Postharvest Fruit Rot Disease Research Articles

Ceratocystis fimbriata Causing Postharvest Fruit Rot of Pear in Punjab, Pakistan.

Pear (Pyrus communis) is an important deciduous fruit cultivated on a worldwide scale including Pakistan. During August 2021, a postharvest fruit rot disease was observed on several pears at various farmers market in Okara- a district of Punjab Province, Pakistan. The incidence of the disease varied from 7 to 20% with 35% disease severity. Necrotic spots (10 to 20 mm diameter) were first observed on the infected pear fruit. The spots enlarged gradually and developed into a brown, water-soaked and rotted lesion. Eventually, the whole fruit became soft, rotted and covered with a gray-brown mycelium. The isolates were obtained from the symptomatic tissues (n = 18) incubated on carrot discs that had been surface sterilized in 100-ppm streptomycin solution. After consistent sporulation of a fungus on the carrot discs, the ascospore masses formed at the tip of perithecia were transferred to malt extract agar (MEA). Primary conidia were cylindrical and hyaline (7 to 11 × 4 to 7 μm) and secondary conidia were hyaline and barrel-shaped (7 to 12 × 5 to 8 μm). Endoconidiophores with primary conidia were (12 to 27 × 2.6 to 5.5 μm). Perithecia produced on carrot discs were dark brown to black, and the base was 157 to 278 μm in diameter. Ascomatal necks were 512 to 656 µm long, dark brown to black, lighter in color at apices, tapering from base (23 to 45 μm diameter) to apex (13 to 24 μm diameter). Ostiolar hyphae were 41 to 79 μm long. Ascospores were hyaline, hat shaped, 3 to 4 μm long, and accumulated in a sticky matrix at the tips of perithecial necks. Mycelium was initially hyaline but became dark greenish brown after 7 days. Dark brown, thick-walled aleuroconidia (13 to 19.5 × 9 to 14 μm) appeared on culture plates after 2 months. Based on morphological characteristics, the fungus was identified as Ceratocystis fimbriata (Engelbrecht, 2005; Suwandi et al. 2021). To further confirm species identification, genomic DNA of two representative isolates (UO-05 and UO-06) was obtained using an extraction kit. The internal transcribed spacer (ITS) region was amplified using ITS1/4 (White et al. 1990). A BLAST search with GenBank accession nos. OR185451 and OR185456 indicated 99 to 100% identity with several C. fimbriata including type species (MH856050.1; KC493160.1; MT560374.1). Pathogenicity tests were conducted by inoculating nine disease-free pear (cv. Concord) fruit after disinfesting in 75% ethanol. A prepared spore suspension (1.0 × 106 spores/ml) was dropped on the wounds (a depth of 1 mm diameter) on the pear surface, which were made by a sterilized needle. 10 μl of a prepared spore suspension was dropped onto nine pears. Sterile water (10 μl) was dropped on the wounded sites of nine pear fruits as negative controls and all fruits were incubated in a growth chamber at 30/26°C (day/night, 90% relative humidity). Symptoms similar to those on the naturally infected fruits began after 4 to 5 days of inoculation, while controls remained healthy. The fungal isolates recovered from inoculated pears were morphologically identical to the C. fimbriata isolates originally recovered from symptomatic fruits fulfilling Koch's postulates. The pathogen has been reported to cause postharvest fruit rot of passion fruit and cucumber (Firmino et al. 2016; Li et al. 2019). To our knowledge, this is the first report of C. fimbriata causing fruit rot of pear in Punjab Province. The detection of this disease will help pear growers to take actions to monitor and prevent disease outbreak as well as develop an effective management practice when it occurs.

Morpho-phylogenetic identification and characterization of new causal agents of Fusarium species for postharvest fruit rot disease of muskmelon in northern Thailand and their sensitivity to fungicides.

A significant global problem affecting muskmelon (Cucumis melo L.) is fruit rot caused by phytopathogenic fungi, which results in unsaleable products and substantial financial losses. In 2022 and 2023, fruit rot on muskmelon was found during the postharvest storage period in Phayao Province of northern Thailand. The aim of the current study was to isolate the species of fungi causing the fruit rot lesions. Out of the rot lesions on muskmelons, nine fungal isolates were received. All isolates of fungi were identified through a combination of morphological characteristics and molecular analyses. Based on their morphological traits, all isolated fungal isolate was assigned to the genus Fusarium. All the fungal isolates were determined to belong to the Fusarium incarnatum-equiseti species complex through multi-gene phylogenetic analysis employing the calmodulin (cam), RNA polymerase second largest subunit (rpb2), and translation elongation factor 1-alpha (tef1-α) genes. These isolates were identified as F. compactum (SDBR-CMU483), F. jinanense (SDBR-CMU484, SDBR-CMU485, and SDBR-CMU486), F. mianyangense (SDBR-CMU487 and SDBR-CMU488), and F. sulawesiense (SDBR-CMU489, SDBR-CMU490, and SDBR-CMU491). Moreover, pathogenicity tests were subsequently carried out, and the results indicated that all fungal isolates caused symptoms of fruit rot on inoculated muskmelon fruits. Notably, this result was consistent with the symptoms observed throughout the postharvest storage period. In the fungicide screening test, all fungal isolates showed sensitivity to copper oxychloride. However, all isolates showed insensitivity to benalaxyl-M + mancozeb, carbendazim, mancozeb, and metalaxy. To the best of our knowledge, the present study is the first to identify F. compactum, F. jinanense, and F. mianyangense as new causative agents of muskmelon fruit rot in Thailand and other regions globally. This is also the first report of postharvest fruit rot on muskmelons caused by F. sulawesiense in Thailand. Furthermore, the fungicide screening results indicate that fungicide resistance can be beneficial in developing potential management strategies against postharvest fruit rot disease of muskmelon caused by these four pathogenic Fusarium species.

First Report of Postharvest Fruit Rot on Gannan Navel Orange Caused by Diaporthe unshiuensis in Jiangxi Province, China.

The Gannan navel orange (Citrus sinensis Osbeck cv. Newhall) is one of the most widely planted citrus fruit cultivars in Ganzhou City, Jiangxi Province, China. A Gannan navel orange was harvested from an orchard in Yudu County, Ganzhou City, Jiangxi Province, China, in October 2022 (25.95N, 115.41E). Approximately 5% of the fruit rotted after being stored at room temperature for about two weeks.Infected fruits appear brown and rotted with slightly indented edges. Initially symptoms of infected fruits was small circular, light brown, which the rot expands, slightly water-stained halo circle with slightly indented edges. The surface of 10 infected fruits was sterilized with 75% ethanol, and the lesion edge was cut into 5-mm-diameter pieces, and the pieces were then placed on potato dextrose agar (PDA) and incubated at 25°C for five days. A total of eight morphologically similar isolates were obtained. PDA results showed dense white and fluffy aerial mycelia in the center of colonies with sparser edges. Two types of conidia were produced; the alpha conidia were hyaline, ellipsoidal or clavate, and aseptate, with 2 oil drops, 4.8 to 7.5 × 2.1 to 2.7 μm (n=30). The beta conidia were hyaline, aseptate, filiform, smooth, straight to sinuous, 16.9 to 27.5 × 1.3 to 1.6 μm (n=30). These isolates exhibit morphological characteristics similar to those of the Diaporthe genus. Genomic DNA of two representative isolates (JFRL-03-1130 and JFRL-03-1131) was extracted for further confirmation. The internal transcribed spacer (ITS) region, beta-tubulin (TUB), calmodulin (CAL), partial translation elongation factor 1-alpha (TEF1-α), and histone H3 (HIS3) genes were amplified and sequenced using primers ITS1/ITS4, Bt2a/Bt2b, CAL228F/CAL737R, EF1-728F/EF1-986R, and CYLH3F/H3-1b, respectively (Udayanga et al. 2015). These nucleotide sequences were deposited into the GenBank database with accession numbers OQ691637-OQ691638 (ITS), OQ701022-OQ701023 (TUB), OQ701016-OQ701017 (CAL), OQ701018-OQ701019 (TEF1-α) and OQ701020-OQ701021 (HIS3). The maximum likelihood analyses were performed for the combined ITS, TEF1-a, TUB, HIS3, and CAL data set using Phylosuite V1.2.2 (Zhang et al. 2020). The phylogenetic tree showed that the two isolates clustered with D. unshiuensis in a clade with 100% bootstrap support. Therefore, the fungus was identified as D. unshiuensis based on morphological and molecular characteristics. To evaluate pathogenicity, a sterile scalpel was used to wound 10 surface-sterilized fruits, and a 5-mm-diameter mycelial plug of the isolate JFRL 03-1130, cultured on PDA at 25℃ for 7-days, was put on the wound. Another set of 10 fruits was similarly inoculated with sterile agar plugs as controls. The fruits were cultured at 25°C and 85% relative humidity, and the test was repeated twice. These fruits inoculated with D. unshiuensis showed similar rot symptoms after 10 days, while the control group remained symptomless. In order to prove Koch's postulates, the pathogen was re-isolated from the inoculated fruits and confirmed as D. unshiuensis by molecular techniques, but never from the control fruits. Diaporthe unshiuensis has been reported as an endophyte associated with citrus and a pathogen that causes melanose disease in citrus (Chaisiri et al. 2020; Huang et al. 2015). However, to the best of our knowledge, this is the first reported case of D. unshiuensis causing postharvest fruit rot on Citrus sinensis. In the past, D. sojae has also been reported causing postharvest fruit brown rot disease on Citrus sinensis in China (Xiao, et al. 2023); Therefore, managers should pay more attention to postharvest fruit rot disease caused by Diaporthe species and implement storage strategies to control and reduce losses.

First Report of Lasiodiplodia pseudotheobromae Causing Postharvest Fruit Rot on Indian Jujube in China.

Indian jujube (Ziziphus mauritiana Lamarck), is one of the most popular fruit crops in South China. In March 2023, a fruit rot of indian jujube with about 5% disease incidence was observed in two supermarkets of Nanchang City, Jiangxi Province, China. Initially, the symptoms appeared as slightly brown spots on the fruit surface, with disease progression, the lesions gradually expanded and covered with a layer of hyphae. Small pieces (3 to 4 mm2) from the periphery of 15 diseased fruit were surface disinfected using 1% sodium hypochlorite for 30 s, rinsed three times in sterilized distilled water, air dried, and then aseptically placed onto potato dextrose agar (PDA) media and incubated at 25°C for three days. A total of ten single spore isolates with similar morphology were obtained. Colonies of these consisted of initially white, gradually turning gray and eventually becoming black, and aerial hyphae were dense and fluffy. Conidiogenous cells were smooth, hyaline, cylinder-shaped, and holoblastic. Conidia were ellipsoidal, top and base-rounded, and thick-walled, immature conidia were colorless, hyaline, and aseptate, compared with dark brown color of the mature conidia, which were one-septate with longitudinal striations, ranging in size from 22.8 to 31.8 (mean 27.6) µm in length and 12.2 to 20 (mean 14.6) µm in width. The morphological characteristics were consistent with the characteristics of the Lasiodiplodia species (Phillips et al. 2013). To accurately identify the strain, three representative isolates, namely JFRL 03-1147, JFRL 03-1148, and JFRL 03-1149, were selected for further identification. The internal transcribed spacers (ITS), translation elongation factor 1-alpha (TEF1-α), and beta-tubulin (TUB2) genes/regions were amplified and sequenced using primers ITS1/ITS4, EF1-688F/EF1-1251R, and Bt2a/Bt2b, respectively (Chen et al. 2021). These nucleotide sequences were deposited in GenBank with accession numbers OQ804425-OQ804427 (ITS), OQ818097-OQ818099 (TEF1-α), and OQ818100-OQ818102 (TUB2). A BLASTn homology search for these nucleotides showed 99-100% identity to ITS (EF622077, 487 nt/487 nt), TEF1-α (EF622057, 306 nt/307 nt) and TUB2 (EU673111, 434 nt/434 nt) sequences of Lasiodiplodia pseudotheobromae CBS 116459 (ex-type). The maximum likelihood analyses were performed for the combined ITS, TEF1-α and TUB2 data set using IQtree web server (Trifinopoulos et al. 2016). The phylogenetic tree showed that the three isolates clustered with Lasiodiplodia pseudotheobromae in a clade with 99% bootstrap support. Therefore, the fungus was identified as L. pseudotheobromae based on morphological and molecular characteristics. To evaluate pathogenicity, 4 healthy fruits of indian jujube were surface sterilized with 75% ethanol and wounded by sterile needle, and a 5-mm-diameter agar with 5-days-old mycelium of the isolate JFRL 03-1148 cultured on PDA at 25°C was put on the wound. Another set of 4 fruits was inoculated with sterile agar plugs as controls. The fruits were cultured at 25℃ and 85% relative humidity, and the test was repeated twice. These fruits inoculated with L. pseudotheobromae showed similar rot symptoms after 3 days, while the control group remained asymptomatic. To fulfill Koch's postulates, the pathogen was re-isolated from the inoculated fruits and confirmed as L. pseudotheobromae by morphological and molecular analysis. L. pseudotheobromae has previously been reported causing fruit rot on citrus, mango and papaya (Alam et al. 2021; Chen et al. 2021; Netto et al. 2014). But to our knowledge, this is the first report of L. pseudotheobromae caused postharvest fruit rot on indian jujube in China. Therefore, managers should pay more attention to postharvest fruit rot disease caused by L. pseudotheobromae, and formulate appropriate disease control measures to reduce its losses.

Early warning of Diaporthe infection in kiwifruit soft rot by plasmonic dimer-enhanced Raman spectroscopy.

Diaporthe genus is the dominant pathogens of kiwifruit soft rot with long incubation period and rapid onset and very hard to detect in advance. It is of great significance to develop point-of-care tests for disease prevention and field management. Here we screen a pair of genus-level primers for constructing plasmonic dimer Rayleigh/Raman spectroscopy, which enables rapid, specific, and sensitive detection of Diaporthe genus in real kiwifruits. Dark-filed Rayleigh scattering clearly visualizes the target-induced dimer assembly of nanoprobes. Plasmonic dimer-enhanced Raman spectroscopy has high specificity for 12 common isolates of Diaporthe genus in one batch samples with highly accurate results and much higher sensitivity than polymerase chain reaction (PCR). It realizes the recognition of Diaporthe infection at an early stage of 24h when the kiwifruits do not have any noticeable symptoms. It demonstrates a bright prospect of point-of-care tests for early warning of kiwifruit soft rot infection and quality control.

First Report of Postharvest Fruit Rot Disease of Yellow Peach Caused by Diaporthe eres in China.

The yellow peach (Amygdalus persica), is a fruit crop native to China with golden peel and pulp that is of particular interest in the fruit markets. In August of 2021, yellow peaches showing fruit rot symptoms were purchased from a commercial market in Linyi city, Shandong province, China. The symptoms included circular, tan to brown in color, rotten, necrotic lesions, and whitish mycelium mass in the center of the lesions. The infected fruit were surface disinfected with 1% NaClO for 30 s and rinsed with sterile distilled water three times. Diseased tissues from the infected fruits were cut into small segments, aseptically shifted onto potato dextrose agar media containing petri plates and incubated at 25℃ for 5 days. Eight isolates were obtained in total from two isolation experiments. Fungal colonies were initially white, aerial, fluffy at first, and gradually turned brown to gray, with black stromata at maturity. Alpha conidia were aseptate，hyaline，fusiform to ellipsoidal，and ranged in size from 4.16 to 7.76 µm × 1.95 to 3.14 µm (n=30). Beta conidia were aseptate, hyaline, filiform, curved to hamate, and 15.91 to 22.55 µm × 0.82 to 1.66 µm (n=30). The morphological characteristics were consistent with those of Diaporthe species (Gomes et al. 2013). For further identification, a multigene phylogenetic analysis was carried out. The internal transcribed spacer (ITS) region, translation elongation factor 1-α (TEF1-α), histone H3 (HIS), calmodulin (CAL), and β-tubulin (TUB) genes of two representative isolates were amplified by using primers ITS1/ITS4, EF1-728F/EF1-986R, CYLH3F/H3-1b，CAL228F/CAL737R, and Bt2a/Bt2b (Chaisiri et al. 2021), respectively. The sequences were deposited in GenBank (Accession No. OL375154 for ITS; OL406409 for TEF1-α; OL406410 for HIS; OL106407 for CAL; OL406408 for TUB). phylogenetic analyses were conducted using the concatenation of multiple sequences (ITS, TEF1-α, HIS, CAL, TUB) with Maximum Likelihood (ML) in IQtree v1.5.6 (Nguyen et al. 2015). Based on the morphological and phylogenetic characters, the isolates were identified as D. eres. A Pathogenicity test was performed by wound inoculation on harvested fruits of A. persica Variety 'Jinxiu'. Mature and healthy yellow peaches purchased from Shandong, Anhui, and Hunan Provinces in China were surface sterilized with 1% NaClO solution for 1 minute, rinsed with sterile water and dried. Each fruit was wounded with a sterile scalpel creating a 2-3 mm incision on the peel. A 5 mm agar disc with mycelium grown on PDA at 28℃ for 7 days was placed on wound and sealed with parafilm. Sterile PDA plugs were used as controls. Ten fruit were used for each treatment and the assays were repeated three times. Inoculated fruit were placed in sterilized transparent plastic cans containing wet, sterile paper towels. After 5 days of incubation at 25℃, the same rot symptoms were observed on fruits inoculated with mycelium and the control remained symptomless. D. eres was re-isolated from the lesions of inoculated fruits and the pathogen identification was confirmed by molecular analysis, thus fulfilling complete Koch's postulates. Although D. eres was previously reported on peach trees of causing shoot blight (Thomidis and Michailides 2009) and stem canker (Prencipe et al. 2017). To our knowledge, this is the first report of D. eres causing postharvest fruit rot of yellow peach in China and it may lead to considerable economic losses in the peach industry should post-harvest disease management practices not be implemented.

IN VITRO EVALUATION OF DIFFERENT LOCALLY AVAILABLE PRESERVATIVES AGAINST THE FUNGUS CAUSING TOMATO FRUIT ROT DISEASE

The post-harvest tomato fruit rot caused by Alternaria solani is a major threat to the tomato fruits. Our main objective of this study was to assess the locally available plant based preservatives against post-harvest tomato fruit rot. The in-vitro antifungal effect of 09 different treatments (07 plants based = black pepper, red pepper, turmeric, clove, garlic, onion and papaya seed and 02 chemical preservatives = Potassium metabi-sulphate, PMS and Sodium benzoate, SB) showed significant (P < 0.05 = 0.0000) variation among the treatment groups. Black pepper followed by red pepper and turmeric remained the best antifungal and preservative after 05, 10 and 15 days after treatment (DAT) in B-group (inoculated and uninjured fruits). However, in A-group (inoculated and injured fruits), lowest infection percent was recorded for black pepper followed by red pepper and turmeric after 05, 10 and 15 DAT. The response of all treatments was obvious after 15 DAT. No infection of test fungus was recorded when black pepper and red pepper were applied. Whereas, with the application of turmeric (8.33%) and garlic (9.33%) lowest infection percent was observed with no significant difference. The response of onion (4.33%), papaya seed (6%), clove (7.66%) and SB (15%) appeared moderate with no significant difference. After 30 days, the lowest number of spoilt fruits were recorded with the treatment of black pepper (01) and red pepper (01) followed by turmeric (02), clove (02), onion (02) and SB (02) in B-group. Based on best physical structure, maximum numbers of tomato fruits were observed in clove (04) followed by black pepper (03) and SB (03) when treated without any injury (B group). While in case of A group, almost all fruits in all treatments become spoilt after 30 days of treatment. It has been proved that locally available plant based preservatives have an excellent antifungal and preservative potential against post-harvest tomato fruit rot disease caused by Alternaria solani. These plant based preservatives increase the shelf-life of tomatoes without any health hazards. To the best of our knowledge, this is the first time conducted study, where antifungal and preservative potential of locally available plant products against post-harvest tomato fruit rot caused by Alternaria solani have been explored.

First Report of Postharvest Fruit Rot Disease of Hardy Kiwifruit Caused by Diaporthe eres in China.

Hardy kiwifruit (Actinidia arguta), as an economically important fruit crop growing in Northeast China with thin, hairless and smooth skin, is susceptible to postharvest decay. In September 2018, infected cultivar Kwilv fruits were obtained from a commercial farm in Liaoning province, northeastern China. The occurring incidence of the rot disease varied from 20% to 90% according to the fruit number in each box during a 7-day-long storage at room temperature, and the initial symptom included a small, soft, chlorosis to light brown lesion and later watery brown lesions. Pure cultures of the same characteristics were obtained from the isolated strains in four rotten fruits on PDA medium. The isolates grew into transparent radial mycelium on PDA in the first two days followed by abundant white, fluffy aerial mycelium. After 14 days, colonies formed white to light brown aerial mycelial mats with gray concentric rings, and they produced gray and embedded pycnidia. Alpha conidia of 4.4 to 8.8 µm × 1.4 to 3.3 µm (n = 50) were abundant in culture, hyaline, aseptate, ellipsoidal to fusiform, while Beta conidia at 20.5 to 28.6 µm × 1.0 to 1.4 µm (n = 50) were hyaline, long, slender, curved to hamate. These morphological characteristics were similar to Diaporthe species (anamorph: Phomopsis spp.) (Udayanga et al. 2014). For identification, DNA was extracted from three single isolates respectively , and the internal transcribed spacer (ITS) region, β-tubulin (BT), and histone (HIS) H3 gene were amplified by using primers ITS1/ITS4 (White et al. 1990), T1/T22 (O'Donnell et al. 1997) and HIS1F/HISR (Gao et al. 2017), respectively. The three isolates produced identical sequences across all three gene regions, which were submitted to NCBI (Genbank accession numbers MT561361, MT561360 and MT855966). Nucleotide BLAST analysis revealed that the ITS sequence shared 99% homology with those of ex-type Diaporthe eres in NCBI GenBank (MG281047.1 and KJ210529.1), so did the BT sequence that had 98% identity to D. eres (MG281256.1 and KJ420799.1) and the HIS 99% identity to D. eres (MG28431.1 and MG281395.1) (Hosseini et al. 2020, Udayanga et al. 2014). Pathogenicity was tested by wound inoculation on the cv. Kwilv fruits. Five mature and healthy fruits were surface-sterilized with 1% NaClO solution, rinsed in sterile distilled water and dried. Every fruit was wounded by penetrate the peel 1-2 mm with a sterile needle, and inoculated with mycelium plugs (5 mm in diameter) of the isolate on PDA, with five inoculated with sterile PDA plugs as controls. Treated fruits were kept in sterilized transparent plastic cans separately under high humidity (RH 90 to 100%) at 28°C. After five days, the same rot symptoms were observed on all fruits inoculated with mycelium while the control remained symptomless. The fungi was re-isolated from the lesions of inoculated fruits and identified as D. eres by sequencing, thus fulfilling Koch's postulates. The pathogenicity experiment was re-performed using D. eres conidial suspension (107 conidia/ml) in sterile distilled water in October 2019 and the same results were obtained. D. eres was recently reported to cause European pear rot in Italy (Bertetti et al. 2018). To our knowledge, this is the first report of D. eres causing a postharvest rot in hardy kiwifruit in China, leading to severe disease and thus huge economic losses in Northeast China. Accordingly, effective measures should be taken to prevent its spreading to other production regions in China.

​Effect of Different Post-harvest Treatments of Fungicides, Botanical Oils, Food Preservatives and Packaging on Black Mould Fruit Rot of Pomegranate

Background: Pomegranate is an important favourite fruit of tropical, subtropical and arid regions. In India, it is grown in almost all the states but a commercial orchard exists in Maharashtra and Gujarat. Some pathogens only attack the fruit from the inside, while the external surface of the fruit remains asymptomatic. Diseased fruit display poor shelf life and flavour quality attributes. Changes include losses in sugars, acids, characteristic aroma and development of off flavours. Therefore, identifying and quick characterization of these disease symptoms is paramount to their effective control and management. Since there is a lack of research work on pomegranate black mould fruit rot diseases caused by Aspergillus niger. Realizing the gaps concerns the environmental factors affecting the prevalence of disease development and management of disease, the present investigation was undertaken with the objective: Management of post-harvest black mould fruit rot diseases of pomegranate by different methods. Methods: The experiments were carried out at Division of Horticulture, UAS, GKVK, Bengaluru by using the variety Bhagwa with 11 treatments which are replicated four times in Completely Randomized Design (CRD). Different chemical, physical methods, food preservatives and packaging were used to control the black mould fruit rot. Result: The epidemiological studies revealed that injury to the pomegranate fruits was found to be a prerequisite for infection. All the fruits inoculated by cork-wounding, pin-pricking, scrapping and rubbing method exhibited symptoms of rot. Cork-wounding proved to be the most efficient method of inoculation. Post-harvest dipping in hot water, fungicides, oils and food preservatives provided effective control of the rot. Oil suspensions were found effective in preventing the rot. Food preservatives were also found effective in preventing rot, potassium metabisulphite (KMS) 0.5 percent followed by sodium benzoate 0.5 percent proved most effective against the rot in both pre-and post-inoculation treatments. Packaging for postharvest storage showed very good performance in maintaining the quality of pomegranate fruits up to 12 weeks.

ANTIFUNGAL ACTIVITY OF SOME ESSENTIAL OILS AGAINST POST-HARVEST FRUIT ROT OF SWEET PEPPER

A laboratory experiment was conducted using some essential oils, i.e., Eucalyptus globulus, Eugenia caryophyllata, Cinnamomum cassia and Zingiber officinale to study their effects at different concentrations (1, 2, 3, 4% v/v) on the radial growth and spore germinations of, Ulocladium chartarum, Aspargillus niger , Fusarium semitectum and Geotrichum candidum , which were Isolated and identified of postharvest rots pepper fruits obtained from local markets using potato dextrose agar. The obtained results generally indicated that the tested essential oils significantly inhibit growth of the tested fungi. Eugenia caryophyllata oil was the most effective one, followed by Eucalyptus globulus of all tested concentrations, flowed by Cinnamomum cassia and Zingiber officinale oils at high concentrations. Moreover, some of the tested fungi were affected by all used essential oils, while Aspargillus niger was the most resistant fungi. Conclusively; it could be concluded that the tested essential oils were effectively inhibited the growth of the tested fungi that cause postharvest fruit rot diseases of sweet pepper at different concentrations tested (1, 2, 3, 4%). The paper aimed to inhibit evaluate the effects of some salts as natural products on postharvest diseases of sweet pepper fruits under the laboratory conditions and evaluate their effects on the development and suppression of these diseases. To achieve these aims, four different salts i.e. sodium carbonate, sodium bicarbonate, potassium nitrate and calcium chloride was applied at four concentrations, 1, 2, 3 and 4% (w/v). Salt solutions were tested on the mycelial growth and spore germination of Ulocladium chartarum, Aspergillus niger, Fusarium semitectum and Geotrichum candidum. Seven days post incubation, mycelial growth of the tested fungi were completely inhibited by the used salt solutions. Generally, the obtained results indicate a clear impact for most used salts solutions especially sodium salts. Finally, the optimum concentrations of salts under the experiments study conditions for inhibiting of growth of Ulocladium chartarum, Aspergillus niger, Fusarium semitectum, and its spores. Geotrichum candidum were 4, 3, 3, 3% (w/v) for sodium carbonate, sodium bicarbonate, potassium nitrate and calcium chloride, respectively. Conclusively, the optimum concentrations of salts in this study for inhibiting growth of some fungi were 4, 3, 3, 3% (w/v) for sodium carbonate, sodium bicarbonate, potassium nitrate and calcium chloride, respectively.

First record of Colletotrichum alienum Causing postharvest Anthracnose disease of mango fruit in China.

Mango (Mangifera indica L.) is one of the world's most significant economic fruit crops, and China is the second-largest producer of mango (Kuhn et al., 2017). Postharvest mango anthracnose is caused by Colletotrichum species and reduce the self-life of mature fruit (Wu et al., 2020). Colletotrichum species also cause postharvest anthracnose and fruit rot disease of Apple, Banana and Avocado (Khodadadi et al., 2020; Vieira et al., 2017; Sharma et al., 2017). In July 2019, mango fruits cv. 'Jin-Hwang' were observed at different fruit markets (39°48'42.1"N 116°20'17.0"E) of the Fengtai district, Beijing, China, exhibiting typical symptoms of anthracnose including brown to black lesions in different size (≤ 2 cm) with identified border on the mango fruit surface. Later, the lesions were coalesced and extensively cover the surface area of the fruit. The lesions were also restricted to peel the fruit and pathogen invaded in the fruit pulp. About 30% of mango fruits were affected by anthracnose disease. The margins of lesions from infected mango fruits (n=56) were cut into 2 × 2 mm pieces, surface disinfected with NaClO (2% v/v) for 30 s, rinsed thrice with distilled water for 60s. These pieces were placed on PDA medium and incubated at 25°C for 7 days. Pure culture of fungal isolates was obtained by single spore isolation technique. Initially, the fungal colony was off white, and colony extended with time, turning light gray at the center. The morphological examination revealed that conidia were hyaline, oblong, and unicellular. The conidia were measured from 10 days old culture and dimensions varied from 13.3 to 15.8 µm in length and 4.6 to 6.1 µm in width. For molecular identification, a multi-locus sequence analysis; the Internal Transcribed Spacers (ITS) region, partial actin (ACT) gene, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene and chitin synthase (CHS-1) gene were amplified by using the primer sets ITS1/4 (White et al. 1990), ACT-512F/ACT-783R (Carbone and Kohn 1999), GDF1/GDR1 (Guerber et al. 2003) and CHS1-79F/CHS-1-354R (Carbone and Kohn 1999) respectively. The partial sequences of MTY21 were deposited to GenBank accessions (MT921666 (ITS), MT936119 (ACT), MT936120 (GAPDH) and MT936118 (CHS-1). All obtained sequences showed 100% similarity with reported sequences of Colletotrichum alienum ICMP.18691 with accessions numbers JX010217 (ITS), JX009580 (ACT), JX010018 (GAPDH) and JX009754 (CHS-1) which represented the isolate MTY21 identified as C. alienum by constructing Maximum Likelihood phylogenetic tree analysis using Mega X (Kumar et al., 2018). For the confirmation of Koch's postulates, the pathogenicity test was conducted on 36 fresh healthy mango fruits for each treatment. Fruits were punctured with the help of a sterilized needle to create 2mm2 wounds and inoculated with 10µL inoculum (107 spores/mL) of MTY21. Control mango fruits were inoculated with 10µL sterilized distilled water and incubated at 25 °C with 90% relative humidity. The lesions appeared at the point of inoculation and gradually spread on the fruit surface after 7 days post inoculation. The symptoms were similar to the symptoms on original fruit specimens. The re-isolated fungus was identified as C. alienum based on morphological and molecular analysis. Mango anthracnose disease caused by several Colletotrichum species has been reported previously on mango in China (Li et al., 2019). Liu et al. (2020) reported C. alienum as the causal organism of anthracnose disease on Aquilaria sinensis in China. C. alienum has been previously reported causing mango anthracnose disease in Mexico (Tovar-Pedraza et al., 2020) To our knowledge, this is the first report of C. alienum causing postharvest anthracnose of mango in China. The prevalence of C. alienum was 30% on mango fruit which reflects the importance of this pathogen as a potential problem of mango fruit in China.

First Report of Ceratocystis changhui Causing Postharvest Fruit Rot of Peach in Kunming, China.

The peach (Prunus persica (L) Batsch) is a predominant commercially grown stone fruit in China (Lee et al. 1990). Ceratocystis changhui is an aggressive pathogen causing typical black rot symptoms on corms of taro (Colocasia esculenta) (Liu et al. 2018), it has not been reported on other hosts. During the summer and autumn of 2013, a postharvest fruit rot disease was observed on several peaches at a farmer's market (N 25°02'; E 102°42') in Kunming City, Yunnan Province, China. The incidence of the disease varied from 5 to 20%. Necrotic spots were first observed on the infected peach fruit (Prunus persica cv. shuimitao). The spots enlarged gradually and developed into a brown, water-soaked and rotted lesion. Eventually, the whole fruit became soft, rotted and covered with a gray-brown mycelium (Fig. 1 A, B). The isolates were obtained from the symptomatic tissues incubated on slices of fresh carrot root (Moller et al. 1968). After 5 to 10 days of incubation, perithecia and mycelium were observed growing on carrot slices. Spore masses were removed from the apices of perithecia, transferred to potato dextrose agar medium (PDA) and incubated at 25°C for 5 to 10 days, followed by single-spore isolation. All eight single-spore isolates from peach fruits obtained in this study were deposited in the State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan Agricultural University, China. In culture, mycelium was initially white, gradually turned to greyish-green or brown (Fig. 1E, F). Measurements were made 7 days after the formation of perithecia. Perithecia (Fig. 1G) were black, globose, 185.71 to 305.56 μm × 142.86 to 264.29 µm and showed a long black neck, 600 to 957.14 µm (Fig. 1H). Ascospores (Fig. 1I) were helmet-hat shaped and 2.86 to 6.67 µm ×3.81 to 4.76 µm. Cylindrical conidia (Fig. 1J) 6.67 to 38. 95 µm × 2.86 to7.62 µm were observed. Chlamydospore (Fig. 1K), 8.57 to 13.33 μm × 5.71 to 9.52 μm, were ovoid or obpyriform, smooth. To further verify pathogen identity the internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS1F and ITS4 (Thorpe et al. 2005), and the total genomic DNA from the mycelia of five isolates was extracted using a CTAB method (Lee &Taylor 1990). The nucleotide sequences have been blasted and deposited in the GenBank database. Analysis of the ITS sequences from the isolates T1-1yp, T1-2yp, T2-1yp (GenBank accession no. KY580895-KY580897) showed 99% to 100% similarity with isolates C. changhui CMW43272 (KY643886), CMW43281 (KY643884), CMW46112 (KY643891) and CMW46113 (KY643892) from taro in China. Phylogenetic trees based on the maximum-likelihood (ML) method were constructed using MEGA 7. ITS sequences of other Ceratocystis spp. were attained from NCBI for comparative analysis (Liu et al. 2018), and Davidsoniella virescens (CMW11164) served as outgroup. The robustness of ML tree was evaluated with 1,000 bootstrap (BS) values. The pathogen was identified as C. changhui based on the phylogenetic analysis (Fig. 2). Three isolates (T1-1yp, T1-2yp, T2-1yp) were used for pathogenicity. Nine Prunus persica cv. yingzuitao fruits at early maturity (8 points out of 10) were wound inoculated with 200μL conidia suspension of the fungus (approximately 2.0 × 106 conidia / mL). Degreasing cotton dipped in sterile water was used to raise the humidity in preservation boxes. Boxes were incubated for 10 days at 25°C. Three peaches as controls were treated only with sterile distilled water in the same way. Symptoms of sunken lesions and fruit rot were observed two days after inoculation, and measured at 1.8 to 3.2 cm from the inoculation point within 5 days (Fig. 1C: right, D). The same pathogen was re-isolated from them confirming Koch's postulates. Control peaches remained symptomless. This fungus was morphologically and phylogenetically identified as C. changhui. To our knowledge, this is the first report of C. changhui on postharvest peach in Yunnan, China. The disease will affect quality and taste of peach, so it is critical to deploy appropriate management strategies to limit the fungus spread.

Isolation and Identification of the Fungi Causing Tomato Fruit Rot Disease in the Vicinity of Tandojam, Sindh

Background: The post-harvest tomato fruit rot disease is common threat to the tomato fruit, causing huge economic loss as revealed by (GOP, 2018). The present study was conducted for isolatation and identification of causative agent of tomato fruit rot in order to formulate the proper management stretegies. Methods: Study was conducted in three phases. Phase one included collection of tomato fruit samples from vicinity of Tandojam. In phase two pathogens were isolated from the samples at laboratory, while in the phase three pathogens were identified using standard procedures. Result: The experimental results indicated Alternaria solani as the main cause of post-harvest tomato fruit rot. The symptoms observed were presence of brown to black rot lesions on tomato fruits with distinct rings ranging from small pin-heads to whole surface of fruit. A total of six different fungi viz., Alternaria alternata, Aspergillus niger, Alternaria solani, Geotrichum candidum, Fusarium oxysporum and Rhizopus stolonifer were found to be associated with post harvest tomato rot. Significantly higher infection was recorded for A. solani (53.667%) followed by A. niger (16.333%) and G. candidum (13.00%). The lowest infection percentage was observed for F. oxysporum (2.333%), followed by A. alternata (4.00%) and R. stolonifer (9.00%). A. solani produced aerial mycelium with yellowish to reddish diffusible pigments. A. niger cultures were typically black and colonies were initially whitish to yellow and later became brown to black in colour. G. candidum produced white and nonaerial colonies. F. oxysporum produced circular, aerial mycelium initially white, later changed to light pink. R. stolonifer produced whitish to grey fuzzy colonies.

Effect of postharvest sodium benzoate treatment on some fruit parameters of two organic apple cultivars

The aim of this study was to investigate the effect of postharvest sodium-benzoate treatment on some fruit parameters of two organic apple cultivars (’Topaz’ and ’Florina’). Assessments were made at three times during storage: 17 November 2019, 20 December 2019 and 23 January, 2020. During every assessment dates, each fruit was observed separately, and determined the proportion of i) healthy fruits (%), ii) post-harvest fruit rot diseases iii) fruits with mechanical injury (%), iv) fruit with russetting (%), v) damage of codling moth and vi) Ca-deficiency symptoms. In addition, fruit weight (g) was measured at each assessment date. Sodium benzoate reduced the fruit decay and the proportion of healthy fruit was higher in this treatment compared to water treated fruit. This effect could be seen in all assessment dates and on both cultivars. Effects on other parameters were various according to cultivar and assessment dates. Loss of fruit weight was similar in both treatments and cultivars compared to control treatment.

Incidence Rates of Major Diseases of Kiwiberry in 2015 and 2016.

Incidence rates of diseases in kiwiberry orchards were investigated monthly from late June to late September in Gwangyang and Boseong in 2015 and 2016. The impact of postharvest fruit rot was investigated during ripening after harvest. Bacterial canker was only observed on one single tree in 2015, but black rot, powdery mildew, leaf spot and blight, and postharvest fruit rot diseases were problematic throughout the study period in both 2015 and 2016. Incidence rates of the diseases varied with kiwiberry cultivar, region and sampling time. Incidence rates of powdery mildew, leaf spot and blight diseases increased significantly during the late growing stages near fruit harvest, while black rot peaked in late August. Incidence rate of postharvest fruit rot on fruit without fruit stalks was less than half of fruit with fruit stalks, regardless of kiwiberry cultivars. Among the four cultivars, Mansu was relatively resistant to black rot and postharvest fruit rot diseases. In our knowledge, this is the first report of various potential pathogens of kiwiberry in Korea.

Occurrence of a Postharvest Fruit Rot Disease of Apples Caused by Boeremia exigua var. exigua in Ptolemaida Kozani, Greece

Occurrence of a Postharvest Fruit Rot Disease of Apples Caused by <i>Boeremia exigua</i> var. <i>exigua</i> in Ptolemaida Kozani, Greece

Neopestalotiopsis vitis sp. nov. causing grapevine leaf spot in China

Neopestalotiopsis species are associated with a post harvest fruit rot and trunk disease of grapevines in China. A new species, N. vitis was isolated from the grape leaf spot in Guangxi Province, China. Initially, small circular, slightly sunken, necrotic spots, developed on the leaf surface, mainly at the inter-vein regions. The spots enlarged rapidly turning irregular, necrotic and brittle with age. Phylogenetic species recognition based on analysis of the combined ITS, TUB2 and tef1 sequence data coupled with morphology distinguished N. vitis from all other known species in the genus. This is the first report of a Neopestalotiopsis species causing a leaf spot on grapevine.

CONTROL OF POSTHARVEST FRUIT ROT DISEASES OF KIWIFRUIT BY ANTAGONISTIC BACTERIUM BACILLUS SUBTILIS

Postharvest fruit rot diseases cause a severe loss of kiwifruit during cold storage, transportation, marketing and in retail stores and Botryosphaeria dothidea, Diaporthe actinidiae and Botrytis cinerea were known as their major pathogens in Korea. Bacillus subtilis SK1-2 which showed the strongest antagonistic effects on B. dothidea, D. actinidiae and B. cinerea in in vitro growth inhibition assays was selected as an antagonist isolated from kiwifruit vines in the course of searching for antagonist candidates for the biological control of the postharvest fruit rot diseases. Cell suspensions of B. subtilis SK1-2 were harvested according to the optimal growth conditions such as pH, temperature and carbon and nitrogen sources defined for its mass production and applied four or six times at intervals of 10 days onto kiwifruit vines grown in rain-proof and open-field orchards to test the control efficacies on the postharvest fruit rot diseases. The control efficacies of B. subtilis SK1-2 on the postharvest fruit rot diseases were 68.2 and 56.5% in rain-proof and open-field orchards treated four times, respectively, and 80.0 and 65.7% in rain-proof and open-field orchards treated six times, respectively, whereas those of the registered chemical fungicide thiophanate-methyl were 72.4 and 67.1% in rain-proof and open-field orchards, respectively. The novel antagonist has the potential to be developed as a potent microbial agent successfully controlling the postharvest fruit rot diseases of kiwifruit vines grown in rain-proof orchards in the future.

REACTIONS OF KOREAN NEW CULTIVAR 'HAEGEUM' AGAINST MAJOR DISEASES OF KIWIFRUIT

‘Hayward’ (Actinidia deliciosa [A. Chev] Liang et Ferguson), which has been cultivated since the late 1970s, is the most popular green kiwifruit cultivar in Korea. However, ‘Hayward’ has been known to be susceptible to problematic foliar diseases such as bacterial canker, bacterial leaf spot, powdery mildew and fungal leaf spot diseases on kiwifruit trees grown at open-field orchards during growing season and postharvest fruit rot diseases during storage and distribution of harvested fruits. ‘Haegeum’ (A. chinensis Planchon), which is one of the new yellow kiwifruit cultivars bred in recent years, has been released at growers’ orchards since 2007 in Korea. Responses of ‘Haegeum’ against the major diseases of kiwifruit were compared with those of ‘Hayward’. The disease severities of the foliar diseases of ‘Hayward’ and ‘Haegeum’ were investigated at six naturally infected orchards during the growing season in 2013. The incidence rates of the postharvest fruit rot diseases on the fruits harvested in 2012 were also investigated at two naturally infected orchards. The disease severities of bacterial canker and bacterial leaf spot diseases were 18.5 and 63.5% on ‘Hayward’ at an orchard, but the disease severity of bacterial canker was 1.2% and no bacterial leaf spot was observed on ‘Haegeum’, respectively. Furthermore the disease severities of powdery mildew ranged from 9.4 to 84.8% on ‘Hayward’ at four orchards, while those on ‘Haegeum’ were less than 20%. The disease severities of leaf spot diseases induced by several fungi were 27.4 and 5.0% on ‘Hayward’ and ‘Haegeum’, respectively. The incidence rates of postharvest fruit rot diseases were 18.5% on ‘Hayward’ and 3.3% on ‘Haegeum’. The overall results reveal that ‘Haegeum’ is relatively resistant to the major diseases of kiwifruit compared with ‘Hayward’.

First Report of a New Leaf Blight Caused by Phacidiopycnis washingtonensis on Pacific Madrone in Western Washington and Oregon.

In recent years, a leaf blight disease, consisting of browned, desiccated leaves occurring mainly in the lower parts of the canopy, has been observed during wet springs on Pacific madrone (Arbutus menziesii) in western Washington and Oregon. In May 2009 and 2011, severe outbreaks occurred and symptomatic leaves from madrones growing in the region were sampled to determine the causal agent. Two symptoms, leaf necrosis or blotching along the edges and tips of the leaves, and leaf spot, were observed. Small segments of diseased tissue were cut from the leaves, surface-disinfected, rinsed, and plated on malt extract agar. Fifty percent of the leaf blotch and 30% of leaf spot samples yielded a fungus that was fast-growing (20 mm diameter in 4 days at 25°C) and produced colonies that were a pale gray with dark gray reverse and a felty texture. On potato dextrose agar (PDA), pycnidia formed and exuded conidia in peach-colored droplets after 2 weeks under room temperature and light conditions. Pycnidia were spherical and 12.5 to 39.8 μm, average 24.2 μm in diameter. Conidia were hyaline, ovoid, and 5.8 to 8.5 × 3.1 to 4.7 μm (average 7.0 × 3.7 μm). The fungus was identified as Phacidiopycnis washingtonensis based on its morphology (1). To confirm the identity, the internal transcribed spacer (ITS) region of the rDNA was amplified with ITS1/ITS4 primers (2) and sequenced (GenBank Accession Nos. JQ743784 to 86). BLAST analysis showed 100% nucleotide identity with those of P. washingtonensis in GenBank (AY608648). The fungus was also isolated from lesions on green shoots and the petiole and leaf blade of dead attached leaves. To test pathogenicity, 3-year-old Pacific madrone seedlings (three for each isolate) were inoculated with five isolates of the fungus and maintained in the greenhouse (25°C); the experiment was conducted twice. Five leaves from each tree were cold injured (-50°C) at a marked 5 × 5 mm2 area with a commercial aerosol tissue freezing product prior to inoculation and five leaves were not cold injured. A 5-mm-diameter mycelial plug cut from the margin of 6-day-old PDA culture was applied to the marked areas on the upper leaf surface. The inoculated area was covered with moist cheese cloth and wrapped with Parafilm. Leaves treated with blank PDA plugs served as control. Leaves were enclosed in plastic bags to maintain moisture for the first 15 h post inoculation and cheese cloths were removed after 15 days. All cold-injured inoculated leaves showed symptoms of blight starting at 2 weeks after inoculation, and no symptoms appeared on the controls. On non-cold injured inoculated leaves, only one isolate caused symptoms (80% of all leaves). The fungus was re-isolated from diseased leaves. These results suggest that P. washingtonensis is able to cause foliar blight on Pacific madrone when leaves are subjected to cold stress. Increased disease severity on madrone observed in spring 2011 in Washington and Oregon may have been due to predisposition of foliage to extreme cold in November 2010 and February 2011. This fungus has previously been reported to cause a postharvest fruit rot disease on apple fruit and a canker and twig dieback disease of apple and crabapple trees in WA (1). To our knowledge, this is the first report of P. washingtonensis causing a leaf blight disease on Pacific madrone in North America.