NSave Nature to Survive

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1 ISSN: NSave Nature to Survive : Special issue, Vol. IV: : 2013 AN INTERNATIONAL QUARTERLY JOURNAL OF ENVIRONMENTAL SCIENCES POST HARVEST DECAY OF PINEAPPLE [(ANANAS COMOSUS L. (MERR)] DUE TO SOME MICRO FUNGI Purabi Sarmah Baruah and T. C. Sarma KEYWORDS Micro-fungi Post harvest Pathogenicity Proceedings of International Conference on Harmony with Nature in Context of Ecotechnological Intervention and Climate Change (HARMONY ) November 11-13, 2013, Gorakhpur, organized by Department of Zoology, Biotechnology and Environmental Sciences DDU, Gorakhpur University, Gorakhpur in association with National Environmentalists Association, India 133

2 NSave Nature to Survive QUARTERLY PURABI SARMAH BARUAH* AND T. C. SARMA Department of Botany, Gauhati University, Guwahati -14, Assam, INDIA ABSTRACT Pineapple [(Ananas comosus L. (Merr)] is an important fruit which is widely cultivated in Nort East India. There are some reports of the farmers and wholesaler regarding spoilage of this useful fruit in the field as well as in the market condition. Post harvest decay of pineapple due to micro fungi in the different store houses as well as in different markets in Barpeta, Assam was investigated by inspecting at regular intervals. Rotting and decay of the pineapple were observed regularly. Pineapple rot was high in the store houses as well as market condition. Total 8 (eight) mico fungi were isolated on decayed fruits. They were Aspergillus flavus, A. niger, Alternaria alternata, Fusarium moniliforme, F. oxysporum, Mucor mucedo, Penicillium funicolosum, Rhizopus stolonifer. Among these micro fungi, A. flavus showed the highest rate of occurrence (90.00 %) whereas Mucor mucedo (30.00 %) showed the lowest rate of occurrence. Pathogenicity test revealed that all the isolated fungi were pathogenic (+ ve) to the different fruits. INTRODUCTION Fruits are the source of vitamins and minerals which are most important for maintaining the good health of human being. Fruits contain high level of sugar, minerals and nutrient elements and their low ph value make them susceptible for fungal attack and are being rotten (Singh and Sharma, 2007). Fungi not only cause rot to a number of fruits but also reduce their market value (Arya, 2004). The post harvest fruits are mainly contaminated with fungi during the transportation and storage condition. Pineapple (Ananas comosus L. Merr) under the family Bromiliaceae is one of the most important commercial fruits of North East India. It is delicious, nutritive and very good source of minerals, vitamin A, vitamin B and vitamin C (Singh and Singh, 1974). Pineapple is considered to be one of the most amazing and important commercial fruit of the world today. It is popular all over the world because of its pleasant taste and flavor. It is also a source of bromelin, a digestive enzyme. In addition to being eaten as fresh, the fruit can also be canned and processed into different forms like jam, jelly, juice etc. Due to its increasing demand, it has become popular among growers. Prolonged period of storage and long distance exports are other factors for decay (Akamine et al., 1975). Degradation of fruits due to microorganisms were reported from Australia (Smith, 1983), Ivory Coast (Teisson et al. (1979), India and Malaysia (Abdulla and Rohaya, 1983), Abdullah et al. (1986) Black Heart disease of pineapple reported by Smith (1983) and Teisson et al. (1979), Pineapple diseases in India and Malayasia were also reported (Abdullah and Rahaya, 1983). Change of taste, alternation of texture of the fruits in post harvest conditions due to spoilage are reported by Singleton P. and Diana S. (1978). Three strains of Penicillium funiculosum viz. non-pigmented reverse strain P1 and two red pigmented strains P2 and P3 were isolated from diseased pineapple fruits (Lim and Rohrbach, 1980). Spoilage of Carica papaya, Citrus sinensis, Ananas comosus and Lycopersicon esculentum due to fungi were investigated by Akinmusire (2011). Keeping in view the above points, an attempt has been done to study fungi responsible for post harvest decaying of pineapple fruit. MATERIALS AND METHODS *Corresponding author The survey was carried out in different markets as well as different store houses of Barpeta town, Barpeta Road and Pathsala of Barpeta district in Assam during the year 2010 and 2011 during June to October. The collected samples wrapped with the cellophane papers and brought to the laboratory for investigation. The fungal organisms associated with diseased fruits were isolated according to the method described by Rawlins (1933) and Aneja (2003). Small pieces of the diseased fruits were surface sterilized with 0.01 per cent HgCl 2 solution and were repeatedly washed with distilled water. These pieces were then transferred to Czapek s Dox Agar High Medium taken in petri plates. After inoculation, the petri plates were incubated at 28 ±1ºC for 5 days and regular observation was carried out. The isolates were sub cultured to get pure culture in the slants and kept for 134

3 POST HARVEST DECAY OF PINEAPPLE Table 1: Showing micro fungi associated with post harvest decay of pineapple with spoilage pattern Sl. No. Spoilage Macroscopic symptoms Microscopic symptoms Organisms 1 Sunken large depression Colonies blue and then light yellow Colonial heads radiate, later splitting into several Aspergillus flavus Link green and ultimately dark yellow green columns conidiophore hyaline, the vesicle is globose and flask shaped that produced chains of rough conidia are borne directly on vesicle 2 Dark brown discolouration, sunken Black colonies with white edge Colonial heads are large, globose and dark Aspergillus niger van. spots, fruit become spongy with brown becoming radiate and splitting into Tieghem gas production several columns edge conidiophore. Stripes are smooth walled, conidia are globose and rough walled. 3 Sunken large depression The rot is characterized by round brown to black. The conidiophores which are relatively short Alternaria alternate Nees The rots become spongy and the affected flesh is and dark coloured. The conidia are dark coloured, streaked with black. Dark coloured muriform and with transverse and longitudinal septa. mycelium may develop in the surface of the lesions. 4 Whitish mycelial growth Colonies are white Macroconidia produced in chains. Fusarium moniliforme Macroconidia delicate, awl-shaped, slightly Sheldon crescent-shaped, tapering at both ends. 3-5 septate. 5 Whitish mycelial growth Colonies are white. The fungus Larger 3-5 septate conidia Fusarium oxysporum produces no aromatic odour Schlechtendahl 6 Water soaked wrinkled appearance The surface colony colour is Non septate hyphae, rhizoids, Mucor mucedo (Linne) with depression initially white becoming grey sporangiophore and sporangia are present. Brefeld to yellowish brown in time. Sporangiphores are usually unbranched, b rown colour. 7 Infected fruits become soft, watery Deep green colony found Conidiophores arise laterally from aerial hyphae. Penicillium funiculosum and discolouration appears The mycelium is well developed and copiously Thom branched. The hyphae are septate. Conidia elliptical, green and smooth. 8 Water soaked wrinkled appearance First colony is white later Non separate mycelium, sporagiophores are Rhizopus nigricans with depression become bluish black at maturity directly opposite to the branched rhizoids. Ehrenberg Sporangia are sub-globose and columella are sub-globose A similar result was revealed by Bagwan N.B. (2010). 135

4 PURABI SARMAH BARUAH AND T. C. SARMA Table 2: Frequency (%) of occurrence of micro-fungi isolated from the infected fruit Fruit sample Total number of samples Infected fruits Micro-fungi Frequency (%) 10 8 Aspergillus flavus Ananas comosus 10 9 Aspergillus niger Alternaria alternata Fusarium moniliforme Fusarium oxysporum Mucor mucedo Penicillium funiculosum Rhizopus nigricans Table 3: Pathogenicity Test Sl. No. Name of Fungal species Fruit inoculated Pathogenicity 1 Aspergillus flavus Ananas comosus + 2 Aspergillus niger + 3 Alternaria alternata + 4 Fusarium moniliforme + 5 Fusarium oxysporum + 6 Mucor mucedo + 7 Penicillium funiculosum + 8 Rhizopus nigricans + further experimental purposes. Isolated microorganisms were identified by microscopic methods [Agrios (2008), Bernett (1992), Ellis (1976), Raper and Thom (1949), Sing (1968), Rangaswami G. and Mahadevan A. (1998). For pathogenicity test, (Koch s Postulation) a loopful of fungal mat was inoculated aseptically to healthy pineapple by making small incision. The inoculated fruit was kept wrapping with cellophane bags. The progress of the spoilage was observed periodically. Frequency of occurrence of the micro fungi associated with the post harvest decay of fruits were ascertained by the following formula as per Van der Plank ( ). Nos. of infected fruits F = X 100 Total number of fruits Where, F = Frequency RESULTS AND DISCUSSION A total 8 (eight) numbers of micro fungi were isolated from infected post harvest fruits of pineapple which were collected from different markets and store houses from time to time. The identified micro fungi were: i. Aspergillus flavus Link ii. A. niger van. Tieghem iii. Alternaria alternata (Nees) iv. Fusarium moniliforme Sheldon v. F. oxysporum Schlechtendahl vi. Mucor mucedo (Linne) Brefeld vii. Penicillium funiculosum Thom viii. Rhizopus nigricans Ehrenberg Pathogenicity Test Pathogenicity of 8 (eight) fungal species isolated from infected pineapples [(Ananas comosus L. (Merr)] wound inoculation. Association of micro fungi with the post harvest decay of pineapple fruits and spoilage pattern as a result of post harvest infection is shown in detail (Table 1). Different micro fungi show different symptoms on the fruits with respect to their association with the collected fruit samples of pineapples. The per cent of frequency of occurrence shown in Table 2 is quite indicative of their highest and lowest frequency of occurrence. i.e. the frequency of occurrence is highest in case of Aspergillus flavus (90.00%) followed by Aspergillus niger and Fusarium moniliforme (80.00%), Fusarium oxysporum and Alternaria alternata (70.00%), Penicillium funiculosum (60.00%). The percentage of occurrence in case of Rhizopus nigricans were accounted as 50.00% each. The lowest percentage of occurrence was observed in case of Mucor mucedo (30.00%). Thus, it can be concluded that the Aspergillus flavus, A. niger and Fusarium moniliforme were more responsible for fruit decaying in store houses as well as in the fruit market than the others. Pathogenicity test revealed that the associated micro fungi were responsible for causing rot of pineapple fruits (Table 3). Akintobi et al. (2011) also revealed a similar result. CONCLUSION Fruits provide good dietary components to our day to day life. From the above study it was observed that a number of micro organisms were responsible for decaying of fruits in the storage as well as in the markets and making the fruits unsuitable for consumption. ACKNOWLEDGEMENT The authors express deep sense of gratitude to Prof. D.K. Jha, Head of the Department of Botany, Gauhati University for providing laboratory facilities for carrying out this piece of research work. The authors are also thankful to the respected faculty members of Botany department, G.U. for their encouragement. REFERENCES Abdullah, H. and Rohaya, M. A The development of black heart disease in Mauritius pineapple Ananas comosus cv. Mauritius during storage at lower temperature, Malayasian Agricultural Research and Development Institute Research Bulletin. 11(3): Abdullah, H. and Rohaya, M. A. and Zaipun, M. Z Storage study of pineapples (Ananas comosus cv. Sarawak) with special emphasis on black heart disorder, Malayasian Agricultural Research and Development Institute Research Bulletin. 14(2): Agrios, G. N Plant Pathology - Basic Procedure in the diagnosis of Plant disease, 5 th Edition, pp Akamine, E. K., Goo, T., Steepy, T., Greidanus and Iwoka, N Control of endogenous brown spot in fresh pineapples in post harvest 136

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