Acceptance and suitability of four plant substrates for rearing Orius sauteri (Hemiptera: Anthocoridae)

Orius sauteri (Poppius) is an important hemipterous predator that has been mass-reared for biological control of numerous pests in protected crop-production systems. To find a good oviposition substrate for mass-rearing this predator under insectary conditions [25°C, 65 ± 5% relative humidity, and a photoperiod of 16:8 (L:D) h], we compared kidney bean, soybean, broad bean sprouts and fresh leaves of kidney bean. We found that O. sauteri made more punctures and laid more eggs in kidney bean sprouts than in the other substrates examined. However, there were no significant differences among substrates in the proportion of punctures receiving eggs. Female O. sauteri laid the most eggs (as many as 68 eggs) in kidney bean sprouts and also had the shortest pre-oviposition period on this plant material. In addition, there were no significant differences in total oviposition durations or female longevity among the four plant substrates. The hatch rates of nymphs in the sprouts and leaves of kidney bean (>90%) were higher than those in soybean and broad bean sprouts. Thus, we found that the kidney bean sprout was the most suitable substrate for mass-rearing of O. sauteri.     

Effect of intercrops on thrips species composition and population abundance on French beans in Kenya

The study aimed at determining thrips species composition and thrips population density on French bean planted as a sole crop and as an intercrop with either sunflower, Irish potato, or baby corn, in various combinations. Field experiments were conducted in two seasons to examine: (1) thrips population development and thrips species composition over time, (2) effect of intercrops on thrips population density and natural enemies, and (3) effect of intercrops on French bean yield. The experiments were conducted at the Kenya Agricultural Research Institute, Embu, Kenya in a randomized complete block design with four replicates. The thrips population on French beans increased with time. It showed a peak at the flowering stage then started declining when the crops were nearing senescence. French beans hosted four thrips species, Megalurothrips sjostedti (Trybom), Frankliniella schultzei (Trybom), Frankliniella occidentalis (Pergande), and Hydatothrips aldolfifriderici (Karny) (all Thysanoptera: Thripidae) in order of decreasing abundance. The main thrips species on Irish potato and sunflower was F. schultzei. Baby corn hosted only Frankliniella williamsi (Hood) and Thrips pusillus (Bagnall). A monocrop of French bean hosted more thrips than a French bean intercrop mix. Thrips natural enemies such as Orius spp. and Ceranisus spp. were recorded in all crop plants but in especially high numbers on French bean and baby corn, respectively. Plots with French bean alone had about 1.4 times higher yields compared to intercropped plots of French bean with sunflower and French bean with baby corn. However, the percentage of pods that could get rejected on the market due to thrips damage was highest on plots with French bean alone (68 and 63%) and lowest on plots with French bean and baby corn (35 and 37%) in the first and second seasons, respectively. This study showed that a complex of thrips is found in the field and its composition varies with crop stage and species. Intercropping French bean with other crops compromises on French bean yield but reduces damage to the French bean pods, thereby enhancing marketable yield.     

Apple pollen as a supplemental food for the western flower thrips, Frankliniella occidentalis: response of individuals and populations

Experiments were performed to investigate the influence of apple pollen, plant diet, and relative humidity on the individual life‐history traits and on the population growth of the western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). In experiments with individual thrips, availability of bean pods plus a mixture of apple pollen and lycopodium shortened larval development time, and hence time from egg to adult, compared to bean pods alone. Similarly, the total number of eggs laid and the mean number of eggs laid per female per day were greater in the presence of pollen plus bean pods than on bean pods alone. Diet did not affect survival or length of the pre‐oviposition and oviposition periods. A diet including bean pods plus pollen shortened the generation time and population‐doubling time, and increased the net reproductive rate and the intrinsic rate of increase, compared to bean pods alone. In experiments on populations, growth was measured with different amounts of pollen on bean pods under two relative humidities, ca. 38% and 66%, and separately on two bean substrates, pods or leaves, at ca. 45% r.h. The presence or amount of pollen had no effect on population growth regardless of the other variables. Greater population growth occurred under the higher relative humidity regardless of substrate. Population growth was greater on bean pods than on bean leaves. Despite reports on the nutritional benefit of pollen for F. occidentalis, this benefit may be affected by pollen type, host plant, and other factors. In situations in which pollen has little influence on the growth of F. occidentalis populations, pollen that benefits a thrips predator could be used in an integrated management plan for controlling thrips.     

Internalization of bioluminescent Escherichia coli and Salmonella Montevideo in growing bean sprouts

AIMS: Investigate the interaction of bioluminescent Escherichia coli and Salmonella Montevideo with germinating mung bean sprouts. METHODS AND RESULTS: E. coli or Salm. Montevideo introduced on mung beans became established both internally and externally on sprouts after the initial 24 h germinating period. In both cases the inoculated bacterium formed the predominant microflora on the sprouted beans throughout. From the bioluminescent profile of inoculated sprouting beans, bacterial growth was found to be in close proximity to the roots but not on the hypocotyls. Clumps (biofilms) of cells with low viability were observed within the grooves between epidermal cells on hypocotyls. Treatment with 20,000 ppm sodium hypochlorite removed the majority of bacteria from the surface of hypocotyls although nonviable single cells were occasionally observed. However, viable bacteria were recovered from the apoplastic fluid, and extracts of surface-sterilized sprouts indicating that the internal bacterial populations had been protected. This was confirmed using in situ beta-glucuronidase staining of surface-sterilized sprouts where cleaved enzyme substrate (by the action of internalized E. coli) was visualized within the plant vascular system. CONCLUSIONS: E. coli or Salmonella present on seeds become internalized within the subsequent sprouts and cannot be removed by postharvest biocidal washing. SIGNIFICANCE AND IMPACT OF THE STUDY: Mung bean production should be carefully controlled to prevent contamination occurring in order to minimize the health risk associated with raw bean sprouts.     

Growth characteristics of mung beans and water convolvuluses exposed to 425‐MHz electromagnetic fields

Effects of high‐frequency, continuous wave (CW) electromagnetic fields on mung beans (Vigna radiata L.) and water convovuluses (Ipomoea aquatica Forssk.) were studied at different growth stages (pre‐sown seed and early seedling). Specifically, the effects of the electromagnetic source's power and duration (defined as power‐duration level) on the growth of the two species were studied. Mung beans and water convolvuluses were exposed to electromagnetic fields inside a specially designed chamber for optimum field absorption, and the responses of the seeds to a constant frequency at various power levels and durations of exposure were monitored. The frequency used in the experiments was 425 MHz, the field strengths were 1 mW, 100 mW, and 10 W, and the exposure durations were 1, 2, and 4 h. Results show that germination enhancement is optimum for the mung beans at 100 mW/1 h power‐duration level, while for water convolvuluses the optimum germination power‐duration level was 1 mW/2 h. When both seed types were exposed at the early sprouting phase with their respective optimum power‐duration levels for optimum seed growth, water convolvuluses showed growth enhancement while mung bean sprouts showed no effects. Water content analysis of the seeds suggests thermal effects only at higher field strength. Bioelectromagnetics 31:519–527, 2010. © 2010 Wiley‐Liss, Inc.     

Damage to lima beans by Helicoverpa zea larvae treated as different instars with a nucleopolyhedrovirus

Larvae of the corn earworm,Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), were caged assecond, third, or fourth instars on lima bean,Phaseolus lunatus L., plants in the fieldwith or without treatment of the plants withlethal concentrations of the H. zeanucleopolyhedrovirus. The virus treatmentprevented over 90% of damage to pods and beansif larvae were second or third instars whenplaced in the cages. Damage to pods and beanswas reduced by 73 and 86%, respectively, ifthe larvae were fourth instars when placed onthe plants. When insects survived to the endof the test (which they did only on controlplants), numbers of pods or beans damaged byeach surviving insect was not affected by thestadium in which the larva was when it wasplaced on the plant. This result indicates thatmost damage was done by fourth or laterinstars. If treatments are to effectivelyprevent damage to lima bean, they should thusbe applied before the insects reach the fourthinstar.     

Natural Spread of Plant Viruses by Birds

Observations made in Mali strongly suggest that Rice yellow mottle virus (RYMV) is spread by weaverbirds (Quelea quelea) below and around baobab trees (Adansonia digitata) in which they nest. Rice leaves in bird nests appeared to be infected. In Spain, an infection of Southern bean mosaic virus (SBMV) in string (climbing) beans (Phaseolus vulgaris) was apparently introduced and spread by sparrows (Passer domesticus) judging from the damage caused on flowers and bean pods. Damaged leaves and pods on SBMV‐infected plants were also found in a screenhouse visited by sparrows and bulbuls (Pycnonotus barbatus) in Morocco. These observations showed that both viruses could be spread by birds when either collecting infected leaves for nesting or feeding on infected plants.     

Hydrolysis of RNA to 5′-nucleotide by seed sprouts,particularly malt sprouts

Conditions for the efficient conversion of commercial RNA to nucleoside 5′-monophosphate by means of a phosphodiesterase in malt sprouts have been determined. A comparison of the enzyme content of the rootlets, stems, and kernels of various plant seedlings, including barley, rye, oat, wheat, rice, and beans shows maximum amounts in the rootlets, and minimum quantities in the ungerminated kernels. Of all the seedlings tested, (mung bean, soy bean, oat, wheat, rice, barley) barley gave the highest conversion of RNA to 5′-nucleotides. Commercial malt sprouts prepared from 6 different malted barleys including 2-rowed and 6-rowed samples all showed about the same amount of phosphodiesterase content. Besides phosphodiesterase, other enzymes capable of hydrolyzing RNA and 5′-nucleotides were found in sprouts. These included 3′-phosphodiesterases, 5′-nucleotidases, and nucleosidases. By carefully pretreating both extracts and the solid sprouts at elevated temperatures for a limited time and by the addition of minimum amounts of Zn⁺², the action of these undesirable enzymes was either effectively destroyed or minimized so that the production of 5′-nucleotides was maximized. It was found that suspensions of appropriately washed and treated barley malt rootlets are substantially more effective than aqueous extracts for converting RNA to 5′-nucleotides.     

True bug (Heteroptera) impact on cocoa fruit mortality and productivity

The real impact of true bug damage on cocoa pods has never been assessed precisely. We conducted a 2-yr study on 1,080 cocoa trees on 36 farms in Cameroon to assess the contribution of true bugs to fruit mortality and production loss. The cocoa fruiting cycle, fruit mortality, and damage caused by true bugs as well as other pests and diseases were monitored on a weekly basis. True bug damage also was described on 2,500 ripe pods per year. Pod weight, bean number, and bean weight were measured and compared for different degrees and types of damage on the ripe pods. Our results showed that true bugs were the main external cause of young fruit abortion. They reduced the abundance of young fruit by up to 10%. In contrast, although one-third of the ripe pods sampled had true bug lesions, only 4% were moderately to heavily damaged. The mean weight of ripe pods was reduced by 12% when there was medium to heavy damage. While the mean weight of wet beans was reduced significantly (by 3-10%), the number of beans per pod was not changed by damage. Despite the reduction in mean weight, the overall weight of beans for the pods sampled was reduced by <2%. Therefore, our study confirmed the common assumption that the economic impact of true bug damage on mature pods is negligible on cocoa farms in Cameroon. However, true bugs have a significant impact on young fruit mortality.     

Anatomical root variations in response to water deficit: wild and domesticated common bean (Phaseolus vulgaris L)

Root anatomical responses to water deficit are diverse and regulation of water uptake strongly depends on plant anatomy. The ancestors of common bean (Phaseolus vulgaris L.) cultivars are the wild common beans. Because wild beans adapt and survive well in the natural environment, it is hypothesized that wild common bean roots are less affected than those of domesticated beans at low substrate water potential (ψW). A wild common bean accession from Chihuahua Mexico and cv. Bayomex were studied. Seedlings with a mean root length between 3 and 4 cm were maintained for 24 h in vermiculite at ψW of -0.03 (well hydrated), -0.65, -1.48 and -2.35 MPa (partially dry). Ten anatomical characteristics of differentiation and cell division in root regions were evaluated. Thickness of epidermis and protoderm diminished similarly in wild and domesticated beans growing at low substrate ψW (between -0.65 and -2.35 MPa). At the same time, parenchymatic cell area diminished by 71 % in the domesticated variety, but by only 32 % in the wild bean at -2.35 MPa. The number of cells in the cortex and the thickness of the xylem wall increased in both wild and domesticated beans at low substrate ψW; nevertheless, the effect was significantly lower in the wild bean. The number of xylem vessels increased in the cultivar (up to 40 %) while in the wild bean it decreased (up to 33 %). The diameter of xylem vessels and transverse root area diminished (15 and 57 %, respectively) in the cultivar, but in the wild common bean were not affected. Anatomical root characteristics and their modifications in both differentiation and cell division in root regions demonstrated that the wild bean reacted quite differently to substrate ψW than the domesticated common bean.     

The distribution of immature thrips among flowers of faba beans in commercial crops and experimental plots

Numbers of eggs and larvae of thrips {Kakothrips pisivorus (Westw.)) infesting pistils of various stocks of faba beans (Viciafaba L.) were recorded in different environments. By the end of flowering in spring beans, 25 times as many flowers were infested with, on average, twice as many thrips, as at the onset of flowering in winter beans. Within each of two seasons, levels of infestation varied up to 30-fold with location. At a single location, some bean stocks were much more heavily infested (greater proportions of flowers contained thrips and/or greater numbers of thrips per infested pistil) than others. Levels of infestation were not clearly associated with the closed-flower character (cf). Stocks with lush growth were more heavily infested than those with either generally restricted growth or with terminal inflorescences (ti). Those with white flowers, long pods or cytoplasmic male-sterility were more infested than those which contrasted for these three characters. The path of the pollen tubes was blocked by eggs of thrips in less than 0–5% of flowers. Infestation had no apparent effect on the fertilisation of flowers.     

Distribution and characterization of diglycosyldiacylglycerol isomers with different anomeric configuration in higher plants.

The novel diglycosyldiacylglycerol (DGDG), galactosyl(beta 1----6)-galactosyl(beta 1----3')-diacylglycerol (B isomer), present in Adzuki beans was found to be distributed together with the well-known galactosyl(alpha 1----6)-galactosyl(beta 1----3')-diacylglycerol (A isomer), in all (10) of the higher plants examined. The highest levels were found in leguminous seeds were the amounts were always less than 33% of the total DGDG of mature seeds. The highest proportion of the B isomer was found in Adzuki bean seed DGDG (26-33%), with the lowest in pea seed DGDG (2%). The amounts of the B isomer in DGDG of Adzuki and kidney beans cotyledons were almost equal to those in mature seeds. Immature seeds and hypocotyls of three kinds of beans also contained the B isomer in small amounts compared with the mature beans, while only trace amounts of the isomer was found in other organs such as leaves, stems, pods, roots and generative organs of plants, except for root from kidney beans. The molecular species composition of the principal diacylglycerol moieties in the A and B isomers of DGDG were found to be significantly different among several plant seeds, although the component diacylglycerol species were qualitatively similar to each other.     

Bean tumbling as a control measure for the common bean weevil, Acanthoscelides obtectus

Recognizing that it takes Acanthoscelides obtectus (Say)(Coleoptera: Bruchidae) larvae over 24 h to bore into a dry red kidney bean (Phaseolus vulgaris L.) and that boring can occur only at particular sites where a bean abuts some other surface, we postulated that this pest might be controlled by periodic tumbling of beans so as to place initiated holes out of register with requisite touching surfaces. Larvae repeatedly forced to initiate holes might die of exhaustion, if not smashed by tumbling beans. Indeed, brief daily tumbling of beans held in half-filled jars, buckets, and gunny sacks reduced A. obtectus populations by 97% relative to stationary controls. We recommend that small lots of beans can be protected indefinitely from the bean weevil when stored in <75% filled cylinders rolled ca 1 circumference every morning and evening. This control method should be immediately useful, particularly among subsistence families, since the only external input is knowledge. Also, this control principle might be broadly applicable to other bean and grain pests having strict spatial and temporal requirements for seed penetration.     

Vitrification of in vitro produced goat blastocysts: effects of oocyte donor age and development stage

This study examines the effectiveness of the cryotop vitrification method for the cryopreservation of goat blastocysts. To determine the effects of embryo development stage and donor age on in vitro survival rates, good-quality blastocysts from adult and prepubertal goats were sorted into non-expanded, expanded, hatching and completely hatched. In vitro produced (IVP) blastocysts were derived from prepubertal goat oocytes by slicing of ovaries from slaughtered animals while adult goat oocytes were collected by the laparoscopic ovum pick up (LOPU) method. Blastocysts were vitrified/warmed using the cryotop technique. Survival rates were determined in terms of blastocoele re-expansion at 3 and 20 h post-warming. For prepubertal goats, survival rates at 3 h post-warming were significantly higher when expanded blastocysts (78.3%) were vitrified/warmed compared to hatched blastocysts (57.4%), whereas non-expanded (62.5%) or hatching blastocysts (71.4%) showed similar rates. For adult goats, survival rates were significantly higher after warming in expanded (36.4%), hatching (75%) or hatched (50%) blastocysts when compared to non-expanded (0%) blastocysts. When survival rates were assessed at 20 h post-warming, no differences were observed when we compared non-expanded (45.8%), expanded (56.5%), hatching (64.3%) and hatched (50.5%) blastocysts from prepubertal goats; and for blastocysts from adult goats, survival rates were only significantly lower for the non-expanded stage (0%) compared to the other stages. For adult versus prepubertal blastocysts at the same developmental stage, our data indicate significantly higher survival rates at 3 h post-warming for non-expanded and expanded blastocysts from prepubertal goats over their counterparts from adult goats. At 20 h post warming, survival rates were only higher for non-expanded blastocysts from prepubertal goats versus adult goats. Collectively, our data reveal that blastocysts produced in vitro from prepubertal goats return similar survival rates regardless of their development stage, whereas blastocysts derived from adult goats are best for vitrification at the expanded, hatching or hatched stage.     

Evaluation of different leguminous seeds as food sources for the bean bug Riptortus pedestris

This study was conducted to determine the effects of six different leguminous seeds (cowpea, white kidney bean, soybean cultivars Cheongja and Daepung, mung bean, and azuki bean) on the life history traits of the bean bug Riptortus pedestris (F.) (Hemiptera: Alydidae). The total developmental time from the first instar nymph to adult ranged from 18 to 25 d; R. pedestris fed with white kidney beans were the slowest to develop. In addition, bugs fed with white kidney beans had the highest mortality (93%) and the shortest longevity (18 d). R. pedestris fed with cowpeas, soybeans, or azuki beans had high reproductive success, with the highest egg hatching success recorded in bugs fed cowpeas. The results suggest that cowpea may be a beneficial food source for the development and reproduction of bean bugs.     

Studies of the infection of the winged bean by Synchytrium psophocarpi in Papua New Guinea

Infection of leaves and stems of Psophocarpus tetragonolobus by Synchytrium psophocarpi only occurred following spray inoculation of motile zoospore suspensions and incubation for a minimum of 12 h in polyethylene bags or a mist chamber. The incubation period was 7 days and generation time 22 days at temperatures of 31 ^oC max, 24 ^oC min and r.h. of 90% max, 70% min. Young, 1–2 day-old leaves were most susceptible; there was no infection on 10 day-old leaves and susceptibility was not increased by the removal of leaf waxes. No infection occurred when plants were grown from seed from infected pods, seed inoculated with zoospores or sporangia and seed sown in soil containing infected leaf debris. Resting spores were not found in infected tissues stored for 12 wk or in plant debris. S. psophocarpi did not infect Arachis hypogaea, Glycine max, Phaseolus aureus, P. coccineus, P. vulgaris, Pisum sativum, Psophocarpus scandens, Vicia faba, Vigna sesquipedalis and V. unguiculata. S. minutum did not infect winged bean. Inoculation confirmed the susceptibility of the winged bean lines UPS 62, UPS 122, UPS 126 and resistance of two Thai winged bean lines 1602/1 and 1611/2.     

Effect of sowing date on incidence of Sudanese broad bean mosaic virus in, and yield of, Viciafaba

Delaying the sowing of irrigated field beans in the Sudan after October greatly lowered the yield of seed, and increased both infestation by aphids and the incidence of Sudanese broad bean mosaic virus (SBBMV). Yield per plant was positively correlated with number of pods, but negatively with percentage infection with SBBMV. In greenhouse conditions, SBBMV was readily acquired from diseased plants and inoculated to healthy plants in 5 min feeding periods by both Aphis craccivora and Acyrthosiphon sesbaniae.     

Fate of Clavibacter michiganensis ssp. sepedonicus, the Causal Organism of Bacterial Ring Rot of Potato, in Weeds and Field Crops

Crops and weeds were tested for their ability to host Clavibacter michiganensis ssp. sepedonicus (Cms), the causal agent of bacterial ring rot in potato. Ten crops grown in rotation with potato in Europe, namely maize, wheat, barley, oat, bush bean, broad bean, rape, pea and onion and five cultivars of sugar beet were tested by stem and root inoculation. About 6–8 weeks after inoculation, Cms could be detected in most crops except onion and sugar beet, in larger numbers in stems (10⁵–10⁶ cells/g of tissue) than in roots (≤10³ cells/g of tissue) in immunofluorescence cell‐staining (IF). Cms was successfully re‐isolated only from IF‐positive stem samples of maize, bush bean, broad bean, rape and pea, but not from roots. Twelve solanaceous weeds and 13 other weeds, most commonly found in potato fields in Europe, were tested in IF as hosts of Cms by stem and root inoculations. Only in Solanum rostratum, a weed present in northern America, Cms persisted in high numbers (10⁸ cells/g tissue) in stems and leaves, where it caused symptoms. In the other solanaceous weeds, Cms persisted at low numbers (approximately 10⁵ cells/g of tissue) in stems but less so in roots. The bacteria could be frequently re‐isolated from stem but not from root tissues. In 2 consecutive years, plants from 14 different weed species were collected from Cms‐contaminated potato field plots and tested for the presence of Cms by dilution plating or immunofluorescence colony‐staining (IFC), and by AmpliDet RNA, a nucleic acid‐based amplification method. Cms was detected in roots but not in stems of Elymus repens plants growing through rotten potato tubers, and in some Viola arvensis and Stellaria media plants, where they were detected both in stems and roots, but more frequently by AmpliDet RNA than by IFC.     

Effects of Nitrate Level on Nitrogen Metabolism in Winged Bean and Soya Bean

The effects of high (15 mM) and low (0.75 mM) solution nitratelevels on nitrogen metabolism in three genotypes (IL 7A, IL13 and IL 21) of winged beans [Psophocarpus tetragonolobus (L.)DC.] and one genotype (Williams) of soya bean [Glycine max (L.)Merrill] were investigated. Plants were grown for 42 days ina greenhouse in solution culture prior to sampling. The 15 mM nitrate treatment resulted in greater growth of allplant parts except roots. Growth of soya beans was more responsiveto nitrate level than was growth of winged beans. The high nitratelevel inhibited nodulation in all plants. The IL 13 and IL 21winged bean genotypes had similar nitrogenase activity (acetylenereduction per plant) as the soya bean and IL 7A winged beangenotype had lower activity. However, the IL 13 winged beangenotype had higher nitrogenase activity (acetylene reductionper unit nodule mass) than the other three genotypes which allhad similar activity. The 15 mM solution nitrate level stimulatedleaf and root nitrate reductase (NR) activity for all plants.All winged bean genotypes had higher leaf NR activity and higherpercentage reduced- and nitrate-nitrogen contents of leavesand stems compared with soya beans. However, total protein (reducednitrogen) was greater in soya beans when sampled indicatingthat more nitrate had been metabolized by soya beans than bywinged beans during the 42-day growth period. Psophocarpus tetragonolobus (L.) DC., winged bean, Glycine max (L.) Merrill, Soya bean, nitrate reductase, nitrogen fixation, nitrogenase activity, nodulation     

Fungal contamination and mycotoxin-producing potential of dried beans

A total of 604 samples of about 7 different types of beans was examined to determine their mycological profiles, and suitability for use as solid substrates for mycotoxin production.All of the samples were collected from bean jam makers in Tokyo by the official food examiners.Genera Penicillium and Aspergillus were predominant, and genus Wallemia was also found commonly in all types of beans.Mycotoxin-producing Aspergillus strains were isolated from 52 samples of beans, approximately 9% of the total. The highest incidence of toxigenic Aspergillus (14.1%) was found in kidney beans. Red beans and peas inoculated with Aspergillus ochraceus were found to produce about 7 to 8 times more toxin than was obtained in a liquid medium, and red beans inoculated with A. versicolor produced more toxin than was obtained in yeast extract sucrose broth. Green peas inoculated with Fusarium graminearum produced about 8 times more T-2 toxin than was obtained in 1% peptone containing Czapek solution under comparable culture conditions.