Pichia fermentans dimorphic changes depend on the nitrogen source

Pichia fermentans DiSAABA 726 is a biofilm-forming yeast that undergoes dimorphic transition. Under yeast-like morphology it controls brown rot caused by Monilia spp. on apple fruit, while under pseudohyphal form, it shows pathogenic behaviour itself on peach fruit. The present study investigates the nutritional factors that induce and separate yeast-like and pseudohyphal morphologies under laboratory conditions. We show that P. fermentans DiSAABA 726 produces mainly yeast-like cells on media containing millimolar concentrations of urea and diammonium phosphate, and forms pseudohyphae at micromolar concentrations of these two salts. With ammonium sulphate, yeast-like or pseudohyphal morphology depends on the N concentration and the pH of the culture media. Amino acids such as methionine, valine, and phenylalanine invariably induce pseudohyphal morphology irrespective of the N concentration and the pH of the culture media. Methionol, 1-butanol, isobutanol, and isopropanol induce pseudohyphal growth, while phenylethanol and isoamyl alcohol fail to induce the formation of filaments. Thus, the morphogenesis of P. fermentans DiSAABA 726 depends more on the nitrogen source than on the N concentration, and is regulated by the quorum-sensing molecules that are generally produced from amino-acid assimilation under nitrogen starvation.     

Effects of a biocontrol agent and methyl jasmonate on postharvest diseases of peach fruit and the possible mechanisms involved

AIMS: To investigate effects of application of 200 micromol l(-1) methyl jasmonate [MeJA (200)] and Cryptococcus laurentii alone or in combination against postharvest diseases (Monilinia fructicola and Penicillium expansum) in peach fruit stored at 25 and 0 degrees C, and to evaluate the possible mechanisms involved. METHODS AND RESULTS: The efficacy of controlling postharvest diseases by resistance induced in peach fruit treated with MeJA (200) and C. laurentii alone or in combination and the relationship between activities of defence-related enzymes in peach fruit and lesions caused by M. fructicola and P. expansum were examined. At the same time, the effects of MeJA (200) on the population of C. laurentii in the peach wounds and on the mycelial growth of M. fructicola and P. expansumin vitro were investigated. The results indicated that treatment of peach fruit with C. laurentii at 1 x 10(8) CFU ml(-1) alone, or combining C. laurentii at 5 x 10(7) CFU ml(-1) with MeJA (200) all resulted in a lower lesion diameter of brown rot and blue mould caused by M. fructicola and P. expansum compared with the controls in peach fruit. MeJA (200) enhanced the population of C. laurentii, and inhibited mycelial growth of P. expansum. However, it had a little effect on M. fructicolain vitro. MeJA and C. laurentii alone or in combination induced higher activities of Chitinase, beta-1,3-glucanase, phenylalanine ammonia-lyase and peroxidase (POD) than applying the yeast alone at both 25 and 0 degrees C. CONCLUSIONS: MeJA (200) not only directly inhibited mycelial spread of postharvest pathogens, but also increased population of C. laurentii, which induced stronger disease resistance in fruit than MeJA or yeast alone, and resulted in a lower lesion diameter of brown rot and blue mould caused by M. fructicola and P. expansum. SIGNIFICANCE AND IMPACT OF THE STUDY: MeJA (200) in combination with C. laurentii was beneficial for controlling brown rot and blue mould caused by M. fructicola and P. expansum in peach fruit. The inhibitory mechanism was mainly because of resistance induced in peach fruit by MeJA and C. laurentii. In addition, direct inhibition of MeJA on P. expansum also played a role in controlling blue mould.     

Response of female oriental fruit moth to volatiles from apple and peach trees at three phenological stages

The attractiveness of peach ( Prunus persica L. Batsch) and apple ( Malus domestica L. Borkh.) (both Rosaceae) tissue to gravid female oriental fruit moth, Grapholita (=  Cydia ) molesta (Busck) (Lepidoptera: Tortricidae), was assessed at three distinct stages throughout the growing season using a dual-choice bioassay. Plant material offered to the female moths consisted of a plant shoot in early spring, before fruit became available. Later, it consisted of a leaf-bearing twig and a fruit, either alone or in combination. The level of attraction of the female moths to the various plant tissues varied substantially over time and according to the plant species. Before fruit became available, female moths were significantly attracted to peach as well as to apple shoots. During the early fruit growth stage, moths were attracted to a leaf-bearing twig originating from a peach tree, but not to that from an apple tree. In peach, it was the vegetative tissue that accounted for the attraction, whereas in apple, it was the reproductive tissue (a developing fruit). During the late fruit growth stage, both peach fruit and apple fruit were highly attractive, whereas a twig with leaves from either an apple or a peach tree was neither attractive nor repellent to the female moths. This changing female olfactory response to volatiles emitted by vegetative tissue and fruits from the two host plant species with progressing season is discussed with respect to the biology and the dispersal of this moth species.     

Combination of antagonistic yeasts with two food additives for control of brown rot caused by Monilinia fructicola on sweet cherry fruit

AIMS: To evaluate beneficial effect of two food additives, ammonium molybdate (NH4-Mo) and sodium bicarbonate (NaBi), on antagonistic yeasts for control of brown rot caused by Monilinia fructicola in sweet cherry fruit under various storage conditions. The mechanisms of action by which food additives enhance the efficacy of antagonistic yeasts were also evaluated. METHODS AND RESULTS: Biocontrol activity of Pichia membranefaciens and Cryptococcus laurentii against brown rot in sweet cherry fruit was improved by addition of 5 mmol l(-1) NH4-Mo or 2% NaBi when stored in air at 20 and 0 degrees C, and in controlled atmosphere (CA) storage with 10% O2 + 10% CO2 at 0 degrees C. Population dynamics of P. membranefaciens in the wounds of fruit were inhibited by NH4-Mo at 20 degrees C after 1 day of incubation and growth of C. laurentii was inhibited by NH4-Mo at 0 degrees C in CA storage after 60 days. In contrast, NaBi did not significantly influence growth of the two yeasts in fruit wounds under various storage conditions except that the growth of P. membranefaciens was stimulated after storage for 45 days at 0 degrees C in CA storage. When used alone, the two additives showed effective control of brown rot in sweet cherry fruit and the efficacy was closely correlated with the concentrations used. The result of in vitro indicated that growth of M. fructicola was significantly inhibited by NH4-Mo and NaBi. CONCLUSION: Application of additives improved biocontrol of brown rot on sweet cherry fruit under various storage conditions. It is postulated that the enhancement of disease control is directly because of the inhibitory effects of additives on pathogen growth, and indirectly because of the relatively little influence of additives on the growth of antagonistic yeasts. SIGNIFICANCE AND IMPACT OF THE STUDY: The results obtained in this study suggest that an integration of NH4-Mo or NaBi with biocontrol agents has great potential in commercial management of postharvest diseases of fruit.     

Calcium ion transport across discs of the cortical flesh of apple fruit in relation to fruit development

Transport of Ca²⁺ through discs of apple fruit tissue was examined in tissue taken at different stages of fruit development. Transport rates decreased with fruit development when cation exchange was the predominant influence on transport (with 10⁻⁶ M ⁴⁵CaCl₂ as the source solution). This decrease was associated with a reduction in relative cell wall surface area, cation exchange capacity and cell wall yield that occurred during fruit growth. When diffusion was the major transport force, and when transport was influenced by solution infiltration of the tissue disc (10⁻² M ⁴⁵CaCl₂ in the source solution), transport rates increased during fruit growth. This increment was related to increases in air space of the tissue. Ca²⁺ transport through apple fruit tissue is influenced by the extent and nature of the cell wall, changing proportions of air space and Ca²⁺ concentration in the extracellular solution.     

Fate of Escherichia coli O157:H7 on Fresh-Cut Apple Tissue and Its Potential for Transmission by Fruit Flies

Pathogenic Escherichia coli O157:H7, as well as nonpathogenic strains ATCC 11775 and ATCC 23716, grew exponentially in wounds on Golden Delicious apple fruit. The exponential growth occurred over a longer time period on fruit inoculated with a lower concentration of the bacterium than on fruit inoculated with a higher concentration. The bacterium reached the maximum population supported in the wounds regardless of the initial inoculum concentrations. Populations of E. coli O157:H7 in various concentrations of sterilized apple juice and unsterilized cider declined over time and declined more quickly in diluted juice and cider. The decline was greater in the unsterilized cider than in juice, which may have resulted from the interaction of E. coli O157:H7 with natural populations of yeasts that increased with time. Experiments on the transmission of E. coli by fruit flies, collected from a compost pile of decaying apples and peaches, were conducted with strain F-11775, a fluorescent transformant of nonpathogenic E. coli ATCC 11775. Fruit flies were easily contaminated externally and internally with E. coli F-11775 after contact with the bacterium source. The flies transmitted this bacterium to uncontaminated apple wounds, resulting in a high incidence of contaminated wounds. Populations of the bacterium in apple wounds increased significantly during the first 48 h after transmission. Further studies under commercial conditions are necessary to confirm these findings.     

Germination and adhesion of fungal conidia on polycarbonate membranes and on apple fruit exposed to mycoactive acetate esters

The adhesion and germination of conidia of nine fungal species were assessed on polycarbonate membranes or on the skin of apple fruit in sealed glass bottles injected or not injected with acetate esters. Adhesion was determined after dislodging conidia from surfaces using a sonication probe. Adhesion and germination of conidia of Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Penicillium citrinum, Penicillium claviforme, or Trichoderma sp. on membranes after 48 h were not increased in a 1.84 microg mL(-1) headspace of butyl acetate (BA), ethyl acetate, hexyl acetate, 2-methylbutyl acetate, pentyl acetate, or propyl acetate. Adhesion and germination of Botrytis cinerea, Penicillium expansum, and Penicillium roquefortii conidia were stimulated by all esters. Only conidia of B. cinerea and P. expansum exhibited increased adhesion and germination on the skin of apple fruit in bottles exposed to 0.92 microg mL(-1) of BA. Only conidia of B. cinerea and P. expansum produced decay in inoculated puncture wounds on fruit. Freshly made puncture wounds or 24-h-old puncture wounds in fruit were more adhesive than the unpunctured skin of fruit to conidia of B. cinerea or P. expansum. Fresh wounds were more adhesive to both fungi than 24-h-old puncture wounds. The skin and wounds of fruit were as adhesive to B. cinerea conidia as they were to P. expansum conidia. A 4-h exposure to 1.43 microg mL(-1) of BA increased adhesion of B. cinerea and P. expansum conidia in 24-h-old wounds. Results suggest that acetate-ester stimulation most likely is not a rare phenomenon in the fungi. For nutrient-dependent decay pathogens of apple fruit, acetate esters may be an alternative chemical cue used to maintain adhesion of conidia to wound surfaces.     

粉红聚端孢侵染新红星苹果的侵入途径与侵染时期

粉红聚端孢Trichothecium roseum引致的苹果霉心病发生严重。对于新红星品种,T. roseum的侵染过程尚未明确。本研究利用T. roseum荧光标记菌株TR45分析其在新红星品种上的侵染时期和侵入通道。发现T. roseum可以侵染花器组织,引起枯萎坏死;落花后10-15 d,受侵染的花器残体被萼片包裹于萼筒内,随后T. roseum从离生花柱合并处的孔口部位入侵进入合生花柱,沿花柱内的多细胞毛状体间扩展;花后30 d左右,花柱缝逐渐开裂,随果实膨大心室腔壁也开裂,逐渐形成一条贯通萼筒与心室开放通道——萼窦;花后60 d左右,T. roseum的菌丝沿着萼窦扩展进入心室腔,从心室上的裂缝进入果肉组织,最终引起果实霉心。本研究结果有助于解析新红星苹果品种发病重以及难防治的原因,对苹果霉心病防控新策略构建具有重要理论意义。     

Functional response of Euseius finlandicus and Amblyseius andersoni to Panonychus ulmi on apple and peach leaves in the laboratory

The functional response of adult females of the predatory mites Euseius (Amblyseius) finlandicus and Amblyseius andersoni to larvae and adult females of the fruit tree red spider mite Panonychus ulmi was determined on apple and peach leaf disks in the laboratory at 25°C and 16:8 (L:D). For adult females of P. ulmi the predation efficiency of E. finlandicus was higher on peach than on apple, whereas that of A. andersoni was higher on apple than on peach. Efficiency of predation on larvae of P. ulmi by either predator did not differ significantly between apple and peach. On both plants, A. andersoni had a higher predation rate than E. finlandicus on larvae of P. ulmi. It is concluded that in the laboratory the host plant has a substantial effect on predation efficiency of A. andersoni and E. finlandicus when they preyed on adults but not when they preyed on larvae of P. ulmi.     

Effects of orchard host plants on the oviposition preference of the oriental fruit moth (Lepidoptera: Tortricidae)

Recently, the oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), has emerged as a major problem on apples (Malus spp.) grown in the mid-Atlantic and midwestern United States, despite its historically important and frequent occurrence as a peach (Prunus spp.) pest. It is possible that host-driven biological phenomena may be contributing to changes in G. molesta population dynamics resulting in outbreaks in apple. Studies were designed to examine the effects of host plants on oviposition behavior, in an effort to clarify the host association status of eastern U.S. populations and also to gain insight into how pest modeling and management efforts may be altered to take into account various host-associated effects. G. molesta adults exhibited ovipositional preference for nonbearing peach trees over nonbearing apple trees in close-range choice tests conducted in the field, regardless of the larval host origin. A significant preference for peach shoots over apple shoots was observed on six of 12 sampling dates with a wild G. molesta population at the interface of adjacent peach and apple blocks. Numbers of eggs found on apple fruit were higher after peach fruit were harvested and apple fruit began to approach maturity (during the flight period for third and fourth brood adults). Possible implications for population modeling and integrated management of G. molesta are discussed.     

Effects of orchard host plants (apple and peach) on development of oriental fruit moth (Lepidoptera: Tortricidae)

Studies were designed to examine the effects of host plants (apple, Malus domestica Borkh., and peach, Prunus persica L.) on the development of oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae). Oriental fruit moth larvae developed faster on peach than on apple, both on fruit as well as on growing terminal shoots. On fruit, these differences were shown to cause significant changes in both the rate (approximately 20-60 degree-days earlier emergence on peach than on apple) and patterns of adult emergence among several cultivars of peaches and apples. Slopes of female emergence plots varied by host in 2003, with emergence occurring over a longer period on peach cultivars than on apple cultivars (with one exception). Slopes of male emergence curves did not differ by cultivar in 2003. These host-driven effects could impact the efficacy of traditional pest management approaches and probably complicate efforts to predictively model G. molesta populations in mixed cultivar orchards. Such developmental effects may help to explain previously observed differences in patterns of pheromone trap captures in peach versus apple orchards. Host-associated effects should be incorporated into future models to develop more realistic predictive tools and thus improve integrated pest management efforts.     

2.5%高效氟氯氰菊酯微乳剂对苹果桃小食心虫的田间应用技术研究

田间应用及安全性评价结果表明,2.5%高效氟氯氰菊酯微乳剂稀释2000～4000倍防治苹果树桃小食心虫Carposina niponensis Walsingham,药后5d各地防效分别为78.14%～99.57%、87.50%～96.58%、80.73%～96.20%,药后10d各地防效分别为84.49%～98.60%、77.78%～94.44%、80.66%～99.29%,药后15d各地防效分别为83.35%～100%、77.55%～98.90%、88.89%～96.58%。2.5%高效氟氯氰菊酯微乳剂稀释2000～3000倍处理与对照药剂2.5%高效氟氯氰菊酯乳油3000倍处理防效相当。最佳用药量以2000～3000倍为宜,施药适期为苹果果实膨大期,且桃小食心虫卵孵化盛末期,虫果率达1%时,用药后5～10d可有效控制虫害,持效期较长。药后苹果树未见异常。     

苏云金杆菌在华北果园土壤中消长动态的研究

本文报道了产生核黄素的苏云金杆菌CH菌株在果园土壤中的消长动态。研究结果表明 ,将CH菌株制剂喷洒于苹果园土壤表面后 1个月内 ,CH菌株的菌数基本上维持在喷洒后的 10 5/ g水平 ,其后菌数急剧减少 ,2个月后从 10 5/ g的水平减少到 10 4 / g的水平 ,并长时间维持 10 4 / g这一水平 ;CH菌株制剂喷洒于果园土壤后对土壤中固有的Bacillus属细菌数的消长没有明显的影响 ,Bacillus属细菌数长时间维持在 10 6～ 10 5/ g之间 ,处于一种相对稳定的状态 ;在土壤透水性和灌水条件良好的土壤条件下 ,喷洒于土壤表面的苏云金杆菌可缓慢向深处移动。     

Larval performance of the oriental fruit moth across fruits from primary and secondary hosts

A common characteristic of many invasive herbivorous insects is their ability to utilize a broad range of host plants. By using various hosts in phenological succession, multivoltine herbivores may increase the number of successful annual generations, at the same time as potentially increasing their overall fitness. To achieve such success, herbivores must be able to develop efficiently on the nutritional resources offered by their hosts. The oriental fruit moth Cydia (= Grapholita) molesta (Busck) (Lepidoptera: Tortricidae) is one of the most damaging invasive insect species. Peach (Prunus persica) is its primary host, whereas the pome fruits apple (Malus × domestica Borkh) and pear (Pyrus communis) are considered as secondary hosts. In many parts of its geographical range, including southern Europe, populations of the moth switch from peach to apple or pear orchards during the growing season. The present study tests whether this temporal switch is supported by the physiological capability of the larvae with respect to developing efficiently on fruits of these taxonomically‐related host plants. Larvae are reared on peach, apple or pear fruits; several life‐history traits are measured; and correlations between the traits are calculated. The results obtained show that larvae do not have the same physiological capability with respect to using apple or pear fruits as hosts compared with using peach fruit. Pear fruit in particular is a sub‐optimal diet. These findings suggest that, in the case of continuous geographical expansion, concomitantly with global warming, apple orchards might support oriental fruit moth populations better than pear orchards, and that the switch onto novel hosts might be accompanied by restricted population growth.     

NaHSO3对桃树的生理效应及产量、品质的影响

于桃树果实膨大期喷施100ppm NaHSO3可获得增产、优质、早熟的效果。此与NaHSO3能增加叶绿素含量、提高光合速率、比叶重、促进希尔反应,抑制硝酸还原酶、过氧化氢酶活性,增加单果重等多重生理效应相关。     

Comparative survival rates of oriental fruit moth (Lepidoptera: Tortricidae) larvae on shoots and fruit of apple and peach

Studies were designed to examine the effects of host plants on oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), survival. G. molesta larval survival in the orchard was comparable between apple (Malus spp.) and peach (Prunus spp.) shoots, indicating that both host species can harbor large larval populations during the early season. G. molesta larvae used multiple shoots for feeding and development on peaches but usually only damaged single shoots in apple. Survival differences were present between peach and apple fruit, but this survival was affected by fruit maturity level. Generally, larval survival higher was in ripening peach fruit than in green, immature apple fruit. Larval survival varied among several apple cultivars, indicating that cultivar-level variability needs to be considered. These host-associated effects may impact efforts to predictively model G. molesta populations in commercial orchards where multiple host plant species, or different cultivars of the same species are often grown in proximity. Thus, host-associated dynamics should be included into future population models that underlie management programs.     

Mycelial yield of<Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> using thinned apple,pear, and peach on submerged culture

The effect of thinned fruits, apple, pear and peach, on the mycelial growth of mushrooms was investigated. The growth of mycelia with the addition of thinned fruit was clearly better than that in the control for all the tested mushrooms. The growth rate ofPleurotus ostreatus was faster than any other mushroom. The optimal concentrations of thinned apple, pear, and peach in a solid culture were 1.0%, 1.0%, and 3%, respectively, while in a liquid culture the optimal concentrations were 5,0%, 3.0%, and 5.0%, respectively. WhenPleurotus ostreatus was incubated in a 20-L pilot scale fermenter with 10 L of a liquid medium containing 3% thinned fruit at 25°C and 6 vvm for 10 days, the mass-production of mycelia was 74.2 g/10 L (apple), 96.2 g/10 L (pear), and 86.3 g/10 L (peach). The mycelial yield ofPleurotus ostreatus in a medium containing thinned fruit was 2≈3 times higher than that in the control.     

Spatial and temporal evolution of quantitative magnetic resonance imaging parameters of peach and apple fruit – relationship with biophysical and metabolic traits

Fruits are complex organs that are spatially regulated during development. Limited phenotyping capacity at cell and tissue levels is one of the main obstacles to our understanding of the coordinated regulation of the processes involved in fruit growth and quality. In this study, the spatial evolution of biophysical and metabolic traits of peach and apple fruit was investigated during fruit development. In parallel, the multi-exponential relaxation times and apparent microporosity were assessed by quantitative magnetic resonance imaging (MRI). The aim was to identify the possible relationships between MRI parameters and variations in the structure and composition of fruit tissues during development so that transverse relaxation could be proposed as a biomarker for the assessment of the structural and functional evolution of fruit tissues during growth. The study provides species-specific data on developmental and spatial variations in density, cell number and size distribution, insoluble and soluble compound accumulation and osmotic and water potential in the fruit mesocarp. Magnetic resonance imaging was able to capture tissue evolution and the development of pericarp heterogeneity by accessing information on cell expansion, water status and distribution at cell level, and microporosity. Changes in vacuole-related transverse relaxation rates were mostly explained by cell/vacuole size. The impact of cell solute composition, microporosity and membrane permeability on relaxation times is also discussed. The results demonstrate the usefulness of MRI as a tool to phenotype fruits and to access important physiological data during development, including information on spatial variability.