臭氧对葡萄灰霉病的抑制效果

研究不同臭氧处理方式对离体及接种在活体葡萄上的灰霉菌的抑制作用,为降低采后葡萄贮藏过程中灰霉病带来的损失提供理论依据。在(20±1)℃条件下用不同剂量的臭氧(0、30、60、90μL/L)对离体灰霉菌分别处理不同时间(5、10、15 min),通过测定菌丝生长长度、孢子形成抑制率、孢子细胞膜完整性及扫描电镜观察来探究臭氧对其抑制效果。用上述4种剂量的臭氧分别对接种了灰霉菌的‘红地球’葡萄处理15 min后(20±1)℃条件下存放5 d。通过测定发病率、病斑直径及相关酶活性来观察臭氧对葡萄灰霉病的控制效果。结果表明:在离体实验中,与对照相比,不同剂量的臭氧对离体灰霉菌处理不同时间后,灰霉菌菌丝的生长均受到了不同程度抑制,随贮藏时间延长抑制效果逐渐减弱;灰霉菌的产孢子率显著下降,部分孢子的细胞膜完整性被破坏,臭氧剂量越高、处理时间越长,效果越好。在接种了灰霉菌的活体实验中,与对照组相比,用60μL/L和90μL/L剂量臭氧处理接种灰霉菌的葡萄果实后,灰霉菌在果实上的生长明显受到抑制;臭氧处理可以使果实丙二醛含量积累减慢,维持较高的抗氧化酶活性。综合分析,臭氧通过抑制灰霉菌菌丝在葡萄果实上的生长和诱导葡萄果实提高抗性来抵抗灰霉菌的侵染,采用90μL/L剂量臭氧处理15 min对离体条件下和接种在‘红地球’葡萄活体上的灰霉菌抑制效果均为最好。     

香茅精油体外和体内对樱桃番茄致病菌灰葡萄孢的抑制性研究

系统研究了香茅精油对灰葡萄孢的抑制作用。以灰葡萄孢为供试菌,研究不同含量的精油在体外PDA培养基上对灰葡萄孢菌丝生长,以及在PDB培养基上对灰葡萄孢孢子萌发、生物量的影响,并在樱桃番茄体内验证其抑菌性,得到最佳抑制含量。用扫描电镜观察精油对灰葡萄孢菌丝微观结构的作用。结果表明:在体外PDA培养基上香茅精油抑菌效果随精油含量的增加而增强,在精油含量1.5μL/m L条件下可完全抑制菌丝的生长。在PDB中精油完全抑制其孢子萌发及菌丝产生的最低含量分别为0.8μL/m L和1.2μL/m L。香茅精油在樱桃番茄体内的最佳抑制发病含量为0.9μL/m L,相比对照,抑制了54.33%的发病。扫描电镜观察精油处理后的菌丝形态变化明显,表现为菌丝短小变形,表面粗糙、褶皱、凹陷。     

臭氧处理对猕猴桃果实采后病害及品质的影响

为了研究臭氧对猕猴桃病害的作用效果,从发病猕猴桃中分离鉴定出病原真菌,研究不同浓度臭氧对此真菌的抑制作用。采用0,90,170,250 mg/m~3臭氧,每天处理1 h,连续处理7 d,观察其对病菌孢子萌发和菌丝生长的影响,同时,分析经臭氧处理的果实在贮藏期间品质的变化趋势。结果表明:猕猴桃主要采后病菌是灰葡萄孢菌和扩展青霉菌,臭氧对这两种病菌的菌丝生长和孢子萌发有明显抑制作用。猕猴桃抗病性试验结果表明:170 mg/m~3臭氧能显著降低由灰葡萄孢菌和扩展青霉菌引起的病害的发病率,抑制率分别为80.5%和60.7%,病斑直径的抑制率为47.6%和56.9%,可维持果实较高的硬度、可溶性固形物、可滴定酸和VC含量。     

SO_2保鲜剂对玫瑰香葡萄灰霉菌的抑制作用

以玫瑰香葡萄果实为材料,分析了SO_2对果实表面微生物活性的影响,并对SO_2的抑菌作用进行了体外实验,以寻求葡萄果实采后保鲜的低浓度SO_2处理方式。研究发现:SO_2可降低葡萄果实表面微生物总量,抑制灰霉菌;SO_2类保鲜剂焦亚硫酸钠能抑制灰霉菌的孢子萌发和菌丝生长,抑菌作用随焦亚硫酸钠浓度升高和作用时间延长而增强,低温能增强抑菌作用;浓度为300和600 mg/L的焦亚硫酸钠,在p H3.00时抑菌强度均高于p H4.00;用300 mg/L焦亚硫酸钠孵育灰霉菌孢子1 h后,4℃培养45 d无可见菌丝。结果表明,一定剂量的SO_2类保鲜剂有效抑制了葡萄果实主要致病菌灰霉菌的发生,采用较低p H、低浓度焦亚硫酸钠处理葡萄果实可对采后贮藏保鲜发挥作用。     

Nα-月桂酰-L-精氨酸乙酯盐酸盐对5种果蔬腐败菌的抑菌活性

通过体外抑菌试验评价N~α-月桂酰-L-精氨酸乙酯盐酸盐(Nα-lauroyl-L-arginate ethylester,LAE)的抑菌活性。以果蔬腐败菌中具有代表性的指状青霉、意大利青霉、灰葡萄孢、链格孢、胡萝卜软腐病坚固杆菌为模式菌,测定LAE对5种腐败菌的最低抑制浓度(Minimum Inhibitory Concentration,MIC),并测定LAE对霉菌的菌丝生长抑制率以及对细菌的杀菌效果。同时探究LAE和其他几种果蔬防腐剂的联合抑菌效果。结果表明:LAE对4种真菌性腐败菌灰葡萄孢、链格孢、意大利青霉、指状青霉的MIC分别为400,200,400,400μg/mL,当LAE浓度达到对各菌的MIC后,能对各霉菌保持较高的菌丝抑制率,分别为100%(16h),78.2%(48h),81.8%(48h),79.1%(48h);LAE对细菌性腐败菌胡萝卜软腐病坚固杆菌的MIC为25μg/mL,且50μg/mL的LAE能在90min内完全杀灭受试菌;LAE与尼泊金甲酯钠共同使用时对模式菌有最佳的联合抑制效果。     

葡萄灰葡萄孢生长及产孢条件研究

灰葡萄孢(Botrytis cinerea)可引起葡萄灰霉病,严重影响葡萄产量和品质。探索不同培养基、温度、光照、碳源、氮源等对葡萄灰葡萄孢菌丝生长和分生孢子产生的影响,以及菌丝与分生孢子的致死温度和时间,旨在探索灰葡萄孢生长及产孢的最佳条件。结果显示,灰葡萄孢菌丝生长的最适培养基为PDA培养基,最适温度25℃;产孢最适培养基为MSM培养基,最适温度23℃;在培养基pH3~11范围内灰葡萄孢均能生长并产生分生孢子,菌丝生长和产生分生孢子的最适pH均为6;灰葡萄孢以葡萄糖和乳糖为碳源时生长速率和产孢量达到最佳,而蛋白胨则为灰葡萄孢生长和产孢的最佳氮源;菌丝的致死条件是50℃处理25 min或55℃处理10 min;分生孢子的致死条件是45℃处理30 min或50℃处理10 min。研究为葡萄灰霉病的有效防治及分子生物学研究提供理论依据。     

洋葱霍尔德氏菌对玫瑰香葡萄采后灰霉病的抑制活性

为研究生防菌洋葱霍尔德氏菌B-1的生防潜力,并为其生产性应用奠定基础,研究洋葱霍尔德氏菌B-1的生物量最大值的培养时间、浓度和发酵时间对玫瑰香葡萄采后病害的抑制效果。结果表明,在接种后24 h,拮抗菌B-1的抑菌效果最好,抑菌圈直径达2.81 cm。在液体培养基中,生防菌可有效抑制灰葡萄孢霉孢子萌发和芽管伸长,稀释5倍发酵液和发酵原液可完全抑制灰葡萄孢霉孢子萌发,稀释40倍发酵液孢子萌发率下降60.39%。在0,16℃和26℃温度条件下,拮抗菌均可显著降低葡萄采后自然腐烂率。菌株B-1不论是在离体条件还是在活体条件下,均有抑制灰葡萄孢霉的作用,能有效防治葡萄采后灰霉病的发生,是一株生防潜力较好的菌株。     

食药同源植物对果实采后常见腐败菌抑制作用

以扩展青霉、灰葡萄孢、链格孢抑菌、大肠杆菌和金黄色葡萄球菌为靶标菌,探究11种食药同源植物提取液的抑菌作用。丁香对3种霉菌的抑制效果显著,抑菌率均达到100%,最小抑菌浓度均为3.13 mg/mL;最小杀菌浓度分别为:扩展青霉3.13 mg/mL,灰葡萄孢6.25 mg/mL,链格孢3.13 mg/mL。3.13 mg/mL浓度丁香提取液对3种霉菌孢子萌发抑制率均为100%。薄荷对扩展青霉和链格孢的抑菌效果仅次于丁香,最小抑菌浓度均为100 mg/mL。丁香和天麻对大肠杆菌有一定抑制作用,抑菌圈分别为17.77 mm和11.67 mm;天麻和鱼腥草对金黄色葡萄球菌抑制效果显著,抑菌圈可达31.77和29.00 mm,最小抑菌浓度分别为30.0 mg/mL和60.0 mg/mL,最小杀菌浓度分别为60.0 mg/m L和120 mg/mL。以蓝莓腐败率为指标验证, 3.13 mg/mL浓度的丁香和30.0 mg/mL浓度天麻混合液处理后蓝莓腐败率显著降低。     

热击对黑曲霉孢子萌发及产木聚糖酶的影响

对黑曲霉孢子进行热击处理,探讨了热击处理促进孢子萌发和木聚糖酶活力的机制。结果显示,42℃下热击4h后转入29℃发酵,可以促使黑曲霉孢子较早萌发和菌丝的较快生长。热击处理后发酵过程中的菌丝干重始终高于恒温发酵处理,其中发酵24h的菌丝干重比恒温发酵高38.5%;热击下变温发酵在72h时达到最大生物量,比恒温发酵提前12h。经过热击处理后的发酵产酶总量和单位质量菌丝产酶量都高于恒温发酵,尤其在发酵86h分别比恒温对照高出46.2%和23.5%。热击处理对于胞内木聚糖酶比例影响不大,酶谱检测发现对木聚糖酶的种类也没有影响。     

茶树精油对灰葡萄孢霉活性氧代谢的影响

为了揭示茶树精油(TTO)对果蔬采后病原真菌灰葡萄孢霉的抑菌作用,以TTO处理的灰葡萄孢霉孢子或菌丝为研究对象,测定真菌活性氧(ROS)积累及相关抗氧化酶类的活性变化,并采用扫描电镜、透射电镜观察精油处理对菌丝形态和超微结构的影响。结果表明:TTO诱导孢子内ROS和菌丝内过氧化氢(H2O2)的大量积累,导致抗坏血酸(ascorbate, AsA)含量下降和超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)和谷胱甘肽还原酶(GR)活性的提高。同时,TTO处理导致灰葡萄孢霉菌丝发生严重的皱缩、干瘪,胞内有空泡状结构,细胞质大量减少,少量存在的细胞质聚缩成块状。结果表明TTO处理导致灰葡萄孢霉ROS的爆发,并严重破坏菌体结构。     

Control of postharvest pear diseases using Rhodotorula glutinis and its effects on postharvest quality parameters

Rhodotorula glutinis was evaluated for its activity in reducing postharvest gray mold decay and blue mold decay of pear caused by Botrytis cinerea and Penicillium expansum respectively, and in reducing natural decay development of pear fruits, as well as its effects on postharvest quality of fruits. There was a significant negative correlation between concentrations of the yeast cells and infectivity of the pathogens. At concentrations of R. glutinis at 5x108 CFU/ml, the gray mold decay was completely inhibited after 7 days incubation at 20 degrees C, while the control fruit had 100% disease incidence and 2.15 cm lesion diameter respectively, at challenged with B. cinerea spores suspension of 1x105 spores/ml; No completely control was got to blue mold, when pear fruits stored at 20 degrees C for 7 d (challenged with P. expansum spores suspension of 5x104 spores/ml), but the decay was distinctly prevented with 20% and 0.60 cm of disease incidence and lesion diameter respectively, while the control fruits were 100% and 2.74 cm, respectively. Rapid colonization of the yeast in wounds was observed during the first 1 d at 20 degrees C, and then the populations stabilized for the remaining storage period. On pear wounds kept at 4 degrees C, rapid colonization of the yeast in wounds was observed during the first 3 d, and then the increase in population density of R. glutinis turned slow, which continued over 6 d after application of the antagonist until it reached a high level. Then, the populations stabilized for the remaining storage period. In the test on PDA plates, R. glutinis significantly inhibit the growth of B. cinerea and P. expansum. Spore germination of pathogens in PDB was greatly controlled in the present of living yeast cell suspensions. R. glutinis significantly reduced the natural development of decay of pear following storage at 20 degrees C for 7 days or at 4 degrees C for 30 days followed by 20 degrees C for 5 days, and did not impair quality parameters, including mass loss, firmness, TSS, ascorbic acid or titratable acidity. Thus, the application of R. glutinis can be an alternative to chemicals for control of postharvest diseases on pear fruits.     

Biological control of postharvest spoilage caused by Penicillium expansum and Botrytis cinerea in apple by using the bacterium Rahnella aquatilis

The epiphytic bacterium Rahnella aquatilis, isolated from fruit and leaves of apples, was tested for antagonistic properties against Penicillium expansum and Botrytis cinerea on Red Delicious apple fruit. In "in vitro" assays, this bacterium inhibited completely the germination of P. expansum and B. cinerea spores, but it needed direct contact with the spores to do it. However the putative mechanism seemed be different for the two pathogens. The bacterium did not produce extracellular antibiotic substances and when the acute toxicity test was performed no mortality, toxicity symptoms or organ alterations of the test animals (Wistar rats) were observed. Assays of biological control of P. expansum and B. cinerea on apple fruit were carried out at different temperatures. At 15 degrees C and 90% RH, the incidence of disease caused by P. expansum on apples stored for 20 days, was reduced by nearly 100% by R. aquatilis (10(6) cells/ml), while in the case of B. cinerea, the reduction of decay severity was nearly 64% but there was no reduction in the incidence of disease. At 4 degrees C and 90% RH the treatment with the bacterium significantly inhibited the development of B. cinerea on apples stored for 40 days and the incidence of disease was reduced by nearly 100%, while the incidence of disease caused by P. expansum at 4 degrees C was 60%. The results obtained show that R. aquatilis would be an interesting microorganism to be used as a biocontrol agent.     

Effects of Ozone Treatment on Botrytis cinerea and Sclerotinia sclerotiorum in Relation to Horticultural Product Quality

ABSTRACT:  Botrytis cinerea  and  Sclerotinia sclerotiorum  are fungal pathogens that cause the decay of many fruits and vegetables. Ozone may be used as an antimicrobial agent to control the decay. The effect of gaseous ozone on spore viability of  B. cinerea  and mycelial growth of  B. cinerea  and  S. sclerotiorum  were investigated. Spore viability of  B. cinerea  was reduced by over 99.5% ( P  < 0.01) and height of the aerial mycelium was reduced from 4.7 mm in the control to less than 1 mm after exposure to 450 or 600 ppb ozone for 48 h at 20 °C. Sporulation of  B. cinerea  was also substantially inhibited by ozone treatments. However, ozone had no significant effect on mycelial growth of  S. sclerotiorum in vitro . Decay and quality parameters including color, chlorophyll fluorescence (CF), and ozone injury were further assessed for various horticultural commodities (apple, grape, highbush blueberry, and carrot) treated with 450 ppb of ozone for 48 h at 20 °C over a period of 12 d. Lesion size and height of the aerial mycelium were significantly reduced by the ozone treatment on carrots inoculated with mycelial agar plugs of  B. cinerea  or  S. sclerotiorum . Lesion size was also reduced on treated apples inoculated with 5 × 10⁶ spores/mL of  B. cinerea , and decay incidence of treated grapes was reduced. The 450 ppb ozone for 48 h treatment had no significant effect on color of carrots and apples or on CF of apples and grapes. Ozone, an environmentally sound antimicrobial agent, inactivates microorganisms through oxidization and residual ozone spontaneously decomposes to nontoxic products. It may be applied to fruits and vegetables to reduce decay and extend shelf life.     

Osmotic dehydration of apple slices with CaCl2 and sucrose limits decay caused by Penicillium expansum, Colletotrichum acutatum, and Botrytis cinerea and does not promote Listeria monocytogenes or total aerobic population growth.

The interaction of Penicillium expansum Link, Colletotrichum acutatum, and Botrytis cinerea Pers.:Fr. with Listeria monocytogenes on osmotically dehydrated apple slices was evaluated. In mineral analyses of the slices, the calcium content of the peel and flesh tissues increased by 4- and 11-fold, respectively, when processed in 2% CaCl2. These slices also exhibited less decay by P. expansum, C. acutatum, and B. cinerea. Inoculation of slices with P. expansum resulted in a decrease in the pH of the flesh tissue at the infection site, while the pHs of slices infected with C. acutatum and B. cinerea increased and remained stable, respectively. Total mold population increased in wounds inoculated with P. expansum or C. acutatum. The presence of L. monocytogenes in the wounds did not significantly affect mold growth. The association of P. expansum and L. monocytogenes on apple slices resulted in a decrease in the bacterial population, whereas L. monocytogenes survived when slices were inoculated with C. acutatum. When associated with B. cinerea, there was a fourfold decrease in the L. monocytogenes population when slices were treated with 2% CaCl2. The total aerobic population was not significantly affected by the type of microorganism added to the wounds or by the osmotic treatment. These data show that osmotic dehydration with 2% CaCl2 combined with 20% sucrose limits decay of apple slices and does not promote bacterial or total aerobic population growth.     

Effect of Hexanal Vapor on Spore Viability of Penicillium expansum, Lesion Development on Whole Apples and Fruit Volatile Biosynthesis

ABSTRACT: The effects of hexanal vapor on spore viability of Penicillium expansum , lesion development on whole apple fruit, and flavor volatile biosynthesis were investigated. Spore viability was reduced by 94% after exposure to a hexanal concentration of 40 μmol/L for 24 h, compared with 50% at 18 μmol/L and 20% at 9 μmol/L. Decay on whole apple fruit inoculated with 5 × 10⁴ spores/mL of P. expansum was reduced with exposure to hexanal vapor for 48 h. Although almost all of the fruit treated with 8 to 12 jmiol/L developed decay lesions, lesion size was reduced compared with the controls. At concentrations of 15 to 19 μmol/L, and 25 to 29 μmol/L, the incidence of fruit with lesions was 44% and 24%, respectively, compared with 100% and 98% in the inoculated control apples and lesion size was further reduced. Apples treated at 4°C with only 5 to 7 jimol/L hexanal vapor also showed a marked reduction in lesion incidence. Hexanal was rapidly converted to high levels of the aroma volatiles hexanol, hexylacetate, hexylbutanoate, and hexylhexanoate, but these decreased to levels similar to the control after 4 to 7 d of being held in air. There was no detectable hexanal after holding fruit in air for 5 h.     

In vitro and in vivo [corrected] activity of eugenol oil (Eugenia caryophylata) against four important postharvest apple pathogens

The activity of eugenol oil was evaluated in vitro and in vivo against four apple pathogens namely Phlyctema vagabunda, Penicillium expansum, Botrytis cinerea and Monilinia fructigena. The minimum inhibitory concentration (MIC) of eugenol incorporated in malt extract agar medium was found to be 2 mg ml(-1). Mycelial growth of the four test pathogens was completely inhibited when treated with 150 microl l(-1) of volatile eugenol whether at 4 or 20 degrees C. Conidia of P. vagabunda, P. expansum, M. fructigena and B. cinerea suspended for 2 min in eugenol solution at 2 mg ml(-1) heated to 50 degrees C germinated at rates of 19, 37, 38 and 39%, respectively. Three different eugenol formulations (Tween 80, ethoxylate and lecithin) were tested for their in vivo efficacy against the tested pathogens on apples. Ethoxylate- and Tween 80-eugenol formulations applied at room temperature were ineffective in reducing disease incidence. When heated to 50 degrees C, both formulations induced phytotoxicity on apple surface and caused cuticle damages as revealed by scanning electronic microscopic observations. A mixture of eugenol at 2 mg ml(-1) and soy lecithin at 50 mg ml(-1) suppressed the phytotoxic symptoms produced by eugenol on apples and reduced the disease incidence of P. expansum, P. vagabunda, B. cinerea and M. fructigena to less than 7, 6, 4 and 2% respectively after 6 months of storage at 2 degrees C. The application of heated lecithin-formulated eugenol could become a successful alternative to the traditional fungicides used in postharvest disease management of apple fruit.     

Biological control of postharvest pear diseases using a bacterium, Pantoea agglomerans CPA-2.

Epiphytic microorganisms isolated from the fruits and leaf surfaces of apples and pears were screened for antagonistic activity against Penicillium expansum on pears. From 247 microorganisms tested for antagonistic properties against P. expansum, a bacterium strain identified as Pantoea agglomerans (CPA-2) was selected. This bacterium was very effective against Botrytis cinerea, P. expansum and Rhizopus stolonifer. Complete control at the three tested concentrations (2 x 10(7), 8 x 10(7) and 1 x 10(8) CFU ml(-1)) was obtained on wounded pears inoculated with 10(3), 10(4) and 10(5) conidia ml(-1) of P. expansum and R. stolonifer. At 8 x 10(7) CFU ml(-1), Pan. agglomerans reduced B. cinerea decay by more than 80% at the three concentrations of the pathogen. In over 3 years of experiments in semicommercial trials, Pan. agglomerans provided excellent control against B. cinerea and P. expansum under cold storage, either in air or in low oxygen atmospheres. Equal control was obtained with Pan. agglomerans at 8 x 10(7) CFU ml(-1), as with the fungicide imazalil at commercial doses, against both pathogens. Pan. agglomerans grew well inside wounds on pears at both room and cold temperatures and under modified atmospheres. In contrast, it grew poorly on the surface of intact fruit.     

采后柠檬酸处理对苹果青霉病的控制及其贮藏品质的影响

以苹果为原料,研究不同质量分数柠檬酸常温浸泡处理对损伤接种扩展青霉（Penicillium expansum）果实病斑扩展的抑制效果及贮藏品质的影响。结果表明：离体条件下柠檬酸能够抑制P. expansum的孢子萌发,其中以1%质量分数效果最佳,1%柠檬酸处理显著降低了损伤接种苹果果实P. expansum病斑直径;柠檬酸处理显著提高了果实过氧化物酶、过氧化氢酶和抗坏血酸过氧化物酶活性。此外,柠檬酸处理还有效延缓了果实质量损失率的升高,抑制果实硬度、抗坏血酸含量、可滴定酸和可溶性固形物质量分数的下降,且推迟了果实呼吸高峰的出现。     

β-氨基丁酸抑制草莓低温贮藏过程中灰霉病的效果及其机理

以‘红颜’草莓为实验材料,研究β-氨基丁酸(β-aminobutyric acid,BABA)对草莓果实采后贮藏在低温((5±1)℃)条件下灰霉病的抑制效果及相关机理。草莓果实经过20 mmol/L的BABA处理后接种灰葡萄孢霉(Botrytis cinerea)孢子,然后在(5±1)℃的条件下贮藏12 d,结果发现BABA显著抑制了灰霉病的发生和病斑直径的扩展,与对照组相比,处理组几丁质酶(chitinase,CHI)和β-1,3-葡聚糖酶(β-1,3-glucanase,GLU)的活力分别提高了9.2%和54.9%,多聚半乳糖醛酸酶(polygalacturonase,PG)和纤维素酶(cellulose,Cel)的活力分别降低了29.3%和24.4%,同时,BABA提高了FaCHI和FaGLU基因的表达,抑制了FaPG和FaCel基因的表达;此外,体外实验的结果发现,BABA能够破坏灰葡萄孢霉孢子的质膜完整性,导致了孢子内部可溶性蛋白质和可溶性糖的泄漏。结果表明,BABA通过诱导抗病相关酶活力来提高抗病性、延缓果实软化并通过对病原菌的直接抑制作用减轻草莓果实采后灰霉病的发生。     

Production of mycotoxins by Penicillium expansum inoculated into apples

We investigated the production of mycotoxins in apple fruits inoculated with spores of 40 strains of apple blue mold, Penicillium expansum. Patulin and citrinin contents in the extracts from apples stored at 25 degrees C for 12 days after inoculation were determined by high-performance liquid chromatography (HPLC) analysis with UV and fluorescence detection. Patulin and citrinin were produced by 90% (36) and 80% (32) of the 40 strains, indicating that P. expansum is a consistent producer of these mycotoxins. The patulin content in the extracts was substantially higher than the citrinin content. Other mycotoxins whose production in pure culture has been reported were simultaneously detected with high-resolution liquid chromatography-mass spectrometry (LC-MS) analysis with the positive ion mode of electrospray ionization. Along with patulin and citrinin, expansolides A and B were identified based on the HPLC and LC-MS spectral data and detected in 88% (35) of the extracts. The results indicate that P. expansum is a consistent producer of expansolides A and B in rotten areas of apple fruits. The findings raise the possibility that products from decayed apples might contain expansolides A and B in addition to patulin and citrinin.