scholarly journals Characterization of Fludioxonil-Resistant and Pyrimethanil-Resistant Phenotypes of Penicillium expansum from Apple

2008 ◽  
Vol 98 (4) ◽  
pp. 427-435 ◽  
Author(s):  
H. X. Li ◽  
C. L. Xiao

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 (EC50 > 1,000 μg/ml) and four pyrimethanil-resistant mutants (EC50 > 10 μg/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 μg/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°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°C. Pyrimethanil and fludioxonil reduced blue mold incited by pyrimethanil-resistant mutants during 12-week storage at 0°C but were not effective at 20°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°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.

Plant Disease ◽  
2017 ◽  
Vol 101 (5) ◽  
pp. 738-743
Author(s):  
Achour Amiri ◽  
Aaron W. Hawkins ◽  
Katie A. Mulvaney

Very little is known about the biology, epidemiology, and best practices to manage Lambertella rot, a newly identified postharvest disease caused by Lambertella corni-maris on apples in the United States. In this study, we investigated the prevalence of L. corni-maris in 92 grower lots throughout Washington State in 2016, evaluated the effect of nutrient availability on L. corni-maris growth, and L. corni-maris sensitivity to pH, osmotic stress, and fungicides in vitro. We assessed pathogen aggressiveness on major apple cultivars and the efficacy of pre- and postharvest fungicides to control L. corni-maris on detached fruit. L. corni-maris was widespread and was found in 40% of the growers lots surveyed at frequencies ranging from 2 to 40% of the total decay. The fungus grew faster on acidic media such as apple juice agar and V8 agar media and was able to grow equally at pH values ranging from 3 to 7. L. corni-maris isolates showed relatively low sensitivity to osmotic stress and grew evenly at 1M KCl. All nine apple cultivars tested were susceptible to L. corni-maris, but the disease severity was significantly higher on some cultivars such as Honeycrisp, Piñata, and Gala. The baseline sensitivity of 100 L. corni-maris isolates was determined based on the effective concentration necessary to inhibit 50% mycelial growth (EC50) for fludioxonil, boscalid, pyraclostrobin + SHAM, pyrimethanil, and thiabendazole. The respective mean EC50 values were 0.07, 0.84, 1.32, 2.45, and 3.68 µg/ml. Fludioxonil and pyrimethanil applied at label rates were the only fungicides able to control L. corni-maris effectively on detached apple fruit. This study is a first step toward understanding the eco-physiological requirements for L. corni-maris to survive and cause decay on apple, assessing potential shifts in fungicide sensitivity, and enhancing Lambertella rot management. We suggest “yellow rot” as the new common name for Lambertella rot.


PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6170 ◽  
Author(s):  
Guangxi Wu ◽  
Wayne M. Jurick II ◽  
Franz J. Lichtner ◽  
Hui Peng ◽  
Guohua Yin ◽  
...  

Blue mold is a postharvest rot of pomaceous fruits caused by Penicillium expansum and a number of other Penicillium species. The genome of the highly aggressive P. expansum strain R19 was re-sequenced and analyzed together with the genome of the less aggressive P. solitum strain RS1. Whole genome scale similarities and differences were examined. A phylogenetic analysis of P. expansum, P. solitum, and several closely related Penicillium species revealed that the two pathogens isolated from decayed apple with blue mold symptoms are not each other’s closest relatives. Among a total of 10,560 and 10,672 protein coding sequences respectively, a comparative genomics analysis revealed 41 genes in P. expansum R19 and 43 genes in P. solitum RS1 that are unique to these two species. These genes may be associated with pome fruit–fungal interactions, subsequent decay processes, and mycotoxin accumulation. An intact patulin gene cluster consisting of 15 biosynthetic genes was identified in the patulin producing P. expansum strain R19, while only a remnant, seven-gene cluster was identified in the patulin-deficient P. solitum strain. However, P. solitum contained a large number of additional secondary metabolite gene clusters, indicating that this species has the potential capacity to produce an array of known as well as not-yet-identified products of possible toxicological or biotechnological interest.


2018 ◽  
Author(s):  
Guangxi Wu ◽  
Wayne M Jurick II ◽  
Franz J Lichtner ◽  
Hui Peng ◽  
Guohua Yin ◽  
...  

Blue mold is a postharvest rot of pomaceous fruits caused by Penicillium expansum and a number of other Penicillium species. The genome of the highly aggressive P. expansum strain R19 was re-sequenced and analyzed together with the genome of the less aggressive P. solitum strain RS1. Whole genome scale similarities and differences were examined. A phylogenetic analysis of P. expansum, P. solitum, and several closely related Penicillium species revealed that the two pathogens isolated from decayed apple with blue mold symptoms are not each other’s closest relatives. Among a total of 10,560 and 10,672 protein coding sequences respectively, a comparative genomics analysis revealed 41 genes in P. expansum R19 and 43 genes in P. solitum RS1 that are unique to these two species. These genes may be associated with pome fruit–fungal interactions, subsequent decay processes, and mycotoxin accumulation. An intact patulin gene cluster consisting of 15 biosynthetic genes was identified in the patulin producing P. expansum strain R19, while only a remnant, seven-gene cluster was identified in the patulin-deficient P. solitum strain. However, P. solitum contained a large number of additional secondary metabolite gene clusters indicating that this species has the potential capacity to produce an array of known, as well as not-yet-identified products, of possible toxicological or biotechnological interest.


Plant Disease ◽  
2014 ◽  
Vol 98 (7) ◽  
pp. 1004-1004 ◽  
Author(s):  
H. J. Yan ◽  
V. L. Gaskins ◽  
I. Vico ◽  
Y. G. Luo ◽  
W. M. Jurick

Apples in the United States are stored in low-temperature controlled atmospheres for 9 to 12 months and are highly susceptible to blue mold decay. Penicillium spp. cause significant economic losses worldwide and produce mycotoxins that contaminate processed apple products. Blue mold is managed by a combination of cultural practices and the application of fungicides. In 2004, a new postharvest fungicide, pyrimethanil (Penbotec 400 SC, Janseen PMP, Beerse, Belgium) was registered for use in the United States to control blue mold on pome fruits (1). In this study, 10 blue mold symptomatic ‘Red Delicious’ apples were collected in May 2011 from wooden bins at a commercial facility located in Pennsylvania. These fruit had been treated with Penbotec prior to controlled atmosphere storage. Ten single-spore Penicillium spp. isolates were analyzed for growth using 96-well microtiter plates containing Richards minimal medium amended with a range of technical grade pyrimethanil from 0 to 500 μg/ml. Conidial suspensions adjusted to 1 × 105 conidia/ml were added to three 96-well plates for each experiment; all experiments were repeated three times. Nine resistant isolates had prolific mycelial growth at 500 μg/ml, which is 1,000 times the discriminatory dose that inhibited baseline sensitive P. expansum isolates from Washington State (1). However, one isolate (R13) had limited conidial germination and no mycelial proliferation at 0.5 μg/ml and was categorized as sensitive. One resistant (R22) and one sensitive (R13) isolate were selected on the basis of their different sensitivities to pyrimethanil. Both isolates were identified as P. expansum via conventional PCR using β-tubulin gene-specific primers according to Sholberg et al. (2). Analysis of the 2X consensus amplicon sequences from R13 and R22 matched perfectly (100% identity and 0.0 E value) with other P. expansum accessions in GenBank including JN872743.1, which was isolated from decayed apple fruit from Washington State. To determine if pyrimethanil applied at the labeled rate of 500 μg/ml would control R13 or R22 in vivo, organic ‘Gala’ apple fruit were wounded, inoculated with 50 μl of a conidial suspension (1 × 104 conidia/ml) of either isolate, dipped in Penbotec fungicide or sterile water, and stored at 25°C for 7 days. Twenty fruit composed a replicate within a treatment and the experiment was performed twice. Non-inoculated water-only controls were symptomless, while water-dipped inoculated fruit had 100% decay with mean lesion diameters of 36.8 ± 2.68 mm for R22 and 38.5 ± 2.61 mm for R13. The R22 isolate caused 30% decay with 21.6 ± 5.44 mm lesions when inoculated onto Penbotec-treated apples, while the R13 isolate had 7.5% decay incidence with mean lesion diameters of 23.1 ± 3.41 mm. The results from this study demonstrate that P. expansum pyrimethanil-resistant strains are virulent on Penbotec-treated apple fruit and have the potential to manifest in decay during storage. To the best of our knowledge, this is the first report of pyrimethanil resistance in P. expansum from Pennsylvania, a major apple growing region for the United States. Moreover, these results illuminate the need to develop additional chemical, cultural, and biological methods to control this fungus. References: (1) H. X. Li and C. L. Xiao. Phytopathology 98:427, 2008. (2) P. L. Sholberg et al. Postharvest Biol. Technol. 36:41, 2005.


Plant Disease ◽  
2011 ◽  
Vol 95 (1) ◽  
pp. 72-72 ◽  
Author(s):  
C. L. Xiao ◽  
Y. K. Kim ◽  
R. J. Boal

Blue mold caused by Penicillium expansum is a major postharvest fruit rot disease of apples (Malus domestica) worldwide. Pyrimethanil was registered in late 2004 in the United States for postharvest use on apples. Since then, pyrimethanil has been increasingly used in Washington State as a postharvest drench treatment for control of blue mold and other postharvest diseases in apples. Baseline sensitivity to pyrimethanil in P. expansum populations from apples in Washington State has been established and all isolates in the baseline population were sensitive to pyrimethanil (1). To monitor resistance to pyrimethanil in P. expansum populations, blue mold-like decayed apple fruit were sampled from May to August 2009 from the fruit that had been drenched with pyrimethanil prior to storage from fruit packinghouses. Isolation of Penicillium species from decayed fruit was attempted. Isolates of Penicillium species were identified to species according to the descriptions by Pitt (2). In total, 186 P. expansum isolates were collected and tested for resistance to pyrimethanil in a conidial germination assay on an agar medium amended with pyrimethanil at the discriminatory concentration of 0.5 μg ml–1 (1). Isolates that were able to germinate were considered resistant to pyrimethanil. Of the 186 isolates tested, one was resistant to pyrimethanil. EC50 (the effective concentration that inhibits fungal growth by 50% relative to the control) of pyrimethanil for the resistant isolate was determined according to a method described previously (1) and the test was done twice. EC50 values of pyrimethanil on mycelial growth and conidial germination for the resistant isolate were 9.9 and 3.1 μg/ml, respectively, which were 7.4-fold and 16.5-fold higher than the means of the baseline population (1). To evaluate whether pyrimethanil at label rate is still able to control this resistant isolate, ‘Fuji’ apples were wounded, inoculated with conidial suspensions (1 × 104 conidia ml–1) of either the resistant isolate or a pyrimethanil-sensitive isolate, treated with either pyrimethanil or sterile water as controls, and stored at 20°C for 10 days following a method described previously (1). There were four 20-fruit replicates for each treatment. The experiment was performed twice. All inoculated fruit in the nontreated controls were decayed. Pyrimethanil applied at label rate completely controlled blue mold incited by a pyrimethanil-sensitive isolate, but 75% of the fruit that were inoculated with the resistant isolate and treated with pyrimethanil developed blue mold. To our knowledge, this is the first report of pyrimethanil resistance in P. expansum from decayed apple fruit collected from commercial packing houses. The pyrimethanil-resistant isolate was obtained from a packing house in which pyrimethanil had been used as a postharvest drench treatment in each of four consecutive years, suggesting that pyrimethanil-resistant individuals are emerging in P. expansum populations in Washington State after repeated use of pyrimethanil. Our results also indicate that pyrimethanil resistance in P. expansum reported in this study can result in failure of blue mold control in apples with pyrimethanil. References: (1) H. X. Li and C. L. Xiao. Postharvest Biol. Technol. 47:239, 2008. (2) J. I. Pitt. A Laboratory Guide to Common Penicillium species. Food Science Australia, North Ryde NSW, Australia, 2002.


2018 ◽  
Author(s):  
Guangxi Wu ◽  
Wayne M Jurick II ◽  
Franz J Lichtner ◽  
Hui Peng ◽  
Guohua Yin ◽  
...  

Blue mold is a postharvest rot of pomaceous fruits caused by Penicillium expansum and a number of other Penicillium species. The genome of the highly aggressive P. expansum strain R19 was re-sequenced and analyzed together with the genome of the less aggressive P. solitum strain RS1. Whole genome scale similarities and differences were examined. A phylogenetic analysis of P. expansum, P. solitum, and several closely related Penicillium species revealed that the two pathogens isolated from decayed apple with blue mold symptoms are not each other’s closest relatives. Among a total of 10,560 and 10,672 protein coding sequences respectively, a comparative genomics analysis revealed 41 genes in P. expansum R19 and 43 genes in P. solitum RS1 that are unique to these two species. These genes may be associated with pome fruit–fungal interactions, subsequent decay processes, and mycotoxin accumulation. An intact patulin gene cluster consisting of 15 biosynthetic genes was identified in the patulin producing P. expansum strain R19, while only a remnant, seven-gene cluster was identified in the patulin-deficient P. solitum strain. However, P. solitum contained a large number of additional secondary metabolite gene clusters indicating that this species has the potential capacity to produce an array of known, as well as not-yet-identified products, of possible toxicological or biotechnological interest.


Plant Disease ◽  
2009 ◽  
Vol 93 (10) ◽  
pp. 1003-1008 ◽  
Author(s):  
C. L. Xiao ◽  
R. J. Boal

Blue mold caused by Penicillium expansum is a major postharvest disease of apples (Malus × domestica). Residual activity of fludioxonil and pyrimethanil in apple fruit against P. expansum was investigated during 2005 to 2008. Fruit of the cultivar Delicious harvested from commercial orchards where fungicides were not used were either not treated or drenched with fludioxonil, pyrimethanil, or thiabendazole prior to storage and then stored in controlled atmosphere at 0°C for 5 or 7 months, after which time the fruit were removed from storage and subjected to washing and brushing, practices that are done at the time of packing. Fruit were then wounded and inoculated with conidial suspensions of P. expansum. Inoculated fruit were treated with either sterile water or fungicides. Fruit were stored at 0°C for 8 weeks and at room temperature for one additional week after cold storage. To determine distribution of fungicide residues in the fruit flesh, fruit were cut horizontally at the equator, sprayed with the conidial suspension of P. expansum, incubated at room temperature, and examined for inhibition of blue mold on the cut fruit 4 days after inoculation. Fungicide residues on/in the fruit were analyzed using a gas chromatograph. Zero to 26% blue mold incidence was observed on fludioxonil-drenched fruit that were inoculated and not treated with fungicides at packing. No decay or <4% blue mold incidence was observed on pyrimethanil-drenched fruit that were inoculated and not treated with fungicides at packing, whereas 65 to 99% blue mold incidence was observed on thiabendazole-drenched fruit that were not treated with fungicides at packing. An average of >32 mm inhibition zone and approximately 5 mm inhibition zone measured from the fruit peel toward the fruit core were observed on pyrimethanil-drenched and fludioxonil-drenched fruit, respectively. Washing and brushing at the time of packing 5 and 7 months after harvest did not remove or only partially removed residues of fludioxonil and pyrimethanil from apple fruit. The results suggest that residues of fludioxonil and pyrimethanil on/in apple fruit are persistent and that residual protection of apple fruit by the two fungicides can last for at least 7 months under apple-storage conditions.


2009 ◽  
Vol 99 (3) ◽  
pp. 258-264 ◽  
Author(s):  
Ting Yu ◽  
Jishuang Chen ◽  
Huangping Lu ◽  
Xiaodong Zheng

Cryptococcus laurentii is a well-known postharvest biocontrol yeast; however, it cannot provide satisfactory levels of decay control when used alone. Here, we evaluated the effects of indole-3-acetic acid (IAA), a plant growth regulator, on the biocontrol efficacy of the yeast antagonist C. laurentii against blue mold rot caused by Penicillium expansum in apple fruit. Results showed that the addition of IAA at 20 μg/ml to suspensions of C. laurentii greatly enhanced inhibition of mold rot in apple wounds compared with that observed with C. laurentii alone. The addition of IAA at 20 μg/ml or lower did not influence the population growth of C. laurentii in wounds, but adverse effects were seen on C. laurentii when the concentration of IAA was increased to 200 μg/ml or above in vitro and in vivo. P. expansum infection in apple wounds was not inhibited when the pathogen was inoculated into the fruit wounds within 2 h after application of IAA; however, infection was reduced when inoculated more than 12 h after IAA application. Treatment of wounds with IAA at 20 μg/ml 24 h before pathogen inoculation resulted in significant inhibition of P. expansum spore germination and host infection. Application of IAA at 20 μg/ml also reduced P. expansum infection when it was applied 48 h before pathogen inoculation in the intact fruit. Thus, IAA could reinforce the biocontrol efficacy of C. laurentii in inhibiting blue mold of apple fruit by induction of the natural resistance of the fruit.


Plant Disease ◽  
2019 ◽  
Vol 103 (2) ◽  
pp. 331-337 ◽  
Author(s):  
Wayne M. Jurick ◽  
Otilia Macarisin ◽  
Verneta L. Gaskins ◽  
Wojciech J. Janisiewicz ◽  
Kari A. Peter ◽  
...  

Penicillium spp. cause blue mold of stored pome fruit. These fungi reduce fruit quality and produce mycotoxins that are regulated for processed fruit products. Control of blue mold is achieved by fungicide application, and in 2015 Academy (active ingredients fludioxonil and difenoconazole) was released for use on pome fruit to manage postharvest blue mold. Baseline sensitivity for fludioxonil but not difenoconazole has been determined for P. expansum. To establish the distribution of sensitivity to difenoconazole before commercial use of Academy, 97 unexposed single-spore isolates from the United States and abroad were tested in vitro. Baseline EC50 values ranged from 0.038 to 0.827 µg/ml of difenoconazole with an average of 0.16 µg/ml. Complete inhibition of mycelial growth for all but three isolates occurred at 5 µg/ml of difenoconazole, whereas 10 µg/ml did not support growth for any of the isolates examined. Hence, 5 µg/ml of difenoconazole is recommended for phenotyping Penicillium spp. isolates with reduced sensitivity. Isolates with resistance to pyrimethanil and to both thiabendazole and pyrimethanil were observed among the isolates from the baseline collection. Academy applied at the labeled rate had both curative and protectant activities and controlled four representative Penicillium spp. from the baseline population. This information can be used to monitor future shifts in sensitivity to this new postharvest fungicide in Penicillium spp. populations.


Plant Disease ◽  
2014 ◽  
Vol 98 (7) ◽  
pp. 924-928 ◽  
Author(s):  
R. Caiazzo ◽  
Y. K. Kim ◽  
C. L. Xiao

Penicillium expansum is the cause of blue mold in stored apple fruit. In 2010–11, 779 isolates of P. expansum were collected from decayed apple fruit from five packinghouses, tested for resistance to the postharvest fungicide pyrimethanil, and phenotyped based on the level of resistance. In 2010, 85 and 7% of the isolates were resistant to pyrimethanil in packinghouse A and B, respectively, where pyrimethanil had been used for four to five consecutive years. In 2011, pyrimethanil or fludioxonil was used in packinghouse A, and 96% of the isolates from the fruit treated with pyrimethanil were resistant but only 4% of the isolates from the fruit treated with fludioxonil were resistant to pyrimethanil, suggesting that fungicide rotation substantially reduced the frequency of pyrimethanil resistance. No pyrimethanil-resistant isolates were detected in 2010 in the three other packinghouses where the fungicide had been used recently on a small scale. However 1.8% of the isolates from one of the three packinghouses in 2011 were resistant to pyrimethanil. A significantly higher percentage of thiabendazole-resistant than thiabendazole-sensitive isolates were resistant to pyrimethanil. Of the pyrimethanil-resistant isolates, 37 to 52, 4 to 5, and 44 to 58% were phenotyped as having low, moderate, and high resistance to pyrimethanil, respectively. Fludioxonil effectively controlled pyrimethanil-resistant phenotypes on apple fruit but pyrimethanil failed to control phenotypes with moderate or high resistance to pyrimethanil and only partially controlled the low-resistance phenotype.


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