Diversity of Marine Fungi Isolated from Nellore Coast, Andhra Pradesh, India

IAETSD JOURNAL FOR ADVANCED RESEARCH IN APPLIED SCIENCES ISSN NO: 2394-8442

Diversity of marine fungi isolated from Nellore coast, Andhra Pradesh, India.

1G. Vidya Sagar Reddy, 2Ch. Vijaya 1Department of Biotechnology, 2 Department of Marine Biology, 1, 2 Vikrama Simhapuri University, Nellore, A.P. [email protected]. 2Corresponding author: [email protected].

Abstract: Biodiversity studies in a region helps to understand about the native species that exist in a particular region. This study aimed to investigate the biodiversity of marine fungi of Nellore coast. Both water and sediment samples were collected from the selected regions followed by serial dilution and isolation on Malt Extract Agar (MEA) medium. Sampling region 1 (SR 1), i.e., Tummalapenta was found to be rich source of marine fungal diversity. Total 21 different fungal isolates were obtained from this coast. Slide cultures were prepared and then stained by using lacto phenol cotton blue. Cultures were identified by morphology using standard fungal keys. Among all the isolates, Phylum Ascomycota was dominant. Sediment samples were found to be rich in fungal diversity than water samples. However, Penicillium, Cladosporium, and Aspergillus sp were found to exist in both water and sediment samples. This study further helps to identify the potential strains by further screening for the production of metabolites and other abilities.

Index Terms - Marine fungi, Morphological identification, diversity, Nellore coast.

I. INTRODUCTION

Ocean is rich in biodiversity. Marine Microbes are responsible for energy transfer, involve in many biological cycles and helps in nutrient recycling. Marine fungal diversity exist along coastal regions mainly, mangroves, sand-beach and estuarine conditions ( Hawksworth, 1991). Marine fungi regarded as a good source of enzymes that have various industrial and other environmental applications (Ebel, 2010, Morrison-Gardiner, 2002). Marine fungi was reported to differ in many chemical aspects but there exists relation with terrestrial fungi (Holler et al. 2000). Hyde et al. 1998 reviewed that Marine fungi grow at a varying conditions than that of terrestrial fungi the conditions that differ are salinity, pH, and water potential, higher concentrations of sodium (Na) ions, low temperature, oligotrophic nutrient environment and more hydrostatic pressure (Calvo, 2002, Chandralata, 1999).

Most of the terrestrial fungi were previously well studied for pharmaceutical purpose such as drug discovery etc., The marine fungi are rich in potential natural products and have attracted attention of researchers as extensive resources solely since the late Eighties (Sridhar and Prasannarai, 2001). It was estimated that 1500 species of Marine fungi exist excluding lichens (Deacon, 2005). However, the potentiality of marine fungi has not yet been fully investigated (Amend, 2014). Up to now less than 10 percent of fungal biodiversity has been discovered and much work has to be done (Mejanelle, 2000, Saravanan and Sivakumar, 2013). The present work is an attempt to study the biodiversity of the fungi of Nellore coast, for biotechnological applications.

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II. MATERIALS AND METHODS

A. Sampling area and sample collection Emphasis was given to choose the unexplored regions for sample collection. Samples were collected from the coast of Nellore District (Andhra Pradesh, Bay of Bengal). The sampling regions (SR) are Tummalapenta (SR1), Mypadu (SR2), Kodur (SR3), Muthukur (SR4), Thupilipalem (SR5) [Figure-1]. Water and shore sediment samples were chosen as the sample matrices. Water samples were collected from chosen area and preserved in labeled 500mL sterile plastic bottles. Sediment samples were collected using the Van Veen Grab by adopting the method used by Uwadiae and Ebonne, 2011. The Sediments were placed in distinctly labeled ziplock polythene bags, maintained in ice-cold conditions and transported to laboratory. Sample collection frequency was once in three months and samples were collected in triplicates during the years 2014 - 2016.

B. Isolation of fungi As the distance between the sampling regions and our laboratory are varying, samples processing was started after 2 hours of sample collection by transferring sample to preprepared nutrient medium (Malt extract agar (MEA) (Himedia, Mumbai) containing, malt extract 30 g/L, peptone 5 g/L, agar 15 g/L in 50 % filtered and sterile marine water (20 ppt)). In the case of sediment samples they were subjected to serial dilutions (10-1 – 10-5) after 2 hours of sample collection. 0.1 mL of the sample (no dilutions performed for water samples) was aseptically transferred by spread plate method. Malt extract agar supplemented with chloramphenicol 100 mg/L was used for isolation and enumeration of fungi. Inoculated plates were kept for incubation at 280C for 5-6 days (Bonugli-Santos et al, 2015).

Fig: 1 Sampling regions (Tummalapenta (SR1), Mypadu

(SR2), Kodur (SR3), Muthukur (SR4), Thupilipalem (SR5).

C. Identification of fungi: The isolated fungi were sub cultured for obtaining pure cultures which were used in the identification studies. Slide culture method was followed for the preliminary identification of fungi. The fungi grown on the slide cultures were stained by tease mounts using lacto-phenol cotton blue. Morphology of the fungal hyphae, sporulation patterns and mycelial structures were studied under a microscope (400 X). These results in combination with fungal identification keys were used in final identification of isolated fungi. The fungal keys used are Barnett and Hunter, 1972, Anisworth et al. 1973 a,b, Kohlmeyer and Kohlmeyer, 1979, 1991, Ainsworth and Bisby’s, 1971 Dictionary of the Fungi- sixth edition.

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III. RESULTS AND DISCUSSION

The fungi belonging to different genera were isolated and enumerated by plating techniques. Extent of dilution was divided by the concentration of fungi present in it and for all the samples it varied from 10-1 – 10-5. Well isolated colonies were obtained on MEA Medium. Colonies were subculture, purified and carried out lacto phenol cotton blue mount for morphological evaluation.

A total of 21 different fungal isolates were obtained from sediment and water samples collected from five sampling regions of Nellore coast, A.P, India. The identified isolates were classified based on the arrangement of genera under their respective orders and families.

Among the 21 isolates, Aspergillums and Penicillium sp. Were dominant in the samples collected from all the regions (SR1 to SR5). The yeast sp were found in the sediment samples of SR2 and SR4. Only a few fungi namely, Penicillium expansum, Cladosporium sp. and Aspergillus sp. Were identified in water samples. Raghukumar (1998) NIO, Goa studied the diversity of east coast of India reported that terrestrial fungi may undergo adaptations in marine or mangrove ecosystem as facultatives or may become indwellers or can be residents (Raghukumar and Raghukumar, 1998). It was also reported that Ascomycetes fungi exist in higher amounts than other filamentous and higher marine fungi (Gnavi et al. 2017). It implicates that the similar diversity spread over the west coast of India. Among the isolates, 19 fungal species were found in sediment samples. Fungi belonging to Ascomycota were dominant in all the sampling regions. Isolates obtained were represented by the taxonomical annotations signifying the distribution of fungi at different sampling points [Table – 1].

Of the 21 isolated fungi, 18 were distributed among 8 different genera and 4 unknown fungi along with a yeast species were yet to be identified. Present study has shown that these filamentous fungi were found in the samples from all the regions indicating that the biodiversity is good with some natural variations [Fig-2]. The sample region 1 (SR 1, Tummalapenta) has rich diversity compared with other regions. Through this study, it might be extrapolated that the observed diversity may exist all along the coast. Marine environment contains high salinity, high pressure and much varying conditions compared to terrestrial environment. Marine Fungi from such environment will definitely have enzymes which can work efficiently in such adverse conditions. Moreover, this rich biodiversity is worth exploiting for isolation of new products of economic importance (Raghukumar, 2017). In this direction, for an attempt is made to extract enzymes like phytases, amylases, lipases, proteases, and chitinases from marine fungi in further studies.

IV. CONCLUSIONS

Our studies clearly indicated that the chosen areas are blessed with good biodiversity. 21 different marine fungi were isolated from water and sediment samples of Nellore coast. Most of the isolated fungal species were known for their pharmaceutical and economic value. This preliminary study suggests that Nellore coast has rich biodiversity and helps to screen the isolated fungi for potential bioactive compounds and industrially important enzymes.

ACKNOWLEDGMENT

Authors are thankful to the authorities of Vikrama Simhapuri University, Nellore for providing financial assistance for this study under FRGS grant.

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TABLE I Fungal biodiversity in the Sampling Regions

Fungal biodiversity in Marine Sampling Regions (SR) System S.No Name of the fungi Taxonomic Classification 1 2 3 4 5 Phylum Family W S W S W S W S W S Ascomyco 1. Aspergillus awamori - + ------+ Trichocomaceae ta Ascomyco 2. Aspergillus clavatus - + - + - + - + - - Trichocomaceae ta Ascomyco 3. Aspergillus fumigatus - + - + ------Trichocomaceae ta Ascomyco 4. Aspergillus granulosus - - - - - + - + - - Trichocomaceae ta Ascomyco 5. Aspergillus nidulans - + - + - - + + - - Trichocomaceae ta Ascomyco 6. Aspergillus terreus + - - + - + - - - - Trichocomaceae ta Ascomyco 7. Aspergillus versicolor + + + - - - - + - + Trichocomaceae ta Ascomyco 8. Cladosporium sp. 1 + + ------+ Davidiellaceae ta Ascomyco 9. Cladosporium sp.2 + + - - + + - - + + Davidiellaceae ta Ascomyco 10. Fusarium sp. - + ------+ Nectriaceae ta Ascomyco 11. Halorosellinia sp. - + - - - + - - - + Xylariaceae ta Ascomyco 12. Leptosphaeria sp. - + - + - + - + - + Phaeosphaeriaceae ta Ascomyco 13. Penicillium sp. - + ------+ Trichocomaceae ta Ascomyco 14. Penicillium expansum + + - - + + - + - - Trichocomaceae ta Ascomyco 15. Savoryella sp. - + - - - + - - - + Hypocreaceae ta Ascomyco 16. Trichoderma sp. - + - + - - - + - - Hypocreaceae ta 17. Yeast sp. - - - + - - - + - - - - 18. Un identified 1 - + ------19. Un identified 2 + ------20. Un identified 3 ------+ - - - - - 21. Un identified 4 ------+ - - (+) Present, (-) Absent

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Fig. 2: Distribution of fungi in sampling regions.

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