Journal of Agricultural Research (2017) Vol. 4, (2) 79-85 79 Effect of Hot Water Treatment on Mango Postharvest Diseases: Stem End Rot and Anthracnose Yi Yi Mon ', Nang Kyu Kyu Win", Seint San Aye’, Yi Yi Soe’, Tin Aye Aye Naing ' Abstract The experiments were conducted at the Department of Plant Pathology, Yezin Agricultural Universi- ty from 2013 to 2016. The occurrences of postharvest diseases were examined on 22 mango cultivars col- lected from 25 mango growing areas of Myanmar during 2013. In this experiment, three fungal diseases - stem end rot (SER) caused by Phomopsis mangiferae, anthracnose (ANTH) caused by Colletotrichum gloeosporioides and aspergillus rot (ASPR) caused by Aspergillus niger - were commonly found. All cul- tivars exhibited SER disease in all the areas of collection. The mean severity of SER disease was 89% while those of ANTH and ASPR were 10% and 4%, respectively. Consequently, on the collected culti- vars, SER disease severity was ranging from 40% to 100%, The severity range of ANTH disease was from 20% to 60% and that of ASPR disease was from 10% to 60%. The effect of hot water treatments on ‘SER and ANTH diseases of mango fruits cv, Sein Ta Lone were investigated by using hot water of differ- ent temperatures, 45°C, 50°C, 55°C and 60°C, for 15 minutes. For SER disease, the treated fruits at 55°C and 60°C inhibited the disease symptom appearance until 12 days after treatment (DAT). For ANTH dis- case, the fruits treated at 55°C and 60°C had no disease appearance until 13 DAT. Therefore, by this study, hot water treatment at 55°C for 15 minutes could be recommended as one of the environmental friendly control measures to suppress common postharvest diseases, SER and ANTH on Sein Ta Lone cultivar. Key words: Stem end rot, anthracnose, aspergillus rot, hot water treatment Introduction Mango is one of the most important and deli- cious seasonal fruits and widely cultivated in the tropics and subtropics (Tharanathan et al., 2006). It is also a popular fruit and can grow well in various climate conditions in Myanmar. About 300 cultivars and 20 kinds of mango species are cultivated throughout Myanmar. Among different cultivars, ‘Sein Ta Lone’, *Yin Kwe’, ‘Shwe Hin Thar’ and “Mya Kyauk’ are of export quality with high sweet- ness level (Naing, 2003; Soe, 2008). However, a considerable amount of mango fruits is lost every year due to a number of biotic and abiotic stresses. Among the biotic stresses, postharvest fungal diseases are the most important causes to the loss of mango production. The post- harvest loss of mango is up to 25-40% in Myanmar (FAO, 2001; Wai, 2004). Among several posthar- vest diseases, stem end rot and anthracnose diseases are usually considered to be the most severe post- harvest disease of mango worldwide (Ko et al., 2009; Malik et al., 2010). The effective disease management is essential for export mangoes. Post- harvest diseases can be limited with the application of chemical fungicide; however, the use of fungi- cides can cause hazardous effects on humans and the environment. Therefore, non-chemical treatment like hot water dipping is the most effective quaran- tine method to prevent anthracnose and stem end rot diseases and widely used in many countries (Aveno and Orden, 2004). In Myanmar, there is little information on the control measures to suppress mango postharvest diseases. Moreover, application of hot water treat- ment is not so familiar a practice for disease control and there is no report concerning the effects of hot water treatment on postharvest diseases and quality of mango. The present study was, thus, carried out to investigate the occurrence of postharvest fungal Department of Plant Pathology, Yezin Agricultural University, Yezin, Nay Pyi Taw, Myanmar *Department of Horticulture and Agricultural Biotechnology, Yezin Agricultural University, Yezin, Nay Pyi Taw, Myanmar *Corresponding author. nangkyukyuwin@gmail.com © 2017 Yezin Agricultural University80 diseases of mango in Myanmar and to determine the effect of hot water treatment on the severity of stem end rot and anthracnose diseases and quality of mango cv. Sein Ta Lone, Materials and Methods The experiments were conducted at the Depart ment of Plant Pathology, Yezin Agricultural Uni- versity from 2013- 2016. Study - 1: Occurrence of Mango Postharvest Fungal Diseases in Myanmar Collection and isolation The occurrences of postharvest diseases were examined on 22 mango cultivars collected from 25 mango growing areas of Myanmar during 2013. Mango fruit samples of 75-80% physiological ma- turity by visual inspection of fruit shoulders were collected and transported to the laboratory of De- partment of Plant Pathology, Yezin Agricultural University, within 24 hours after harvesting. The fiuits were washed and surface sterilized by dipping, in 1% sodium hypochlorite solution for 5 minutes and then washed 3 times with running water, and allowed to dry at room temperature (27 + 5°C). Three disinfected fruits were incubated in each plas- tic box (30 em x 20 em x 20 cm) at high moisture (> 90%) for 2-3 days. Then the boxes were kept at room temperature (27 + 5°C) for 7-10 days. Posthar- vest fungal diseases were observed for their symp- tom appearance for about 10 days afier incubation. The infected tissues were taken by cutting small pieces and examined under a microscope to check spores and spore fruits. Then, the causal pathogens were isolated by tissue culturing method on 2% water agar medium and the plates were incubated at room temperature for 2-3 days. After the incubation period, a bit of growing hyphal colony was trans- ferred with a sterilized needle to potato dextrose agar (PDA : 200 g potato, 20 g dextrose, 20 g agar, IL water) medium to get pure cultured pathogens. Identification and confirmation of postharvest fungal diseases The characters of colonies grown on potato dextrose agar media were recorded and spore and spore fruits were identified using a microscope. The fungal pathogens identified were inoculated onto the healthy fruits to confirm the causal agent and its symptoms. Mycelial plugs of the causal fungi were used for inoculation. The symptoms and pathogens were checked again at 3-5 days after inoculation, that was, when symptoms appeared. Journal of Agricultural Research (2017) Vol. 4, (2) 79-85 Data collection Numbers of postharvest fungal diseases found on collected cultivars and the percent of postharvest disease severity were recorded on about 20 days after incubation. Disease progress of treated fruits was daily assessed by visual estimation. For the estimation of fruit area diseased, the whole fruit surface area was taken as 100% and thereby the infected area were determined by eye estimation to calculate severity. Percent of disease severity (DS %) was calculated by using the following formula (Cooke, 2006). sm) = Afe2 of infected fruits Total area of fruits X 100 Study - 2: Effect of hot water treatment on stem end rot and anthracnose diseases of ngo The experiment was laid out on a completely randomized design (CRD) including 6 treatments with 3 replications. Each treatment contained 3 fruits. Fruits preparation ‘The fruits of the tested mango cy. Sein Ta Lone were harvested from an orchard in Myittha Township, Mandalay Region and were transferred to the laboratory of Plant Pathology, Yezin Agricul- tural University, within 24 hours after harvest. The fruits were prepared as described in study 1. The fruits were sorted for uniformity in size and absence of mechanical damage or disease. Inoculum preparation and inoculation The causal fungi, Phomopsis mangiferae and Colletotrichum gloeosporides that were isolated from infected fruits, were freshly prepared for inoc- ulation of stem end rot and anthracnose diseases, respectively. The mycelial plug (5 mm) from active- ly growing area of 3-5 days old culture of each fun- gus was used. Before inoculation, the surface steri- lized fruits were made wounds of 10 mm in length and Imm in depth each at 3 places on the skin by using a sterile scalpel. One mycelial plug was placed in inverted position on each wound, After that, the inoculated fruits were incubated in steri- lized plastic box with high moisture (> 90%) to allow pathogen establishment, Three hours after incubation, the mycelial plugs were removed. Different temperatures for hot water treatment The inoculated fruits were treated by dipping in a temperature controlled hot water bath at different © 2017 Yezin Agricultural UniversityJournal of Agricultural Research (2017) Vol. 4, (2) 79-85 temperatures, 45°C, 50°C, 55°C and 60°C, for 15 minutes. The fruits were turned over manually dur- ing treatment time to ensure even distribution of heat. The inoculated fruits dipped in water at room temperature (27 + 5°C) were used as control. After giving treatment, the fruits were cooled down by placing them in running tap water for 10 minutes. Then the treated fruits were placed in sterilized plastic boxes with high moisture (> 90%) and incu- bated at room temperature (27 + 5°C) for 13-18 days. Data collection All the fruits were kept at 80-90% relative hu- midity and room temperature, 27°C, for two weeks. Disease progress of treated fruits was daily assessed by visual estimation and weight loss and total solu- ble solid contents were recorded at 13 days after treatment. Percent of disease severity (DS %) was calculated as described in study 1 and percent of weight loss (WL %) was calculated by using the following formula (Angasu et al., 2014). Intl weight of mango (@)-Fial weight of MANO) 5 jeg wie ‘nial weight oF mango) The total soluble solid content (TSS) of the pulp for each treatment was measured using digital pocket refractometer of 0-85°Brix range (ATAGO PAL-1) and expressed as percent TSS of the fruit. For measuring the TSS (‘Brix), three fruit juice samples each of 0.3 ml were placed into the well of refractometer and the data were recorded. Statistical Analysis The data was statistically analyzed by using GenStat software (version 9) and means were sepa- rated by least significant difference (LSD) at 5% level. Results and Discussion Occurrence of mango postharvest fungal diseas- es in Myanmar The mango postharvest fungal diseases were found to be stem end rot caused by Phomopsis man- giferae, anthracnose caused by Colletotrichum gloe- osporioides and aspergillu rot caused by Aspergillus niger. Disease severities of the mango fruits in the man- go growing areas of collection Mean severities of stem end rot, anthracnose 81 and aspergillus rot in all 25 areas were 89%, 10% and 4%, respectively. The range of severity of three diseases varied in the collection areas as shown in Table 1. ‘The severity of stem end rot disease ranged from 50% to 100% in all areas and 100% severit were found on the fruits in 15 areas. The fiuits from Meiktila showed the least severity, 50%. ‘As for anthracnose disease, the maximum se- verity, 50%, was found on the fruits of Meiktila and the minimum severity, 5%, on those of Zeyathiri. This disease did not occur on the fruits of 13 areas; Danuphyu, Hlegu, Kyaukpadaung, Lashio, Mag- way, Myaungmya, Natogyi, Nyaungshwe, Palate, Popa, Pyawbwe, Wundwin and Yetsout. Aspergillus rot disease severity was 20% on the fruits of 3 areas; Kyaukse, —Natogyi and Pyawbwe followed by that of Magway with 15%. The fruits from Nganzun showed the least severity, accounting 5%, In the remaining 17 areas, this disease was not observed. The present investigation stated that stem end rot and anthracnose were the more prevalent diseas- es in Myanmar during 2013. Malik et al., (2010) reported that anthracnose, stem end rot and bacterial canker were the commonly found major postharvest diseases of mango wherever it is grown, In Myan- mar, Htein (2011) found that the most occurring diseases of mango were anthracnose, stem end rot, powdery mildew and sooty mold. In India, the com- mon postharvest fungal diseases of mango were aspergillus rot, anthracnose, black rot, fusarium rot, alternaria black spot, stem end rot, rhizopus rot and scab (Sharma, 2014). The diseases occurring in My- anmar and in India were mostly similar. It might be due to the same environmental conditions of Indo- ‘Myanmar regions. Disease severities of the collected mango culti- vars ‘The highest severity. 85%, of stem end rot dis- ease was observed on all collected cultivars fol- lowed by that of anthracnose with 16% severity and aspergillus rot with 13% severity. The range of the severity of three postharvest diseases varied among, the collected cultivars (Table 1). Stem end rot disease was the most severe on nine cultivars; Aung Din, Be, Ma Ta Yar, Ma Thi, Sa Mate Sein Sar, Sein Ta Lone, Shwe Hin Thar, U ‘Thar Zan and War Soe with 100% severity. Thone Lone Ta Taung cultivar showed the minimum se- © 2017 Yezin Agricultural University82 Journal of Agricultural Research (2017) Vol. 4, (2) 79-85 ‘Table 1. Severity of three postharvest fungal diseases in the mango growing areas of collection and on the collected mango cultivars in 2013, Location Belin Danuphys Hlega Kyaukpadaung Kyaukse Kyaunggon Lashio 9 Magway 10 Meiktita TL Myaungmya 12) Myittha 13. Natogyi 14 Nganzun 15, Nyaungshwe 16 Palate 17 Phyo 18 Popa 19° Pyawbwe 20. Shwebo 21 Waw 22 Wundwin 23 Yetsout 24 Zabutiti “Mean SER= “Ayadaw Disease severity) SE ANT ASP R oF 100300 Ca) fo 0 0 oo 0 6 0 0 10 wo 1s 20 % 30 0 mo 0 0 oo 0 45 30 500 9% 0 100 1s oo 0 20 10 10 65 wo 0 0 0 0 Oo 7m Ww 8 0 0 0 0 9 2% 7m 30 10 so 2 oO 0 0 0 wo 0 0 m0 Oo Discase severity (74) SE ANF ASP R HR o 2 Be 10 9 10 3. Be War % 0 6 4 MaChitSu 80-2026 5 MaNawNwe 70 600 6 MaTaYar 100 30 7 Ma Thi 10 0 8 MyaTaBat = 60030 9 NamDocMai 80 0 0 10 PaTaMyar = 800 Nga Mout 11 Pan Swal 90 0 20 12 PhoeThaKyar 90 600 1p SAMMSIN ggg Sar 14 Sein Sar er) 15 ScinTaLone 100 20 0 16 ShweHin Thar 100 0 10 17 TaSuMae = 70-2010 18 ThoneHnit 8-200 19 Thone LoneTa 40 0 0 Taung 20 UTharZan = 100 21 War Soe 10 0 0 2 YinKwe 8 20 40 Mean se 6 © 2017 Yezin Agricultural UniversityJournal of Agricultural Research (2017) Vol. 4, (2) 79-85 ‘Stem end rot B30 2» 1 Boalddalcl™ lnlellideee 12345678 91011213 83 Anthracnose Control ease soc sessc “2-600 ‘ 12345678 910111213 Day after treatment (DAT) Figure 1. Effect of different hot water temperatures on the disease severities of stem end rot and an- thracnose on mango ev. Sein Ta Lone verity with 40%. The highest severity, 60%, of anthracnose dis- ease occurred on three cultivars; Aung Din, Ma ‘Naw Nwe and Phoe Tha Kyar. Five cultivars - Ma Chit Su, Sein Ta Lone, Ta Su Mate, Thone Hnit and Yin Kwe - showed the lowest severity, 20%, The fruits of 12 collected cultivars did not show anthrac- nose disease symptoms. ‘The maximum severity, 60%, of aspergillus rot disease was recorded on Aung Din and Be War cul- tivars. The least severity, 10%, was found on three cultivars - Be, Ta Su Mate and Shwe Hin Thar. There was no aspergillus rot disease on the fruits of 13 collected cultivars. Srivastava (1998) reported that there was a wide difference in cultivars for susceptibility de- pending upon the type of cultivars, age of the plant and agro climatic conditions of the area. In the pre- sent study, Thone Lone Ta Taung cultivar was at- tacked only by stem end rot pathogen with the Ieast severity. The difference observed in the severity of diseases among the cultivars could be due to the different resistant capabilities of the mango culti- vars. Although the severity of the three diseases un- der study was significantly different among the mango growing areas and cultivars, stem end rot was found to have maximum severity on all collect- cd cultivars and in all collection areas. Therefore, stem end rot can be regarded as the most predomi- nant disease in Myanmar. This result confirmed that stem end rot disease was found in various mango growing countries of the world as a major posthar- vest disease (Ko et al., 2009; Malik et al., 2010) Effect of hot water treatment on stem end rot and anthracnose diseases and quality of mango ev. Sein Ta Lone The initial symptom of stem end rot disease was observed as the irregular brown spots at the stem end of the untreated fruits at 3 days after treat- ment (DAT). The treated fruits at 45°C, 50°C, 55° and 60°C showed the initial disease symptoms at 7, 8, 11 and 12 DAT, respectively (Figure 1). At 13 DAT, the untreated fruits showed 100% disease severity while the fruits treated at 45°C, 50°C, 55 and 60°C appeared 97%, 85%, 16% and 4% of s verity, respectively. Disease severity was not signif icantly different between the untreated fruits and treated fruits at 45°C and 50°C, Hot water treatment at 55°C and 60°C showed good results in relation to the lower level of disease severity. For anthracnose disease, the untreated fruits showed circular or itregular, dark brown to black spots as the first visible symptoms of an- thracnose disease at 5 DAT. The fruits treated at 45°C and 50°C produced anthracnose symptoms at 7 and 8 DAT, respectively. The disease was not ob- served on the treated fruits at 55°C and 60°C until 13 DAT (Figure 1). At 13 DAT, the treated fruits at 50°C showed 20% of severity. The disease severity of 45 °C hot water treated fruits was not significant ly different from that of control. By this experiment, the hot water treatment could be proved as a reliable treatment to prevent the progress of stem end rot and anthracnose diseas- es of mango cy, Sein Ta Lone. It was found that dipping in hot water at 55°C and 60°C for 15 © 2017 Yezin Agricultural University‘Stem end rot 6 L Weight toss (%) Journal of Agricultural Research (2017) Vol. 4, (2) 79-85, Anthracnose Control muse sore assc s6orc Hot water temperature (©) Figure 2, Effect of different hot water temperatures on weight losses of Sein Ta Lone mango inoculated with stem end rot and anthracnose pathogens at 13 days after treatment minutes reduced the severities of both stem end rot and anthracnose diseases at 13 DAT. However, dip- ping at 60°C for 15 minutes caused black patches on the fruit skin immediately after treatment. There- fore, this temperature could not be used in practice on the mango fruits of ev, Sein Ta Lone although disease severity was lower. Tandon and Singh (1968) reported that hot wa- ter treatment at 55°C and 60°C for 15minutes checked anthracnose disease completely in cultivar Dashehari, Nelson (2008) reported that using hot water dip of mango fruits for 15 minutes at 49-55°C was effective for controlling postharvest diseases depending on the variety. Therefore, the effective temperature and dipping duration of hot water treat- ‘Stem end rot Total soluble solid (Brix) ment might vary depending on the cultivars. In stem end rot disease, weight losses were highly significantly different between the untreated fruits and treated fruits at 50°C, 55°C, 60°C. At 13 DAT, the weight loss ranged from 4% to 11% among the treatments. The untreated fruits showed the highest weight loss, 11%, compared to other treatments (Figure 2). The lowest weight loss, 4%, was recorded in the treated fruits at 55°C and 60°C. In anthracnose disease, at 13 DAT, the highest weight loss was found on the treated fruits at 45°C and the untreated fruits. The fruits treated at 50°C showed 5% of weight loss and the lowest weight Joss, 3%, was recorded in those treated at 55°C and 60°C. Although the weight loss was not significant- Anthracnose ‘uty —ab—ab—ab = Control — m4s'c 50°C a55°C 560°C Fo e é s Hot water temperature ('C) Figure 3 Effect of different hot water temperatures on total luble solid contents of Sein Ta Lone mango inoculat- ced with stem end rot and anthracnose pathogens at 13 days after treatment © 2017 Yezin Agricultural UniversityJournal of Agricultural Research (2017) Vol. 4, (2) 79-85 ly different between the treatment, 45°C, and con- trol, significant difference was found with other two treatments, 55°C and 60°C (Figure 2). For stem end rot disease, there were highly significant differences in total soluble solid content (TSS) between the treated and untreated fruits. At the end of storage (13 DAT), the fruits treated at 55°C had the highest TSS value, 19°Brix while the control mango fruits showed the lowest TSS value, 14’Brix, However, TSS values were not significant- ly different among three treatments, 45°C, 50°C and 60°C (Figure 3). For anthracnose disease, the total soluble solid content (TSS) of mango fruits was not significantly different among all treatments (Figure 3). At the end of storage (13 DAT), the mango fruits treat- ed at 55°C showed the highest TSS values, 20°Brix, followed by those at 45°C, 50°C and 60°C with the 19°Brix of TSS. The lowest TSS value, 17°Brix was observed in the untreated fruits (control). ‘Treating the fruits at 55°C for 15 minutes re~ duced the weight loss and maintained total soluble solid content (TSS). This result was in accordance with the finding of Angasu et al. (2014), who re- ported that hot water treated mangoes had higher TSS than the control. Similarly, the final TSS value of Keitt mango was higher in fruits treated with hot water at 50°C for 30 minutes (Djioua et al., 2009) and Kensington mango treated at 45°C for 30 minutes or 47°C for 15 minutes increased “Brix Glacobi et al., 2000). It can be concluded that hot water treatment at 55°C for 15 minutes could be suggested as one of the most suitable control measures to suppress the stem end rot and anthracnose diseases in “Sein Ta Lone’ mango fruits References Angasu, O.N., 0.G. Dessalgne and T.N, Tadesse. 2014, Effect of hot water Treatment on quality and incidence of postharvest disease of mango (Mangifera indicia L.) fruits. Asian Journal of Plant Sciences, 13 (2): 87-92. Aveno, J.L, and M.E.M. Orden. 2004. Hot water ‘treatment of mango: A study of fruit export comporations in the Philippines, KMITL Sci- ence Journal, 4 (1): 1685-2044. Cooke, B.M. 2006. Disease assessment and yield loss. In: The epidemiology of plant diseases, 2 Edition. Cooke, B. M., D. G. Jones and B. 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