Characterization of fludioxonil-resistant and pyrimethanil-resistant phenotypes of Penicillium expansum from apple

Penicillium expansum is the primary cause of blue mold, a major postharvest disease of apple. Fludioxonil and pyrimethanil are two newly registered postharvest fungicides for pome fruit in the United States. To evaluate the potential risk of resistance development in P. expansum to the new postharvest fungicides, one isolate of each of thiabendazole-resistant (TBZ-R) and -sensitive (TBZ-S) P. expansum was exposed to UV radiation to generate fungicide-resistant mutants. Four fludioxonil highly-resistant mutants (EC(50) > 1,000 microg/ml) and four pyrimethanil-resistant mutants (EC(50) > 10 microg/ml) were tested for sensitivities to thiabendazole, fludioxonil, and pyrimethanil, and fitness parameters including mycelial growth, sporulation on potato dextrose agar (PDA), sensitivity to osmotic stress, and pathogenicity and sporulation on apple fruit. The stability of resistance of the mutants was tested on PDA and apple fruit. Efficacy of the three fungicides to control blue mold incited by the mutants was evaluated on apple fruit. Six fungicide-resistant phenotypes were identified among the parental wild-type isolates and their mutants based upon their resistance levels. All four fludioxonil highly-resistant mutants were sensitive to pyrimethanil and retained the same phenotypes of resistance to TBZ as the parental isolates. All four pyrimethanil-resistant mutants had a low level of resistance to fludioxonil with a resistance factor >15. The two pyrimethanil-resistant mutants derived from a TBZ-S isolate became resistant to TBZ at 5 microg/ml. After 20 successive generations on PDA and four generations on apple fruit, the mutants retained the same phenotypes as the original generations. All mutants were pathogenic on apple fruit at both 0 and 20 degrees C, but fludioxonil highly-resistant mutants were less virulent and produced fewer conidia on apple fruit than pyrimethanil-resistant mutants and their parental wild-type isolates. Compared with the parental isolates, all four fludioxonil highly-resistant mutants had an increased sensitivity to osmotic stress on PDA amended with NaCl, while the pyrimethanil-resistant mutants did not. Pyrimethanil was effective against blue mold caused by fludioxonil-resistant mutants at both 0 and 20 degrees C. Pyrimethanil and fludioxonil reduced blue mold incited by pyrimethanil-resistant mutants during 12-week storage at 0 degrees C but were not effective at 20 degrees C. TBZ was not effective against pyrimethanil-resistant mutants derived from TBZ-S wild-type isolates at room temperature but provided some control at 0 degrees C. The results indicate that: (i) a fitness cost was associated with fludioxonil highly resistant mutants of P. expansum in both saprophytic and pathogenic phases of the pathogen but not pyrimethanil-resistant mutants; (ii) pyrimethanil possessed a higher risk than fludioxonil in the development of resistance in P. expansum; and (iii) triple resistance to the three apple-postharvest fungicides could emerge and become a practical problem if resistance to pyrimethanil develops in P. expansum populations.     

Multiple preharvest treatments with harpin reduce postharvest disease and maintain quality in muskmelon fruit (cv. Huanghemi)

Decay caused by fungal pathogens is responsible for significant economic loss of muskmelon fruit (Cucumis melo L.) in China. However, postharvest treatment is not effective in controlling all of these pathogens, owing to the ability of some of them for latent infection. Therefore, preharvest treatment has been considered as a promising method to control postharvest decay in muskmelon fruit. The effect of multiple preharvest treatments with harpin on postharvest disease and quality of cv. ‘Huanghemi’ muskmelon fruit was investigated in this study. Results showed that harpin treatment significantly decreased the incidence of latent infection caused by Alternaria alternata and Fusarium spp. in harvested fruit. Meanwhile, the postharvest pink rot in treated fruit inoculated with Trichothecium roseum was suppressed noticeably. Moreover, preharvest treatment increased the activities of peroxidase, phenylalanine ammonia lyase, 4-Coumarate:CoA ligase and β-1,3-glucanase compared with control fruit. Harpin treatment also contributed to the accumulation of total phenolic compounds and lignin. In addition, harpin treatments were able to maintain the quality of harvested fruit. These results suggest that multiple treatments with harpin before harvest may be a promising technology to control postharvest disease and delay quality losses in Huanghemi muskmelon fruits.     

Epidemiological effect of gene deployment strategies on bacterial blight of rice

ABSTRACT Experiments were conducted in farmers' fields at two locations of the irrigated lowlands of Laguna province in southern Luzon island, Philippines, during the wet seasons of 1993 and 1994. Nine rice populations were studied including pure stands, two-component mixtures, two-gene combinations of backcrossed lines containing varying combinations of the bacterial blight resistance genes Xa-4, xa-5, and Xa-10, and a non-isogenic cultivar containing Xa-4 and partial resistance to bacterial blight. The area under the disease progress curve (AUDPC) of both gene combinations studied was significantly less than the single most effective gene of each combination deployed singly. A mixture of a susceptible and a resistant line expressed an AUDPC significantly less than the mean of its component pure stands, but two other mixtures did not. The cultivar IR20, which contains both Xa-4 and partial resistance, reduced the AUDPC by about two-thirds as compared with IR-BB4, which contains Xa-4 and little or no partial resistance.     

Control of postharvest pathogens and colonization of the apple surface by antagonistic microorganisms in the field

ABSTRACT Selected isolates of Aureobasidium pullulans, Rhodotorula glutinis, and Bacillus subtilis reduced the size and number of lesions on wounded apples caused by the postharvest pathogens Penicillium expansum, Botrytis cinerea, and Pezicula malicorticis. Combinations of the antagonistic microorganisms were applied to apple trees in the field late in the growing season of two consecutive years. The population dynamics of the introduced microorganisms and the incidence of fruit decay were determined. Population sizes of introduced antagonists on apple surfaces increased in the field following application of treatments until harvest. After transfer of the fruit from the field into cold storage, the populations of the introduced antagonists remained higher than in the control treatments. Identification of the applied isolates of A. pullulans and R. glutinis during the experiments was achieved by isolate-specific DNA probes generated from random amplified polymorphic DNA. A combination of two strains of A. pullulans and one strain of R. glutinis suppressed rotting of apple to the same extent as the commonly used fungicide Euparen. Our data demonstrate that the application of antagonistic microorganisms in the field represents a promising alternative to fungicide treatments to control post-harvest diseases of apple.     

Rhizobacteria-Mediated Growth Promotion of Tomato Leads to Protection Against Cucumber mosaic virus

ABSTRACT We evaluated combinations of two strains of plant growth-promoting rhizobacteria (PGPR) formulated with the carrier chitosan for the ability to induce growth promotion of tomato plants and resistance to infection by Cucumber mosaic virus (CMV). Each PGPR combination included GB03 (Bacillus subtilis) and one of the following PGPR strains: SE34 (B. pumilus), IN937a (B. amyloliquefaciens), IN937b (B. subtilis), INR7 (B. pumilus), or T4 (B. pumilus). The PGPR combinations formulated with chitosan are referred to as biopreparations. Tomato plants treated with each of the biopreparations appeared phenotypically and developmentally similar to nonbacterized control plants that were 10 days older (referred to as the older control). When plants were challenged with CMV, all plants in the biopreparation treatments and the older control treatment had significantly greater height, fresh weight, and flower and fruit numbers than that of plants in the CMV-inoculated same age control treatment. CMV disease severity ratings were significantly lower for biopreparation-treated and older control tomato plants than for that of same age control plants at 14 and 28 days postinoculation (dpi). CMV accumulation in young noninoculated leaves was significantly less for all biopreparation-treated plants and those in the older control than for the same age control plants at 14 dpi and for four of the five biopreparation treatments at 28 dpi. In those tomato plants shown to be infected, the amount of CMV in noninoculated leaves was significantly lower for three of the biopreparation treatments and the older control treatment at 14 dpi and biopreparation G/INR7 treatment at 28 dpi when compared with the control treatment. These data show that treatment of tomato plants with biopreparations results in significant enhancement of growth and protection against infection by CMV.     

Resistance of postharvest biocontrol yeasts to oxidative stress: a possible new mechanism of action

ABSTRACT We detected the generation of the reactive oxygen species (ROS) superoxide anion ( O.(-) (2)) and hydrogen peroxide (H(2)O(2)) in apple wounds 2 immediately after wounding, and assessed the relationships between (i) timely colonization of apple wounds by biocontrol yeasts, (ii) resistance of these microorganisms to oxidative stress caused by ROS, and (iii) their antagonism against postharvest wound pathogens. We analyzed a model system consisting of two yeasts with higher (Cryptococcus laurentii LS-28) or lower (Rhodotorula glutinis LS-11) antagonistic activity against the postharvest pathogens Botrytis cinerea and Penicillium expansum. LS-28 exhibited faster and greater colonization of wounds than LS-11. In contrast to LS-28, the number of LS-11 cells dropped 1 and 2 h after application, and then increased only later. In vitro, LS-28 was more resistant to ROS-generated oxidative stress. The combined application of biocontrol yeasts and ROS-deactivating enzymes in apple wounds prevented the decrease in number of LS-11 cells mentioned above, and enhanced colonization and antagonistic activity of both biocontrol yeasts against B. cinerea and P. expansum. Polar lipids of LS-11 contained the more unsaturated and oxidizable alpha-linolenic acid, which was absent in LS-28. Resistance to oxidative stress could be a key mechanism of biocontrol yeasts antagonism against postharvest wound pathogens.     

Characterizing the mechanism of biological control of postharvest diseases on fruits with a simple method to study competition for nutrients

ABSTRACT Biocontrol agents may compete with pathogens for nutrients and space to delay or prevent decay of fruits after harvest. These mechanisms of biological control have been difficult to study because no method has been available to determine the significance of each of the components of competition. We developed a nondestructive method using tissue culture plates with cylinder inserts containing defusing membrane at one end to study competition for nutrients without competition for space. Other biocontrol mechanisms in which direct contact between an antagonist and a pathogen is not required also can be studied. The method was used to determine the competition between the yeastlike biocontrol agent, Aureobasidium pullulans, and Penicillium expansum for limited nutrients in apple juice during 24 h incubation, simulating a fruit wound. The antagonist depleted amino acids and inhibited germination of P. expansum conidia. Exposing these conidia to fresh apple juice increased conidial germination to the level comparable to that exhibited by conidia which were not exposed to the antagonist. Because the culture plate method was nondestructive, follow-up experiments in an agar diffusion test were conducted. Juice in which the antagonist grew did not inhibit germination of P. expansum conidia that were seeded on the plates. This corroborates findings from the culture plate method that inhibition of the conidia germination resulted from competition for nutrients. The new method can be coupled with existing techniques to improve understanding of antagonist-pathogen interaction for biological control of postharvest diseases.     

RETRACTED ARTICLE: Effect of calcium chloride on the biocontrol efficacy of two antagonistic yeasts against <Emphasis Type="Italic">Penicillium expansum</Emphasis> on apple fruit

Two antagonistic yeasts, Candida membranaefaciens and Pichia guilliermondii, were evaluated for the control of the blue mold of apple caused by Penicillium expansum. Dual culture, cell-free metabolite and volatile tests were used for in vitro assay. Yeast strains of two genera inhibited growth of P. expansum; inhibition varied from 30.27% to 44.19% in dual culture, from 79.40% to 90.57% in the volatile metabolite test, and from 72.99% to 88.77% in the cell-free metabolite test. Calcium chloride (2% w/v) significantly inhibited the growth of the pathogen P. expansum, but did not affect the colony-forming units (CFU) of the yeasts C. membranaefaciens and P. guilliermondii in potato dextrose broth. The concentration of yeast suspension influenced spore germination and germ tube growth of P. expansum in vitro, as well as disease incidence and lesion development in fruits. There were significant negative relationships between the suspension concentrations of the yeasts and the growth as well as infectivity of the pathogen. The addition of calcium resulted in lower spore germination rates and slower growth of germ tubes in vitro, as well as in lower disease incidences and smaller lesion diameters compared with treatments with yeast antagonists alone. When yeast cell suspensions reached a concentration of 10⁷ CFU ml^-1, growth of the pathogen was completely limited in vitro, and no infection was found in apple fruits treated with or without calcium. This article has been retracted because part of the data shown has already been published before, by different authors. An erratum to this article can be found at     

超敏蛋白对烟草黄瓜花叶病毒的诱导抗性及生长、品质的影响

为明确超敏蛋白对烟草黄瓜花叶病毒(Cucumber mosaic virus,CMV)的诱导抗性及其生长、品质的影响,采用半叶枯斑法和田间试验研究了10%超敏蛋白对枯斑三生烟和普通烟的诱导抗性及促生作用,并分析了对烤烟经济性状和内在质量的影响。结果表明,10%超敏蛋白在活体外对CMV无钝化作用,但预防、治疗效果显著,接种前48 h喷施预防效果达75.70%,接种后24 h喷施治疗效果达65.45%,均显著高于对照;10%超敏蛋白处理后烟草的株高、茎围、最大叶面积均高于其它处理,单位产量比20%吗啉胍·乙铜可湿性粉剂处理增产211.8 kg/hm2,增值3 519.2元/hm2;经10%超敏蛋白处理后的烤烟化学成分均处于优质烟的适宜质量分数范围内。表明超敏蛋白不仅有利于提高烟草抗性和促进烟株生长,且可以明显提高烟叶经济效益和烟叶内在质量。     

Factors affecting the efficacy of post-harvest fungicide applications for the control of blue mould (Penicillium expansum) in stored apples

The effects of apple fruit maturity, temperature of fruit or dip solution, period of time in fungicidal dip, storage conditions, and spore inoculum concentration, on the efficacy of fungicides for control of blue mould ( Penicillium expansum ) were examined in various experiments.
Iprodione and imazalil were only effective when inoculum concentration was low, whilst prochloraz was highly effective in controlling rot on fruit inoculated with 3 × 10⁶ spores/ml. There was no consistent effect of dip temperature or fruit maturity on the efficacy of the fungicides. Iprodione was more effective on warm fruit (19°C) than cold fruit (6°C) whilst the reverse was true of imazalil. Extended periods of immersion in the fungicides slightly reduced the incidence of rotting but not to any useful degree.
The incidence of rotting in fruit treated with prochloraz and etaconazole was less in fruit stored under controlled-atmosphere cold storage conditions than in fruit stored in air cold-storage. Both fungicides were also effective for short-term storage at 20 C in air. Captan, benomyl, captan plus benomyl or vinclozolin were either ineffective or of poor efficacy under all storage conditions.     

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).     

Stability and fitness of pyraclostrobin- and boscalid-resistant phenotypes in field isolates of Botrytis cinerea from apple

Phenotype stability, fitness, and competitive ability of pyraclostrobin- and boscalid-resistant isolates of Botrytis cinerea from apple were investigated. Stability of resistance was determined after consecutive transfers on potato dextrose agar (PDA) or being cycled on apple fruit. In vitro fitness components mycelial growth, osmotic sensitivity, conidial germination, and sporulation were evaluated on agar media. Pathogenicity, virulence and sporulation on apple fruit were evaluated at both 20 and 0°C. Competition between fungicide-resistant and -sensitive isolates on apple fruit also was evaluated. Resistance to the two fungicides was retained at levels similar to that of the initial generation after 20 and 10 transfers on PDA and five and three disease cycles on apple fruit at 20 and 0°C, respectively. Great variability in individual fitness components tested was observed among isolates within the same phenotype groups either sensitive or resistant to the fungicides but, when compared as phenotype groups, there were no significant differences in the mean values of these fitness components between resistant and sensitive phenotypes except that the phenotype resistant only to boscalid produced fewer conidia in vitro than sensitive isolates. Resistant isolates were as pathogenic and virulent on apple fruit as sensitive isolates. There was no significant correlation between the values of individual fitness components tested and the level of resistance to pyraclostrobin or boscalid, except that virulence at 20°C positively correlated with the level of resistance to the two fungicides. The final frequency of pyraclostrobin-resistant individuals in the populations was significantly decreased compared with the initial generation and no boscalid-resistant individuals were detected after four disease cycles on apple fruit inoculated with a pair mixture of a dual-sensitive isolate and one isolate each of the three phenotypes resistant to pyraclostrobin, boscalid, or both. The results suggest that resistance of B. cinerea to pyraclostrobin and boscalid was stable in the absence of the fungicides and that resistance to the two fungicides did not significantly impair individual fitness components tested. However, both pyraclostrobin- and boscalid-resistant isolates exhibited competitive disadvantage over the dual-sensitive isolate on apple fruit.     

Disease Development and Symptom Expression of Xanthomonas axonopodis pv. citri in Various Citrus Plant Tissues

ABSTRACT Experimental inoculations of Xanthomonas axonopodis pv. citri in different tissues of Tahiti lime and Pineapple sweet orange were conducted monthly under natural conditions on Réunion Island. The interactions between a set of environmental and epidemic variables associated with disease expression and 184 different factor combinations were investigated to determine the parameters needed to explain Asiatic citrus canker (ACC) disease expression. Area under the disease progress curve (AUDPC), inoculation date (Id), fruit and leaf age ratings (FAR and LAR), and number of days during the first 2 weeks postinoculation for which the temperature was less than 14 degrees C (T(min)) or more than 28 degrees C (T(max)) were retained by principal component analysis and canonical correlation analysis as the most meaningful epidemic and environmental variables, respectively. AUDPC as the strongest dependent variable and combinations of the environmental variables as independent variables were used in multiple regression analyses. Tissue age rating at the time of infection was a good predictor for disease resulting from spray inoculation on fruits and leaves and also on fruits following a wound inoculation. Temperature, as expressed by T(min) or T(max), was also a significant factor in determining disease development described by AUDPC. Mature green stems were highly susceptible after wounding, similarly to leaves, but buds and leaf scars expressed the lowest susceptibility. These variations in disease expression according to the tissues will have different impacts on ACC epidemiology.     

Prestorage heat treatment reduces pathogenicity of Penicillium expansum in apple fruit

Penicillium expansum is one of the main postharvest pathogens of apples in Israel. Heating apple fruit inoculated with P. expansum for 96 h at 38°C completely inhibited decay development. Fruit held for 24 h at 42°C or 12 h at 46°C had significantly reduced decay after an additional 14 days incubation at 20°C, compared with unheated inoculated control fruit. Mycelial growth and percentage spore germination in vitro were inversely proportional to length of time of exposure to various temperatures. The ET₅₀ for spore germination was 42, 34 and 20 h at 38, 42 and 46°C, respectively, while the ET₅₀ for mycelial growth was 48, 44 and 36 h at those temperatures. When Penicillium spores were incubated on crude extract prepared from the peel of apple fruits held 4 days at 38°C, germ tube elongation was significantly reduced, while the walls of the tubes were thicker, compared with germ tubes from spores incubated on crude extract prepared from peel of non-heated fruit. The evidence presented here supports the hypothesis that the effect of heating on the decay of apples caused by P. expansum is not only the result of direct inhibition of fungal germination and growth by high temperature, but is also partly due to the formation of an inhibitory substance in the heated peel.     

小麦品种混种对白粉病的调控及其对部分产量性状及蛋白质含量的影响

对品种混合种植控制小麦白粉病的效果以及对小麦穗重、千粒重、蛋白质等指标的影响进行了研究,为混合种植控制白粉病提供理论依据和品种组合。在对37个小麦品种进行SSR多样性分析的基础上,选用8个生产品种（后备品种）,按品种数3、4、5、6、8进行组合,在田间以随机区组设计种植混播组合和单播小区,人工接种白粉菌,比较各小区中小麦白粉病的AUDPC值、穗重、千粒重和粗蛋白差异。结果表明,8个品种亲缘关系较远;4品种混种时AUDPC最小,穗重最重。26个品种混种组合中有防治效果的组合占到73.08%,相对防效为1.23%~56.65%;混种未对穗重和粗蛋白含量造成负面影响。品种多样性种植可以用做调控小麦白粉病的一项措施。     

Impact of intercropping on epidemics of groundnut leaf spots: defining constraints and opportunities through a 7‐year field study

Plant diversity can have a profound impact on disease dynamics, with important applications for enhancing sustainability. Disease is often reduced by intercropping, but variability can be high. This study investigated integration of several management approaches to stabilize this variability for early leaf spot (ELS) and late leaf spot (LLS) of groundnut, over seven seasons in three phases. In phase 1, monocrops and alternating row and strip intercrops with maize were artificially inoculated with ELS in an area with little groundnut production. Reductions in AUDPC of 37–73% in strip treatments compared to monocrops prompted testing of the efficacy of intercropping in intensive production areas for phases 2 and 3. Additional treatments included cotton strip intercrops, and integration of intercropping with reduced fungicide treatments and partial resistance to leaf spots. In phase 2, the use of cotton strip intercrops lowered natural ELS epidemics by 25–41% (AUDPC) through delayed disease onset, but maize had inconsistent effects. Intercropping was not effective against LLS, which dominated in phase 3. Reduced fungicide regimes and partial resistance lowered disease, and in one case interacted with intercropping to enhance disease suppression. Groundnut yields generally were inversely proportional to disease levels and not significantly reduced by intercropping. Separate studies to determine maize impacts on ELS infection implicated disruption of dispersal as the mechanism of disease reduction. This work demonstrates that intercropping may be most effective where low levels of ELS are present, using strip patterns with cotton, and combined with other tools such as resistance and reduced fungicide application.     

套袋苹果黑点病的发病诱因、机制与条件

 “套袋苹果斑点病”是套袋果实的重要病害,每年导致3%~10%损失,而“套袋苹果黑点病”是套袋苹果斑点病的一种类型。本研究采用病原菌分离、接种、诱导发病等方法,揭示了黑点病发病的诱因、机制与条件,可为病害的防治提供科学依据。结果表明,诱发套袋苹果黑点病的病原菌是粉红单端孢(Trichothecium roseum);苹果谢花30 d后,幼果上就潜带有大量病原菌;自未喷施过杀菌剂的果园内摘取苹果幼果,在20℃~30℃、100%的相对湿度条件下保湿培养3 d可以诱发黑点病斑;在2周时间内,保湿培养的时间越长,诱发的黑点病斑数量越多;黑点病菌侵入果肉组织后能诱发寄主细胞木栓化,木栓化细胞进一步阻止了病菌的生长与扩展;5月下旬(谢花后30 d)采摘的幼果可诱发产生黑点病斑,但病斑较小;7月上旬(谢花后60 d)苹果果实对黑点病菌最敏感,保湿后诱发的病斑数量最多,形成的病斑大;进入8月份,果实的抗病性明显增强;所测试的9种杀菌剂都能有效抑制黑点病斑的形成,降低果实发病率,但不能完全阻止黑点病斑的形成。苹果果实套袋前,喷施杀菌剂降低果实的带菌量是防治黑点病的重要技术措施。     

套袋苹果黑点病的发病诱因、机制与条件

 “套袋苹果斑点病”是套袋果实的重要病害,每年导致3%~10%损失,而“套袋苹果黑点病”是套袋苹果斑点病的一种类型。本研究采用病原菌分离、接种、诱导发病等方法,揭示了黑点病发病的诱因、机制与条件,可为病害的防治提供科学依据。结果表明,诱发套袋苹果黑点病的病原菌是粉红单端孢(Trichothecium roseum);苹果谢花30 d后,幼果上就潜带有大量病原菌;自未喷施过杀菌剂的果园内摘取苹果幼果,在20℃~30℃、100%的相对湿度条件下保湿培养3 d可以诱发黑点病斑;在2周时间内,保湿培养的时间越长,诱发的黑点病斑数量越多;黑点病菌侵入果肉组织后能诱发寄主细胞木栓化,木栓化细胞进一步阻止了病菌的生长与扩展;5月下旬(谢花后30 d)采摘的幼果可诱发产生黑点病斑,但病斑较小;7月上旬(谢花后60 d)苹果果实对黑点病菌最敏感,保湿后诱发的病斑数量最多,形成的病斑大;进入8月份,果实的抗病性明显增强;所测试的9种杀菌剂都能有效抑制黑点病斑的形成,降低果实发病率,但不能完全阻止黑点病斑的形成。苹果果实套袋前,喷施杀菌剂降低果实的带菌量是防治黑点病的重要技术措施。