Nutritional Enhancement of Biocontrol Activity of Candida sake (CPA-1) against Penicillium expansum on Apples and Pears

Pome fruits are poor in nitrogenous compounds and the addition of nitrogen can improve colonisation of the fruits by antagonists. Twenty-two nitrogenous compounds were evaluated for their effect on Candida sake (CPA-1) growth in vitro. Ten compounds that induced greater growth were applied with the antagonist to wounded fruits to evaluate their effect on enhancing control of Penicillium expansum. Calcium chloride and 2-deoxy-D-glucose were also tested. L-serine and L-aspartic acid enhanced biocontrol by C. sake against P. expansum on apples. On apples and pears, ammonium molybdate, calcium chloride and 2-deoxy-D-glucose improved the capacity of the antagonist to control P. expansum. The addition of ammonium molybdate at 1 mM allowed C. sake to be used on apples and pears at a lower concentration without diminishing control. Similar results were observed with the addition of calcium chloride to the antagonist. 2-deoxy-D-glucose at 6 and 18 mM enhanced biocontrol on pears by over 81%, but on apples the improvement of biocontrol was observed only at 6 mM. In cold storage, the combination of ammonium molybdate and C. sake completely eliminated the incidence of blue mould on pears, and reduced its severity and incidence by more than 80% on apples.     

Liquid Formulation of the Postharvest Biocontrol Agent Candida sake CPA-1 in Isotonic Solutions

ABSTRACT Viability of the postharvest biocontrol agent Candida sake CPA-1 stored as liquid formulation was evaluated by studying the effect of growth, preservation medium, and temperature. C. sake was grown in molasses medium with unmodified water activity (a(w)) and in the same with a(w) modified to 0.98 with the addition of several solutes. Cells were preserved with isotonic solutions of different substances. Efficacy of liquid formulations stored for different periods was tested against infection by Penicillium expansum on apples. The best growth media were the (unmodified one and those modified to 0.98 a(w) with the addition of glycerol or sorbitol. For all growth media, the best preservation medium was the isotonic solution prepared with trehalose. When the effect of trehalose concentration in the preservation medium was studied, generally, at trehalose concentrations below the isotonic one, C. sake viabilities increased with increased trehalose. However, the best results were obtained when cells were preserved with the trehalose solution which was isotonic with cells. After 7 months of storage at 4 degrees C, cells that were grown in the sorbitol-modified medium and preserved with the isotonic solution of trehalose (0.96 M) maintained their viability and efficacy against P. expansum infection of apples.     

Control of Postharvest Decay of Apple Fruit with Candida saitoana and Induction of Defense Responses

ABSTRACT The ability of Candida saitoana to induce systemic resistance in apple fruit against Botrytis cinerea was investigated. To separate the antagonistic activity of C. saitoana from its ability to induce resistance, the antagonist and the pathogen were applied in spatially separated wounds. In fresh apples, C. saitoana applied 0 or 24 h before inoculation with B. cinerea showed no effect on lesion development caused by B. cinerea. When applied 48 to 72 h preinoculation with B. cinerea, however, C. saitoana reduced lesion diameter by more than 50 and 70%, respectively, compared with wounding. C. saitoana had no effect on lesion development on stored apples, regardless of the lag period between yeast treatment and inoculation with B. cinerea. In addition to inducing systemic resistance, C. saitoana increased chitinase and beta-1,3-glucanase activities with a higher accumulation in fresh than in stored apples. In fresh apples, the onset of systemic resistance to B. cinerea coincided with the increase in chitinase and beta-1,3-glucanase activity in systemically protected tissue. These studies show that C. saitoana is capable of inducing systemic resistance in apple fruit and indirectly suggest that antifungal hydrolases are involved in the observed systemic protection.     

Improving Control of Green and Blue Molds of Oranges by Combining Pantoea Agglomerans (CPA-2) and Sodium Bicarbonate

The potential of using Pantoea agglomerans (strain CPA-2) alone, or in combination with sodium bicarbonate or sodium carbonate solutions, for control of Penicillium digitatum (green mold) and Penicillium italicum (blue mold) on oranges was investigated under ambient (20 °C) and cold storage (3 °C) conditions. P. agglomerans controlled both pathogens on oranges at 2 × 10⁸ cfu ml^-1. The biocontrol agent was found to be completely tolerant to 2% sodium bicarbonate at room temperature, although its culturability was reduced by > 1000-fold after 30 min in 2% sodium carbonate. The efficacy of P. agglomerans for control of green mold was improved when combined with sodium bicarbonate, resulting in complete and 97.6% reduction of decay incidence at 3 °C and 20 °C, when compared to untreated controls. Satisfactory results were also obtained with the combined treatment for control of blue mold. P. agglomerans grew well inside wounds on oranges at both 20 °C and 3 °C. In contrast, it showed a reduced growth on the surface of intact fruit. Sodium bicarbonate at 2% concentration did not noticeably affect antagonist population development. Thus, use of bicarbonate treatment at 2% followed by the antagonist P. agglomerans CPA-2 could be an alternative to chemicals for control of postharvest diseases on oranges.     

Effect of the antagonist Candida sake on apple surface microflora during cold and ambient (shelf life) storage

This study examined the impact of the application of a biocontrol yeast, Candida sake CPA-1 (3 × 10⁶ colony forming units (cfu) ml^-1) on the resident microbial populations just prior to harvest, during 7 months cold storage and subsequent ambient shelf-life in two seasons on apples untreated or treated with a preharvest pesticide programme. The changes in populations of the antagonistic yeast ( C. sake) were also monitored. Generally, application of the antagonist had little effect on the total bacterial populations which remained very low both prior to harvest and subsequently during cold storage. White yeasts were predominant on the apples during the experimental period, with lower populations of pink yeasts. When apples were removed after 7 months to ambient conditions the yeast populations increased quickly but those apples treated with C. sake had significantly less white yeasts than untreated controls. The dominant filamentous fungi isolated were Cladosporium, Alternaria and Penicillium spp. Penicillium spp. which is responsible for major postharvest diseases was seldom isolated at preharvest but it became important during later cold storage and shelf life period. Populations of Cladosporium and Penicillium were significantly reduced by the C. sake treatment when removed from cold storage during the ambient shelf-life. In contrast, the Alternaria spp. were unaffected by the antagonist. The actual populations of C. sake applied decreased significantly immediately after application (24 h). However, they subsequently increased to a maximum after one month cold storage (10³ cfu g^-1), and populations increased again under ambient shelf-life conditions. The C. sake populations, and the resident microbial populations, were unaffected by preharvest fungicide applications. This study demonstrates that preharvest application of an antagonistic yeast such as C. sake has an impact on the resident mycoflora both during 7 months cold storage and during ambient shelf-life storage.     

Control of Blue Mold of Apples by Preharvest Application of Candida sake Grown in Media with Different Water Activity

ABSTRACT Unmodified and low water activity (a(w))-tolerant cells of Candida sake CPA-1 applied before harvest were compared for ability to control blue mold of apples ('Golden Delicious') caused by Penicillium expansum under commercial storage conditions. The population dynamics of strain CPA-1 on apples were studied in the orchard and during storage following application of 3 x 10(6) CFU/ml of each treatment 2 days prior to harvest. In the field, the population size of the unmodified treatment remained relatively unchanged, while the population size of the low-a(w)-modified CPA-1 cells increased. During cold storage, the populations in both treatments increased from 10(3) to 10(5) CFU/g of apple after 30 days, and then declined to about 2.5 x 10(4) CFU/g of apple. In laboratory studies, the low-a(w)-tolerant cells provided significantly better disease control as compared with the unmodified cells and reduced the number of infected wounds and lesion size by 75 and 90%, respectively, as compared with the non-treated controls. After 4 months in cold storage, both unmodified and low-a(w)-tolerant cells of C. sake were equally effective against P. expansum on apple (>50% reduction in size of infected wounds).     

Biological control of botrytis bunch rot in organic wine grapes with the yeast antagonist Candida sake CPA‐1

The aim of this research was to confirm the efficacy of the yeast antagonist Candida sake CPA‐1 in suppressing botrytis bunch rot development, in an organic vineyard under Mediterranean conditions for two seasons, and compare its performance with that of two biologically based products currently registered for botrytis bunch rot control in New Zealand. In 2009, treatments applied were: commercial formulations of Ulocladium oudemansii (BOTRY‐Zen^®) and chitosan (ARMOUR‐Zen^®), C. sake CPA‐1 combined with the fatty acid‐based additive Fungicover^® and combinations of these products. All treatments were applied six times between early flowering and harvest and compared with an unsprayed control. In 2010, the treatments focused on C. sake and Fungicover and the number of applications was reduced from six to four. The population dynamics of U. oudemansii and C. sake were measured and wine quality tests were carried out in both seasons. Disease control achieved by C. sake treatments in 2009 were comparable to those achieved by BOTRY‐Zen and ARMOUR‐Zen. Applications of C. sake plus Fungicover between flowering and harvest significantly (P < 0·05) reduced botrytis bunch rot incidence and severity by 64% and 90%, respectively, compared with the untreated control in 2009, and by 67% and 89%, respectively, in 2010. Treatments did not adversely affect wine quality parameters after treated grapes were processed. Candida sake consistently provided effective control of botrytis bunch rot in grapes under different meteorological and disease pressure conditions, thereby improving its potential for future commercial applications.     

PP 1/304 (1) Control of suckers in orchards

Specific scope

This Standard describes the conduct of trials for the efficacy evaluation of plant growth regulators used to control suckers in orchards. The principal requirement is to remove sucker growth without a negative impact of the plant protection product on the parent plant.

Specific approval and amendment

First approved in 2017‐09.     

不同作物套栽对枯草芽孢杆菌控制石榴枯萎病效果的影响

本文室内测定了枯草芽孢杆菌Bacillus subtilis对石榴枯萎病菌Ceratocystis fimbriata的抑菌作用,检测其在石榴Punica granatum、番茄Lycopersicum esculentum、大豆Glycine max、青菜Brassica chinensis、芫荽Coriandrum sativum、大葱Allium fistulosum var. giganteum和大蒜Allium sativum根际的定殖动态及其结合作物套栽对石榴枯萎病的盆栽控制效果。结果表明,芽孢杆菌在不同供试植物根际的定殖能力差异大,在石榴根际的定殖力弱,在番茄、大葱和大蒜根际的定殖力强;作物套栽结合施用芽孢杆菌明显地减轻了石榴枯萎病的发病程度,其中芽孢杆菌结合套栽番茄、大葱和大蒜的平均防效分别达78.14%、80.99%和78.91%。综上所述,套栽番茄、大葱和大蒜有望成为一种提高枯草芽孢杆菌控制石榴枯萎病的有效手段。     

Ultrastructural and Cytochemical Aspects of the Biological Control of Botrytis cinerea by Candida saitoana in Apple Fruit

ABSTRACT Biocontrol activity of Candida saitoana and its interaction with Botrytis cinerea in apple wounds were investigated. When cultured together, yeast attached to Botrytis sp. hyphal walls. In wounded apple tissue, C. saitoana restricted the proliferation of B. cinerea, multiplied, and suppressed disease caused by either B. cinerea or Penicillium expansum. In inoculated apple tissue without the yeast, fungal colonization caused an extensive degradation of host walls and altered cellulose labeling patterns. Hyphae in close proximity to the antagonistic yeast exhibited severe cytological injury, such as cell wall swelling and protoplasm degeneration. Colonization of the wound site by C. saitoana did not cause degradation of host cell walls. Host cell walls in close contact with C. saitoana cells and B. cinerea hyphae were well preserved and displayed an intense and regular cellulose labeling pattern. In addition to restricting fungal colonization, C. saitoana induced the formation of structural defense responses in apple tissue. The ability of C. saitoana to prevent the necrotrophic growth of the pathogen and stimulate structural defense responses may be the basis of its biocontrol activity.     

Control of Colorado beetle (Leptinotarsa decemlineata) in Poland1

Leptinotarsa decemlineata (Colorado beetle) was found in Poland for the first time in 1945 and a great invasion of this pest took place in 1950. It now plays a very important role in Polish plant protection, mainly because the acreage of potato in Poland amounts to about 1.8 million ha. The pest is controlled by repeated insecticide applications to a yearly total of 2 million ha, which is the highest amount of insecticides used in Poland. Research has been carried out in Poland on the following subjects: biology and ecology of Colorado beetle, influence of pest appearance and control on the biocenosis of fields, methods of control, biochemical factors influencing the life cycle, forecasts and warning of appearance, artificial diet, resistance of potato cultivars and reaction of leaf blades of potato to the eggs of Colorado beetle. Research is also carried out on chemical control, modernization of pesticide recommendations and resistance of Colorado beetle to pesticides. The influence of biotic and abiotic factors on the reduction of Colorado beetle numbers has been studied. With a view to possible biological control, predatory bugs (Perillus bioculatus and Podisus maculiventris) were introduced into Poland in 1959 and 1979. Research is also conducted on entomopathogenic fungi, parasitic nematodes, protozoans, spiroplasms and Bacillus thuringiensis.     

风力除虫机对假眼小绿叶蝉的控制效果

吹吸式风力除虫机对假眼小绿叶蝉成虫和若虫具有显著的控制作用,风力越大,控制效果越好.当风速为30m/s(±3%±0.1)时,处理后1d校正虫12减退率达76.25%,处理后7 d校正虫口减退率为61.49%,产卵量减退率为41.38%.吹吸式风力除虫机对假眼小绿叶蝉春季和秋季高峰期的虫口数量具有显著的控制效果,但其效果显著低于0.3%印楝素EC2.0 mg/L.     

胡瓜新小绥螨对B型烟粉虱的控制能力

为明确胡瓜新小绥螨Neoseiulus cucumeris Oudermans对烟粉虱Bemisia tabaci Gennadius的控害潜能,在（26±1）℃、相对湿度65%±5%、光周期16L：8D条件下,研究了各螨态胡瓜新小绥螨对不同发育阶段烟粉虱的捕食作用。结果表明,胡瓜新小绥螨幼螨期较短,不取食烟粉虱,其他不同螨态的胡瓜新小绥螨可捕食除4龄若虫和成虫以外的其他各虫态的烟粉虱,捕食功能反应符合HollingⅡ型;胡瓜新小绥螨雌成螨对烟粉虱卵的日捕食量最大（8.5粒/雌）,对烟粉虱3龄若虫的日捕食量最小（2.1头/雌）;雌成螨对烟粉虱卵具有较高的瞬间攻击率（0.038）和寻找效应（0.025）。研究表明,胡瓜新小绥螨对烟粉虱卵及低龄若虫的捕食能力较强,是烟粉虱发生初期的有效天敌。     

Control of Cirsium arvense (L) Scop, by herbicides and mowing

The efficacy of mowing and of herbicides (applied by spot spraying or boom spraying) for the selective control of Cirsium arvense (L) Scop, in Victorian pastures was investigated. The pastures were based on Trifolium repens L. and Lolium perenne L. A general decline in the number of thistle shoots in untreated plots at three out of five sites was recorded.‘Tordon 50-d' (5% a.i. picloram and 20% a.i. 2,4-D as triisopropanolamine salts), MCPB, 2,4-DB and mowing resulted in suppression, but not eradication, during the experimental period. MCPB and 2,4-DB are the most suitable herbicides for selective control. The use of‘Tordon 50-d' is restricted by its effect on clovers, soil persistence and cost. Variable growth of C. arvense, associated with variable summer rainfall in Victoria, increases the difficulty in controlling this weed and necessitates a persistent long term approach.     

Enhancement of Postharvest Disease Resistance in Ya Li Pear (Pyrus bretschneideri) Fruit by Salicylic Acid Sprays on the Trees during Fruit Growth

The Ya Li pear (Pyrus bretschneideri) trees were sprayed three times with 2.5 mM salicylic acid (SA) around 30, 60 and 90 days after full flowering. The fruit were harvested at commercial maturity (about 120 days after full flowering), inoculated with Penicillium expansum, and incubated at 20 °C, 95–100% RH. The results showed that resistance to the pathogen of the mature pear fruit was remarkably enhanced by the SA sprays. Disease incidence in the SA-treated fruit was 58.0% or 26.5%, and lesion diameter on SA-treated fruit was 58.4% or 29.0% lower than that in/on fruit without SA treatment (control) on day 12 or 17 after incubation, respectively. The SA spray applied to the trees around 30 days after full flowering notably enhanced accumulation of hydrogen peroxide in the young fruit. Meanwhile, activities of defense enzymes, including peroxidase, phenylalanine ammonia-lyase (PAL), chitinase or β-1,3-glucanase in the young fruit from SA-treated trees was 29.5%, 60.0%, 24.4% or 35.7% higher than that in the control fruit 4 days after the SA spraying. Furthermore, after harvest, activities of PAL, chitinase and β-1,3-glucanase were still significantly higher in the mature pear fruit from the trees sprayed three times with SA than those of the control fruit. Activities of the antioxidant enzymes including catalase and ascorbate peroxidase in the young fruit were significantly reduced by SA spraying. However, the activity of another antioxidant enzyme, glutathione reductase in the young fruit was significantly enhanced by SA spraying. These results suggest that enzymes exerting their functions in different ways may be coordinately regulated by SA in the pear fruit. Our study indicates that treatment of SA sprays on the trees may provide further protection against postharvest disease of Ya Li pear fruit in practice and could be used as an alternative and economical approach to reduce application of chemical fungicides.