Effects of low temperature on mycelial growth and spore germination of Botrytis allii in culture and on its pathogenicity to stored garlic bulbs

Botrytis allii was incubated at 20, 10,4, 2,0, – 2 and –4° to investigate effects of temperature on growth, sporulation and germination on potato dextrose agar (PDA), and to estimate incidence and severity of disease on garlic bulbs inoculated with the pathogen during storage. B. allii-was capable of growing to a colony diameter of 17 mm after 20 weeks, sporulating and germinating on PDA at temperature as low as – 4°C. After 12 weeks at –4 and -2°C the mycelial growth was observed only on 45–54% of bulbs wound-inoculated with conidial suspension, and infection was limited to 5 mm in diameter on the surface of inoculation sites without producing any symptoms of the disease. Temperature responses were similar for mycelial growth, conidial germination and infection, but mycelial growth and sporulation was slower and later on garlic bulbs than in culture. Wounds were readily colonized by B. allii. No disease was found on unwounded bulbs that were inoculated with conidial suspensions and with mycelial plugs at various temperatures. Only 16–95% of cloves became infected by contact with other cloves within the same bulb after 12 weeks of storage at temperatures of 4–20 C.     

Metabolic alteration induced by Botrytis allii in two onion cuitivars with different resistances to neck rot

Disease index, dry weight loss and leakage of K⁺ ions in the tissues of two onion cultivars demonstrated that the red cultivar Tropeana had a greater susceptibility to neck rot caused by Botrytis allii than the white cultivar Rovato. The comparison between the cultivars showed that the less susceptible one was characterized by lower polygalacturonase activity (with fewer isoenzyine forms), more rapid accumulation of phenolic compounds in the early stages and faster activation of peroxidase. Polyphenoloxidase was not found in either the healthy or the diseased tissues of the two cultivars. The significance of these findings is discussed in relation to possible host defence mechanisms.     

The effect of temperature and water potential on the production of conidia by sclerotia of Botrytis squamosa

The rate of conidiogenic germination of Botrytis squamosa was highest at 16°C and the greatest numbers of conidia per sclerotium (up to 5 × 10⁴) were produced at temperatures of 5–10°C. At temperatures above 20°C, the percentage of sclerotia producing conidia declined rapidly. Decreasing water potential reduced the rate at which conidia were produced and also resulted in fewer conidia produced per sclerotium. However, conidia were produced at water potentials as low as −2 MPa, at which sclerotial germination was at least 60%. A simulation model that included effects of both temperature and water potential was developed from laboratory and field data obtained for conidial production in sclerotia exposed for periods of 1, 2, 3 or 4 weeks during an entire year. There was good agreement between conidiogenic germination predicted by the model and conidial production observed in onion plots artificially inoculated with sclerotia. Temperature and water potential were therefore considered to be the principal microclimatic factors affecting conidial production by B. squamosa. The role of sclerotia in the context of UK onion production is discussed.     

Distribution of benzimidazole-resistant strains of the onion gray-mold neck rot pathogens,Botrytis aclada and Botrytis allii,in Hokkaido,Japan

Botrytis aclada and Botrytis allii, associated with onion gray-mold neck rot and isolated in Hokkaido, were tested for sensitivity to benzimidazole. Of the B. aclada strains, 59% were highly resistant and the remaining 41% were sensitive; all strains of B. allii were sensitive. Resistant strains were widespread in Hokkaido. We analyzed the sequences of the β-tubulin gene of resistant strains and detected the replacement of glutamic acid (GAG) by lysine (AAG) at codon 198. This is the first report of benzimidazole resistance in B. aclada. This study revealed a difference in fungicide sensitivity between the two Botrytis species.

     

The composition of wall alterations and appositions (reaction material) and their role in the resistance of onion bulb scale epidermis to colonization by Botrytis allii

Ultrastructural examination of deposits of reaction material (RM) showed them to be composed of paramural granules (osmiophilic to varying degrees) lying within amorphous electron–dense material. Epidermal cell walls per se became increasingly electron–dense around the short infection hyphae produced by Botrytis allii. Fluorescence microscopy, histochemical tests and microautoradiography showed the accumulation of phenolic compounds at infection sites. Use of phenylalanine and cinnamate for E.M. autoradiography demonstrated the accumulation of phenolics within paramural granules, amorphous aggregates and within the penetrated cell wall, Paramural deposits were soluble in organic solvents and probably represented reservoirs of low MW phenolics which were progressively incorporated into insoluble polymers within the cell wall. Initial synthesis or accumulation of phenolics appeared to take place in rough endoplasmic reticulum during the first 2 h after cuticle penetration. Suppression of RM deposition and wall alterations was achieved by treatment of bulb scales with cycloheximide. Tissues treated with the protein synthesis inhibitor became susceptible to colonization by B. allii. Accumulation of phenolics at infection sites rendered the onion cell walls resistant to attack by a mixture of cell wall–degrading enzymes, Methanolic extracts of epidermal strips containing RM deposits contained very weakly active inhibitors of B. allii germ–tube growth. The role of RM deposits and wall alterations in the resistance of onion epidermis to colonization by Botrytis is discussed.     

Bacteria associated with internal breakdown of onion bulbs and their possible role in disease expression

Bacteria of the genera Pseudomonas. Erwinia. Klebsiella, Enterobacter and Escherichia were isolated from internal swollen leaf bases of naturally infected onion bulbs and were pathogenic to onion tissue on reinoculation. In pathogenicity tests, inoculation of slices from symptomless bulbs with one bacterium or with a sterile needle often resulted in isolation of P. aeruginosa, Serratia marcesvens and Bacillus cereus. It is postulated that the bacteria isolated form part of the endophytic microflora of an onion bulb, and that they may become opportunistic pathogens when changes in host physiology are triggered by high temperatures at bulb maturity.     

Effect of inoculum composition on infection of French bean leaves by conidia of Botrytis cinerea

Inoculation of leaves of French bean (Phaseolus vulgaris) with sprays or small drops of a suspension of conidia ofBotrytis cinerea gave rise to spreading lesions, lesions remaining restricted in size or to no visible necrosis. The type of reaction depended on the composition of the inoculum. In studies with drop inoculations with buffered inocula some of the factors involved were analyzed. The formation of spreading lesions depended on pH, type and molarity of the buffer, presence of glucose, and concentration of conidia in the inoculum. If the phosphate buffer used in most of the inocula was replaced by monobasic phosphate, similar results were obtained. The reactions were not influenced by the proportion of K⁺ or Na⁺ ions in the phosphate buffer. Inoculations with conidia suspended in a solution of 0.067 M phosphate buffer (pH 5.0) or monobasic phosphate and 0.11 M glucose always evoked a susceptible reaction, i. e. the formation of spreading lesions.     

Wind dispersal of conidia of Botrytis spp. pathogenic to Vicia faba

Increasing numbers of conidia were blown from sporulating cultures of Botrytis fabae and B. cinerea as windspeed increased up to 10 m/s. Partial drying of cultures increased the number of spores blown away at low and intermediate windspeeds. Release of spores at a constant windspeed was sustained over a prolonged period. Different patterns of release from colonies of the two species when windspeed was gradually increased, or at a constant windspeed. may be related to differences in spore size affecting the drying rate. Many conidia of both species were released as clumps. A higher proportion of B. cinerea than B. fabae conidia were clumped, partly because the mean number of spores per clump was greater. Individual conidia fell more slowly in still air than did clumps. The humidity in a bean crop was more favourable to development of Botrytis lesions than that above the crop. Low windspeeds measured within crops may restrict dispersal of conidia and may result in uneven distribution of chocolate spot lesions.     

Effect of interrupted leaf wetness periods on suppression of sporulation ofBotrytis allii andB. cinerea by antagonists on dead onion leaves

Saprophytic antagonists were evaluated for suppression of sporulation ofBotrytis allii andB. cinerea on artificially killed segments of onion leaves that were pre-inoculated with the pathogens. During incubation of the antagonisttreated leaf segments in moist chambers, periods of leaf wetness and leaf dryness were alternated to simulate conditions in the field. Interruption of humid conditions with dry periods had a differential effect on antagonists.Alternaria alternata, Chaetomium globosum, Ulocladium atrum andU. chartarum suppressed sporulation ofB. allii almost completely under continuously wet conditions, and when the leaf wetness periods were interrupted with drying periods of 9h imposed 16, 40, and 64 h after the antagonists were applied. When leaf wetness was interrupted 16 h after antagonist application, the number of conidia ofB. allii produced cm^–2 leaf surface after eight days was under the detection limit of 5.2 × 10³ conidia on leaves treated with these antagonists compared to 3.7 × 10⁵ conidia on leaves that were not treated. On the other hand,Gliocladium roseum, G. catenulatum andSesquicillium candelabrum, all highly efficient under continuously wet conditions, were of low to moderate efficiency when leaf wetness periods had been interrupted 16 h after application of the antagonists. The antagonists showed the same differentiation and sensitivity to interrupted wetness periods when tested withB. cinerea.     

Effects of temperature,relative humidity and age of conidia on the incidence of spotting on gerbera flowers caused by Botrytis cinerea

In recent years, spotting of ray florets of gerbera flowers has become an important problem. This type of small necrotic lesions may occur before, but especially shortly after harvesting the flowers.Botrytis cinerea was easily isolated from such lesions. Inoculation withB. cinerea only gave typical necrotic lesions, when dry conidia were dusted on the flowers with a short period of high rh after inoculation. At 18–25 °C a high rh for at least 5 hours was necessary. Rotting of ray florets and receptacles byB. cinerea occurred when inoculated flowers were kept wet for a few days. Spots consist of one to several necrotic, usually epidermal cells. A single conidium could give rise to a necrotic lesion after germination. Germination of conidia and lesion formation occurred between 4 and 25 °C; at 30 °C, germination and lesion formation did not occur. Between 18 and 25 °C, many lesions became visible within 1 day after inoculation; at 4 °C it took 2 to 3 days before lesions could be seen. If kept dry, conidia ofB. cinerea remained ungerminated on ray florets of gerbera flowers and could be removed from the ray florets. Within 1 day at high rh, germination occurred and lesions were produced. Conidia ofB. cinerea, stored dry, were able to survive much longer than the lifetime of a gerbera flower. Even after storage at room temperature for up to 14 months, some conidia were able to germinate in vitro and on ray florets and induce the formation of lesions. Addition of gerbera pollen diffusate stimulated germination and lesion formation.     

Retention of iprodione on salad onion leaves in relation to the control of Botrytis spp.

Using bioassay and fluorescent tracer techniques it was demonstrated that more fungicide was retained on mature leaves of summer salad onion crops if 1 kg/ha iprodione wettable powder (w.p.) formulation plus wetting agent was sprayed at 280 1/ha than at 560 1/ha or 1120 1/ha.
The lowest spray volume (280 1/ha) with four sprays of iprodione w.p. (0.5 kg in October, November, December and January) gave effective control of Botrytis cinerea and significantly increased plant stand and yield. More fungicide was required to obtain equivalent results using a mixed programme of two thiram (4 kg/ha total) and two iprodione w.p. (2 kg a.i./ha total) sprays at the same application volume and intervals. Where the mixed programme was applied at 1 1 20 1/ha, 1 2 kg a.i. (total fungicide) was necessary to achieve equivalent disease control.
At 280 1/ha, retention and persistence of iprodione on onion foliage was greater with the oil-based than with the wettable powder formulation. In summer especially this resulted in improved Botrytis control and greater numbers of marketable onions. In winter onions, oil and w.p. formulations were equally effective and gave equivalent improvements in plant stand and yield.     

Degradation of stilbene-type phytoalexins in relation to the pathogenicity of Botrytis cinerea to grapevines

The ability of eight isolates of Botrytis cinerea to degrade the stilbene phytoalexins, resveratrol and pterostilbene, was compared with their pathogenicity to grapevines. All strains which degraded resveratrol and pterostilbene were highly or moderately pathogenic to in vitro cultures of grapevines ( Vitis rupestris ) after inoculation with agar disks containing mycelium, while those which were unable to degrade phytoalexins were non-pathogenic. In all cases, the hydroxystilbene-degrading activity was related to the presence of laccase activity in the culture filtrates, as shown by using syringaldazine as substrate. The role of laccase-mediated degradation of phytoalexins in relation to pathogenicity of B. cinerea to grapevines in discussed.     

Adsorption of14C-labelled exudates by conidia of Botrytis fabae on Vicia faba leaves

Samenvatting TuinboonplantenVicia faba werden na een 24-urig verblif, in een¹⁴CO₂-milieu geïnoculeerd met conidiën vanBotrytis fabae. Zes uur na de inoculatie werden de conidiën verzameld. De adsorptie van met¹⁴C gemerkte bladexudaten door de conidiën kon microautoradiografisch worden vastgesteld.     

Some factors influencing the outgrowth of Botrytis tulipae from lesions on tulip bulbs after planting

Lesions ofBotrytis tulipae on tulip bulbs do not give rise to new infections during storage but may do so after planting. Compared with storage at 20°C and a relative humidity (r.h.) of 40%, storage at 20°C and 95% r.h. reduces, and at 5°C and 95% r.h. tends to increase the rate of successful isolations from lesions during the storage period and the number of new infections after planting. The bulbs show more new infections during growth in soil at 9°C than at 18°C.Samenvatting Lesies, veroorzaakt doorB. tulipae op de buitenste rok van tulpebollen, veranderen niet van uiterlijk tijdens de bewaarperiode. Na het planten kan de schimmel tot aan het bloeitijdstip (waarna geen waarneming aan de moederbol meer mogelijk is) op elk moment, zelfs vlak voor het bloeitijdstip, vanuit de lesies uitgroeien en nieuwe infecties veroorzaken vlak naast de oude lesies of op enige afstand daarvan (Tabel 1). Het is niet bekend of bij deze late uitgroei een remmende factor in of rond de lesies een rol speelt. Bij een aantal lesies groeit de schimmel niet uit, hoewel hij uit enkele daarvan wel kon worden geïsoleerd, zelfs nog omstreeks de bloeitijd.Hoewel in vitro een temperatuur van 20°C en een hoge relatieve vochtigheid (rv) voor de schimmel optimale groeiomstandigheden vormen (Válaková, 1963), bleek dat na bewaring van de bollen bij 20°C en een hoge rv (95%), het uitgroeien vanB. tulipae vanuit de lesies na het planten geringer was dan na bewaring bij 20°C en 40% rv, terwijl bewaring bij 5°C de uitbreiding bevorderde (Tabel 3). Na het planten vond bij lage bodemtemperatuur (9°C) meer uitbreiding van de lesies plaats dan bij hogere. De oorzaak van de verschillen tussen het uitgroeien van de schimmel in vitro en in vivo is niet bekend; misschien blijft de schimmel levenskrachtiger in het bolweefsel bij lage temperatuur of is bij hoge temperatuur een remmende factor meer actief.     

Laboratory evaluation of temperature effects on the germination and growth of entomopathogenic fungi and on their pathogenicity to two aphid species

As part of an approach to select potential mycoinsecticides for aphid biocontrol, we investigated the effects of temperature on the growth, germination and pathogenicity of some hyphomycete fungi. Commercially available mycoinsecticides (based on Beauveria bassiana (Balsamo) Vuillemin and Verticillium lecanii (Zimmermann) Viegas) and other isolates of B bassiana, V lecanii, Metarhizium anisopliae (Metschnikoff) Sorokin and Paecilomyces fumosoroseus (Wize) Brown & Smith were evaluated. The rate of in vitro conidial germination of all isolates was slower at 10 and 15 degrees C than at 20 and 25 degrees C. Similarly, in vitro growth of most isolates was adversely affected at 10 and 15 degrees C. The greatest reduction at 10 degrees C in rates of conidial germination and colony growth, compared with other temperatures, was for M anisopliae isolates. Germination of V lecanii (isolate HRI 1.72) was fastest at 10 degrees C compared with the other fungi. It was also the most pathogenic of three isolates tested against Aphis fabae Scopoli and Myzus persicae Sulzer at 10, 18 and 23 degrees C. Generally, A fabae was more susceptible than M persicae to infection by the fungal isolates tested. A significant interaction between aphid species and temperature indicated that the pathogenic nature of an isolate was dependent not only on the target aphid species but also the temperature conditions of the bioassay. The series of studies, detailed above, allowed a temperature profile to be formed for the different isolates. Verticillium lecanii isolate HRI 1.72 (commercialised as Vertalec) was the most promising isolate selected from results of the series of experiments. Temperature profiles in conjunction with infectivity assays can be useful in selecting appropriate isolates for a particular thermal environment.     

Effect of storage duration and temperature on the survival of Rhizoctonia solani in tulip and iris bulbs

In general, the viability ofRhizoctonia solani in lesions of stored tulip and iris bulbs decreased rapidly after harvest. However, some of the mycelium remained viable and acted as source of infection after planting. Temperature had no influence on the rate of survival, as estimated by isolation on an agar medium, but after planting moreRhizoctonia grew out on iris bulbs which had been stored continuously at 30°C for 20 or 32 weeks than on bulbs kept at lower temperatures. The surviving fungus was also able to infect neighbouring iris plants. The presence of healthy iris bulbs close to similar but diseased bulbs promoted the development ofRhizoctonia on the latter after planting.Samenvatting De levensvatbaarheid vanRhizoctonia solani in lesies op tulpe- en irisbollen nam tijdens de bewaring na de oogst in het algemeen snel af. Er bleef echter enig mycelium in level, dat na het planten als infectiebron fungeerde. De temperatuur had geen invloed op de mate van overleving wanneer de mate van slaging van isolaties op een agarmedium als criterium werd genomen. Na planten groeide echter meerRhizoctonia uit op irisbollen die permanent gedurende 20 en 32 weken bij 30°C waren bewaard dan bij bollen die bij lagere temperaturen waren opgeslagen. De overlevende schimmel was ook in staat om naburige irissen aan te tasten. De aanwezigheid van gezonde irissen in de nabijheid van aangetaste bevorderde de ontwikkeling vanRhizoctonia op geplante zieke bollen.     

Effects of postharvest onion curing parameters on bulb rot caused by Pantoea agglomerans,Pantoea ananatis and Pantoea allii in storage

Crop loss of onion bulbs during storage carries an exceptionally high economic impact because a large portion of the production expenses has been expended before storage. Because of this, it is important to define practices that can reduce onion bulb losses caused by storage rots. This study investigates the impact of various curing parameters on disease development resulting from infection by Pantoea agglomerans, P. ananatis and P. allii on onion bulb cultivars Vaquero and Redwing, during storage. Overall, both the incidence and mean rot severity were similar amongst the bulbs under comparable conditions regardless of the species of Pantoea inoculated, although a significant difference was detected between the two onion bulb cultivars. In addition, a significant reduction of storage rot was observed when curing temperatures were ≤35°C. At temperatures >35°C, a shorter curing duration (2 days vs 14 days) decreased the severity of bulb rot due to Pantoea. This increased understanding of the inter‐relationships between the parameters used for curing, and the incidence and severity of bulb rot caused by Pantoea helps provide guidance towards using the curing process as a means to reduce the level of damage resulting from post‐harvest storage rot.     

The effects of direct harvesting and drying systems on the incidence and control of neck rot (Botrytis allii) in onions

Direct harvesting, with mechanical removal of the foliage (topping), of onion crops followed by post-harvest drying at ambient temperatures ( c . 18°C) resulted in an increase in the incidence of onion neck rot ( Botrytis allii ).
The disease was substantially reduced if topped onions were dried at 30°C with an airflow of 425 m³ air/h/tonne. The treatment was most effective if the crop was removed from the field for drying within 48 h of topping thus avoiding severe infection of the damaged green tissues of the necks of onions.     

温度调控对番茄灰霉病菌侵染的影响

从群体的角度研究了番茄灰霉病菌对温度的适应性,结果表明灰霉病菌孢子的萌发适温在15～25℃,最适温度为20℃。15～25℃适宜菌丝生长、侵染与扩展,20℃为最适温度。30℃以上病害发展受到抑制。低温有利于病菌产孢,最适温度为15℃。变温研究表明,在一天中,随着30℃以上高温时间的增长,病菌(菌丝)的致病力呈下降趋势,6 h以上高温能够限制病害发展。     

Influence of environmental conditions on dispersal of Botrytis cinerea conidia and on post-harvest infection of gerbera flowers grown under glass

Dispersal of Botrytis cinerea in a gerbera crop grown in two glasshouses 30 km apart was studied over a period of 18 months, in 1988 and 1989. Conidia were caught in spore traps consisting of agar in petri dishes exposed at different heights in the crop in each glasshouse. No seasonal patterns could be identified in the spore catches, assessed as colonies on the agar traps after incubation. The number of lesions caused by conidial infection of gerbera flowers following incubation, however, showed a distinct pattern. In spring and early summer few lesions were recorded whereas at other times of the year many lesions appeared. In linear regression analysis, variation in numbers of colonies (spore catches) could not be explained by environmental factors recorded during the experiments. Linear regression accounted for 77% and 81% of the variation in the number of lesions on flowers in the two glasshouses, in terms of relative humidity (postively correlated), global radiation outside the glasshouse (negatively correlated) and age of the crop (positively correlated). Despite differences in the systems by which the gerbera crop was produced and in the spore catches, the numbers of lesions on gerbera flowers in the two glasshouses were significantly correlated though not significantly different from each other.