhalothrin with high toxicity to the predators with 73.83% mortality on average (highest 98.85% and lowest 41.70%). The susceptibility of the pest and its predators to GCSC-BtA and the insecticides is discussed. GCSC-BtA as a biological control agent is recommended for use in the integrated pests control programs in the vegetable fields.     

Suppressive effect of potassium silicate on powdery mildew of strawberry in hydroponics

From 1996 to 1997, potassium silicate (SiO₂) was tested at 0, 25, 50, and 100mgl^–1 in hydroponics to control powdery mildew. Other elements were added in the usual amounts, and the strawberries were cultivated hydroponically in a greenhouse for 4 months (from October to January). The powdery mildew spread in the control plot, but little mildew developed in the plot with 25mgl^–1 silicate, and none in plots with more than 50mgl^–1 silicate. The suppressive effect lasted for about 4 months on fruits and even longer on leaves. On analysis of mineral content in the leaves, only the silicate content differed markedly between the control and treated plants. Nitrogen, phosphate, potassium, and calcium contents did not differ greatly. The maximum silicate content was about 24 times that of the control, and disease severity decreased significantly when the content was more than 1.5% in the leaves. The hardness of the strawberry leaves, measured by a creep meter, was increased by the silicate treatment.     

Prey consumption during development as well as longevity and reproduction of Typhlodromus pyri Scheuten (Acari, Phytoseiidae) at higher temperatures in the laboratory

The predatory mite Typhlodromus pyri Scheuten (Acari, Phytoseiidae) has been reported as an important predator of the European red mite, Panonychus ulmi (Koch) (Acari, Tetranychidae) in apple culture and vineyards at below 25°C. However, sufficient biological data was lacking on its efficiency at temperatures above 25°C. Therefore, the purpose of the present laboratory work was to obtain experimental data on prey consumption during development as well as longevity and reproduction of T. pyri on apple leaf discs and in Plexiglas cells at constant temperatures of 25±2°C and 30±2°C with P. ulmi as prey.The results showed that mean daily and total prey consumption by both the nymphs and adults of T. pyri decreased significantly on both the arenas as the temperature was increased from 25°C to 30°C, whereby adult prey consumption, both mean daily and total, was higher than that of nymphs. Prey consumption by both the nymphs and adults was significantly higher in the Plexiglas cells than on the leaf discs at both temperatures. Mean total prey consumption during nymphal development was 16.1 () and 12.8 () at 25°C compared to 7.0 () and 5.8 () preys at 30°C on the apple leaf discs and 46.0 () and 38.5 () at 25°C compared to 25.2 () and 20.3 () preys at 30°C in the Plexiglas cells. Mean duration of nymphal development was similar for the two sexes at the same temperature, but it was longer at 25°C than at 30°C. It was 6.0 and 4.0days on the apple leaf discs while 7.0 and 6.0days in the Plexiglas cells at 25°C and 30°C, respectively. Mean daily and total prey consumption by both male and female adults also decreased with the increasing temperature, whereby the females consumed more than double the mean total number of prey than the males on both the arenas of observation and at both temperatures: 355.4 versus 149.7 preys at 25°C and 192.2 versus 85.6preys at 30°C on the leaf discs and 826.8 versus 374.5 preys at 25°C and 488.9 versus 187.9 preys at 30°C in the Plexiglas cells. Longevity of the females was longer than males on both arenas and at both temperatures and it was longer at 25°C than at 30°C. Mean total longevity on the apple leaf discs was 68.3 () and 50.8 () days at 25°C compared to 52.5 () and 36.8 () days at 30°C, while in the Plexiglas cells it was 91.0 () and 65.8 () days at 25°C compared to 75.3 () and 48.5 () days at 30°C. Reproduction in females also decreased significantly with increasing temperature. It decreased from 62.0 to 39.0 eggs/female on the leaf discs and 75.0 to 47.1 eggs/female in the Plexiglas cells. The females laid significantly higher numbers of eggs at both temperatures in the Plexiglas cells than on the leaf discs. Oviposition period in females was 30days at 25°C on both the arenas, while at 30°C it was 26days on the apple leaf discs and 27days in the Plexiglas cells.     

Treatment of rice with diatomaceous earth and effects on the mortality of the Red flour beetle Tribolium castaneum (Herbst)

Diatomaceous earth has been demonstrated in Australia, Germany, and the USA to protect stored products from infestation by insect pests. However, application of diatomaceous earth has rarely been used in Asian countries for the protection of stored grain from insect damage. These facts and the paucity of existing data from the tropics prompted the present study of storage with diatomaceous earth. We exposed rice treated with the diatomaceous earth product Fossil Shield® to infestation by Tribolium castaneum Herbst. During the dry season, when tests were conducted, the mean temperature was 29±2°C and relative humidity was 80±3%. The population of T. castaneum in rice was significantly reduced after treatment with Fossil Shield®. Diatomaceous earth at 1g and 2g per kilogram rice reduced population growth of T. castaneum to a coefficient of 0.27, and 0.02 respectively after 42 days. Fossil Shield® at 0.5g per kilogram rice increased the mortality of T. castaneum, but did not stop population growth.     

Pathogenic variation in poplar rust <Emphasis Type="Italic">Melampsora larici-populina</Emphasis> from England

Using a leaf disc method, 19 isolates of the poplar rust, Melampsora larici-populina , and one isolate of M.populnea from England were inoculated on to 25 poplar clones belonging to Populus nigra and P.trichocarpa, and hybrids between P. deltoides and P. nigra, P. deltoidesand P. trichocarpa, P.tacamahaca and P.trichocarpa, and P. alba and P. tremula. Disease was scored based on the pustule area and inoculum density. In terms of whether sporulating uredinia formed, the 19 isolates showed seven different patterns to the tested poplar clones. The majority of the rust isolates infected P. nigra P3090 and Vereecken, P.nigra×P. deltoides Casale and Tasman, P. tacamahaca×trichocarpa 36 and Balsam Spire, and P.trichocarpa Blom. Populus trichocarpa×P. deltoides 69039/4 was infected by only three isolates collected from southern England. No visible symptoms appeared on P. alba ×P. tremulaTower and P.trichcarpa×P. deltoides×P. deltoides76028/5 in inoculations with M. larici-populina isolates. Populus alba×P.tremula Tower was infected only by M. populnea. When M. larici-populina isolates were tested using AFLP, no differences were found either between isolates from different geographical regions or between those having narrow spectrum of virulence and those showing wide spectrum of virulence on the tested clones. The results suggest that the UK rust populations possess virulences which were found in races E1, E2, E3 and E4 in continental Europe and that rust having virulence patterns similar to race E4 has occurred in UK poplar plantations since 1996.     

Loss of Viability of Dematophora necatrix in Solarized Soils

To study the relationship between temperature regimes and loss of viability of Dematophora necatrix in soil, two field experiments were conducted to determine the effectiveness of soil solarization on reducing the population of D. necatrix colonizing avocado root segments buried at a depth of 15–60cm. Increase of maximum hourly temperatures attributable to soil solarization reached, depending on depth, 6.7–4.6°C in unshaded areas and 3.9–1.5°C for shaded areas in the first experiment (starting in early June, 1995). The better environmental conditions in the second experiment (starting by mid-July, 1995) led to higher temperature increases (8.6–5.6°C, depending on depth) when solarization was conducted in unshaded areas. One, 4, 5 and 6 weeks of solarization were required to eliminate the viability of D. necatrix at 15, 30, 45 and 60cm depths in the first experiment, whereas only 8, 10, 15 and 22 days of solarization were needed for the loss of viability of D. necatrix at the same depths in the second experiment. In shaded areas, however, soil solarization attained significant effectiveness at 15cm depth.Regression analyses of fungal viability (ln-transformed data) over accumulated temperature–time showed best fits when the minimum threshold temperature was 30°C. Although eradication of D. necatrix in soil can be achieved down to 60cm depth in solarized plots, and at 15cm depth in unsolarized unshaded plots, the accumulation of temperature–time appeared less effective in reducing inoculum viability in the latter.     

Conjugation of δ-endotoxin from Bacillus thuringiensis with abamectin of Streptomyces avermitilis as a new type of biocide, GCSC-BtA, for control of agricultural insect pests

Conjugation of -endotoxin from Bacillus thuringiensis with abamectin, a toxin of Streptomyces avermitilis, was carried out to form a new type of biocide, GCSC-BtA based on Germany-China Scientific Cooperation research, for the control of agricultural insect pests. The strategy for biochemical linkage was designed by conjugating an amino group in B.t. protoxin with a carboxyl group in carboxylated abamectin under the treatment of conjugator EDC [1-Ethyl-3-(3-dimethylaminopropyl carbodiimide Hydrochloride)]. The formation of B.t. protoxin was processed by solubilizing B.t. crystal in 25mM dithiothreitol (DTT) at 37°C for 2h. The carboxylated abamectin was formed by carboxylating the NaH-activated abamectin with 10mg/ml butyric anhydride at 111°C in a water-circumfluent condensation device for 2h. The conjugating reaction, consisting of 5mg/ml B.t. protoxin, 10mg/ml carboxylated abamectin and 19.17mg/ml EDC, was successfully conducted at room temperature for 24h. Significant differences were found between pure abamectin, carboxylated abamectin and the conjugated BtA by means of UV-photo absorptions recorded at wavelengths 354, 438, 518, 600nm (P<0.01). LT₅₀ of the conjugated GCSC-BtA to the 3^rd instar larvae of Plutella xylostella (L.) (Lep., Plutellidae) was 35.27 g a.i./ml, about 62% and 76% of that caused by the B.t. protoxin and the caxboxylated abamectin, respectively. The conjugated GCSC-BtA caused 87.14% mortalities in larvae of P. xylostella, 93.75% in adult Myzus persicae (Sulzer) (Hom., Aphididae) and 89.33% in adult Phyllotreta vittata Fabricius (Col., Chrysomelidae) as compared to 48.33% by the B.t. crystal only in P. xylostella. The symptoms caused by conjugated GCSC-BtA in the 3^rd instar of P. xylostella were black color in the head part and white-yellow in the abdomen of dead larvae, which differed from the black color or the white-yellow all along the body caused by either the B.t. crystal or the abamectin, respectively. It was concluded that the conjugated GCSC-BtA biocide had a broader host spectrum and a faster killing speed than either the B.t. crystal or abamectin alone for the control of agricultural pests.     

Infection of Tomicus piniperda (Col., Scolytidae) with Canningia tomici sp. n. (Microsporidia, Unikaryonidae)

Canningia tomici sp. n. (Microsporidia, Unikaryonidae) infects the midgut epithelium, the gut muscules, Malpighian tubules, connective tissues, adipose tissues and the gonads of the pine shoot beetle, Tomicus piniperda (L.) (Coleoptera, Scolytidae). The infection is present in populations of Tomicus piniperda in Europe and in the United States. Uninucleate oval single spores occur in two sizes: 2.8±0.4× 1.4±0.4m and 3.8±0.3×2.0±0.2m. The polar filament of this microsporidium is fixed subapically in a flat anchoring disc. The thick posterior lamellae of the binary polaroplast are asymmetric due to the lateral fixation of the polar filament.     

Investigations on side-effects of the mixed biocide GCSC-BtA on different predators of Plutella xylostella (L.) (Lep., Plutellidae) in southeastern China

The present paper deals with laboratory studies on side-effects of the mixed biocide GCSC-BtA on Plutella xylostella (L.) (Lep., Plutellidae) and its predators, e.g. Amblyseius longispinosus (Evans) (Acari, Phytoseiidae), Erigonidium graminicola (Sundvall) (Araneae, Linyphiidae), Orius similis Zheng (Het., Anthocoridae) and Coccinella septempunctata L.(Col., Coccinellidae), in comparison to the commercial insecticides, e.g. Abamectin, Tebufenozide, Dichlorvos, Cartap and Lambda-cyhalothrin.The results showed that GCSC-BtA was highly toxic to the 3^rd instar of P. xylostella with 91.18% mortality, followed by Cartap with 84.38%, Abamectin with 78.00%, Tebufenozide with 75.57%, Lambda-cyhalothrin with 63.75% and Dichlorvos with 50.86% mortality. On the other hand, GCSC-BtA was found to be comparatively less toxic to the predators, causing 31.11%, 13.33%, 11.54% and 6.00% mortalities in A. longispinosus, E. graminicola, O. similis and C. septempunctata, respectively. For comparison, the mortalities recorded for Abamectin, Tebufenozide, Dichlorvos, Cartap, Lambda-cyhalothrin were 72.94%, 55.55%, 70.00%, 53.26% and 98.85% in A. longispinosus, 46.51%, 55.10%, 60.00%, 46.00% and 73.68% in E. graminicola, 22.00%, 16.00%, 35.71%, 26.78% and 81.03% in O. similis, 15.55%, 19.64%, 28.00%, 16.66% and 41.79% in C. septempunctata, respectively.     

The Role of Trichoderma harzianum Protease in the Biocontrol of Botrytis cinerea

The role of protease of Trichoderma harzianum in the biocontrol of Botrytis cinerea was examined. Two isolates of T. harzianum were compared for their ability to produce protease in liquid culture medium and on the surface of bean leaves. The biocontrol agent T. harziaum T39 produced 58mU/ml of protease and T. harzianum NCIM1185 produced 54mU/ml on the 5th day of growth in liquid culture medium. On bean leaves, combinations of B. cinerea and T. harzianum isolates were examined for the synthesis of protease. The protease activities were 0.9 and 0.6mU/ml for T. harzianum T39 and NCIM1185, respectively, and 0.5mU/ml for B. cinerea alone after 48h of incubation. In the presence of T. harzianum T39 culture liquid containing protease, a 55% reduction in B. cinerea germination and a 80% reduction in the germ tube length were observed after 17h of incubation in vitro. When T. harzianum isolates were added to B. cinerea on bean leaves, increased synthesis of protease was observed (1.0 and 1.2mU/ml for T39 and NCIM1185, respectively). In the presence of T. harzianum NCIM1185 protease, although the rate of germination was reduced, B. cinerea attained 98% germination after 17h of incubation. The hydrolytic enzymes produced by B. cinerea, endo-polygalacturonase (PG) and exoPG were partially deactivated by protease from the T. harzianum isolates. Carboxymethyl cellulase was deactivated only by protease of NCIM1185. On the surface of bean leaves, the protease (obtained from liquid culture medium of T. harzianum isolates) resulted in 56–100% reduction of disease severity. The culture liquid containing protease synthesized on the surface of bean leaves treated with B. cinerea and with T. harzianum was collected and added to fresh leaves infected by B. cinerea. There was 56–100% and 30–75% reduction of disease severity with liquid droplet collected from the leaves treated with T. harzianum T39 and NCIM1185, respectively. Increased control of disease was obtained by combining the conidia of T. harzianum isolates with protease obtained from culture media. Protease inhibitors, trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E64), antipain hydrochloride, and a mixture of inhibitors, but not pepstatin A, fully or partially nullified the biocontrol effect of T39. T39 was found to be a poor producer of chitinase and -1,3-glucanase in vitro. These enzymes were not detected on leaves treated with T39. Involvement of protease in biocontrol of B. cinerea is suggested.     

Induced Resistance to Cyst and Root-knot Nematodes in Cereals by DL-β-amino-n-butyric Acid

Foliar sprays and soil drenches with DL--amino-n-butyric acid (BABA) reduced the number of Heterodera avenae and H. latipons cysts on wheat and barley. Foliar sprays of wheat with 8000mgl^–1 BABA reduced the number of H. avenae cysts by 90%, whereas 2000mgl^–1 BABA was enough to reduce the number of H. latipons cysts by 79%. Multiple spray treatments with 2000mgl^–1 BABA at 10-day intervals reduced the number of H. avenae cysts on wheat and barley. A soil drench of wheat with 125mgl^–1 BABA reduced the number of H. latipons cysts by 93% and H. avenae cysts by 43%. Second-stage juveniles of these nematodes penetrated and formed syncytia in wheat roots soil-drenched with BABA. More adult males of H. avenae were produced in BABA (<250mg1^–1)-treated wheat roots (~76%) than in untreated roots (27%). Soil drenches with higher concentrations of BABA inhibited development of adult males and females. Several chemical elicitors of induced resistance were tested for their ability to reduce the number of H. avenae cysts on wheat. Only BABA was found to be an effective resistance inducer. The number of egg masses of an unidentified Meloidogyne sp. root-knot nematode, which infects only monocots, was also reduced by 95% by a soil drench of wheat with 500mgl^–1 BABA. Development of this nematode inside the BABA-treated roots was also inhibited.     

Effects of soil solarization on nematodes in Croatia

For the first time soil solarization was investigated in Croatia both in the field and in the greenhouse in 1991, 1992, 1993 and 1994. For two months (July and August), the soil was mulched with transparent polyethylene (PE) sheets of 0.015 or 0.050mm thickness. Soil temperatures at depths of 5, 10 and 20cm were recorded daily. In order to assess nematode population densities, soil samples were analysed before mulching and at the end of the mulching treatment. The results of these experiments showed that soil solarization drastically reduced the population of plant-parasitic nematodes (Meloidogyne, Pratylenchus, Paratylenchus, Tylenchus, Tylenchorhynchus spp.) by about 97–100% at a depth of 10cm and 92–97% at a depth of 20cm in the field, while in the greenhouse, the population of plant-parasitic nematodes was reduced by about 89–100% at a depth of 10cm and 98–100% at a depth of 20cm.In the same experiments, the population of saprophytic nematodes in the field was reduced by about 86–90% at a depth of 10cm and 72–89% at a depth of 20cm. In the greenhouse, the population of saprophytic nematodes was reduced by about 87–97% at a depth of 10cm and 87–93% at a depth of 20cm. This data shows that soil solarization was less effective in the control of saprophytic nematodes, which is considered to be an advantage.     

Detection of Xanthomonas sp., the Causal Agent of Onion Bacterial Blight, in Onion Seeds Using a Newly Developed Semi-selective Isolation Medium

Onion bacterial blight, caused by Xanthomonas sp., is a potentially severe disease in several tropical and subtropical areas. Although little research has been undertaken on this pathosystem, seed transmission of the pathogen has been hypothesized. Because of an important bacterial microflora naturally associated with onion seeds, detection of the pathogen is difficult using non-selective agar media. A new semi-selective medium, whose selectivity was obtained by a combination of four antibiotics, was developed. The new NCTM1 medium contained (per liter) yeast extract 7g, peptone 7g, glucose 7g, agar 15g, neomycin 10mg, cephalexin 30mg, trimethoprime 3mg, pivmecillinam 100mg and propiconazole 20mg. Plating efficiencies, using 16 pure cultures of the pathogen, ranged from 79% to 142%, with an average of 110% compared to the basal medium. All onion Xanthomonas sp. strains from several countries grew on NCTM1 medium. The pathogen was repeatedly isolated using this medium from seed samples containing approximately 10⁶ saprophytic bacteria per gram, as well as from symptomless plant material. Xanthomonas sp. was detected only in seeds originating from one infected seed production site. This is the first report of selective isolation of Xanthomonas sp. from onion seeds. NCTM1 medium should be a valuable tool to study the ecology and epidemiology of Xanthomonas sp. causing onion bacterial blight.     

Effects of Potato-cyst Nematodes (Globodera pallida) and Soil pH on Root Growth, Nutrient Uptake and Crop Growth of Potato

Potato-cyst nematodes (Globodera pallida) cause severe yield losses in potato. Plants infected with potato-cyst nematodes generally have reduced concentrations of nitrogen, phosphorus and potassium in the foliage. This study investigated whether reduced growth of nematode-infected potato is caused by nutrient limitation.Experiments in the field and in containers showed that phosphorus concentration correlated best with total crop biomass at early stages of growth. The role of phosphorus in nematode damage was further investigated in the field and in the Wageningen Rhizolab. The experimental field was infested with potato-cyst nematodes and two levels of nematode density were established by fumigation with a nematicide. Prior applications of calcium carbonate resulted in pHKCl levels of 4.8 and 6.1. Two levels of phosphorus fertiliser were applied: either 0 or 225kgPha–1. In the Wageningen Rhizolab, soil of both pH levels from the field was used after treatment with 1MRad gamma irradiation to kill the nematodes. Subsequently, half of the soil was inoculated with cysts to give a nematode density of 30 viable juveniles per gram of soil.In the field, nine weeks after planting, the total crop biomass ranged from 107gm–2 for the treatment with nematodes at pHKCl 6.1 without phosphorus fertiliser to 289gm–2 for the fumigated treatment at pHKCl 4.8 with phosphorus fertiliser. The differences in total biomass for the various treatments were explained by differences in foliar phosphorus concentration. Nematodes induced or aggravated P deficiency and reduced total biomass. This was not the only damage mechanism as at high, non-limiting levels of foliar phosphorus concentration, nematodes still reduced total biomass.In the Wageningen Rhizolab, directly after planting, the number of roots visible against minirhizotrons was reduced by nematodes. However, the increase of root number in the nematode treatment continued longer than in the control, until root number was higher than that of the control. The compensary root growth of the nematode treatment was restricted to the top 30cm and nematodes reduced rooting depth.High soil pH reduced growth, mainly by reducing the availability of phosphate. Both nematodes and high soil pH reduced nutrient uptake per unit root length. Our results lead us to suggest an interaction between nematodes and soil pH, with nematode damage being higher at pHKCl 6.1 than at pHKCl 4.8.