ISSN – 0215-1162
International Journal on Coconut R & D - Vol. 31 No. 1, 2015
Antimicrobial Properties of Cocos nucifera: A Review - Lalitha Ramaswamy, R. Rajendran, U. Saraswathi, R. Sughanya and C. Geethadevi
Formulation of Zinc Rich Coconut Nutribar Designed for Athletes - Anusha Priyadarsini. K and Dr. Lalitha Ramaswamy
Ecofriendly Organosolv Process for Pulping of Tender Coconut Fibre - Jincy P.J., Anita Das Ravindranath and U.S. Sarma
Low Temperature Grafting of MMA on to Coir Fibre - Lakshmi N.S., Sarika Babu, Sumy Sebastian and P.K. Ravi
Characterisation of Silver Deposited Coir Fibers by Magnetron Sputtering - Melvi Chandy, U.S. Sarma, M.S. Latha and K. Shreekrishna Kumar
An Investigation of the Tender Nut Potential of Diverse Coconut (Cocos nucifera L.) Varieties/Forms in Sri Lanka - S.A.C.N. Perera, G.K. Ekanayake and H.M.N.B. Herath
Pollen Dispersal and Pollination Patterns Studies in Pati Kopyor Coconut using Molecular Markers - Siti Halimah Larekeng, Ismail Maskromo, Agus Purwito, Nurhayati Anshori Matjik and S. Sudarsono
Asian and Pacific Coconut Community Cord
Cord is a semi-annual Journal of the Asian and Pacific Coconut Community (APCC) devoted to coconut research and development (R & D). The APCC is the first commodity based organization established under the auspices of United Nations-Economic and Social Commission for Asia and the Pacific (UN-ESCAP) in 1969. The APCC is an independent intergovernmental organization, currently consisting of eighteen member countries, namely: Federated States of Micronesia, Fiji, India, Indonesia, Kiribati, Malaysia, Marshall Islands, Papua New Guinea, Philippines, Samoa, Solomon Islands, Sri Lanka, Thailand, Tonga, Vanuatu, and Vietnam. Jamaica and Kenya are associate members of the APCC. The objectives of the APCC are to promote, coordinate and harmonize all activities of the coconut industry to achieve the maximum socio-economic development of the industry. In addition to Cord, the APCC publishes The Cocommunity (monthly newsletter), Coconut Statistical Yearbook (yearly) and Cocoinfo International (semi-annual popular Journal on the coconut industry) and other ad-hoc publications. Cord welcomes original research articles on any aspect of the coconut industry. The views expressed in Cord do not necessarily represent those of the editors or the APCC. Although the editors are responsible for the selection and acceptance of articles, the responsibility for the opinions expressed and for the accuracy of statements rests with the authors.
The annual subscription rates (including postage) for two issues are as follows:
APCC Member Countries US$ 40.00 Non-APCC Member Countries US$ 50.00
For further particulars, please write or call:
Asian and Pacific Coconut Community P.O. Box 1343 Jakarta, Indonesia Tel. No. : (62-21) 5221712 to 13 Fax No. : (62-21) 5221714 E-mail : [email protected] Home page : http://www.apccsec.org
C o r d International Journal on Coconut R & D EDITORIAL BOARD
URON N. SALUM Editor-in-Chief and Executive Director Asian and Pacific Coconut Community (APCC)
D.L. JAYARATNE H.A. TILLEKERATNE Department of Microbiology Consultant in Development of Coconut Processing & Faculty of Science Marketing University of Kelaniya (Former Director General, Coconut Development Kelaniya 11600, Sri Lanka Authority, Sri Lanka) Email: [email protected] No. 18D, Temple Road Kalubowila, Sri Lanka DINA B. MASA Email: [email protected] Manager Product Development Department HENGKY NOVARIANTO Philippine Coconut Authority Coconut Breeders 152 Chico St., Project 2 Indonesian Palm Research Institute (IPRI) Quezon City, Philippines P.O. Box 1004, Manado 95001 Email: [email protected] North Sulawesi Province, Indonesia Email: [email protected] DIVINA D. BAWALAN Free Lance Consultant on Coconut Processing and H.P. SINGH Utilization Deputy Director General (Horticulture) B-44 Far East Asia Village Indian Council of Agriculture Research (ICAR) Marcos Highway, Bo. Mambugan Krishi Anusandhan Bhavan-II 1870 Antipolo City, Philippines Pusa, New Delhi 110012, India Email: [email protected] Email: [email protected]
DOMINIQUE BERRY HUGH HARRIES Director Moderator Tree Crops Department Coconut Information Exchange and Time Line CIRAD-Cultures Perennes 2 Beech Road, Broadway TA 80/PS3, Boulevard de la Lironde Weymouth, Dorset DT3 5NP 34398 Montpellier United Kingdom Cedex 5, France Email: [email protected] Email: [email protected] JAMES V. KAIULO ERLINDA P. RILLO Managing Director Chief, Tissue Culture Division KOKONAS Indastri Koporesen (KIK) P.O. Box. 81 Philippine Coconut Authority Port Moresby Albay Research Center Papua New Guinea Banao, Guinobatan Email: [email protected] Albay 4503, Philippines Email: [email protected] JOCELYN E. EUSEBIO Director GEORGE V. THOMAS Crops Research Division (CRD) Formerly Director Philippine Council for Agriculture, Forestry and Natural Central Plantation Crops Research Institute Resources Research and Development (PCARRD) (CPCRI) Department of Science and Technology (DOST) Vettimoottil House, Vakayar P.O. Paseo de Valmayor, Los Banos Konni 689698, Kerala, India Laguna 4030, Philippines Email: [email protected] Email: [email protected]
K.V.A. BAVAPPA NANTARAT SUPAKAMNERD Karooth Villa Senior Scientist (Soil Science) P. O. KAPPUR Horticulture Research Institute Kumaranellur, Palakkad - 679 552 Department of Agriculture Kerala, India 50 Phaholyothin Road Email: [email protected] Bangkok, Thailand Email: [email protected] L.C. PRIYANTHIE FERNANDO Additional Director and Head NARONG CHOMCHALOW Crop Protection Division Chairman of the Conservation and Development of Coconut Research Institute Coconut Oil Forum of Thailand and Lunuwila 61150 Former FAO-RAP Sri Lanka Production Officer (Industrial Crops) Email: [email protected] 770 Phahonyothin 32, Chatuchak Bangkok 10900 LEX A.J.THOMSON Thailand EU-FACT Team Leader Email: [email protected] Agri-Forestry Export Production Specialist Secretariat of the Pacific Community (SPC) P.K. THAMPAN Private Mail Bag, Suva President Fiji Islands Peekay Tree Crops Development Foundation Email: [email protected] M.I.G. 141, Gandhi Nagar Cochin 682 020, India LUISITO J. PENAMORA Email: [email protected] Division Chief lll Timber Utilization Division PEYANOOT NAKA Zamboanga Research Center Senior Scientist (Post Harvest and Processing Specialist) San Ramon, Zamboanga City Horticulture Research Institute Philippines Department of Agriculture Email: [email protected] Ministry of Agriculture and Cooperatives Phaholyothin Road, Chatuchak M. SYAKIR Bangkok 10900, Thailand Director Email: [email protected] Central Research Institute for Estate Crops Jl. Tentara Pelajar No. 1 PISAWAT BUARA Bogor 1611 Former Director Indonesia Horticulture Research Institute Email: [email protected] Department of Agriculture 50 Phaholyothin Road MICHEL DOLLET Bangkok, Thailand CIRAD Email: [email protected]
Campus International de Baillarguest PONCIANO A. BATUGAL TA-A-98/F Chairman, 34398 Montpellier Cedex 5 APCC Technical Working Committee (TWC) on Climate France Change and Poverty Reduction Project Proposal Email: [email protected] Block 5 Sacay Grand Villas
Los Banos, Laguna 4031 MIKE FOALE Philippines Honorary Research Consultant Email: [email protected] The School of Land Crops and Food Sciences
The University of Queensland PONNIAH RETHINAM 4 Carinya Close, Maleny Plantation Crops Management Specialist Queensland 4552 18, Lakshmi Nagar S. N. Palayam Australia Coimbatore 641007 Email: [email protected] Tamil Nadu, India
Email: [email protected] MILLICENT WALLACE
Botanist/Plant Breeder PRASERT ANUPHAN Coconut Industry Board Former Deputy Director General Ministry of Agriculture Department of Agriculture 18 Waterloo Road, Kingston 50 Phaholyothin Road Jamaica Bangkok, Thailand Email: [email protected] Email: [email protected] RAMON L. RIVERA TEVITA KETE Deputy Administrator for Research and Export Processing and Marketing Officer Development Branch (Value-Added Coconut Products) Philippine Coconut Authority Land Resources Division Don Mariano Marcos Ave., cor Elliptical Road Secretariat of the Pacific Community (SPC) Diliman, Quazon City Private Mail Bag, Suva Philippines Fiji Islands Email: [email protected] Email: [email protected]
RICHARD MARKHAM U.S. SARMA Research Program Manager - Pacific Crops Director Australian Centre for International Agricultural Central Coir Research Institute Research (ACIAR) P.O.-Kalavoor, Dist.-Alappuzha C/o Secretariat of the Pacific Community (SPC) Kerala-688 522 Private Mail Bag, Suva India Fiji Islands Email: [email protected] Email: [email protected] VIVENCIO C. GALLEGO ROLAND BOURDEIX Scientist I COGENT Coordinator Crop Protection Division Bioversity HRF, Cirad Umr CEFE Philippine Coconut Authority Davao Research Center Montpellier. 1990, Bd de la Lironde Bago Oshiro, Davao City 8000 Parc Scientifiques Agropolis II Philippines 34397 Montpellier Email: [email protected] France Email: [email protected] WALTER L. BRADLEY Distinguished Professor of Mechanical Engineering ROSA S. ROLLE School of Engineering and Computer Science Senior Agro-Industry and Post Harvest Officer Baylor University FAO Regional Office for Asia and the Pacific Waco, TX 76798-7356 Texas 39 Phra Atit Road USA Bangkok 10200, Thailand Email: [email protected] Email: [email protected] YUPIN KASINKASAEMPONG SOMCHAI WATANAYOTHIN Agricultural Senior Scientist Senior Agriculture Scientist Chumphon Horticultural Research Center Department of Agriculture 70, Moo 2, Wisaitai Subdistrict Phaholyothin Road, Chatuchak Sawee District, Chumphon 86130 Bangkok 10900, Thailand Thailand Email: [email protected] Email: [email protected]
EDITORIAL STAFF Associate Editors ……………………………………….... DEEPTHI NAIR, S. Assistant Director, APCC
MUHARTOYO Librarian / Documentalist, APCC
Editorial Assistant………………………………………. TATI M. KABIB Office Assistant, APCC
Cord 2015, 31 (1)
Table of Contents
Page:
Antimicrobial Properties of Cocos nucifera: A Review - Lalitha Ramaswamy, R. 1-6 Rajendran, U. Saraswathi, R. Sughanya and C. Geethadevi
Formulation of Zinc Rich Coconut Nutribar Designed for Athletes - Anusha 7-12 Priyadarsini. K and Dr. Lalitha Ramaswamy
Ecofriendly Organosolv Process for Pulping of Tender Coconut Fibre - Jincy P.J.,., 13-23 Anita Das Ravindranath and U.S. Sarma
Low Temperature Grafting of MMA on to Coir Fibre - Lakshmi N.S., Sarika Babu, 24-31 Sumy Sebastian and P.K. Ravi
Characterisation of Silver Deposited Coir Fibers by Magnetron Sputtering - Melvi 32-38 Chandy, U.S. Sarma, M.S. Latha and K. Shreekrishna Kumar
An Investigation of the Tender Nut Potential of Diverse Coconut (Cocos nucifera 39-45 L.) Varieties/Forms in Sri Lanka - S.A.C.N. Perera, G.K. Ekanayake and H.M.N.B. Herath
Pollen Dispersal and Pollination Patterns Studies in Pati Kopyor Coconut using 46-60 Molecular Markers - Siti Halimah Larekeng, Ismail Maskromo, Agus Purwito, Nurhayati Anshori Matjik and S. Sudarsono
Cord 2015, 31 (1)
Antimicrobial Properties of Cocos nucifera: A Review
Lalitha Ramaswamy1, R. Rajendran2, U. Saraswathi3, R. Sughanya4 and C. Geethadevi4
Abstract
Coconut is known as the ―wonder food‖ and is regarded as perfect diet because it contains almost all essential nutrients needed by the human body. The various products of coconut include tender coconut water, copra, coconut oil, raw kernel, coconut cake, coconut toddy, coconut shell and wood based products, coconut leaves, coir pith etc. Coconut is a unique source of various natural products for the development of medicines against various diseases and also for the development of industrial products. Almost all the parts of the palm have medicinal properties such as antibacterial, antifungal, antiviral, antiparasitic, antidermatophytic, antioxidant, hypoglycemic, hepatoprotective and immunostimulant property. The various medicinal properties of coconut and its products are summarized in this paper.
Keywords: coconut, antimicrobial properties
______1Head of the Department and Associate professor, Department of Nutrition and Dietetics, PSG College of Arts and Science, Coimbatore, India. Email: [email protected] 2Principal, PSG College of Arts and Science, Coimbatore, India. 3Associate professor, Department of Biochemistry PSG College of Arts and Science, Coimbatore, India. 4Junior Research Fellow, Department of Nutrition and Dietetics, PSG College of Arts and Science, Coimbatore, India.
1 Cord 2015, 31 (1)
Introduction All parts of coconut tree besides being used as food and commercially, are also rich in Plants have always played a major role in medicinal properties. Most of the parts of the treatment of human and animal diseases. coconut tree such as endosperm, coconut oil, Medicinal plants can be used in different forms: tender coconut water, inflorescence and root are (i) as raw materials for extraction of active being used in the Ayurvedic medicine for the compounds or (ii) for extraction of abundant but treatment of several clinical conditions. In view inactive constituents which can be transformed of its utilitarian value of the entire tree it is often by partial synthesis into active compounds as referred as Kalpavriksha (Pushpan et al., 2013). such as extracts or as traditional preparations (Mukherjee, 2008). It is the unique source of various natural products for the development of medicines Coconut is a drupe borne by the coconut against various diseases and also for the palm (Cocos nucifera), a member of the development of industrial products. The parts of monocotyledonous family Palmae. It is known as its fruit like coconut kernel and tender coconut the ―wonder food‖ and is regarded as perfect diet water have numerous medicinal properties such because it contains almost all essential nutrients as antibacterial, antifungal, antiviral, needed by the human body. Coconut palms are antiparasitic, antidermatophytic, antioxidant, grown in more than 80 countries of the world, hypoglycemic, hepatoprotective and with a total production of 61 million tonnes per immunostimulant property. Coconut water and year (FAO, 2010). India is the third largest coconut kernel contain microminerals and coconut producing country, having an area of nutrients, which are essential to human health, about 1.78 million hectares under the crop. In and hence coconut is used as food by the peoples India, the four south Indian states namely Kerala, in the globe, mainly in the tropical countries Tamilnadu, Karnataka and Andhra Pradesh (Deb Mandal and Mandal 2011). account for around 90% of the coconut production in the country (NMCE, 2007). Anti-bacterial activity of coconut Coconut is a very versatile and indispensable The antibacterial activities of coconut fruit for most people under the tropical belt. It is endocarp extracts using methanolic and aqueous a complete food is rich in calories, vitamins, and extracts showed a strong activity against Bacillus minerals. It is nourishing, strengthening and subtilis, Pseudomonas aeruginosa, fattening food. It has high oil content. The Staphylococcus aureus, and Micrococcus luteus protein is of high quality and contains all amino (Singla et al., 2011). The antimicrobial property acids essential for the growth and maintenance of coconut shell showed strong antibacterial of the body. It is rich in K, Na, Mg and S. The activity on Ecsherichia coli and Salmonella typhi energy value of the dried coconut is 662 calories (Verma et al., 2012). The antibacterial potentials per 100 g (Bakhru, 2000). The nutrient content of crude aqueous and n-Hexane extracts of the of nuts varies by species, but in general they husk of Cocos nucifera against forty-five strains provide rich sources of vegetable protein, of Vibrio pathogens and twenty-five other monosaturated and polyunsaturated fatty acids, bacterial isolates those normally implicated in dietary fiber, vitamins E & K, folate, food and wound infections were studied. The magnesium, copper, selenium and potassium. aqueous extract was active against 17 of the Nuts are also naturally low in saturated fatty tested bacterial and 37 of the Vibrio isolates; acids and sodium (O’Neil et al, 2012). Nuts also while the n-Hexane extract showed antibacterial provide phenols, phytosterols, flavanoids, activity against 21 of the test bacteria and 38 of proanthocyanidins, resveratrol and arginine; the test Vibrio species (Akinyele et al., 2011). these bioactive compounds, coupled with micronutrients such as vitamin E and selenium, Tender coconut water is given to cholera serve as antioxidants and are anti-inflammatory patients because of its saline and albumen (Bolling et al., 2010). content (Effiong et al., 2010). It is generally used to treat urinary infection and diarrhea. Three
2 Cord 2015, 31 (1) peptides lower than 3kDa were purified and proved to have potential cure for oral diseases identified from green coconut water by using (Alviano et al., 2008). reversed phase-high performance liquid Lauric acid is a natural compound that is chromatography (RP-HPLC), showing molecular the main acid in coconut oil and also resides in masses of 858Da, 1249Da and 950Da. These human breast milk. Studies have been done to peptides have remarkable potential to contribute prove the potentiality of lauric acid against many in the development of novel antibiotics from harmful pathogens. Batovska et al., in 2009 natural sources (Mandal et al., 2009).Bacterial worked on antibacterial activity of medium chain isolates obtained from fermented toddy were fatty acids and their 1- monoglycerids. In the checked for the activity against B. cereus, study they have reported that among all Listeria monocytogenes and E. coli which are monoglycerides, monolaurin displayed the common food borne pathogens that infect the greatest anti bacterial activity towards various gastro intestinal tract. The results showed that Gram positive strains. The antimicrobial two of the ten isolates could inhibit the indicator property of lauric acid against organisms, however, at different inhibition levels Propionibacterium acnes both in vitro and in (Krishnamoorthy and Arjun 2012). vivo was studied. The in vitro studies proved that Antimicrobial property of coconut leaf lauric acid inhibited the growth of P. acnes, and extracts against Acinetobacter spp., B. cereus, did not have much effect on resident flora of E. coli, S. typhi, Shigella dysenteriae, S. aureus, skin. The cytotoxicity effect of lauric acid Aspergillus flavus and A. niger were reported that higher concentrations of lauric acid investigated. The results showed that the leaf also did not affect viability of the cells, instead extract were active against all the organisms killed the acne causing bacteria. (Nakatsuji et al., except S. aureus and A. flavus (Ifesan et al., 2009). 2013). The phytochemical screening of coconut Anti-fungal activity flowers demonstrated the presence of alkaloids, flavonoids, phenols, phytosterols, tannins, Heating the coconut shells gives oil that is aminoacids and carbohydrates (Dyana and used against ringworm infections in the popular Kanchana, 2012). These phytochemicals present medicine of India. The alcoholic extract of ripe in coconut flowers have well known curative dried coconut shell has antifungal activity activity against several human pathogens. against Microsporum canis, M. gypseum, M. Antibacterial properties of aqueous and audouinii, Trichophyton mentagrophytes, T. methanolic extracts of 26 medicinal plants used rubrum, T. tonsurans and T. violaceum. The in Mexico to treat gastrointestinal disorders were extract showed antifungal activity against all studied. The results showed that coconut being dermatophytes tested with twice the one of the plants which possessed a strong concentration needed against Epidermophyton bactericidal activity against tested species flocossum (200 ug/ml. The activity was mainly (Alanis et al., 2005). attributed to the high content of phenolic compounds (Venkataraman et al., 1980). An The effects of oil-pulling against oral extensively study carried out by Ogbolu et al., microorganisms in biofilm models using (2007) revealed that the anti- fungal activity of different edible oils were investigated. The study coconut oil against 6 Candida sp. obtained from proved that oil-pulling using coconut oil clinical settings. They further reported that exhibited antimicrobial activity against S. coconut oil was active against species of mutans and C. albicans which are considered to Candida at 100% concentration compared to be the predominant microorganisms found in fluconazole. dental caries (Thaweboon et al., 2011). Owing to the high Lauric acid content in coconut flour, it Antiviral activity has used as a medication for oral sores (Taheri et Coconut oil is very effective against a al., 2010). Husk fibers extract of coconut was variety of viruses that are lipid-coated. The
3 Cord 2015, 31 (1) medium chain fatty acids in coconut oil destroy Conclusion the viruses by disrupting their membranes, Coconut has a potent antimicrobial activity interfering virus assembly and maturation (Arora against various microflora. Besides being et al., 2011). Coconut oil as an anti- HIV antimicrobial in nature, coconut and its products medication was administered to 15 HIV positive have other health benefits such as improving patients at different concentrations for 6 months. heart health, digestion, management of diabetes By the end of three months 50% of the patients mellitus and some other diseases. The utilization showed reduced viral load and by the end of 6th of coconuts on a regular basis can be month, eight patients showed reduced viral load encouraged. Food formulations with coconut and and favorable CD4/ CD8 count (Conrado 2000). its products can be made commercially viable Extraction of polyphenols from husk fiber was and popularized as a functional food. Drugs and done and the extract was checked for supplements can be formulated with coconut antimicrobial and antiviral activities. The products for prophylactic and therapeutic selective antibacterial activity of C. nucifera purposes. against S. aureus and Herpes Simplex Virus - 1(HSV-1) suggests that this plant may be useful References for topical application in wound healing Akinyele, T.A., 2011. Assessment of the (Esquenazi et al., 2002). antibacterial properties of n-Hexane Antiprotozoan activity extract of Cocos Nucifera and its interactions with some Conventional The antihelmintic assay was performed on antibiotics. Masters Dissertation. chironomus larvae by Mariselvam et al., 2013. University of Fort Hare, Alice. The crude extract of the coconut inflorescence not only confirmed inactivation of helminthes, Alan´ıs, A.D., Calzada, F, Cervantes J.A., but also caused death in the shorter time as Torres, J., and Ceballos, G.M. 2005. compared to standard drug albendazole. The in Antibacterial properties of some plants vitro leishmanicidal effects of coconut husk used in Mexican traditional medicine for extracts on Leishmania amazonensis were the treatment of gastrointestinal disorders, evaluated. These results showed that extract of Journal of Ethnopharmacology. 100, 153– coconut husk at 10 µg/ml was a strikingly potent 157. leishmanicidal substance which inhibited the Alviano, W.S., Alviao, D.S., Diniz, C.G., growth of both promastigote and amastigote Antoniolli, A.R., Alviano, C.S., Frias, developmental stages of L. amazonensis after 60 L.M. 2008. In vitro antioxidant potential of min, presenting no in vivo allergenic reactions or medicinal plant extracts and their activities in vitro cytotoxic effects in mammalian systems. against oral bacteria based on Brazilian A combination of specially prepared folk medicine. Arch Oral Biol. 53:545- extracts of onion (Allium cepa) and coconut was 552. tested against the organisms causing Arora, R, Chawla, R, Marwah, R, Arora, P, gastrointestinal infection in animals. The sheep Sharma R.K., Kaushik, V, Goel, R, Kaur, with gastrointestinal helminthic infection were A, Silambarasan, M, Tripathi, R.P., and fed with extract for 8 days containing each 60 g Bhardwaj, J.R. 2011. Corporation. coconut and onion extract. The results showed Evidence-Based Complementary and that the worm stages disappeared from the feces Alternative Medicine. Hindawi Publishing. and were also not found 9 and 20 days after the end of the feeding with the extract (Mehlhorn et Bakhru, H.K. 2000. Foods That Heal. Orient al., 2011). Paper Backs, New Delhi. Batovska, I.D., Todorova, I.T., Tsvetkova I.V., and Najdenski H.M. 2009. Antibacterial Study of the Medium Chain Fatty Acids
4 Cord 2015, 31 (1)
and Their 1- Monoglycerides: Individual bacteria from fermented toddy of Cocos Effects and Synergistic Relationships. nucifera, J. Acad. Indus. Res. Vol. 1(3). Polish Journal of Microbiology. 58: 43- Mandal S.M., Dey, S, Mandal, M, Sarkar, S, 47. Maria-Neto, S. and Franco, O.L. 2009. Bolling, B.W., McKay, D.L., Blumberg, J.B. Identification and structural insights of 2010. The phytochemical composition and three novel antimicrobial peptides isolated antioxidant actions of tree nuts. Asia Pac J from green coconut water. Peptides. 30. Clin Nutr; 19: 117-23. 633-637. Conrado S. Dayrit. 2000. Read at the XXXVII Mariselvam, R, Ranjitsingh, A.J.A., Nandhini, Cocotech Meeting, Chennai, India U.R.A. and Kalirajan, K. 2013. Antihelmintic and antibacterial activity of Deb Mandal M, Mandal. S 2011. Coconut Cocos nucifera tree inflorances crude (Cocos nucifera L.: Arecaceae): In health extract. IJSID, 3 (2), 311-316. promotion and disease Prevention. Asian Pacific Journal of Tropical Medicine, 241- Mehlhorn H, Al-Quraishy S, Al-Rasheid KAS, 247. Jatzlau A, and Abdel-Ghaffar F. Addition of a combination of onion (Allium cepa) Dyana, J.P., and Kanchana, G. 2012. Preliminary and coconut (Cocos nucifera) to food of Phytochemical Screening of Cocos sheep stops Gastrointestinal Helminthic Nucifera L. Flowers, International Journal infections. Parasitol Res (2011) 108:1041– of Current Pharmaceutical Research, Vol 1046. 4, Issue 3. Mendonça-Filho R.R, Rodrigues I.A, Alviano Effiong, G.S., Ebong, P.E., Eyong, E.U., Uwah, D.S, Santos A.L.S, Soares R.M.A, Alviano A.J., and Ekong, U.E. 2010. Amelioration C.S, Lopes A H.C.S., Rosa M.S. 2004. of Chloramphenicol Induced Toxicity in Leishmanicidal activity of polyphenolic- Rats by Coconut Water, Journal of rich extract from husk fiber of Cocos Applied Sciences Research, 6(4): 331-335. nucifera Linn. (Palmae) Research in Esquenazi D, Wigg M.D, Miranda M.M.F.S., Microbiology 155: 136–143. Rodrigues H.M., Tostes J.B.F., Rozental Mukherjee PK, Kumar SN and Heinrich M S, da Silva A.J.R. and Alviano, C.S 2002. (2008). Plant Made Pharmaceuticals Antimicrobial and antiviral activities of (PMPs)- Development of Natural Health polyphenolics fromCocos nucifera Linn. Products from Bio-Diversity. Indian J. (Palmae) husk fiber extract. Research in Pharm Educ. Res 42(2), 113-121. Microbiology 153: 647–652. Nakatsuji, T, Kao M.C., Fang, J.Y., Zouboulis, Food and Agricultural Organization of the C.C., Zhang, L, Gallo R.L. and Huang United Nations, Economic and Social C.M. 2009. Antimicrobial Property of Department. Statistics division (September Lauric Acid against Propionibacterium 2, 2010). FAOSTAT- Production- Crops acnes: Its Therapeutic Potential for [Selected annual data]. Retrieved April 14, Inflammatory Acne Vulgaris, Journal of 2011 from the FAOSTAT database. Investigative Dermatology, Volume 129. Ifesan, B.O.T., Fashakin, J.F., Ebosele, F, and NMCE. Report on Copra. National Multi- Oyerinde, A.S. 2013. Antioxidant and Commodity Exchange of India Limited; Antimicrobial Properties of Selected Plant 2007, 1-14. Leaves, European Journal of Medicinal Plants 3(3): 465-473. O’Neil, C.E., Keast, D.R., Nicklas, T.A. and Fulgoni V.L. 2012. Out of-hand nut Krishnamoorthy, M. and Arjun, P. 2012. consumption is associated with improve Probiotic and antimicrobial activity of nutrient intake and health risk markers in
5 Cord 2015, 31 (1)
US children and adults: National Health and Nutrition Examination Survey 1999- 2004. Nutr Res 32: 185–194.
Ogbolu D.O., Oni AA, Daini OA, and A.P.
Oloko 2007. In Vitro Antimicrobial
Properties of Coconut Oil on Candida
Species in Ibadan, Nigeria, J Med Food 10
(2), 384–387.
Pushpan R, Kumari H, Nishteswar K and N Vishwanathan. 2013. Preliminary Phytochemcial Screening of Narikelapushpa (Flower of Cocos nucifera L.) Global Journal of Traditional Medicinal Systems, 2(2): 1-3. Singla R.K., Jaiswal N, Bhat V and Hitesh Jagani 2011. Antioxidant & Antimicrobial Activities of Cocos Nucifera Linn. (Arecaceae) Endocarp Extracts Indo Global Journal of Pharmaceutical Sciences; 1(4): 354-361.
Taheri J.B., Espineli F.W., Lu H, Asayesh M, Bakshi M, Nakhostin M.R 2010. Antimicrobial effect of coconut flour on oral microflora: An in vitro study. Res J Biol Scs. 5(6): 456-459. Thaweboon S, Nakaparksin J, Thaweboon B.2011. Effect of Oil-Pulling on Oral Microorganisms in Biofilm Models, Asia Journal of Public Health, Vol. 2 No. 2.
Venkataraman S, Ramanujam T.R, Venkatasubbu V.S. 1980. Antifungal activity of the alcoholic extract of coconut shell—Cocos nucifera Linn. J. Ethnopharmacol. 2: 291–293. Verma V, Bhardwaj A, Rathi S. and Raja R.B 2012. A Potential Antimicrobial Agent from Cocos nucifera mesocarp extract; Development of a New Generation Antibiotic. ISCA Journal of Biological Sciences. Vol. 1(2), 48-54.
6 Cord 2015, 31 (1)
Formulation of Zinc Rich Coconut Nutribar Designed for Athletes
Anusha Priyadarsini. K1 and Dr. Lalitha Ramaswamy2
Abstract
The main dietary goal for athletes is to obtain competent nutrition to improve health, fitness and sports performance. Athletes require nutritious convenient foods like nutribars to provide instant energy and other nutrients. Zinc supplementation has been shown to increase anabolic hormone status to meet catabolic activity in athletes which is profound during training. In the current study nutribar was formulated using zinc rich food sources namely dehydrated coconut meat, cashewnuts, pumpkin seeds and cocoa. Two variations of the nutribar were prepared variation I using dark chocolate and variation II with milk chocolate. The samples were subjected to organoleptic evaluation using a 9 point Hedonic scale by semi trained panel members. The nutribars were also analyzed for selected nutrients using standardized procedures. The mean score obtained for over all acceptability was 8.0±0.632, 8.7±0.458 and 8.5±0.5 by the control, variations I and II respectively. Variations I and II had obtained a mean score of more than 8.0 for flavour and taste, while the control sample had obtained lesser scores. The zinc content of variations I and II was 7.38 mg & 5.2 mg being greater than the control.
Keywords: Dehydrated coconut meat, Cashew nuts, Cocoa, Pumpkin seeds, Athletes, Zinc, Nutribar
______1AnushaPriyadarsini. K, Research Scholar, Department of Nutrition and Dietetics, PSG College of Arts & Science, Avinashi road, Civil Aerodrome, Coimbatore-641014, Tamilnadu, India. Email: [email protected].
2Dr. LalithaRamaswamy, Associate professor and Head, Department of Nutrition and Dietetics, PSG College of Arts & Science, Avinashi road, Civil Aerodrome, Coimbatore-641014, Tamilnadu, India. Email: [email protected].
7 Cord 2015, 31 (1)
Introduction deficiency can compromise muscle function (Lukaski HC, 2000). Dietary zinc intake and Sports nutrition is a part of nutrition plasma zinc each have a positive association science that relates to the interaction of nutrition with BMD in men leading to osteoporosis and physical activity (Maughan, 2001). Nutrition (Tiasun H Hyun, 2004). Therefore in order to plays a vital role in sports performance. Dawn augment zinc deficiency in athletes and to Jackson, RD (2003) spokeswoman for the enhance their endurance activity level, this American Dietetic Association, concurs; nutribar was designed. The objective of the study nutribars are convenient, especially for is to formulate zinc rich coconut nutribar for physically active individuals. The coconut has athletes and finding out its organoleptic been a traditional food in almost all the countries acceptability and nutrient content. where it is grown. It enters the diet of the people many ways, the water from the tender coconut is Materials and methods drunk and the mature nuts are used in cooking in Selection of ingredients the preparation of sweetmeats and for house hold oil production (Grimwood, 1975). Increasingly, There are several zinc rich food sources there is an emphasis on the synergistic relation like pumpkin seeds, sesame seeds, squash seeds, between diet and exercise for well being and a watermelon seeds, dark chocolate, wheat germ growing awareness of the beneficial role that (Edward, 2013). The researcher selected mineral element nutrition may play in achieving dehydrated coconut meat, cashew nut and good health and enhanced physiologic function. pumpkin seeds as zinc sources as they have 5mg, In contrast with the role of macro nutrients in the 6mg and 6.6mg/100gm respectively. These three body, micronutrients despite their relative ingredients were also found to blend well in the paucity in the diet and the body, perform final product after a pilot study. The chocolate important roles in regulating whole body coating given to the product also enhanced the metabolism, including energy utilization and zinc content dehydrated coconut meat is dried work performance. The importance of these coconut flesh which is widely used for extracting micro nutrients is revealed by the diversity of coconut oil and the remaining coconut meal is metabolic process they help to regulate. Zinc is a used for cattle feeding. In India, dehydrated ubiquitous element, that plays a fundamental role coconut meat is used for cooking food, apart in many cellular reactions. It is also an from using coconut oil. Dehydrated coconut intracellular cation, and is required to serve meat meets 42% of daily requirement of Zinc for either a catalytic or structural role by 300 an adult. The recommended dietary allowance of enzymes in mammals. (Ralf, 2003). Zinc Zinc for an adult per day is 12 mg (Sesikeran, containing enzymes participate in many 2010). The nutrient content of dehydrated components of macro nutrient metabolism and coconut meat is presented in Table 1. cell replication. In addition, some zinc Table 1. Nutrient Content of Dehydrated containing enzymes, such as carbonic anhydrase coconut meat per 100gm and lactate dehydrogenase, are involved in intermediary metabolism during exercise (Henry, S. No Nutrients Values 2000). Zinc is essential in keeping the immune 1 Energy 662 kcal system strong. For athletes, this is especially 2 Carbohydrate 18.4 gm important, since being side lined with a viral infection, or other illness, can force one to miss 3 Protein 6.8 gm valuable workout time. Zinc deficiency is 4 Fat 62.3gm problematic as plasma testosterone is regulated 5 Crude fibre 6.6 gm in part by zinc. Therefore, zinc deficiency may 6 Calcium 400 mg adversely affect this hormone, causing muscular 7 Phosphorus 210 mg mass and strength to suffer. Exercise increases 8 Iron 7.8 mg losses from the human body, and severe zinc 9 Zinc 5.0 mg Source: Nutritional value of Indian Foods, National Institute of Nutrition
8 Cord 2015, 31 (1)
Preparation of nutribar Nutrient analysis Dehydrated coconut meat, cashew nut and pumpkin seeds were purchased in a pure form The nutribars were analyzed for their fiber, from a reputed shop known for selling these protein, zinc calcium and iron. These nutrients products. Other ingredients namely cocoa were selected for analysis as they affect the powder, sugar and honey were purchased from a bioavailability of zinc. Protein was estimated by reputed departmental store. 100gm of the Kjeldahl method, fiber by AOAC method, zinc dehydrated coconut meat is taken and the brown by atomic absorption spectrophotometer, iron by outer skin was peeled and discarded as it was Wongs method and calcium was estimated by found to interfere with the organoleptic precipitating it as calcium oxalate. The characteristics. The pulp was flaked and toasted estimations were carried out in triplicates. in a frying pan till a fine aroma emanated from Sensory evaluation it. 20 gm of pumpkin seeds and 10 gm of cashew nuts were also dry roasted separately. All the Sensory evaluation is defined as a ingredients were coarsely powdered and kept scientific discipline used to evoke, measure, aside. Sugar syrup was prepared up to the thread analyze, and interpret those responses to stage in 1:1 ratio. The other ingredients were products that are perceived by the senses of added to the syrup and cooked in a low flame sight, smell, touch, taste, and hearing (Stone and with constant stirring till the thick mass did not Sidel,1993). The nutribars thus prepared were stick to the sides of the pan. The mass was then evaluated for the sensory attributes. Each of the allowed to set on a greased plate and cut into qualities namely appearance, texture, flavour, uniform sized pieces each in the shape of a bar taste and over all acceptability was assessed weighing approximately 50gm each and allowed using a 9 point Hedonic scale by a team of 10 to cool. One half (variation I) was coated with semi trained panel members. The scores given dark chocolate and other half (variation II) with by the panelists were statistically analyzed by milk chocolate as the preference for the dark and ANOVA. milk chocolate varies between individual. Results and discussion (Figure 1 shows a display of the formulated products). A control sample was also prepared Nutritional composition of Nutribar replacing dehydrated coconut meat, cashew nut, The nutrient content of the prepared pumpkin seeds and chocolate with oats and the nutribar is presented in Table 2. The nutribar had same method of preparation was followed. a protein content of 13.86, 15.60 and 15.82g% Figure 1. Formulated Coconut nutribar for the control, variations I and variation II respectively. The protein is essential for muscle building, and wear and tear of the athletes and it also enhances the absorption of zinc. The Zinc content of the nutribar was higher in variation I (7.38 mg %) due to the addition of dark chocolate, as it has more cocoa in it. Cocoa is one of the zinc sources. In dark chocolate, the amount of cocoa added being greater has enhanced the zinc content. The amount of zinc in variation II was 5.32mg of which was nearly 2mg lesser than the variation I while that of control was even lesser (3.08 mg %), as there was no zinc food source added to it. The fiber content of the both the variations was almost same (7.10g and7.14g% for variations I and II respectively) and was higher than the control
9 Cord 2015, 31 (1)
Table 2. Nutrient composition of Nutribar per 100gm S. No Nutrients Control Variation I Variation II (Dark chocolate) (Milk Chocolate) 1 Protein 13.86gm 15.60gm 15.82mg 2 Zinc 3.08mg 7.38mg 5.32mg 3 Fibre 6.70gm 7.10gm 7.14gm 4 Calcium 310.0mg 330.0mg 342.0mg 5 Iron 4.80mg 6.0mg 6.20mg
Table 3. Mean Scores of the organoleptic evaluation of nutribars
Sensory Control Variation I Variation II P value attributes (Dark Chocolate) (Milk Chocolate) Colour 8.4±0.663 8.0± 0.4 8.5± 0.670 0.342774 Texture 8.7 ± 0.458 8.5 ±0.670 8.3± 0.640 0.377798 Taste 7.8 ± 0.6 8.4± 0.663 8.4± 0.663 0.091761 Flavour 7.9 ± 0.7 8.7± 0.458 8.5± 0.670 0.028525 Over all 8.0 ± 0.632 8.7 ±0.458 8.5± 0.670 0.0282674 acceptability
Figure 2. Mean Scores of the organoleptic evaluation of nutribars
9 8.8 8.6 8.4 8.2 Control(Varies ±0.458 to 0.7) 8 7.8 Variation I ( Dark Chocolate) 7.6 varies from ± 0.4 to 0.67 7.4 Variation II( Milk 7.2 Chocolate)varies ± 0.5 to 0.67
10 Cord 2015, 31 (1)
(6.7g %). Addition of dehydrated coconut meat Consumption of the formulated nutribar added to the variation may be the reason for this. (100 gm) will help in contributing to the zinc The fiber concentration is high in dehydrated requirement, thereby enhancing the muscle coconut meat and palm kernel products power and endurance capacity of athletes. The (Stephaine, 2013). Calcium in the control group designed nutribar is organoleptically accepted was found to be 310mg whereas in variations I and nutritious in terms of protein, zinc, iron, and II it was 330.0mg and 342.0mg respectively. calcium and fiber. The formulated nutribar can The iron content was found to be 4.80mg in the be recommended as healthy nurturing snack for control and comparatively high in variations I athletes to increase their endurance performance. and II (6.0mg and 6.20mg respectively). Acknowledgments Sensory evaluation of nutribar My in-depth thanks to my guide Dr. The sensory attributes and the mean scores Lalitha Ramaswamy M.Sc., Ph.D Associate of the nutribar are indicated in Table 3. The Professor and Head, Department of Nutrition and mean scores obtained for over all acceptability Dietetics for her guide ship and support was 8.0±0.632, 8.7±0.458 and 8.5±0.5 throughout this research and Dr. R. Rajendran respectively by the control, variation I and II M.Sc., PGDEM., MBA., Ph.D Principal PSG respectively. Variations I and II had obtained a Arts & Science for permitting me to perform this mean score of more than 8.0 for flavour and research. I acknowledge PSG Institute of taste, while the control sample had obtained Medical Science & Research Institutional lesser scores. Results of ANOVA indicate no Human Ethics Committee recognized by significant difference (p≥0.05) in colour, texture SIDCER for approving my proposal. and taste between the three samples, while it was References significant (p≤0.05) for flavour and over all acceptability. This is due to the addition of Brain. E Grimwood., (1975) “Coconut palm chocolate and dehydrated coconut meat as it has products, Their processing in developing good flavour (Gohl, 1982) in the variations countries. Food and agricultural which have mutually enhanced the acceptability organization of united nations”. of the nutribar. Graphical representation of Dawn Jackson, RD., (2003). Speaker for the organoleptic scores are shown in figure 2. American Dietetic Association. Conclusion Edward., (2013) “8 Foods high in zinc”, Global The control sample had obtained highest healing centre. www.globalhealingcenter. mean score in the texture criteria compared to com/natural-health/foods-high-in-zinc the variations I and II. This is because the texture Göhl., (1982) FAO, Division de Production et of the oats being crispy and crunchy, whereas the Santé Animale, Roma, Italy. variations I and II had obtained higher mean scores in over all acceptability compared to the Gopalan. C, Rama Sastri B.V and control sample. Statistically it was found out that Balasubramaniam S.C, Narasinga Rao, there was no significant difference (p≥0.05) in Deosthale Y.G, Pant K.C., (2012) the colour, texture and taste between the three “Nutritive value of Indian Foods”. samples, while it was significant (p≤0.05) for National Institute of Nutrition, Indian flavor and over all acceptability. The addition of council of Medical Research. Hyderabad- dehydrated coconut meat, pumpkin seeds, 500007, India. cashew nuts and chocolate in the variations are not only nutritious, but also highly acceptable. Henry C Lukaski., (2000) “Magnesium, zinc, The nutribar provide almost one half of the daily and chromium nutriture and physical zinc requirement of athletes. activity”. American Society for Clinical Nutrition, Vol. No.72 (6), 585-593.
11 Cord 2015, 31 (1)
Lukaski HC., (2000) “Magnesium, zinc, and chromium nutriture and physical activity”.
American Society for Clinical Nutrition Vol. No.72 (6), 585-593. Maughan R.J., (2001) “Clinical sports nutrition” Roseville, NSW: McGraw-Hill Book company Australia Pty Lit. (pp.369-390).
Ralf R. Henkel1, Kerstin Defosse1, Hans-Wilhelm Koyro2, Norbert Weissmann3, Wolf- ll1 Bernhard Schi ., (2003) “Estimate of oxygen consumption and intracellular zinc concentration of human spermatozoa in relation to motility” Asian. J. Androl Mar; 5: 3-8
Sesikeran B., (2010). Revised RDA for Indians, National Institute of Nutrition ICMR, Hyderabad. Stephanie Henry., (2013) “Establishing nutritional value in dehydrated coconut meat and palm products fed to pigs”. http://www.journalofanimalscience.org/co ntent/91/3/1391. Stone Herbert, Sidel Joel., (1993) “Sensory Evaluation Practices”, Academic Press Incorporated, Jan 1, Science pp. 338. Taisun H Hyun, Elizabeth Barrett-Connor, and David B Milne., (2004) “Zinc intakes and plasma concentrations in men with osteoporosis: the Rancho Bernardo Study”. American Journal of Clinical Nutrition, vol. (80) no. 3, pp. 715-721.
12 Cord 2015, 31 (1)
Ecofriendly Organosolv Process for Pulping of Tender Coconut Fibre
Jincy P.J.1, Anita Das Ravindranath2 and U.S. Sarma3
Abstract
The huge biomass generated by vendors of tender coconut is the broken husks refuse dumped along roads and highways in Kerala. These dumps become breeding grounds for diseased causing germs and carrier mosquitoes causing threat to human life. In order to avoid pollution and find use of the rejected biomass of tender husks, a study was carried out on pulping of tender coconut husk fibre which could be used for papermaking. The optimum pulping condition, the quality of the pulp and its yield was evaluated using different variables like time and temperature. It was observed that the organosolv process could efficiently remove lignin from the tender coconut fibre yielding maximum cellulose. During the traditional pulping processes such as Kraft pulping to isolate the cellulose fibers for the production of paper, the hemicellulose and lignin fractions are degraded, limiting their valorization possibilities. Organosolv pulping has been advocated as the environmentally benign version of the kraft process. Unlike other pretreatment methods, organic solvents can easily be recycled and reused. The lignin dissolved by organosolv pulping is easily recovered by dilution and is unsulphonated and relatively unmodified. Products like handmade paper, egg cartons, handicraft items, garden articles like paper pots could be made from the organasolv pulp of tender coconut husk fibre.
Keywords: tender coconut, organosolv pulping, morphology, handmade paper, organosolv lignin
______1Department of Environmental Engineering, National Chung Hsing University, Taichung 402, Taiwan, ROC. Email: [email protected] 2Microbiology Department, Central Coir Research Institute, Kalavoor P.O, Alleppey Dist, PIN-688522 Kerala, India. Email: [email protected] 3Ex-Director, Research (RDTE), Central Coir Research Institute, Kalavoor P.O, Alleppey Dist, PIN-688522 Kerala, India. Email: [email protected]
13 Cord 2015, 31 (1)
Introduction polymer composed of mainly xylose (five- carbon) and further arabinose (five- carbon), The coconut palm (Cocos nucifera), an galactose, glucose and mannose (all six carbon); important member of the family Arecaceae it also contains smaller amounts of non-sugars (palm family) is a tropical tree that is cultivated such as acetyl groups. Hemicellulose, because of in nearly 90 different countries (Pires et al. its amorphous nature, is relatively easy 2004). It is estimated that out of total production tohydrolyzes. Hemicellulose attaches weakly to of coconuts in the country 10% are plucked as both cellulose and lignin and fills the intervening tender coconuts. The water of the tender spaces. Extractives are non-structural coconut, technically called the liquid endosperm, components of biomass samples that is soluble is enjoyed by people in tropical regions of the either in water or ethanol. It includes, but is not world, especially in Tropical Asia as well as limited to sucrose, nitrates, nitrites, protein, Central and South America. It was discovered chlorophyll, and waxes (fatty acids). The ash that this water has medicinal characteristics as an content is a measure of the mineral content and antioxidant, replenish fluid hydration, provide other inorganic matter in biomass. parenteral nutrition, etc. Despite its benefits, the market for coconut water causes solid waste Regardless of the application of environmental problems and is one of the major lignocellulosic materials, it is required a agro-industrial waste generators in some preliminary processing to separate the three developing countries. It is a lignocellulosic macromolecular fractions, particularly the lignin, material which is resistant to easy decomposition which can be considered the main physical due to its high lignin content and causes barrier for making the fibers (mainly cellulose) environmental pollution. The husk containing cemented together. These processes modify the about 80-85% moisture acts as breeding sites for lignocellulosic material by disruption of cell wall insectsmosquitoes, flies etc. With the objective structure of plant biomass, removing, of utilization of the tender coconut solubilizing or depolymerizing lignin. The kinds huskagrowaste and reducing the environmental of processes depend on the material used and the impact of its disposal, an ecofriendly organosolv proposed purpose of lignocellulosic fractions process for pulping of tender coconut fibre could utilization and may be mechanical, physical, be developed. biological or chemical. The development of pretreatment processes strong enough as to Tender coconut fibre is a lignocellulosic separate the cell wall arrangement and mild material and is composed of carbohydrate enough as to avoid a significant chemical polymers (cellulose and hemicellulose), lignin degradation of biomass components is a and a remaining smaller part (extractives, acids challenge for today’s chemical industry and salts and minerals). Cellulose is a polymer (Canetieri et al, 2007). For the novel composed of chains of six carbon sugars pretreatment methods it is advisable to use cheap (primarily glucose). These chains are bundled and easily recoverable chemicals and low-cost into strong fibers that on close inspection are equipment. The use of environmentally friendly seen to have an organized crystalline structure. and low energy-intensive approaches is highly Lignin is a complex, three dimensional polymer desired. The chemical cooking process is the composed of linked six-carbon phenolic rings most efficient and most used to perform the with various carbon chains and other chemical separation of lignocellulosic components of functionalities. Lignin is non-crystalline, and its vegetal biomass (Fernandez, 1996). It results in structure has been described as analogous to a enlargement of the inner surface area of substrate gel or foam. The lignin serves to bind the particles, accomplished by solubilization and/or cellulose fibers. It is degradable by only few degradation of hemicellulose and lignin. organisms, into several higher value products such as organic acids, phenols and vanillin. Via The shortage of raw materials and the chemical processes valuable fuel additives may large energy and water consumptions are be produced. Hemicellulose is a very complex presently the main concerns of pulp and paper
14 Cord 2015, 31 (1) industry. Considerable research effort has been availability. In the present study, a 70% aqueous made to introduce alternative species as raw solution of acetic acid was selected for tender materials, like biomass from agricultural and coconut fibre pulping. Dilute sulfuric acid was forestry residues. Organosolv processes were applied as the catalyst for organosolv pulping. experimented based on the use of organic The effect of cooking time and temperature on solvents as delignification agents, to degrade the pulp properties like yield, Kappa number and % lignocellulosic biomass to obtain cellulose fibers delignification were investigated. The bleaching for paper making, high quality hemicelluloses ability of organosolv pulp by totally chlorine free and lignin degradation products avoiding (TCF) bleaching using alkaline hydrogen emissions and effluents. Organosolv peroxide was also examined. pretreatment has been evaluated as an effective Conventional methods of bleaching pretreatment method for high lignin involve treatment with molecular chlorine or lignocellulosic biomass. The treatment can break chlorine based chemicals. Effluent from these down internal lignin and hemicellulose bonds processes produces large amounts of chlorinated and thus remove all of the lignin from biomass. organic compounds and releases into the A strong inorganic acid is usually applied in the environment. The interest in totally chlorine free organosolv pretreatment as a catalyst to bleaching processes has led to the development hydrolyze the lignin-lignin and lignin- of peroxide based bleaching. One of the biggest carbohydrate bonds in biomass. Inorganic acids advantage of hydrogen peroxide is that it is including hydrochloric acid, sulfuric acid and environmentally friendly throughout its whole phosphoric acid have been frequently chosen. life cycle. The perhydroxyl anion (HOO-) is the Generating high quality lignin is one of the principal active agent in peroxide bleaching. unique advantages of the organosolv This anion is a strong nucleophile which during pretreatment over alternative methods, such as bleaching converts electron rich chromophores steam explosion, dilute acid treatment and hot to their non chromophoric counterparts. The water treatment, where the only proposed use for reaction of lignin with peroxide is not reversible the lignin is as a boiler fuel. The organosolv and lead to the permanent removal of most of the spent liquor mixed with water could precipitate chromophoric groups in the lignin molecule the dissolved lignin. In contrast to lignin (Presley and Hill, 1996). produced by other technical processes, such as Kraft pulping, organosolv lignin is a sulfur free, Materials and methods low molecular weight product of high purity. Substrate preparation High quality lignin can be used as a substitute for polymeric materials, such as phenolic powder Tender coconut husk used in this work was resins, polyurethane foams and epoxy resins (Li collected from the local vendors in Alleppey et al, 2012). The other main advantages of the district. Husk was depithed using shredder and organosolv processes over the conventional ones the fibers were separated. After the separation of are the following: low environmental impact, fiber from pith, the fibers were chopped into higher pulp yield, ease of bleaching, easy solvent uniform sized pieces and air dried. The fibre had recovery, low capital for a new plant and an average moisture content of 11.7%. recovery of lignin and sugars for profitable Chemical analysis of tender coconut fibre utilization (Saberikhah et al, 2011). The specimens were sampled and In this investigation, the feasibility of an characterized according to the standard methods. organosolv pulping process for the production of Chemical composition, given on an oven dry pulp from depithed tender coconut husk fiber weight basis, was the following: 27.02% was evaluated. Acetic acid was selected as an Cellulose (Uppdegraf, 1969), 56.82% organic solvent for tender coconut husk fibre Holocellulose (ASTM D1104-56(1978), 36.9% pulping, considering the results obtained in Acid insoluble lignin ( TAPPI T 222 om-98), preliminary experiments and due to its easy 3.62% extractives in alcohol-benzene (TAPPI
15 Cord 2015, 31 (1)
T204 cm-97) and 3.1% Ashes at 5250C (TAPPI Fiber morphology using Scanning electron T211 om-02). microscopy (SEM) Organosolv pulping of tender coconut fiber The changes in the morphology of the fibers were examined microscopically. For Organosolv pulping involves contacting a scanning electron microscopy (SEM) analysis, lignocellulosic feedstock with an aqueous fiber was cut into small samples, mounted on organic solvent at temperatures ranging from stubs with adhesive, and then they were placed 140°C to 220°C. This causes lignin to break under vacuum, evacuated and sputter-coated down by hydrolytic cleavage of alpha aryl-ether with platinum using auto fine coater (JEOL JFC- links into fragments that are soluble in the 1600). After preparation of samples, the samples solvent system. So in order to achieve high were observed under the Scanning Electron temperature, acetic acid cooking was performed Microscope (JEOL JSM-6380LV). under different conditions in a laboratory pulping unit provided with six autoclaves, each Thermal stability using thermogravimetric of 2.5 liter rotate in a heated poly glycol bath. analysis (TGA) The ratio of liquor/ fibre, acetic acid and H SO 2 4 The thermal stability and decomposition catalyst concentration was maintained constant analyses of the fibre and pulp were evaluated by at 10:1, 70% and 0.5% (v/v) respectively. thermogravimetric analysis (TGA) using a Pulping was done at four temperatures (1500C, Mettler Toledo TGA/SDTA 851e 1600C, 1700C and 1800C) and five cooking times Thermogravimetric Analyser. A sample mass of 5,10,15,20 and 25 min respectively at each 2-3 mg were heated from 50 º C up to 700 º C at temperature. After the digester was loaded with a rate of 5º C/min under nitrogen atmosphere in tender coconut fibre and cooking liquor, the a constant flow of 80 cm3 min-1. temperature was allowed to rise. The rising period to the operating temperature is maintained Results and discussion constant as 70 minutes. After pulping for different time intervals the digester was kept for Optimization of organosolv pulping of tender 10 min before opening. The spent liquor was coconut fibre immediately separated from the pulp by filtering The pulp yield, Kappa number, and % in order to avoid lignin precipitation. The delignification of organosolv pulp obtained from resultant pulp was then washed with fresh tender coconut husk fibre by different cooking corresponding acetic acid and finally with water conditions have been furnished in Figures 1,2 and air- dried overnight. The performance of the and 3 respectively. pulping process was evaluated by Kappa number and yield. The Kappa number (TAPPI T236 cm- The first assessment after any stage of 85) is the volume (mL) of 0.1 N potassium lignocellulosic material treatment is the yield permanganate solution consumed by 1 g of oven measurement (which is a parameter for the dry fibrous material under the conditions process classification). The total yield is specified in this standard method. measured as the ratio between the mass of material obtained after the treatment stage and Bleaching of organosolv pulp the initial mass used to perform the same. Bleaching of organosolv pulp was carried (Candido et al, 2012). out using 15% hydrogen peroxide by weight of The results indicate that the temperature the dry pulp at a pH of 11.5 at 1: 15 solid to and reaction time had a significant influence on liquid ratio. 3% Sodium silicate was added to delignification. It was found that with increase in stabilize hydrogen peroxide. The material was temperature and time there was a decrease in o treated for 4 hours at 80 C. After treatment, the pulp yield, Kappa number and residual lignin. pulp was taken out, washed with cold water and However, it was seen that at all temperatures the air dried in the shade.
16 Cord 2015, 31 (1) delignification reactions were slow down after It was seen that the pulping at 1800C the initial 15 minutes reaction time. resulted in an increased delignification. However the pulp yield was lowered to an unacceptable level. From the studies carried out it could be concluded that pulping at 1700C for 15 minutes was the most suitable condition for digestion of the fibre. Pulping at these conditions gave higher pulp yield and % delignification with low Kappa number. When the reaction time increased from 15 to 25 minutes the delignification didn’t increase significantly and the pulp yield was reduced from 39.94% to 34.49%.The pulp yield at 180oCis very low compared to the pulp yield at170oC which is undesirable for the pulp and paper production. Increase in reaction time from 5 to 25 minutes resulted in the decrease of pulp yield from 30.97 to 21.08. This indicates that at Figure 1. Effect of Temperature and Reaction higher temperature the loss of weight of tender Time on Pulp Yield coconut fiber was more due to the loss of more amounts of cellulose and hemicellulose content along with lignin. Therefore, the optimum cooking conditions for a bleachable pulp of tender coconut husk fibre are as follows: acetic acid 70 %, temperature 1700C, cooking time 15 minutes and catalyst concentration 0.5%. The pulp yield, % delignification and Kappa number values at this condition were 39.94%, 5.2 and 85.91 respectively. Tender coconut fiber before and after organosolv pulping Figure 2. Effect of Temperature and Reaction Time on Kappa Number
Figure 3. Effect of Temperature and Reaction Time on Delignification Figure 4. Raw fiber
17 Cord 2015, 31 (1)
bleaching the pulp yield, % delignification and Kappa number of the organosolv pulp at 1700C for 15 minutes was 92.6, 94.31 and 16.2 respectively. Surface morphology of tender coconut fibre and pulp Microscopic examination reveals that a single fiber comprises a bundle of multiple cells called ultimates. These ultimates in aggregate bundle are bound together by cementing materials like lignin and oriented roughly parallel to one another (Figure 6). The fibre shows globular structures in regular intervals (porous) and other constituents, probably organic Figure 5. Fiber after organosolv treatment residues (Mohanty and Nayak 2004; Ameida et at 1700C and 15 min al 2006) of the extraction of the fibres. The fibrils are covered by a cuticle layer, i.e. wax of The material tends to get a dark colour aliphatic origin (Ratta 1999). Each cell is after organosolv treatment (Figure 5) in contrast roughly polygonal in shape, with a central hole to raw fiber which is brown in colour (Figure 4). or lumen, comprising about 10% of the cell area Therefore, the pulp has to be subjected to of cross section. bleaching for acceptable brightness. In addition the structure of tender coconut fiber after organosolv treatment changed dramatically compared to the raw material (from fibers to a cardboard like appearance). The cellulose and hemicelluloses present in lignocellulosic fibers do not contribute significantly to coloration, due to their naturally white color characteristics. On the other hand, lignin and other extractives contribute to its darkness and therefore can be removed by bleaching. After pulping, acetic acid can be regenerated from the black liquor by simple distillation and can be re-used in the cooking. The dissolved lignin and hemicelluloses in the acetic acid are collected as concentrated slurry after the distillation. The concentrated slurry is Figure 6. Cross–section of Raw coir fiber shown mixed with water, lignin is separated as in SEM precipitate and the hemicelluloses remain in water solution. The hemicelluloses are precipitated in ethanol. Therefore, we can utilize After Organosolv treatment, the impurities all the fractions of tender coconut in the acetic and wax cuticle layers of the fibres surface were acid pulping. removed. The fibrils separate from each other due to removal of the lignin, the cementing Bleaching of the organosolv pulp was component, by the action of the organic solvent, done by hydrogen peroxide. Physical and optical leading to an increase of the surface area (Figure properties of the organosolv pulp were improved 7a, b&d). during hydrogen peroxide bleaching. After
18 Cord 2015, 31 (1)
(a) (d) Figure 7. (a) & (b) micrograph of fiber after organosolv pulping, (c) surface of the fiber after organosolv pulping, (d) micrograph of pulp after blending The treatment also leads to changes in the hydrogen bonding interactions of hydroxyl groups of cellulose, resulting in the deformation of individual micro fibrils (Ganan and Mondragoni 2005). The treatment with acetic acid showed a more effective result in the lixiviation of the cuticle layer in removal of bonding material. Under this condition, the most external layer of the fibre was eliminated, revealing the fibrillar structures (Figure 7a&b). Grinding separates the individual fibers (figure 7 d). The surface morphology of the unbleached (b) pulp (Figure 7 c) showed globular protrusions, identified as silicate stegmata. The cavities became more defined, and the surface fibres became rougher than in the untreated fibre.
(c) (a)
19 Cord 2015, 31 (1)
resulting from organosolv treatment which also indicates that the macrocomponents were removed by the organosolv treatment. (Esmeraldo et al 2010). The degradation in the treated fibres is attributed to the pyrolysis of cellulose and to residual hemicelluloses and this is shifted to lower temperature when the amount of lignin is reduced. Reported by Mitra et al (1998), Varma et al (1986) and Van Dam et al (2004) state that weight losses above 2000C can be attributed to the oxidation and condensation of carbohydrates, while the phenolic components of lignin are (b) considered to be more stable. Also Vazquez- Figure 8. (a) & (b) micrograph of the organosolv Torres et al (1992) after extraction and pulp after hydrogen peroxide characterization of lignin samples from coconut bleaching fibre, verified that their decomposition starts from a temperature of approximately 3800C. After hydrogen peroxide treatment the surface of the organosolv pulp was free of silica bodies, exposing the visible pits (Figure 8a&b). The surface became smoother than the unbleached pulp. The pits measured about 2.44 µm in diameter. Thermo gravimetric Analysis / Simultaneous Differential Thermal Analysis (TGA/SDTA) of raw fibre and pulp The thermo gravimetric (TG) and derivative of the thermogravimetry curve(DTG) curves for the raw tender coconut fibres and organosolv treated fibres are shown in Figure 9 Figure 9. TG/DTG curve of raw tender coconut and 10. The results indicate that the range of fibre thermal degradation decreased after organosolv treatment ( Karnani 2004, Mohanty 2000). The DTG curves show the first peak below 1000C as a result of evaporation of residual moisture. (Pothan et al. 1997; Pothan and Thomas 2003; Eichorn et al. 2001).
Thermal degradation in N2 atmosphere was characterized by an overlapping of processes that may occur in the same range of temperature, which can be separated only by observation of one of the peaks in the DTG output. The treated fibre showed a single degradation peak in contrast to the double peaks observed in untreated fibres results. This is a further indication of a higher homogeneity Figure 10. TG/DTG curve of organosolv pulp treated at 1700 C, 15 min
20 Cord 2015, 31 (1)
Handmade papers and garden articles made The pulp can also be moulded into various from tender coconut husk organosolv pulp disposable articles like handmade paper, egg cartons, paper plates, cups, glass covers, garden The organosolv pulp was diluted with articles like paper pots for seedling, and water and then poured uniformly over a screen. packaging materials. The wet pulp was then transferred onto a cloth to remove the excess water. As the water drains Conclusions out, the fibers come closer and closer together The potential of tender coconut fibre for forming a tightly bonded mesh. The wet sheet making diversified products by an ecofriendly was pressed to remove the remaining moisture. organosolv process could be confirmed. The pressed sheets were then laid on a smooth Optimum digestion conditions to obtain a surface and dried under the sun. After the drying, bleachable pulp of tender coconut husk fibre calendaring of the handmade paper was done were as follows: using acetic acid 70 %, at a between stainless steel sheets. It was observed temperature of 1700C and cooking time 15 that blending bleached organosolv pulp with minutes in presence of a catalyst (0.5%). The waste paper pulp in suitable proportions pulp yield, % delignification and Kappa number improves strength and surface smoothness of the values at this condition were 39.94, 85.91 and 40 handmade paper. respectively. After bleaching the % delignification and Kappa number of the organosolv pulp has become 94.31 and 16.2 respectively. The drawbacks of kraft pulping relate to bad-odor problems, sulfur usage and bleaching problems could be eliminated considerably by applying the ecofriendly organosolv pulping processes. The recovery of the solvent from black liquor is easy and effective. Lignin is precipitated and can be separated from the black liquor after distillation. Therefore, complete utilization of the tender coconut husk fiber is possible by this (a) environment friendly pulping method. Acknowledgment The authors gratefully acknowledge the financial support of the Coconut Development Board, Ministry of Agriculture, Government of India. Thanks are due extend to Prof. G. Balachandran, Ex-Chairman, Coir Board for his keen interest and guidance in this work. References Acid-insoluble Lignin in Wood and Pulp. TAPPI standard T 222 om-98. Ameida, J.R.N.D., Aquino, R.C.M.P., and (b) Monteiro, S.N. 2006. Tensile mechanical Figure 11. (a) Seedling pots made from properties, morphological aspects and unbleached organosolv plp, (b) chemical characterization (Attaleafunifera) Handmade paper made from fibres, Composite Part A. Applied Science bleached organosolv pulp and Manufacturing 37 (9): 1473-1479.
21 Cord 2015, 31 (1)
Ash in wood, pulp, paper and paperboard: composites. Polymer Engineering and combustion at 525°C. TAPPI standard T Science 37(2): 476-483. 211 om-02 Laly, A. Pothan., Sabu Thomas. 2003. Polarity Candido, R.G., Godoy, G.G., Goncalves, A.R. parameters and dynamic mechanical 2012. Study of sugarcane bagasse behaviour of chemically modified banana pretreatment with sulfuric acid as a step of fiber reinforced polyester composites. cellulose obtaining, World Academy of Composites Science and technology 63(9): Science, Engineering and Technology 1231-1240. 61:101-105. Li, Yang ., Arthur, J. Ragauskas. 2012. Ethanol Canetieri, E., Rocha, G.J.M., de Carvalho, J.R., organosolv lignin-based rigid polyurethane Silva, J.B. 2007. A. Optimization of acid foam reinforced with cellulose hydrolysis from the hemicellulosic nanowhiskers. RSC Adv 2:3347-3351. fraction ofEucalyptus grandis residue Method of test for holocellulose in wood, USA. using response surface methodology. 1978. ASTM D: 1104-56. Bioresource Technology 98(2):422-8. Mitra B.C., R.K. Basak, and M. Sarkar 1998. Eichhorn, S. J., Baillie, C.A., Zafeiropoulos, N., Studies on jute-reinforced composites, its Mwaikambo, L.Y., Ansell, M.P., limitations and some solutions through Dufresne, A., P.J., Escamilla, G. C., chemical modification of fibers. Journal of Groom, L., Hughes, M., Hill, C., Rials, T. Applied Polymer Science 67(6):1093– G. and Wild, P. M. 2001. Review: Current 1100. international research into cellulosic fibres and composites, Journal of Materials Mohanty, K., M. Misra and G. Hinrichsen. 2000. Science 36 (9):2107-2131. Biofibres, biodegradable polymers and biocomposites: An overview. Esmeraldo M.A., Antonio C.H. Barreto., Jose E. Macromolecular materials and B. Freitas., Pierre B., A. Fechine., S.B. engineering 276/277:1-24. Sombra., Elisangela Corradini., Giuseppe Mele., Alfonso Maffezzoli and Selma, E. Mohanty, S., and Nayak, S.K. 2004. Effect of Mazzetto. 2010. Dwarf-green coconut MAPP as coupling agent on the fibers: a versatile natural renewable raw performance of sisal-PP composites. bioresource. Treatment, morphology, and Journal Reinforced Plastics and physicochemical properties, Bio Resources Composites 23(18):2047-2063. 5 (4): 2478-2501. Pires, M., Costa, R.S., Jose´, A.S., Badaro´, M. Fernandez, N. 1996. Pulp and paper M., Midlej, C., Alves, J.M. 2004. A development from sugarcane bagasse. In: cultura do coco: umaana´ liseeconoˆ mica Third Internacional Non-wood Fiber (The coconut culture: an economical Pulping and Papermaking Conference, evaluation). Rev. Bras. Frutic. 26(1): 173- Proceedings, Pequim (1):231-240. 176. Ganan, P., and Mondragon, I.2005. Effect of Pothan L.A., Thomas, S., N.R. Neelakantan. fibre treatments on mechanical behavior of 1997. Short Banana Fiber Reinforced short fique fibre- reinforced polyacetal Polyester Composites: Mechanical, Failure composites. Journal of Composites and Aging Characteristics. Journal of Science and Technology 59(9): 1303-1309. reinforced plastics and composites 16(8): “Kappa Number of Pulp.” TAPPI Method T 236 744-765. cm-85 Pothan, L.A., Oommen, Z., Thomas, S., Karnani R., Krishnan M., Narayan, R. Dynamic mechanical analysis of banana 2004. Biofiber reinforced polypropylene fiber reinforced polyester composites
22 Cord 2015, 31 (1)
2003. Compos. Sci. Technol. 63 (2), 283– Vasquez-Torres, H., Canche-Escamilla, G., and 293. Cruz-Ramos, C.A.1992. Coconut husk lignin. 1. Extraction and characterization. Presley, J.R., R.T. Hill. 1996. The Technology of Journal of Applied Polymer Science 45: Mechanical Pulp Bleaching. In: Peroxide 633-644. Bleaching of Chemimechanical Pulps, Pulp Bleaching: Principles and Practice, Zhiqiang. Li, Zehui Jiang, Bengua Fei, Xunjun Carlton W.D. and D.W. Reeve (Eds.). Pan, Zhiyong Cai, Xing’e Liu and Yan Yu. Chap. 1. TAPPI Press, Atlanta: 457-512. 2012. Bamboo organosolv pretreatment, Bioresources 7 (3): 3452-3462. R.C. Leitão, A. M. Arau´ jo, M.A. Freitas-Neto, M.F. Rosa, S.T. Santaella. 2009. Anaerobic treatment of coconut husk liquor for biogas production. Water Science & Technology—WST, 59(9):1841- 1846. Ratta, V. 1999. Thesis on the crystallization, morphology, thermal stability and adhesive properties of novel high performance semi crystalline polymides. Faculty of Virginia Polytechnic Institute and State University, Virginia.
Saberikhah E., J. Mohammadi Rovshandeh., P.
Rezayati-Charani. 2011. Organosolv
pulping of wheat straw by glycerol.
Cellulose Chemistry and Technology,
45(1-2):67-65.
Solvent extractives of wood and pulp. (Proposed revision of TAPPI T 204 cm-97) Uppdegraf . 1969. Semimicro Determination of Cellulose in Biological Materials. Analytical Biochemistry 32:420-424
Van Dam, J.E.G., Van Oever, M.J.A., Wouter Teunissen, Edwin R. P Keijsers, Aurora G Peralta. 2004. Process for production of high density/high performance binderless boards from whole coconut husk: Part 2: Coconut husk morphology, composition and properties. Industrial Crops and Products 19(3): 207-216. Varma, D.S.,Varma, M., and Varma, I.K.1986. Thermal behavior of coir fibres. Thermochimica Acta 108:199-210.
23 Cord 2015, 31 (1)
Low Temperature Grafting of MMA on to Coir Fibre
Lakshmi N. S.1, Sarika Babu2, Sumy Sebastian3 and P.K. Ravi4
Abstract
Low temperature grafting of methyl methacrylate (MMA) on to coir fibre was carried out in aqueous medium using Potassium per sulphate (PPS) as an initiator under the catalytic influence of Ferrous ammonium sulphate (FAS). Optimization of various parameters of grafting viz. monomer, initiator and catalyst concentration, time and temperature was carried out to obtain the maximum tensile properties. Evidence of grafting was characterized from Fourier Transform Infrared spectroscopy (FTIR), Scanning Electron Microscopy (SEM) and Thermal analysis (TGA). The maximum breaking stress (BS) of control and grafted coir fibre were 213.08 and 365.00 N/mm2 respectively. Hence the percentage of improvement of grafted coir fibre was found to be 71.30%. Increase in tensile properties with maximum BS observed under monomer (25%), initiator (0.75%) and catalyst (0.75%) concentration, time (150min) and temperature (500C) respectively. The t-test and Analysis of variance (ANOVA) were studied for statistical significance and the P values obtained were less than 0.05 which revealed that the value was highly significant for the improvement of mechanical strength on coir fibre by graft Co- polymerization.
Keywords: Methyl Methacrylate, Potassium persulphate, Ferrous ammonium sulphate, grafting, Co- polymerization.
______1Incubation Assistant, Department of Chemistry, Central Coir Research Institute (Coir Board), Alleppey, Kerala. Email: [email protected] 2Project Assistant, Department of Chemistry, Central Coir Research Institute (Coir Board), Alleppey, Kerala. Email: [email protected] 3Scientific Assistant, Department of Chemistry, Central Coir Research Institute (Coir Board), Alleppey, Kerala. Email: [email protected] 4Senior Scientific Officer, Department of Chemistry, Central Coir Research Institute (Coir Board), Alleppey, Kerala. Email: [email protected]
24 Cord 2015, 31 (1)
Introduction chemicals. In order to make modification process more economic, effort has been made to find out Natural fibers are abundant, versatile, the optimum modification condition depending renewable, cheap and biodegradable raw on the concentration of monomer, initiator and material which can be used for making wide catalyst, reaction time and temperature. Graft variety of products. Natural fibers have a number yield was determined on the basis of the weight of advantages such as low cost, less toxicity, non increase of the coir fibre treated. Some abrasive nature and easy process ability. Among instrumental analysis such as Infrared the natural fibres, Coir occupies a prominent spectroscopy (IR), Scanning Electron position owing to its qualities such as natural Microscopy (SEM) and Thermo Gravimetric resilience, biodegradability, salt resistance, Analysis (TGA) were done to identify graft on mechanical strength and toughness which coir fibre. enhances its commercial and industrial usefulness. (Singha and Raj, 2010) Materials and methods Coir is a multicellular biopolymer in Materials which crystalline cellulose is arranged helically Mechanically extracted coir fibres were in a matrix consisting of non crystalline collected from Alleppey, Kerala. Monomer: lignocellulosic complex. (Ott et. al., 1954). Due Methyl Methacrylate (MMA), Initiator: to hydrophilic nature and low thermal stability Potassium per sulphate (PPS) and Catalyst: coir fibers have got certain limitations. There are Ferrous ammonium sulphate (FAS) used was of several methods such as acetylation, analytical grade. benzoylation, grafting, silane treatment, plasma treatment etc. which have been employed to Methods improve the properties of the natural fibers Purification of materials through surface modification. Graft copolymers are assuming increasing importance because of The fibres were cleaned first in Willowing their tremendous industrial potential. Graft machine and sorted using combing board. The copolymerization is one of the best techniques fibres were soaked in water over night. Then it for modifying the properties of natural polymers. was washed with water under stirring for 8 hrs It is a convenient and clean means for altering and dried in hot air oven at 700C. the properties of numerous polymer backbones. MMA was washed with 5% NaOH As the natural fibres bear hydroxyl groups from solution followed by water and dried over cellulose and lignin, they are amendable to anhydrous sodium sulphate. Catalyst ferrous chemical modifications. Chemical modification ammonium sulphate was recrystalised from hot may activate these groups or can introduce new water. Initiator potassium per sulphate was used moieties that can effectively interlock with as received. matrix. Modifying the properties of natural polymers through graft copolymerization has Graft Co-polymerization been reported by various workers. (Renu et. al., Fibres were immersed in distilled water for 2010). Desirable and targeted properties can be 24 hrs prior to the reaction. The material to imparted to the natural polymers through graft liquor ratio was maintained at 1:50. A known copolymerization in order to meet out the amount of the initiator, catalyst and monomer requirement of specialized applications. Grafted were then added to the reaction vessel fibers have been found to increase the strength maintained at required temperature. Grafting of and thermal stability than raw fibers. MMA on to coir fibre was carried out under In the present study coir fibre was grafted constant stirring. At the end of the desired with Methyl Methacrylate (MMA) for improving reaction period, the coir fibre was thoroughly its properties such as tensile strength, thermal washed with acetone to remove any stability, light fastness and resistance to homopolymer generated during the reaction.
25 Cord 2015, 31 (1)
Fibres were then washed with distilled water and Results and discussion dried in a hot air oven at 700C till a constant Mechanism weight was obtained. Polymerization reaction were done at varying conditions of 5-75% It has been observed that C2, C3, C6 monomer, 0.0-1.5% initiator, 0.0-1.25% catalyst hydroxyls and C-H sites of cellulose back bone based on the weight of the fibre, 300C-800C of are the active sites of coir fibre for grafting of temperature and 30-210 minute reaction time on MMA. PPS takes part in the redox reaction with 2+ - to the coir fibre. The reaction conditions were Fe to produce SO4 *. optimized so as to get maximum Grafting Yield Fe2+ + -O S-O-O-SO - Fe3+ + SO 2- + SO -* (2) (GY) and BS. Percentage GY was calculated by 3 3 4 4 - the equation Interaction of SO4 * with H2O generates OH free radicals (OH*) and these free radicals GY = (W2-W1/W1) 100 (1) are responsible for free radical generation on coir
Where W1 and W2 are the weights of coir and the monomer as well as further chain fibre and grafted fibre respectively. propagation there by resulting in the formation of graft copolymer. Scanning Electron Microscopy (SEM) Optimization of different reaction parameters Scanning Electron Microscopy studies of for grafting Methyl Methacrylate on to Coir ungrafted coir fibre and grafted coir fibre was Fibre carried out on Scanning Electron Microscope (SEM) (JEOL JSM-6390 LV, Tokyo, Japan) Optimization of various reaction system with an accelerating voltage of 10 kV. parameters such as concentration of the The SEM images are studied under 10µm monomer, initiator, catalyst, reaction time and magnification. temperature were carried out for graft co- polymerization of methyl methacrylate on to coir Infrared Spectroscopy (IR) fibre. Infrared spectra of the ungrafted and i. Effect of monomer concentration grafted coir fibre were taken on Perkin Elemer Spectrophotometer. Grafting increases with increase in monomer concentration up to 25% and further Thermo Gravimetric Analysis (TGA) increase in monomer concentration results in The Thermo Gravimetric Analysis (TGA) decrease of grafting yield (fig 1). This may be of the coir fibre was done using Mettler Toledo due to the formation of more homo polymer as TGA/SDTA 851e, Japan. The test was carried compared to graft co-polymer at higher out in an inert atmosphere under nitrogen flow monomer concentration. Due to homo rate at 10ml/min and heating rate throughout the polymerization, viscosity of the reaction medium test was 100C/min. The weight change was increases which creates hindrance in the recorded as a function of the heating movement of the free radical towards the active temperature. sides, there by resulting in decrease of grafting yield (Singha et.al., 2007 ). Tensile properties ii. Effect of initiator Tensile properties of coir fibres were determined using UTM (Universal Testing The initiator plays an important role in Machine) Shimadzu AG-X/R. At test obtaining higher graft yield. It is observed (fig 2) parameters; strain rate = 10mm/min, gripping that the percentage of grafting increases with length=5cm at atmospheric temperature. increase in initiator concentration PPS up to 0.75% and there after the percentage of grafting
starts decreasing.
26 Cord 2015, 31 (1)
decreasing. Decrease in grafting may also be explained due to the termination of growing 300 14 grafted chain by primary free radical resulting 12 from the decomposition of excess initiator.
250 10 iii. Effect of catalyst ) 2 8 It is observed that (fig 3) grafting yield 200 6 increased with increasing catalyst concentration. As the catalyst concentration increases, more 4 (%) GY BS (N/mmBS numbers of Fe (III) ions as well as more reactive 150 BS 2 site on the coir fibre surface are produced, GY thereby increasing the grafting up to a certain 0 concentration of catalyst. But beyond this 100 -2 concentration the grafting yield declined. This is 0 5 15 25 35 45 55 65 75 probably due to the formation of large number of Fe (III) ions which promote the premature Monomer concentration (%) Figure 1. termination of the growing grafted chain.
350 4 300 2 2 1 300 0 0 250 ) -1 2 250
) -2 2 -2
200 -3 200 (%) GY BS (NmmBS -4 -4 GY(%) BS GY
BS (N/mmBS 150 -5 -6 150 BS -6 GY -7 100 -8 100 -8 0 0 0.25 0.5 0.75 1 1.25 Catalyst concentration (%) 0 0 0.25 0.5 0.75 1 1.25 1.5 Figure 3. Initiator concentration (%) Figure 2. iv. Effect of reaction time
Initiator PPS generates activated The percentage of grafting increases with monomers, which show a sufficient affinity to increase in reaction time up to 150min for MMA cellulose macro radicals that are produced by the and there after the percentage of grafting starts action of catalyst FAS on the surface of coir decreasing (fig 4). The initial sharp increase in fibre and enhances the grafting up to a certain grafting is due to the high co-polymerization initiator concentration (Arifuzzaman Khan et. reaction between the reactive sites of coir fibre al., 2009). But at a high initiator concentration and the activated monomer molecules. After large number of activated monomers 150min the homo polymerization reaction accumulates and forms the homo polymer, prevails over the co polymerization possibly due thereby reducing the active monomers available to the decrease in reactive sites on the coir fibre in the vicinity of cellulose macro radicals and (Salam, M.A. 2005). So the percentage of hence the percentage of grafting starts grafting becomes steady. On prolonged exposure
27 Cord 2015, 31 (1) to heat, the percentage of grafting decreases due The t-test was studied for statistical to the partial dissolution of grafted fibre. significance and all the P values obtained were less than 0.05 (table). Since it shows that the 400 2 values were statistically significant. Analysis of 1 variance (ANOVA) of BS of grafted Coir fibre 350 0 (P<0.0021) was also calculated and the p value is (p<0.05) revealed that this was highly significant
) 300 -1 2 for the improvement of mechanical strength on -2 250 Coir fibre by graft Co-polymerization. -3 (%) GY
BS (N/mmBS Characterisation Study 200 -4 BS -5 Scanning Electron Microscopy (SEM) 150 GY -6 It is quite evident from figures that there
100 -7 has been a sufficient deposition of MMA on to
0 30 60 90 120 150 180 210 coir fibre. Comparison of the Scanning electron Reaction time (min) micrographs of ungrafted coir fibre Fig. 6(a) and MMA grafted coir fibre; Fig. 6(b) reveals a clear Figure 4. cut distinction between the ungrafted and grafted fibers. v. Effect of temperature Figure 6. SEM images of (a) Ungrafted It is observed that at ordinary temperature, Coir fibre (b) MMA grafted Coir fibre 0 the grafting is slow. But at 50 C the percentage of grafting is high (fig 5). The increasing percentage of grafting up to optimum temperature may be due to increase in the rate of production of active free radical which increase the number of grafting sites at a higher rate, the rate of initiation by coir fibre radicals is thereby increased. The increase in temperature increases the rate of diffusion of monomers into the coir fibre matrix. The decrease in percentage of grafting beyond optimum temperature may be attributed to the increase in the rate of homo polymerization. Figure 6(a)
400 3 2.5 350 2
) 300 2 1.5 250 1
0.5 (%) GY BS (NmmBS BS 200 GY 0 150 -0.5
100 -1
0 30 40 50 60 70 80 Reaction temperature (0C)
Figure 5. Figure 6(b)
28 Cord 2015, 31 (1)
The pits observed in ungrafted coir fibre The peak assignments of the absorption are no longer observed in grafted fibres. The band corresponding to various groups are surface becomes more or less uniform owing to summarized in table 1 (Zuraida et. al., 2011) deposition of PMMA grafts along the direction Table 1. FTIR spectral data of ungrafted Coir of fibre axis. This orderly arrangement of fibre PMMA changes the porous surface of coir fibre
resulted in an appreciable increase in the strength Position / cm Possible Assignment of grafted fibres compared with ungrafted fibres ~3600-3200 ν (OH) broad, strong band from (Khullar et. al., 2008). the cellulose, hemicellulose and FTIR Spectra lignin of coir ~3000-2900 ν (C-H) in aromatic rings and Figure 7. FTIR spectrums of the (a) ungrafted alkanes coir and (b) MMA grafted Coir fibre ~1732.7 ν (C=O) most probably from the lignin and hemicelluloses
97.9 ~1608.5 ν (C=C) aromatic in-plane ~1512.6 ν (C=C) aromatic skeletal ring 90 vibration due to lignin ~1464.2 δ (C-H); δ (C-OH) 10 & 20 alcohol 80 ~1426.6 δ (C-H) ~1375.4 δ (C-H) 70 ~1267.4 δ (C-OH) out-of-plane % Transmittance % ~1046.0 ν (C-OH) 20 alcohol ~898.5 ν (C-O-C) in plane symmetric 60
FTIR spectrum of ungrafted coir and 52 4000 3000 2000 1500 1000 400 MMA grafted coir fibre are shown in fig.7 (a-b). -1 Wavenumbers (cm ) The ungrafted coir fiber shows the characteristic absorption of -OH group ~3600-3200cm-1 Figure 7(a) (Zuraida et. al., 2011). But in MMA grafted coir fibre, broadening of absorption band takes place at ~3600-3200cm-1. The intensity of –OH peaks
97.9 in MMA grafted fibres reduced as a result of grafting, since it is the probable site of grafting. Grafted fibre shows an additional peak at
85 ~1732cm-1 may be due to the presence of (C=O) of MMA.( Balamurugan et. al., 2004; Jogeswari et. al., 1999a). 75 Thermo gravimetric analysis (TGA)
% Transmittance % 65 Thermal behavior of ungrafted coir fibre and its MMA grafted coir fibre were examined by a study of their TGA thermograms. From the 55 results, listed in Table 2, it is evident that initial 4000 3000 2000 1500 1000 400 Wavenumbers (cm-1) and final decomposition temperature of ungrafted coir fibre are 80 and 4500C where as in Figure 7(b) the case of MMA grafted coir fibre the initial and final decomposition temperature are 900C and 6700C respectively (Fig. 8 (a & b ). From the
29 Cord 2015, 31 (1) results it is observed that the thermal stability of Tensile Properties MMA- grafted fibre is higher than that of The tensile properties of ungrafted and ungrafted fibre. (Jayaraj et. al., 2014) grafted coir fibre were determined by taking a
minimum of 50 fibres from each optimization
fig. 1 to 5.
It clearly shows that there is an appreciable change in the tensile properties under maximum stress at break. Grafting of coir fibres with MMA from control to complete optimization brings about a substantial increase in maximum stress at break. The increase in maximum stress in the case of completely optimized fibre from 213.08 N/mm2 to 365 N/mm2 may be due to the increase in the degree of crystallinity due to the orderly arrangement of the short grafts of the PMMA units on the cellulosic backbone. In all the cases of grafting it has been found that the maximum
stress at break is higher than ungrafted fibre Fig. Figure 8(a) 9 (a). (Jogeswari et. al., 1999b)
Figure 9. Tensile properties of Optimized Condition
Maximum stress at break 400 365 350 ²) 360 300 325 250 284.96 289 200 213.08 150
100 Break load load Break (N/mm 50 0 C MGf Igf CGf TGf °CGf Optimized conditions
Figure 8(b) Figure 9(a)
Table 2. TGA of ungrafted and MMA grafted Conclusion Coir fibre Low temperature Graft Copolymerization of MMA on to coir fibre was done at an T10 T50 T90 Sample optimum temperature of 50OC. Percent of (°C) (°C) (°C) grafting increases with the increase in each of Untreated coir fibre 80 310 450 the reaction parameters, such as monomer concentrations, initiator concentrations, catalyst concentrations, polymerization time and MMA grafted coir 270 350 630 temperature up to a limited extent and then fibre decreases. During graft copolymerization
30 Cord 2015, 31 (1) keeping all other variables constant, better graft bamboo (Dendrocalamus strictus). yield were obtained at 50OC, 2.30 hr, 25% eXPRESS Polymer Letters 2 (1): 12-18. monomer, 0.75% initiator, 0.75% catalyst. The Ott E, Spurline H M & Graffin M W, 1954. polymer grafted on to coir fibre improves Cellulose and cellulose derivatives, Part II magnificently the tensile strength. FTIR, SEM (Interscience, Newyork), 863 and TGA measurements characterized the extent of grafting. Renu S., S Vashistha and S. Kalia. 2010. Biologically and chemically modification Acknowledgments of ramie fibers: A comparative study. The authors are very much grateful to Coir International Conference on Advance in Board, Cochin and Central Coir Research polymer Technology.239 Institute (CCRI), Kalavoor for permitting to Salam, M.A. 2005. Graft co polymerization of conduct the studies and publication of this methyl acrylonitrile monomer on to article. sulfonated jute- cotton blended fabric. J. References Textile and Apparel technology and Management 4(4). Arifuzzaman Khan, G.M., Sheruzzaman., S.M. Abdur Razzaque., Sakinul Islam and Singha, A.S., S. Anjali and V.K. Thakur. 2007. Shamsul Alam. 2009. Grafting of Pressure induced graft copolymerization of acrylonitrile monomer on to bleached okra acrylonitrile on to Saccharum cilliare fibre bast fibre and its textile properties. Indian and evaluation of some properties of J. Fibre and Textile Research 34: 321-327. grafted fibre. Bull. Master. Sci. 31(1):7- 13. Balamurugan, A., Kannan, S., Selvaraj, V. and Rajeswari, S. 2004. Development and Singha A.S and Raj K.R. 2010. Graft Spectral Characterization of Poly (Methyl Copolymerization of Methyl Methacrylate Methacrylate) /Hydroxyapatite Composite (MMA) onto Agave americana Fibers and for Biomedical Applications. Trends Evaluation of their Physicochemical Biomater. Artif. Organs, Vol. 18 (1), 41- Properties. International Journal of 45. Polymer Analysis and Characterization 11(1):27-42. Jayaraj A.P., Anitha Das Ravindranath and U.S. Sarma. 2014. Nanocellulose from diseased Zuraida, A., S. Norshahida., Sopyan and H. coconut wood. Cord 30 (1), 1-10. Zahurin. 2011. Effect of fibre length variation on mechanical and physical Jogeswari R., Manjusri M. and Amar K.M. 1999. properties of coir fibre reinforced cement- Surface Modification of Coir Fibers I: albumin composite (CFRCC). J. IIUM Studies on Graft Copolymerization of Engineering 12(1). Methyl Methacrylate on to Chemically
Modified Coir Fibers. Polym. Adv. Technol. 10: 336-344.
Kaith, V.S and S. Kalia. 2008. Graft
Copolymerization of MMA on to flax
under different reaction conditions: a
comparative study. eXPRESS Polymer
Letters 2(2): 93-100. Khullar, R., V.K. Varshney., S. Naithani and P.L. Soni. 2008. Grafting of acrylonitrile on to cellulosic material derived from
31 Cord 2015, 31 (1)
Characterisation of Silver Deposited Coir Fibers by Magnetron Sputtering
Melvi Chandy1, U.S. Sarma2, M.S. Latha3 and K. Shreekrishna Kumar1*
Abstract
Silver thin films are extensively used due to their superior optical, electrical and antimicrobial properties. Recent development in the incorporation silver thin films on natural fibers makes it possible to utilize its excellent physical and chemical properties in the field of textiles. Present study focuses on the surface functionalization of natural coir fiber with silver thin film by magnetron sputtering. This will help to widen the use of natural coir fibers. The surface morphology of the coated coir fibers are investigated by employing a scanning electron microscope. The results show that the surface functionalization of silver-coated coir fibers are highly versatile, and it possess excellent protection against ultraviolet radiation, exhibit excellent hydrophobicity (contact angle=105.2°) and good antibacterial effects. This study demonstrates that treatment, which uses silver thin films by magnetron sputtering, is a promising method for achieving multifunctional coir fabrics.
Keywords: Silver thin film/magnetron sputtering/hydrophobicity
______
1School of Technology and Applied Sciences, Mahatma Gandhi University, Malloossery P.O, Kottayam District, Kerala-686 041, India. E-mail: [email protected] 2 Central Coir Research Institute, Kalavoor Post Office, Alapuzha District, Kerala-688522, India.
3S.N. College, Neduvaramcode P. O., Alappuzha District, Kerala-689 508, India. * To whom correspondence should be addressed. E-mail: [email protected]
32 Cord 2015, 31 (1)
Introduction water. On the other hand, as eco friendly and dry technology, sputtering is an attractive alternative Natural fibres are environmentally friendly by adding new functionalities such as materials, with many advantages like eco- hydrophobicity, UV protection and antibacterial friendliness, biodegradability, user friendly etc. properties, making it a promising technique for Even though they have some markable future applications. The ability to deposit a well- advantages, inherent drawbacks of the natural controlled silver coating on textiles would extent fibres are high hydrophilic nature Bledski & the applications of coir fibers, based on changes Gassan (1999), low resistance to microbial to both the physical and chemical properties of attack, the tendency to form aggregates during textiles. Not much study has been reported on processing and poor surface adhesion for the surface functionalisation of coir fibers. association with a polymer matrix Cho et.al (2002). Various physical and chemical methods This investigation is directed towards have been employed to improve the properties of coating of coir fibers with silver thin films by natural fibers. The chemical method involves magnetron sputtering technique. Their dewaxing, delignification, bleaching, acetylation, morphological characterisation is studied by mercerization, salinization and chemical grafting using SEM. Moreover the coated coir fibers are Mohanty et.al (2001) and physical methods studied to characterise the functional properties involves plasma treatment, corona treatment, such as hydrophobicity and antibacterial activity. thin film coating and irradiation Seong and Hae The thermal stability of the coated fibers are also (2010). studied. The coir (Cocos nucifera) is an important Experimental lignocellulosic hard and stiff fibre obtained from Silver target with high purity (purity: coconut husk, by the process of retting or 99.99%) is used to deposit silver thin films onto mechanical combing. Because of its hardwearing coir fiber by using a magnetron sputtering quality, durability and other advantages, it is system. The dimensions of the silver target are used for making a wide variety of floor 50 mm in diameter and 5 mm in thickness. In the furnishing materials, yarn, rope etc Silver et. al present study acetone is used for removing the (2001). However, these traditional coir products fats and oils on the coir fiber substrates. Prior to consume only a small percentage of the total deposition, the coir fiber samples are first world production of coconut husk and these immersed into an acetone solution for 2 min, and products are facing stiff completion with the then washed thrice with deionised water. synthetic fiber alternatives. Hence, research and Following that, the coir samples are dried at a development efforts have been underway to find temperature of 80°C for 24 hours. some diversified application areas for coir fibres including utilization of coir fibres as The pressure before evaporation is kept at reinforcement in polymer composites Moteiro et. 2* 10-6 Torr and then it is increased to about al (2008), desiccant materials Ratanakaran et. al 7*10-6 Torr when argon is introduced into the (2010) geotextiles, and sound absorbents. vacuum chamber as the bombardment gas. The Wassilief (1996). deposition rate is 1Ǻ/sec for all coating. The thickness of the coating was monitored by using In recent years, natural fiber industries a quartz crystal monitor. The sputtering current have been focussing on improving the functional and voltage are maintained at 0.15A and 115V properties of fibers such as UV protection, fire respectively. Since the size and shape of the coir retardency, and antibacterial properties. fibers may vary from fiber to fiber, we had taken Tremendous effort has been made to apply silver a set of 20 samples to produce reliable results. thin films on fibers, including electroless plating, The deposition time is 10min and 20 min on the electroplating, and vacuum deposition. However, samples. such conventional techniques generally use wet- chemical processing and produce much waste
33 Cord 2015, 31 (1)
The surface morphology and chemical composition analysis of the silver coated coir fibers are performed by a scanning electron microscope (SEM). The thermal stability of the uncoated (UC) and silver coated (SC) are evaluated by means of a thermo gravimetric analyser. Water contact angle (WCA) of the samples is measured at ambient temperature using a SEO Phoenix optical contact angle system. A volume of 5µl water is applied in this (a) untreated test. The hydrophobicity of the silver coated coir fabrics are further confirmed by water drops test, floatation and swelling tests. Antibacterial tests on UC and SC coir fibers are carried out following the Standard ‘SNV 195920-1992’. The antibacterial activity of the samples is assessed through a diffusion test in Agar Aldrich). In this method, an E. Coli colony is positioned in a petri dish filled with agar gel. UC and SC fibres are placed over the (b) silver coated (10min) colony and the whole dish is incubated in oven at 37oC for 24 h. After this period of time, the dish is removed from the oven and the area covered by the bacterial colony with respect to the samples is evaluated. If a growth inhibition zone is observed close to the sample (characteristic zone size ≥1 mm), antibacterial property is indicated as ‘good’. If the sample is totally rehabitated by the bacteria, the antibacterial property is indicated as ‘not sufficient’. (c) silver coated (20min) Results and discussion The surface morphology of the coated and Figure 1. uncoated coir fibers is shown in the Fig.1 The SEM study confirms that silver particles are The hydrophobicity of the silver coated and grafted on to the fiber suraface. The surface of uncoated coir fibers is examined by WCA uncoated coir fiber shows the presence of measurement. Fig. 2 shows the microscopic view micro pores. Metal coated fiber, on the other of the contact angle. Generally coir fibers are hand is smooth and without any pores highly hydrophilic due to the presence of indicating the formation of metal layer on the abundant hydroxyl groups on the surface. When surface. After sputtering for 10min, a thin layer metal particles are coated on the surface of the of silver is obtained on the fiber surface. With fabric, the fabric becomes hydrophobic. The easy an increase in sputtering time homogenous silver rolling of the water droplet on the fabric surface is depositions are observed on the coir fibers. the primary evidence for that. The hydrophobisation occurs due to the layer of metal coated on the surface of the coir fiber. The WCA on the surface of the coir fabric coated with silver for ten selected areas is measured as 105.2.2o +/- 2.5oC for a 5µl water droplet on the silver coated
34 Cord 2015, 31 (1) fabric. The WCA measurement of silver coated penetrate the fabrics are measured. The results are fabric (20 minutes) is 106.3o C. All the samples shown in Table 1, in which the absorption time is showed almost same WCA measurements. After recorded for different metal coated coir fabrics. 5 washes the metal coated coir fabrics shows 25% From the results it is clear that the water decrease in the WCA as compared to the uncoated repellency of the coir fabric is increased one. The reason for this decrease in WCA of considerably with the metal coating. In the case of coated fabric is due to the slow removal of metal uncoated coir fabric the water droplet disappears particles from the surface of the coir fabric during immediately within seconds, with water spreading washing. over large areas. In the case of coated fabric the droplet remained spherical even after 30min with no spreading, indicating that all the modified fabrics are water repellent. From the floatation tests (Fig 3) it is found that the coated fabric floated on the surface of water for more than 100 minutes, whereas the uncoated coir fabrics gradually sink to the bottom, when they are placed on the water surface. Fig 4 shows the variation of percentage water absorption coated and uncoated coir fabric with time. More absorption of water is obtained in untreated coir fabric and lesser absorption in metal coated coir fabrics. The samples with 20minutes sputtering time showed lesser absorption of water at ambient temperature than the respective untreated sample.
Figure 2. Microscopic view of water contact angle
The water repellent property is further confirmed by water drops test and floatation tests. In water drops tests, the drops of controlled size are placed at a constant rate upon the fabric Figure 3. Floatation tests for a) uncoated and b) surface and duration of time required for them to silver coated coir fabric
35 Cord 2015, 31 (1)
degradation behaviour of the coir fibers, promoting an increase in the temperature at which
the thermal degradation take place These changes may be attributed to the quick distribution of heat on the surface of the coated coir fabric when
compared to uncoated one.
Figure 4. Water absorption versus time untreated coir fiber Silver coated (10min sputter coated Silver coated (20 min sputter coated (a)
Table 1. Absorption time of untreated and silver coated coir fibers
Samples Absorption time
Untreated 3s Ag coated sample for 10min >100min Ag coated sample for 20min >100min (b) The thermal stability of the uncoated and coated fibers is found out by using TGA. The Figure 5. TGA analysis of a) untreated fiber b) TGA of all the coated and uncoated samples are silver coated (10min) shown in Fig 5a & 5b. From the figure it is evident that the thermal stability increased by Table 2. Thermogravimetric analysis of coir metal coating. In the case of untreated coir fabrics fibers and silver coated coir fibers there are three stages of degradation. The first one is attributed to the evaporation of water and No. Sample Transition Temperature Weight occurs between room temperature and 1500 C. temperature of maximum loss range (0C) (%) The second step, which corresponds to the hemicelluloses degradation, starts at about 1900 C; 1 Untreated 30-150 53 5 0 the third step occurs between 290 and 360 C, 150-297 286 26 corresponding to thermal degradation of cellulose. Lignin presents a broad peak throughout the 297-500 334 45 0 range, degrading between 280 and 500 C. The untreated coir fibers are completely oxidised to 0 2 Silver 30-170 55 5 form char residue at 700 C Alvaraz & Wasquez coated (2004), Tomzac et. al (2007). Metal coating is 170-320 301 26 found to have a considerable effect on the thermal 320-500 360 49
36 Cord 2015, 31 (1)
Figure 7. Antibacterial Activity Test
Table 3. Characteristic antibacterial zone for coated and uncoated coir fiber
No. Sample Sputtering Bacteria Inhibition Level of Time (min) Zone antibacterial capacity. Pollini et. al (2009) 1 Untreated E. Coli ------Not sufficient (3845)
2 Silver Coated 10 E. Coli <1mm Fairly good (3847 & 3849)
3 Silver Coated 20 E. Coli >1mm Good (3846 & 3848)
In literature it is reported that, either antibacterial zone of uncoated and coated coir polyester or cotton fibers coated by silver exhibit fibers are given in Table 3. Silver in its metallic antibacterial capability against E. coli. In this state is inert, but it reacts with moisture on the study antibacterial tests are performed on an skin and fluid on a wound, and become ionized. uncoated and silver coated coir fiber. The effect The ionized silver is highly reactive as it binds to of silver compared with the control samples is tissue proteins and brings structural changes to the determined by measuring the size of the area of bacterial cell wall and nuclear membrane leading inhibition growth close to each sample. As clearly to cell distortion and death. Jiang et. al (2010). It shown in Fig. 3, the E. coli colony seeded in the can be observed a clearly defined bacterial free petri dish filled with agar grows on the untreated zone around each coated sample which confirms coir sample after incubation in oven at 370C for the growth inhibition effect induced by silver 24 h: untreated coir fiber does not display any ions. antibacterial activity. The characteristic
37 Cord 2015, 31 (1)
Conclusion Ratanakorn Rawangkula, Joseph Khedaria, Jongjit Hirunlabhb, Belkacem Zeghmatic. Silver thin films on coir fibers are prepared 2010. Characteristics and performance by magnetron sputtering technique. SEM anlysis analysis of a natural desiccant prepared shows a uniform layer of silver over the coir from coconut coir, Science Asia, 36; 216- fiber surface. The results obtained from water 222. contact angle confirms that the hydrophobicity of the silver coated coir fabric is higher than that Seong OH, Hae YC. 2010. Morphology and of the uncoated fabric. Silver coated coir fabrics surface properties of natural fibers treated show excellent antibactrial activity against E. with electron beam. Microscopy: Science Coli. as confirmed by the antimicrobial tests. Tachnology, Applications and education, Such silver coated coir fabric could therefore be 1880-1888 used in water repellent coating and antibacterial Silva GC, Souza DA, Machado JC, Hourston DJ. applications. The investigation described 2001. Mechanical and thermal suggests that the silver coated coir fabrics will be charecerization of Brazilia coir fibers, promising multifunctional material for military Journal of Applied Polymer science, 76, and medical application. 1197-1206 References S.X. Jiang, W.F. Qin, R.H. Guo, L. Zhang. 2010. Alvaraz V.A, Vazquez. 2004. Thermal Surface functionalisation of degradation of cellulose derivatives/starch nanostructured silver coated polyester blends ans sisal fiber biocomposites, fabric by magnetron sputtering, Surface Polymer degradation and stability, 84; 13- Coating and Technology, 204, 3662-3667. 21. Tomzac, F., Satyanarayana, K.G., Sydenstricker, Bledski AK, Gassan J. Composites reinforced T.H.D. 2007. Studies on lignocellulosic with cellulose based fibers. 1999. Progress fibers of Brazil: Part II- morphology and in Polym. Sci.; 24: 221-274. properties of Brazilian coconut fibers, Composites: Part A, Applied Science and Cho A, Lee SG, Park WH, Han SO. 2002. Eco- Manufacturing, 38, 1710-1721. friendly biocomposite materials using biofibrils. Polym. Sci. Technol. I; 13; 460- Wassilief C.1996. Sound absorption of wood 476. based materilas, Applied acoustics, 48; 339-356. Mohanty AK, Misra M, Drsal LT. 2001. Surface modification of natural fibers and performance of the resulting composites; an overview, Comp Interfaces, 8: 313-43.
Moteiro SN, Terrones LAH, Almeida JRM. 2008. Mechanical performance of coir fiber/polyester composites, Polymer Testing, 591-595.
M. Pollini, M. Russo A. Licciulli, A. Sannino, A.
Maffezzoli. 2009. Characterization of
antibacterial silver coated yarns, Journal
of Material Science, 20, 2361–2366.
38 Cord 2015, 31 (1)
An Investigation of the Tender Nut Potential of Diverse Coconut (Cocos nucifera L.) Varieties/Forms in Sri Lanka
S.A.C.N. Perera1, G.K. Ekanayake2 and H.M.N.B. Herath3
Abstract
There is a rising demand in the world for coconut water as a healthy natural beverage. Different coconut varieties are used in different countries to be processed as a natural drink. The coconut form “King coconut” has long been used in Sri Lanka as the ideal coconut variety for this purpose. However, with the expansion in the local and export beverage coconut market, the supply does not meet with the demand at present. Therefore, this study was conducted to identify potential coconut varieties/forms mainly relating to the yield of beverage. The study was conducted in the main coconut triangle and Southern Sri Lanka. Eleven coconut forms, namely, King coconut and Bothal thembili in the variety Aurantiaca, Red, Yellow, Green and Brown dwarf and Murusi in the variety Nana, Bodiri, Dothalu, and Sri Lanka tall. These are included in the variety Typica along with Nipol, which is a natural hybrid being identified to be suitable as beverage coconuts. Out of them the yielding ability was high in King coconut, Red dwarf and Yellow dwarf while Bodiri, Nipol and Dothalu fared well. Many coconut forms suitable for beverage purpose were observed to be having seasonality in nut production. Ensuring adequate soil fertility and prevention of water deficit over prolonged periods were observed to help sustain yield and reduce seasonality in bunch emission. There were no large scale plantations for beverage coconuts in Sri Lanka, and the entire collection was from home gardens and very small scale holdings. It is recommended to establish medium scale holdings with coconut forms having high yield potential as identified in the current study, with proper management guidance to ensure a steady supply of beverage coconuts to the local as well as export market.
Keywords: Beverage, Coconut, Cocos nucifera, King coconut, Varieties
______Coconut Research Institute, Lunuwila, Sri Lanka.
[email protected] [email protected] [email protected]
39 Cord 2015, 31 (1)
Introduction consumption of coconut in the world. The estimated per capita consumption of coconut in Currently, there is a worldwide demand for Sri Lanka is around 110 nuts/year (Anon, 2009) coconut as a natural beverage. Coconut water and 80% of the production is consumed became a popular natural beverage firstly in the domestically. Much of the production is used in coconut producing countries in Asia and in kernel-derived products: oil; copra; and Brazil and later in western countries, where a desiccated coconut; in addition to the use of huge demand has been created at present. There mature green nuts in daily cuisine. However, is a very high international demand for coconut consumption of both tender and mature coconut water, especially in the United States of water as a beverage has been a tradition in Sri America, and spreading in the Europe and Lanka over a long period of time and there are Middle Eastern Countries as well. certain varieties preferred for this purpose. Coconut water has gained recognition as a The semi tall coconut variety King healthy non-alcoholic natural beverage, suitable Coconut (locally known as Thembili or Rath as a restorative energy and sports drink. Coconut thembili) has long been known in Sri Lanka as water is rich in ions and minerals containing, the ideal variety for tender coconut beverage Potassium (at high concentration) and purpose (Liyanage, 1958; Bandaranayake and Magnesium, Calcium, Phosphorous, Iron, Fernando, 1999). King coconut is intermediate in Copper, Sulphur, Chlorides, Sodium as well as form between tall and dwarf. It is semi tall, has amino acids and vitamins B and C. Coconut a medium height growth rate, produces a root water is equally beneficial for everyone bole and is predominantly self pollinating. It including children and the aged. It is of pleasant bears profusely fruit having an orange epicarp. taste and has the added bonus of a low King coconut production in Sri Lanka is almost concentration of sugar in comparison to many exclusively for tender coconut water. However, other natural beverages. there are several other varieties which are less Almost all of the main coconut growing commonly used for beverage purposes and yet countries are in Asia except Brazil. Indonesia, with a higher potential productivity as beverage the Philippines, India, Brazil and Sri Lanka are coconuts. Therefore this study was conducted the major coconut growing countries. Out of with the objective of identifying varieties having them the main economic product in Brazil is a potential for beverage production and studying tender coconut water, while kernel and copra some of their specific characteristics which make products have been economically more them suitable for tender nut purposes. important in Asia. Despite this, there are Materials and methods expanded tender coconut industries in Asian countries such as India and Thailand. In these A field survey was conducted in Puttalum, countries different varieties and hybrids are used Chillaw and Gampaha in the main coconut for beverage purposes. In India, Chowghat triangle in the North Western province and Orange Dwarf has been found to be the best Galle, Unawatuna area in the Southern Sri tender nut variety followed by Malayan Orange Lanka. The coconut varieties which were used Dwarf and Philippine Ordinary (Dhamodaran et for beverage purposes were identified. al., 1993). An aromatic and other dwarf varieties Approximately 30 palms (except for are popular in Thailand and other South East Dothalu and Nipol in which the sample sizes Asian countries. Although there may be several were smaller) of each of the identified varieties varieties suitable as tender coconuts there is were used for recording the variables: number of competition for them to be used in other bunches; number of nuts in each bunch; nut industries especially kernel based products such water volume at 7-8 month maturity stage; as oil and copra. seasonality in bunch emission; and nut setting. Sri Lanka is the 5th largest coconut Observations were made over a period of one producer and has the highest per capita year and data were analysed by deriving means
40 Cord 2015, 31 (1)
Table 01. Coconut varieties used as beverage coconuts at the tender nut stage
Common name Variety/form (in Sri Distribution and occurrence of palms Lankan classification)
Thembili Aurantiaca/King coconut Throughout the coconut growing areas.
Bothal Thembili Aurantiaca/Bothal thembili Only in the Southern Province of Sri Lanka
Red Dwarf Nana/Red Dwarf Throughout the country. More common in the Southern Province.
Green Dwarf Nana/Green Dwarf Throughout the country. More common in the Southern Province.
Brown Dwarf Nana/Brown Dwarf Throughout the country. More common in the Southern Province.
Yellow Dwarf Nana/Yellow Dwarf Throughout the country. More common in the Southern Province.
Murusi Nana/Green Dwarf „Murusi‟ Only in the Southern Province of Sri Lanka
Bodiri Typica/Bodiri Throughout the country. More common in the Southern Province.
Dothalu Typica/Dothalu Only in the Southern Province of Sri Lanka
Nipol* A natural hybrid Throughout the country
Sri Lanka Tall Typica/Typica Throughout the country
*Nipol is a natural hybrid between King coconut and Sri Lanka tall
Figure 01. Coconut forms used as tender nut beverage in Sri Lanka
41 Cord 2015, 31 (1) and performing an analysis of variance and mean of the Tall along with King variety in home separation in statistical software SAS v8. gardens has simultaneously given rise to Nipol, a natural hybrid. While the King coconut is Results and discussion naturally self-pollinating there is some out- Varieties used for beverage purpose in Sri crossing. Therefore, when the farmers collect Lanka seednuts of the King coconut there would be a The results revealed the presence of low percentage of Nipol hybrid seedlings due to several varieties/forms which are used for pollen shed by the surrounding Tall palms. beverage at the tender nut stage (Figure 01). Out Evaluation of the potential of different of them, King Coconut was identified to be the varieties/forms for beverage purpose dominant variety for beverage purpose in Sri The yield variables related to quantity of Lanka. The other varieties were used in more nut water measured at the 7-8 month limited quantities at the tender nut stage (Table developmental stage of the nuts are presented in 01). Table 02. Thembili (King coconut) exceeded 75% of Number of bunches emitted during a given the tender nut market in Sri Lanka. Also the period, number of nuts produced in each bunch main purpose of growing Thembili was to be and the volume of nut water at the appropriate used as beverage at the tender nut stage; being 7- stage are the main yield components. These 8 months of developmental from flowering. The variables were significantly different among the coconut form Red Dwarf was also highly valued studied coconut forms (Table 02). Bodiri as beverage coconuts. The general public could recorded the highest values for both bunch not distinguish the fruit of Thembili and Red emission and the nut production. However, the dwarf due to the close morphological similarity nuts of Bodiri were smaller resulting in a lesser of the fruit forms. The pericarp colour of both amount of water per nut. It was observed that was orange while the only visual difference was when the number of nuts per bunch was higher the unique button-like protrusion at the tip of the size of the nuts became smaller and vice- Thembili fruit. The remaining dwarf forms; versa in prolific bearers: Green Dwarf; Brown green, yellow and brown, were used in limited Dwarf; Murusi; Bodiri; and Dothalu, resulting in scale as beverage being less common, especially lesser volume of water per nut. This factor was in the main coconut triangle of the country. The not prominent in the other phenotypes. Clearly main commercially grown cultivar, Sri Lanka the above values need be considered in tall was occasionally used for beverage purposes combination rather than in isolation to determine all over the country. The other listed two tall the yield potential of different varieties and forms, Dothalu and Bodiri were also used forms for beverage purpose. The total volume of because their smaller nut size made them less nut water calculated in this manner in each desirable for processing into kernel products. coconut form/variety is presented in Figure 02. Both were rare and found only in the Southern Province of Sri Lanka. Murusi has the phenotype As presented in Figure 02, Red Dwarf, King of the Green Dwarf very common in the home Coconut and Yellow Dwarf were the highest gardens of Southern province. It was a very yielders in that order, each recording volumes popular beverage form in the south of Sri Lanka. exceeding 50 litres per palm in the annum. Bothal thembili was also found only in the south Bodiri, Nipol, Sri Lanka Tall and Dothalu were being a form related to the variety Aurantiaca, also higher yielders while the rest of the dwarf but with a different nut shape (Figure 01). Bothal varieties along with Bothal thembili recording thembili was also a source of beverage. Among volumes in the range of 30-40 litres of nut water the various forms used for beverage in Sri per palm in the annum. Lanka, are several different phenotypes which are not found in the main coconut triangle, but only in the south. The wide occurrence together
42 Cord 2015, 31 (1)
Table 02. Beverage related yield variables of different coconut varieties/forms Common name Mean number of Total nut No. of Volume of bunches in the production in the nuts/bunch water/nut (mL) year year Thembili 15.6b 156c 10.0 352bc
Bothal thembili 13.6cd 108.8e 8.0 316d
Red Dwarf 15.5b 164.3c 10.6 346c
Green Dwarf 14.6bc 150.2cd 10.3 231ef
Brown Dwarf 13.2d 137.3d 10.4 236ef
Yellow Dwarf 14.1c 138.2d 9.8 378b
Murusi 14.2c 149.1cd 10.5 252e
Bodiri 16.5a 221.1a 13.4 224f
Dothalu 14.2c 184.6b 13.0 218f
Nipol 13.5d 140.4d 10.4 336c
Sri Lanka Tall 13.5d 91.8e 6.8 448a
Figure 02. Annual volume of nut water in diverse coconut varieties/forms
43 Cord 2015, 31 (1)
Seasonality in nut production potential as beverage coconuts. This will avoid competition with processors seeking supplies of Seasonality in bunch emission was mature. Collection of coconut water from the observed to varying degrees in all the coconut mature nut at the processing plant of kernel forms studied. In general bunch emission was products is also done. Alternative techniques are low in the months of December to February in needed in processing this mature nut water for all the forms including the widely grown beverage purpose instead of discarding it. In this commercial Sri Lanka Tall, which is known to method a smaller amount of water will be be hardy and more tolerant to variation in obtained per nut but there will be the twin environmental conditions. During this period the advantage of deriving kernel products and saving number of bunches, number of nuts per bunch, the mature nut water for a separate beverage as well as the nut size, were observed to be less. market. In addition, the dwarfs (except Yellow Dwarf), Bodiri and Dothalu, showed clear lean periods in The composition of nut water is an both bunch emission and nut production. This important quality issue in beverage coconuts that seasonality was less common in King Coconut, has not been considered in the current study. Red Dwarf and Nipol when they were under However, earlier studies have reported that the good management and when the palms were not composition of nut water depended on the stage subjected to prolonged periods of water deficit. of maturity of the nut (Nathanael, 1970). The desirable palatability of King coconut and other Potential varieties/forms as tender nut beverage forms mainly depends on the sugars beverage coconuts and mineral components present. The maximum In Sri Lanka the predominantly grown concentration of sugar and optimum levels of tender nut beverage coconut variety is King minerals and vitamins are found at the 7-8 Coconut or Thembili, in local dialect. The months stage from the pollination present study provided evidence for the equal (Gunawardena, 1973; Ediriweera, 1996). The potential of Red Dwarf and Yellow dwarf with sugar component comprises a larger portion of respect to yielding ability and hence need to be reducing sugars compared to non-reducing promoted in the development of tender nut sugars (Mohandas, 1982). With the ripening of beverage industry within Sri Lanka as well as to the nuts major changes in composition of sugars provide for the export market. takes place. There may be genetic differences in In the current study Nipol is the only these factors in the potential beverage varieties hybrid that was tested. Nipol is not a identified in this study and future research efforts recommended coconut hybrid in Sri Lanka. on the biochemical aspects could help in However, this natural hybrid proved to have a developing the quality of beverage coconuts. high potential for beverage production. Beverage coconuts are not grown in Similarly, the recommended hybrids between tall plantation scale in Sri Lanka but only in home and dwarf coconuts have been known to have a gardens or as small blocks adjacent to larger high potential or beverage (Dhamodaran et al., plantations. However, with the huge increase in 1993) However, in Sri Lanka, there is a very local and international beverage there is an high competition for the nuts of these urgent need for establishing larger scale coconut recommended hybrids for different uses mainly holdings for beverage coconuts. With the for kernel products, limiting their availability for information derived in the current study suitable beverage. This situation is more so for the most varieties for this purpose can be selected to expansively grown commercial coconut cultivar, provide a generically diverse population of nut Sri Lanka Tall, which is almost entirely water producing palms. harvested at the mature stage and used in kernel The coconut forms King coconut, Red based products. Therefore, in Sri Lanka it is Dwarf and Yellow dwarf are the best suited advisable to concentrate on the coconut forms forms throughout the country. In addition, Bodiri which have been identified as having a high
44 Cord 2015, 31 (1) and Nipol will also be very high yielders. Using Gunawardena, M. 1973. A study of the free Sri Lanka tall and recommended coconut hybrids amino acids in the liquid endosperm of will not be practical due to the demand for kernel coconut. Ceylon Coconut Quarterly products. There is a need to rationalise 24:102-106. management of the tender nut beverage coconut Liyanage, D.V. 1958. Varieties and forms of the holdings. Regional diversity could avoid coconut varieties grown in Ceylon. Ceylon localised periods of water deficit and overcome Coconut Quarterly 11:43-51. seasonality and to stabilise the supply of nuts. With adequate soil fertility and soil moisture Mohandas, S. 1982. Report of the coconut supply, all the dwarf forms can be promoted as processing division. Ceylon Coconut suitable for tender nut beverage purposes in Sri Quarterly 33:48-56 Lanka. Nathanael, W.R.N. 1970. Non-conventional uses Acknowledgments and processing techniques for coconut products. Ceylon Coconut Quarterly This research was conducted with the 21:99-106. financial assistance from the Coconut CESS
Fund in Sri Lanka. Authors are grateful to Dr. L. Perera, Head of the Genetics and Plant Breeding Division and Dr. J.M.D.T. Everard, former Deputy Director of the Coconut Research Institute of Sri Lanka, for their assistance and comments. References Anonymous 2009. Annual Report of the Central Bank of Sri Lanka.
Bandaranayake, C.K. and Fernando, W.M.U. 1999. Genetic improvement of King coconut, Cocos nucifera var. aurantiaca in Sri Lanka. Plant Genetic Resources Newsletter No. 118:30-33.
Dhamodaran, S., Ratnambal, M.J., Chempakam, B., Pillai, R.V. and Viraktamath, B.C. 1993. Evaluation of tender nut water in coconut cultivars. In: Proceedings of the International Symposium on Advances in Coconut Research and Development ISOCRAD II (Eds: M.K. Nair, H.H. Kan, P.
Ediriweera, N.D. 1996. King coconut. CORD, XII No.2:43-47. Gopalasundaram and E.V. Bhaskara Rao). Oxford & IBH Publishing Co. Pvt. Ltd., New Delhi. pp. 123-128.
45 Cord 2015, 31 (1)
Pollen Dispersal and Pollination Patterns Studies in Pati Kopyor Coconut using Molecular Markers
Siti Halimah Larekeng1,2, Ismail Maskromo1,3, Agus Purwito1, Nurhayati Anshori Matjik1, and S. Sudarsono1
Abstract
Parentage analysis has been used to evaluate pollen dispersal in Kopyor coconut (Cocos nucifera L.). Investigations were undertaken to elucidate (i) the dispersal of pollen, (ii) the rate of self and out- crossing pollination, and (iii) the distance of pollen travel in Pati kopyor coconut population. The finding of this activities should be beneficial to kopyor coconut farmers to increase their kopyor fruit harvest and to support breeding of this unique coconut mutant. As many as 84 progenies were harvested from 15 female parents. As many as 95 adults coconut provenances surrounding the female parents were analyses as the potential male parents for the progenies. The adult coconut palms were mapped according to their GPS position. All samples were genotyped using six SSR and four SNAP marker loci. Parentage analysis was done using CERVUS version 2.0 software. Results of the analysis indicated that evaluated markers were effective for assigning candidate male parents to all evaluated seedlings. There is no specific direction of donated pollen movement from assigned donor parents to the female ones. The donated pollens could come from assigned male parents in any directions relative to the female parent positions. Cross pollination occured in as many as 82.1% of the progenies analyzed. Outcrossing among tall by tall (TxT), dwarf by dwarf (DxD), hybrid by hybrid (HxH), TxD, DxT, TxH, DxH, and HxD were observed. Self-pollination (TxT and DxD) occurred in as many as 17.9% of the progenies. The dwarf coconut was not always self pollinated. The presence of DxD, TxD, and HxD outcrossing was also observed. The donated pollens could come from pollen donor in a range of at least 0-58 m apart from the evaluated female recipients. Therefore, in addition to the wind, insect pollinators may have played an important role in Kopyor coconut pollination.
Keywords: Coconut mutant, abnormal endosperm, parentage analysis, pollen movement, outcrossing rate, self pollination, SSR marker, SNAP marker
______1PMB Lab., Department of Agronomy, Fac. of Agriculture, IPB, Bogor, Indonesia. 2Fac. Of Forestry, Hasanuddin University, Makasar, Indonesia. 3Indonesian Palm Research Institute, Agency for Agricultural Research and Development (AARD), Manado, Indonesia. Corresponding author: S. Sudarsono, PMB Lab., Department of Agronomy, Fac. of Agriculture, IPB, Jl. Meranti – Darmaga Kampus, Bogor 16680, Indonesia. Ph./Fax. 62-251-8629353; Email: [email protected]
46 Cord 2015, 31 (1)
Introduction strategy. Tall coconuts are generally outcrossing since male flower mature earlier than the female Kopyor coconuts are natural coconut counterpart in the same inflorescence. Dwarf mutants having abnormal endosperm and only coconut tends to self-pollinate because of an exist in Indonesia. The endosperm is soft, overlapping maturation period between male and crumbly and detached from the shell, forming female flowers (Deb Mandal and Shyamapada flakes filling up the shell (Maskromo et al. 2007; 2011). Novarianto et al. 2014). The Makapuno coconut grown in the Philipines and other Asian countries Pollination in coconut most probably is is another example of coconut mutant exhibiting assisted by insect pollinators or by the wind endosperm abnormality (Samonthe et al. 1989; (Ramirez et. al. 2004). The family of Diptera, Wattanayothin, 2010). This mutant has been used Coleoptera and Hymenoptera are reported as as parent for hybridizations in coconut breeding effective pollinators of coconut (Ramirez et al. (Wattanayothin, 2005). The Macapuno coconut 2004). Distances of pollen transfer between male exhibits a soft and jelly-like endosperm (Santos, and female parents may be used to predict the 1999) that is phenotypically different to type of pollinator assisting pollination in coconut. Indonesian Kopyor coconut. Such question may be answerred by studying pollen dispersal. The kopyor coconut mutant phenotype is genetically inherited from parents to their Evaluating pollen dispersal in various progenies (Sukendah 2009) and most probably is plant species usually use an approach based on controlled by a single locus (K locus) regulating the parent – progeny genotype genotype the endosperm development of coconut (Austerlitz et al. 2004). Evaluations have been (Sudarsono et al. 2014). However, the identity of done in pines (Schuster and Mitoon 2000), the regulatory locus has not yet been resolved. Dinizia excels - Fabaceae (Dick et al. 2003), Quercus garryana - Fagaceae (Marsico et al. The abnormal endosperm phenotype in 2009) and teak (Prabha et al. 2011). Availability kopyor coconut is controlled by the recessive k of molecular markers capable of identifying allele; therefore, the genotype of kopyor fruit of genotype of parents and their progenies should coconut would be homozygous kk for the zygotic assist the pollen dispersal studies. Using such embryos and homozygous kkk for the endosperm. markers, it should also be possible to estimate On the other hand, the genotype of the normal the self-pollination and outcrossing rates in a fruit of coconut would either be a homozygous certain population (Milleron et al. 2012). KK or a heterozygous Kk for the zygotic embryo and either a homozygous KKK, heterozygous To our understanding, pollen dispersal KKk, or heterozygous Kkk, respectively. analysis has not been evaluated in coconuts. With the development of kopyor coconut in The origin of Kopyor coconut mutant is Indonesia, availability of information associated not well documented; however, currently the with pollen dispersal should be beneficial kopyor palms are found in a number of areas in considering the recessive nature of the kopyor Java and southern part of Sumatera (Novarianto character. Such coconut pollen dispersal and Miftahorrachman 2000). The district of Pati, evaluation requires availability of some coconut Central Java Province is recognized as one of the progeny arrays and polymorphic loci for Kopyor coconut production centers. Kopyor molecular markers of coconut genome. coconuts have existed in this region for generations, especially the dwarf type of Kopyor Co-dominant markers, such as SSR and coconuts. Although only in a limited numbers, SNAP markers for coconut have been developed Kopyor Tall and Kopyor Hybrid coconut types and routinely evaluated at PMB Lab, Department also exist along side of the dwarf one. of Agronomy and Horticulture, Faculty of Agriculture, Bogor Agricultural University The tall and dwarf coconut have different (IPB), Bogor, Indonesia for a number of plant morphological characters and pollination species. These include coconut (Sudarsono et al.
47 Cord 2015, 31 (1)
2014), cacao (Kurniasih 2012), and nut meg – coconut type, the sampled population consisted Myristica sp. (Soenarsih 2012). Moreover, the of 68 dwarf, 14 tall and 13 hybrid coconuts. gene specific SNAP markers have also been Moreover, based on their phenotype, they were developed and used successfully in coconut recognized as 22 normal homozygous KK and (Sudarsono el al. 2014). 73 kopyor heterozygous Kk coconuts. Map of the existing coconuts in the research site was The SSR markers have successfully been generated using the GPS position of all used in gene flow analysis of pines (Lian et al. individuals. 2001; Burczyk and Koralewski. 2005). SNAP marker have also been reported as an effective Six dwarf, seven tall, and two hybrid co-dominant marker for plant analysis (Morin et coconuts among the kopyor heterozygous Kk al. 2004, Sutanto et al. 2013) and proven to trees were selected as female parents. They were generate better data quality for the majority of selected using purposive random sampling to samples on plant genetic studies (Brumfield et represent different sites in the sampled al. 2003) and population structure analysis population. A single fruit bunch from each (Herrera et al. 2007). female parent containing 2-10 fruits/bunch was harvested 10-11 months after pollination. The The objectives of this research were is to total harvested fruits were collected and evaluate (i) the dispersal of pollen, (ii) the rate of identified as either kopyor or normal fruits. The self and out-crossing pollination, and (iii) the identified normal fruits were germinated and distance of pollen travel in Pati kopyor coconut DNA was isolated from leaf tissue of the population. The finding of these activities should germinated seedlings (63 seedlings of normal be beneficial to kopyor coconut farmers to fruits). The kopyor fruits are not able to naturally increase their kopyor fruit yield and to support germinate since this character is lethal. Zygotic breeding and cultivar development of this unique embryos were isolated from the identified mutant. kopyor fruits and DNA was isolated directly Materials and methods from the whole zygotic embryo tissues (21 zygotic embryos). Among the 84 DNA samples, Time and Location of Research 26 samples were from tall, 45 from dwarf, and This research was conducted during the 13 from hybrid female parents. period of July 2012 up to January 2014. The field activities were at the Kopyor coconut Genotyping of Parents and Progenies plantation belonging to local farmer’s at DNA isolation was conducted using the Sambiroto, Pati District, Central Java, Indonesia. CTAB method (Rohde et al. 1995). Either young The research site was at the following GPS coconut leaf or zygotic embryo (0.3-0.4 g) was location: S 6 32.182 E 11 03.354. The soil in the homogenized in 2 ml of lysis buffer, containing evaluated Kopyor coconut plantation is sandy 0.007 g PVP and 10 μl2-mercaptoetanol. The soil. The laboratory activities were done at Plant homogenized tissues were then incubated in Molecular Biology Laboratory (PMB Lab), 65°C waterbath for 60 minutes and the mixtures Department of Agronomy and Horticulture, were centrifuged at 11000 rpm for 10 minutes Faculty of Agriculture, Bogor Agricultural using using the Eppendorf 5416 centrifuge. University, Bogor, Indonesia. Supernatant was then transferred to an Eppendorf tube and an equal volume of Selection of Parents and Progeny Arrays chloroform:isoamyl-alcohol (24:1) was added. There were 164 adult coconut trees in the The mixtures were mixed well; centrifuged at field research site, consisted of a mixture of both 11000 rpm for 10 minutes and the supernatant kopyor heterozygous Kk and normal was transferred into new microtube. homozygous KK coconut trees. Only 95 out of 164 adult coconut trees in one block of 100x100 Cold isopropanol (0.8 volume of m2 were sampled in this evaluation. Based on the supernatant) and sodium acetate (0.1 volume of supernatant) were added into the supernatant.
48 Cord 2015, 31 (1)
After overnight incubation, the mixture was Identification of the Candidate Male Parents centrifuged at 11000 rpm for 10 minutes and Each sample of the progeny arrays has a DNA pellet was retained. The DNA pellet was known female parent but unknown pollen donor washed using 500 μl of cold 70% ethanol, (the male parent). The candidate male parents centrifuged and air dried before it was diluted could be any one of the sampled adult population into100 μl aquabidest. RNA contaminants were including the female parents. Studies were remove using RNase treatment following conducted to determine the assigned male parent standard procedures (Sambrook and Russel donating pollen to generate any fruit in the 2001). progeny arrays. SSR marker at 37 loci (Lebrun et al. 2001) Identification of the assigned male parent were evaluated for their polymorphism 6 was done by analyzing genotype of progeny and polimorphic loci were selected. In addition, four the respective female parent versus the genotype SNAP marker loci developed based on of all adult trees in the selected samples. The ID nucleotide sequence variabilities of both SUS of the potential male parent for any progeny was and WRKY genes were also used to genotype all determine based on the results of parentage of the parents and progeny arrays. To generate analysis. Simulation was conducted to determine markers, PCR amplifications were conducted the threshold level for confidence interval of using the following reaction mixtures: 2µl of 80% (relax) and 95% (strict) levels before the DNA; 0.625µl of primers, 6.25µl PCR mix final parentage analysis steps. Parentage analysis (KAPA Biosystem), and 3µl ddH20. using the genotype of progenies, female parents, Amplifications were conducted using the and potential male parents was done using following steps: one cycle of pre-amplification at CERVUS version 2.0 software (Marshall et al. 95°C for 3 minutes, 35 cycles of amplification 1998). Most likely approach (potential male steps at 95°C for 15 seconds (template parent with the highest LOD score) based on the denaturation), annealing temperature for 15 matching genotype of progeny, female parent seconds (primer annealing), and 72°C for 5 and potential male parent were used as the basis seconds (primer extension), and one cycle of for assigning certain adult individual as the final extention at 72°C for 10 minutes as potential male parent or pollen donor of a suggested by KAPA Biosystem kit. progeny. The progeny and female parent The generated SSR markers were genotype were compared with those of other separated using vertical 6% polyacrilamide gel adult trees and the assigned male parent was electrophoresis (PAGE) using SB 1x buffer selected based on the output of CERVUS version (Brody and Kern 2004) and stained using silver 2.0 analysis results (Marshall et al. 1998). staining. The silver staining was done following Pattern of Pollen Dispersal methods developed by Creste et al. (2001). Electrophoregrams were visualized over the light The location of the female and the table and used to determine the genotype of the assigned male parents were plotted in the map of evaluated samples. adult individuals generated by Garmin MapSource GPS mapping software version The generated SNAP markers were 76C5x. The distance between the known female separated using 1% agarose gel electrophoresis parent and the assigned male parent was using TBE 1x buffer and stained using standard calculated using the same software. The DNA staining procedures (Sambrook and Russel distances and positions of both female and male 2001). The electrophoregrams were visualized parents in the generated map was then used to over the UV transluminescence table and ilustrate pattern of pollen dispersal in the recorded using digital camera. The recorded location. Self pollination was defined if the pictures were used to determine the genotype of assigned male parent was the same as the female the evaluated samples. parent. Otherwise, they were assigned as
outcrossing. The outcrossing were further
49 Cord 2015, 31 (1) grouped as outcrossing between either dwarf allele (Fig. 3). All individuals were genotyped (dwarf parent pollinated other dwarf), tall (tall using the same approaches. The summary of parent pollinated other tall), or hybrid (hybrid genotping results for a total of 179 individuals parent pollinated other hybrid); outcrossing using six SSR and four SNAP marker loci are between dwarf and either tall or hybrid (either presented in Table 1. The marker loci generated tall or hybrid parent pollinated by dwarf); a range of 2-4 alleles per locus (Table 1). outcrossing between tall by hybrid coconuts (tall Mean number of alleles per locus is 3.4 parent pollinated hybrid) or vice versa. The and mean PIC for all marker loci was 0.47. The numbers of both self pollination and the polymorphic information content (PIC) for SSR respective cross pollination were calculated. marker loci ranges from 0.31-0.68 while that of Results and discussion SNAP markers ranges from 0.28-0.37 (Table 1). The PIC values represents measures of The Parents and Progeny Arrays polimorphism between genotypes in a locus Map of the existing coconut palms in the using information of the allele numbers (Sajib et research site are presented in Fig. 1. As al. 2012). Total exclusionary power using the ten indicated, the sample coconut population marker loci is either 0.85 (first parent) or 0.97 consists of a mixture of normal homozygous KK (second parent), indicating the SSR and SNAP and kopyor heterozygous Kk individuals and a markers should be informative enough for mixture of dwarf, tall and hybrid coconuts. All analyzing the evaluated coconut population. of these adult trees were used as potential male parents capable of donating pollens to and pollinating the selected female parents and generating the evaluated progeny arrays. The position of the selected female parents (6 dwarf, 7 tall, and 2 hybrid kopyor heterozygous Kk coconuts) are indicated in Fig. 1. The harvested progenies from selected female parents ranged from 2-10 progenies per female parent. Out of 84 selected progenies, 21 were kopyor nuts and 63 were normal ones. They were harvested from tall (26 progenies), dwarf (45 progenies), and hybrid (13 progenies) female parents, respectively.
Genotyping of Parents and Progeny Arrays The selected SSR and SNAP marker loci Figure1. Map of study site with the existed generated polymorphic markers in the evaluated coconut palms at Pati, Central Java, coconut population. An example of the Indonesia. The marks in this map polymorphic marker generated by either the indicated the positions of the coconut selected SSR (CnCir_56 locus) and SNAP (SUS provenances. The sampled 1_3 locus) primer pairs producing polimorphic provenances in an approximately one markers is presented in Fig. 2. and 3. In Fig. 2, hectare area are in the square box. The the evaluated individuals are either homozygous flags indicate the positions of the cc (sample #1), bb (sample #7-10), heterozygous selected female parents. bc (sample # 2-6, and 11), or heterozygous ab Note: The marks indicate the position (sample # 12) for the CnCir_56 SSR locus. On of ( ) normal tall, ( ) kopyor tall, ( ) the other hand, the evaluated individuals (sample normal dwarf, ( ) kopyor dwarf, ( ) # 1, 3, 4, 6) are heterozygous for reference and normal hybrid, and ( ) kopyor hybrid alternate SNAP alleles and the other two (sample coconuts, respectively. # 2 and 5) are homozygous for the reference
50 Cord 2015, 31 (1)
1 2 3 4 5 6 7 8 9 10 11 12 M
Figure 2. Polymorphism of SSR markers generated by PCR of the genomic DNA sample #1-12 with a pair of CnCir_56 SSR primers. M: 100 bp DNA ladder markers. The a, b, and c are the three specific alleles of the CnCir_56 locus
1 2 3 4 5 6 R A R A R A R A R A R A M
Figure 3. Polymorphism of SNAP markers generated by PCR of the genomic DNA samples 1-6 with two pairs (the R: reference and the A: alternate primer pairs) of the SUS 1_3 SNAP locus. M: 100 bp DNA ladder markers. The R– PCR product of the R primer pairs and the A – PCR product of the A primer pairs. The occurrences of both R and A PCR products indicating the evaluated individuals are heterozygous, while if either onlyA or B indicating they were homozygous
51 Cord 2015, 31 (1)
Table 1. Number of alleles and individuals, heterozygous and homozygous individuals, observed (O) and expected (E) heterozygosity, and polymorphic information content (PIC) at 10 molecular marker loci of Kopyor coconut No. Of Heterozygosity PIC No. of No. of Locus Name Hetero- Homozy- alleles individual O E zygous gous CnCir_87 2 179 27 152 0.15 0.39 0.31 CnCir_86 4 179 100 79 0.56 0.72 0.67 CnZ-18 4 179 74 105 0.41 0.61 0.57 CnZ_51 5 179 72 107 0.40 0.58 0.54 CnCir_B12 6 176 64 112 0.37 0.72 0.68 CnCir_56 5 179 78 101 0.44 0.64 0.57 CnSus1#14 2 179 143 36 0.80 0.49 0.37 CnSus1#3 2 174 147 27 0.85 0.50 0.37 WRKY19#1 2 176 111 65 0.63 0.47 0.36 WRKY 6#3 2 179 76 103 0.43 0.34 0.28
Table 2. Crossing schemes and pollination types identified based on results of pollen dispersal analysis of the progeny arrays Crossing Scheme Pollination types Event numbers Percentage TxT Self 2 2.4% DxD Self 13 15.7% HxH Self 0 0 Sub-total Self 15 17.9 TxT Outcross 4 4.8% TxD Outcross 16 19.3% TxH Outcross 4 4.8% DxD Outcross 15 18.1% DxT Outcross 6 7.2% DxH Outcross 11 13.3% HxH Outcross 2 2.4% HxT Outcross 0 0 HxD Outcross 11 13.1% Sub-total Outcross 69 82.1% Total progenies 84 100.0% Note: T - tall coconut, D - dwarf coconut, and H - hybrid coconut
52 Cord 2015, 31 (1)
Identification of the Candidate Male Parents probably always cross pollinated (Ramirez et al. 2004; Maskromo et al. 2011). However, our data Results of simulation analysis using indicated there are at least 2.38% of self 10.000 iterations, 95 candidate male parents, and pollination among the tall coconut (Table 2). the known female parent for each progeny, predicted the rate of success in identifying male Self pollination is characterized by the parents at 95% (strict) was 32% and at 80% pollination of female flower by male pollen of (relax) confidence interval was 62%. Parentage the same parent. Self pollination produces full- analysis was able to resolve the identity of the sib progenies. Total numbers of self pollination male parent for every individual in the 84 are observed in as many as 15 events (17.9 %) in progeny arrays using the most likely parent the evaluated progeny arrays (Table 2). They approach. Moreover, the results of analysis also consist of two self pollination events in the tall indicated that assignment of the predicted male kopyor coconut (2.38 %) and 13 self pollination parents for the 20% (17 individuals) progenies events in the dwarf kopyor one (15.48%). Based are at least in the minimum of 95% confidence on 13 progeny arrays harvested from the hybrid and 43% (36 individuals) were at least in the parents, no self pollination in the hybrid coconut minimum of 80% confidence. The assignment is recorded (Table 2). for the male parents of other 57% (48 The general understanding stated that individuals) progenies were at the level of less because of the overlapping period between male than 80 % confidence. Although the confidence and female flower maturation, dwarf coconut is level was below 80 %, the male parent always self pollinated (Maskromo et al. 2011). assignment for these progenies shows LOD However, our data indicated the dwarf coconut is (likelihood of odds) value higher than 0. A not always self pollinated. Contrary to the basic positive LOD value indicates the suspected male understanding, our data indicated the presence of parent might be the true parents. According to more dwarf to dwarf (15 events, 17.86%), dwarf Marshall et al. (1998), the higher the LOD value to tall (6 events, 7.14%) and dwarf to hybrid (11 the higher the possibility for the assigned male events, 13.1%) outcrossing (Table 2). parent to be the actual parent (Marshal et al. 1998). Finding by Rajesh et al. (2008) has previously indicated that cross pollination did Cross pollination is pollination of female occur in dwarf coconuts. Availability of new flower by male pollen from different parents. tools, such as molecular markers, for analyzing Cross pollination produces half-sib progenies. outcrossing rate may change the previous The tall, dwarf and hybrid coconuts could understanding. Such changes have been shown reciprocally donate their pollens. Based on the in Hymenaea coubaril which was previously assigned male parent of the 84 progeny arrays, reported as more cross pollinated because of self cross pollination occured in as many as 69 incompatibility (Dunphy et al. 2004). However, events (82.1 %). Among those identified as more recent pollen dispersal studies indicated outcrossing, 4 events are cross pollination that H. coubaril is more self pollinated (Carneiro between tall x tall (TxT), 16 tall by dwarf (TxD), et al. 2011). and 4 tall by hybrid (TxH) parents. Moreover, outcrossing among DxD (15 events), DxT (6 Other alternative explanation for this events), DxH (11 events), HxH (2 event) and findings is that it is just a special case in the HxD coconuts (11 events) were also observed. evaluated site. In the study site, coconut palms Complete scheme and pollination types were planted in high density. Moreover, identified based on results of pollen dispersal population of honey bees exist in the coconut analysis are presented in Table 2. plantation. Honey bees are known to roam around the male and female flowers and function The general understanding stated that as effective pollinators for coconuts. The high because of the open flower morphology and the density planting and the availability of differences in flower maturation, tall coconut is pollinators may have caused the unexpected
53 Cord 2015, 31 (1)
Numberof male parent
Number of donated pollen Figure 4. Numbers of assigned male parent donating different numbers of pollen to evaluated female parents
Numberof female parent
Number of assigned male parents
Figure 5. Numbers of female parent receiving donated pollens from different number of assigned male parents
vents Number of e of Number
Class distance between male to female (m) Figure 6. Numbers of pollination events for each distance class from the assigned male to the female coconut parents
54 Cord 2015, 31 (1) outcrossing rate. However, those are subjects of a single female recipient parent are presented in further investigations. Figures 7-11. One assigned male parent may donate one As the female parent, Hybrid kopyor # 059 or more pollens to the evaluated female coconut (Fig. 7) received 6 donated pollens from six parent, with a range of 1-5 pollens per assigned different assigned pollen donors. The pollen male one. Number of assigned male parents contributors to the progeny array harvested from donating certain numbers of pollen to the Hybrid kopyor # 059 female parent were all evaluated female parents is presented in Fig. 4. kopyor heterozygous Kk coconuts. However, the The data indicate that most of the assigned male seven progenies harvested from this female parents contribute only one pollen to the parent were all phenotypically normal, i.e. evaluated female parents. Only three assigned genetically either a normal heterozygous Kk or male parents (two dwarf, and one hybrid homozygous KK. The positions of the assigned coconuts) donated 4 or 5 pollens to the male parents relative to the female parent # 059 surrounding female parents. in the study site are presented in Fig. 7. The same female parents may receive donated pollens from different numbers of assigned male parents, with a ranged of 1-7 assigned male parents donated pollen to the same female one. The numbers female parents receiving donated pollens from different number of assigned male parent iss presented in Fig. 5. The data indicated a single female parent most frequently received pollens from 2, 4 or 5 different assigned male parents. Only three female parents evaluated in this experiment (two dwarf and one hybrid coconuts) are found receiving pollens from at least 6 assigned male parents (Fig. 5).
Pattern of Pollen Dispersal Figure 7. Pattern of pollen movement to female The distances between female to the parent #059 inferred from parentage assigned male parents have been determined analysis. The mark indicates position based on their GPS positions. The distance of of ( ) Dwarf kopyor, ( ) Hybrid pollen travel between assigned male to female kopyor as the assigned male (pollen parents as measured in this evaluation ranged donor) parents, and ( ) hybrid from 0 - 58 m. Numbers of pollination events of kopyor #59 as the female recipient, each distance class from the assigned male to the respectively female coconut parents are presented in Fig. 6. The Dwarf kopyor # 067 (Fig. 8) received The assigned male parents are distributed almost 10 donated pollens from eight different assigned evenly in the different class distances from the male parents. The assigned pollen contributors to female parents. The 0 m distance between the Dwarf kopyor # 067 female parent were all parents indicates self pollination events. kopyor heterozygous Kk coconuts. Only one out To evaluate pattern of pollen dispersal of the 10 progenies harvested from this female among the assigned male parent to the female, parent was phenotypically kopyor. The assigned the positions of assigned male parents as pollen male parent for the harvested kopyor fruit was donors to one female parent are plotted to a map the tall kopyor # 089. The positions of the using their GPS positions. Representative assigned male parents relative to the female samples of the assigned male parent positions to parent # 067 are presented in Fig. 8.
55 Cord 2015, 31 (1)
kopyor as the assigned male parents (pollen donors), and ( ) Dwarf kopyor #068 as the donor pollens and female recipient, respectively Dwarf kopyor # 084 (Fig. 10) received 8 donated pollens from surrounding pollen donors. The pollen contributors to the Dwarf kopyor # 084 female parent were all kopyor coconuts. Only two out of the 8 progenies harvested from Dwarf kopyor # 084 were phenotypically Figure 8. Pattern of pollen movement to female kopyor. These two kopyor fruits received parent #067 inferred from parentage donated pollens from either The two assigned analysis. The marks indicate position male parents, either dwarf kopyor # 056 (one of ( ) Dwaf kopyor, ( ) Hybrid pollen) and hybrid kopyor # 057 (one pollen), kopyor, ( ) Tall kopyor as the each contributed a one pollen to the evaluated assigned male (pollen donor), and progenies. Moreover, assigned male parent # 32 ( ) Dwarf kopyor #067 as the donor is the most distance pollen contributor among the pollens and female recipient, evaluated trees. The positions of the assigned respectively male parents (pollen contributors) relative to the female parent # 084 are presented in Fig. 10. Dwarf kopyor # 068 (Fig. 9) received 9 donated pollens from four assigned male parents. The four progenies were the result of outcross with either hybrid (# 59) or dwarf (#87 or # 90) and from self pollination. The assigned pollen contributors to the Dwarf kopyor # 068 were all kopyor heterozygous Kk coconuts. Three out of the 9 progenies harvested from Dwarf kopyor # 069 were phenotypically kopyor. These kopyor fruits received one donated pollen from either the tall kopyor # 059, dwarf kopyor # 68 or # 87. The positions of the assigned male parents relative to the female parent # 68 are presented in Fig. 9. Figure 10. Patterns of pollen movement to female parent #084 inferred from parentage analysis. The marks indicate positions of ( ) Dwarf kopyor, ( ) Hybrid kopyor, ( ) the Tall kopyor as the assigned male - pollen donors, and ( ) Dwarf kopyor #84 as the female recipient, respectively Dwarf kopyor # 089 (Fig. 11) received 7 donated pollens from surrounding pollen donors. Figure 9. Pattern of pollen movement to female The pollen contributors to the Dwarf kopyor # parent #068 inferred from parentage 089 female parent were all kopyor coconuts. analysis. The marks indicate position None of the 7 progenies harvested from Dwarf of ( ) Dwarf kopyor and ( ) Hybrid kopyor # 089 was phenotypically kopyor. The
56 Cord 2015, 31 (1) positions of the assigned male parents relative to Kopyor coconut pollination, especially from the female parent # 089 are presented in Fig. 11. closely spacing male pollen donors. This might have been the first report of using molecular marker to study pollen dispersal in coconut. Results of this study point to new finding about pollen dispersal and pollination, selfing and out-crossing rates among dwarf, hybrid, and tall coconuts, respectively. However, further research and evaluation are necessary to generalize the finding since the present study is specific for the current study site. Conclusion The evaluated markers were effective for Figure 11. Pattern of pollen dispersal plotted assigning candidate male parents to all evaluated based on the assigned male parent seedlings. There is no specific direction of as pollen donor to female parent donated pollen movement from assigned donor #089. The marks indicate position parents to the female ones. The donated pollens of ( ) Dwarf kopyor, ( ) the Tall could come from assigned male parents in any kopyor as the assigned pollen directions relative to the female parent positions. donor, and ( ) Tall kopyor #89 as Based on the assigned male parent of the 84 the donor pollens and female progeny arrays, cross pollination occured in as recipient, respectively many as 69 events (82.1%) including one among tall by tall (TxT), dwarf by dwarf (DxD) and Fig. 7-11 indicated that there is no specific hybrid by hybrid (HxH) cross pollination events. direction of donated pollen movement from Moreover, outcrossing among TxD, TxH, DxH assigned male parents to the female parents. The and vice-versa were also observed. This finding donated pollen could come from assigned male also indicated the dwarf coconut is not always parents in any directions relative to the female self pollinated. The presence of 17.86 DxD, parent positions. 19.05% TxD and 13.10% HxD were also In the reseach location, wind blows from observed. In Kopyor coconut, the pollens could left to right during the night and from right to travel from pollen donors as far as 58 m apart left during the day. If the wind is the major from the evaluated female recipients. Therefore, pollinators, there should be a specific pattern of insect pollinators may have played an important pollen movement. Moreover, the distance of role in long distance pollen dispersal in Kopyor pollen dispersals should be close to the pollen coconut. donors. Our data did not support the wind as the Acknowledgment only major pollinator in Kopyor coconut since pollens disperse in random directions and the This research was supported by DGHE, assigned male parents are as far as 58 m apart MoEC, Republic of Indonesia through “HI- from the evaluated female recipients. Our data LINK Kopyor Coconut Project – 2012-2014” also indicated that insect pollinators may play an entitled “Increasing Kopyor Fruit Yield with the important role in Kopyor coconut pollination. Help of Honey Bee Pollinators and Production of Numbers of insects are associated with Kopyor true-to-type Seedlings through Control inflorescence of kopyor coconuts. Such insects Pollination” coordinated by SDR. This research may aid pollination and promote cross is used by SHL for her PhD dissertation. SHL pollination in kopyor coconuts, as it happens to was also supported by Graduate Scholarship other plant species (Bown 1988). These findings, Program (BPPS) and Hibah Doktor Program, however, do not rule out the role of wind in the DGHE, MoEC, Republic of Indonesia.
57 Cord 2015, 31 (1)
References and fragmented Amazonian rainforest. Mol Ecol. 12:753-764. Austerlitz F, Dick CW, Klein EK, Muratorio SO, Smouse PE, Sork VL. 2004. Using a Dunphy BK, Hamrick JL, Schwagerl, J. 2004. A genetic marker to estimate the pollen comparison of direct and indirect dispersal curve. Molecular ecology. measures of gene flow in the bat- 13:937-954. pollinated tree Hymenaea courbaril in the dry forest life zone of south-western Bown D. 1988. Aroid: Plants of the arum family. Puerto Rico. Int J Plant Sci. 165: 427-436. Oregon (US): Timber Press Portland. Herrera MM, Alan WM, James WB, David NK, Brody Jonathan R, Scott E Kern. 2004. History Raymond JS. 2007. Usefulness of WRKY and principles of conductive media for gene-derived markers for assessing standard DNA electrophoresis. Analytical genetic population structure: An example Biochemistry 333 (1): 1–13. with Florida coconut cultivars. Scientia Brumfield RT, Beerli P, Nickerson DA, Edwards Horticulturae 115:19–26. SV. 2003. The utility of single nucleotide Kurniasih Surti. 2012. Pemanfaatan marka polymorphisms in inferences of molekuler untuk mendukung perakitan population history. Trends Ecol. Evol. kultivar unggul kakao (Theobroma cacao 18:249-256. L.) [disertasi]. Bogor (ID): Institut Burczyk J, Koralewski T. 2005. Parentage versus Pertanian Bogor. two-generation analyses for estimating Lebrun P, Baudouin L, Bopurdeix T, Konan JL, pollen-mediated gene flow in plant Barker JHA, Aldam C, Herran A, Ritter E. population. Mol Ecol. 14:2525-2537. 2001. Construction of linkage map of the Carneiro FS, Lacerda AEB, Lemes MR, Gribel Rennell Island Tall coconut type (Cocos R, Kanashiro M, Wadt LHO, Sebbenn nucifera L.) and QTL analysis for yield AM. 2011. Effects of selective logging on characters. Genome. 44:962-970. the mating system and pollen dispersal of Lian C, Miwa M, Hogetsu T. 2001. Outcrossing Hymenaea courbaril L. (Leguminosae) in and paternity analysis of Pinus densiflora the Eastern Brazilian Amazon as revealed (Japanese red pine) by micro-satellite by microsatellite analysis. Forest Ecology polymorphism. Heredity 87:88–98. and Management. 262:1758-1765. Marshal TC, Slate J, Krilek LEB, Pemberton JM. Creste S, Tulmann AN, Figueira A. 2001. 1998. Statistical confidence for likehood Detection of single sequence repeat based paternity inference in nature polymorphism in denaturing populations. Mol Ecol. 7:639-655. polyacrylamide sequencing gels by silver staining. Plant Mol Biol Reporter 19:299- Marsico, T.D., Jessica, J.H. and Romero- 306. Saverson, J. 2009. Patterns of seed dispersal and pollen flow Quercus Deb Mandal M, Shyamapada M. 2011. Coconut garryana (Fagaceae) following post- (Cocos nucifera L. - Arecaceae): In health glacial climatic changes. J. Biogeogr. promotion and disease prevention. Asian 36(5):929-941. Pacific Journal of Tropical Medicine. 4(3):241-247. Maskromo I, H. Novarianto dan N. Mashud. 2007. Potensi pengembangan kelapa Dick, C. W., Etchelecu, G. and Austerlitz, F., kopyor di Indonesia. Warta Penelitian dan 2003. Pollen dispersal of tropical tree Pengembangan Tanaman Industri Vol 13 (Dinizia excels: Fabaceae) by native No. 1. insects and African honeybees in pristine
58 Cord 2015, 31 (1)
Maskromo I, Novarianto H, Sudarsono. 2011. 2004. Mixed mating strategies and Fenologi pembungaan tiga varietas kelapa pollination by insects and wind in coconut genjah kopyor pati. Di dalam: Roedhy P, palm (Cocos nucifera L. (Arecaceae)): Slamet S, Anas D, Nurul K, Dewi S, importance in production and selection. Sintho WA, editor. Prosiding Seminar Agricultural and Forest Entomology PERHORTI Kemandirian Produk 6:155-163. Hortikultura untuk Memenuhi Pasar Rohde W, Kullaya A, Rodriguez MJB, and Domestik dan Ekspor; 2011. Nov 23-24; Ritter E. 1995. Genetic analysis of Cocos Lembang, Indonesia. Bogor (ID): nucifera L by PCR amplification of spacer Perhimpunan Hortikultura Indonesia. Hlm sequences separating a subset of copia-like 1002-1010. EcoR1 repetitive elements. J Genet Breed. Milleron Matias, Unai Lopez de Heredia, Zaida 49 : 179-186. Lorenzo, Ramon Perea, Aikaterini Sajib AM, M.M. Hossain, A.T.M.J. Mosnaz, H. Dounavi, Jesus Alonso, Luis Gil, Nikos Hossain, M.M. Islam, M.S. Ali, SH. Nanos. 2012. Effect of canopy closure on Prodhan. 2012. SSR marker-based pollen dispersal in a wind-pollinated molecular characterization and genetic species (Fagus sylvatica L.). Plant. diversity analysis of aromatic landreces of Ecology. 213:1715-1728. rice (Oryza sativa L.). J. BioSci. Biotech Morin PA, Luikart G, Wayne RK, The SNP 1(2): 107-116. working group. 2004. SNPs in ecology, Sambrook J. and Russel DW. 2001. Molecular evolution and conservation. Trends Ecol cloning: a Laboratory Manual. Third Evol. 19:208-216. Edition. Cold Spring Harbor Laboratory Novarianto H, Maskromo I, Dinarti D, and Press, Cold Spring Harbor, New York. Sudarsono. 2014. Production technology Samonte LJ, Mendoza EMT, Ilag LL, De La for Kopyor coconut seednuts and Cruz ND, and Ramirez DA. 1989. seedlings in Indonesia. International Galactomannan degrading enzyme in Journal on Coconut R & D. 30(2):31-40. maturing normal and Makapuno and Novarianto, H dan Miftahorrachman. 2000. germinating normal coconut endosperm. Koleksi dan konservasi jenis-jenis kelapa Phytochemistry 28:2269-2273. unik. Makalah poster dalam Simposium Santos GA. 1999. Potential use of clonal Pengelolalan Plasma nutfah dan propagation in coconut improvement Pemuliaan Bandung 22-23 September. program. In Oropeza C, Verdiel JL, Perhimpunan Ilmu Pemuliaan Indonesia. Ashburner GR, Cardena R, and Samantha Prabha SS, Indira EP, Nair PN. 2011. JM (eds.) Current Advances in Coconut Contemporary gene flow and matting Biotechnology – Current Plant Science system analysis in natural teak forest using and Biotechnology in Agriculture. Kluwer microsatellite markers. Current Science Acad. Publ., London. Pp. 419-430. 101 (9):1213-1219. Schuster WSF, Mitoon JB. 2000. Paternity and Rajesh MK, Arunachalam V, Nagarajan P, gene dispersal in limber pine (Pinus Lebrun P, Samsudeen K, Thamban C. flexilis James). Heredity. 84:348-361. 2008. Genetic survey of 10 Indian coconut Soenarsih Sri. 2012. Pala (Myristica spp.) landraces by Simple Sequence Repeats Maluku Utara berdasarkan keragaman (SSRs). Scientia Horticulturae 118:282– morfologi, kandungan atsiri, pendugaan 287. seks tanaman dan analisis marka SSR Ramirez VM, Tablat VP, Kevan PG, Morillo IR, [disertasi]. Indonesia (ID): Institut Harries H, Barrera MF, Villareal DZ. Pertanian Bogor.
59 Cord 2015, 31 (1)
Sudarsono, Sudrajad, Novarianto H, Hosang MLA, Dinarti D, Rahayu MR, Maskromo I. 2014. Produksi bibit kopyor true to type dengan persilangan terkontrol dan peningkatan produksi buah kopyor dengan polinator lebah madu. Laporan Akhir Program Hi Link. Bogor (ID): Institut Pertanian Bogor.
Sukendah 2009. Teknologi pembiakan kultur in
vitro dan analisis molekuler pada tanaman
kelapa kopyor. [disertasi]. Indonesia (ID):
Institut Pertanian Bogor
Sutanto A, D. Sukma, C. Hermanto, and Sudarsono. 2014. Isolation and characterization of Resistance Gene Analogue (RGA) from Fusarium resistant banana cultivars. Emirates Journal of Food and Agriculture. 26(6):508-518.
Wattanayothin, S. 2005. The study on curd coconut hybrids. J. TNCEL 1(3):6-7. Wattanayothin, S. 2010. Variety improvement of Makapuno. Proceedings of the XLIV
COCOTECH Meeting, 5-9 July 2010, Samui Island, Thailand. pp. 96-108.
60 Guidelines to the Contributors of CORD
The Asian and Pacific Coconut Community (APCC) publishes semiannually (in April and October) an international journal on coconut research and development known as Cord. High standard articles written in English are published in Cord. The manuscript, complete in all respects, should be sent through email (high resolution scanned tables, photographs, graphs, etc may be sent in text boxes), addressed to the Chief Editor, Cord, Asian and Pacific Coconut Community, 3rd Floor, Lina Building, J1. H.R. Rasuna Said, Kav. B-7, Kuningan, Jakarta 12920, Indonesia. Email address: [email protected]. Hereunder are the guidelines/editorial style in preparing the article for CORD: Type the manuscript in A4 size paper with double line spacing using MS word. The title of the article should be short and specific. It should be typed in bold letters, for scientific name use italics. Below the title, type author's (s) name (s), each name should be superscripted as 1, 2 .… Each superscripted number should be explained by providing the complete mailing address of the author (s) with all details including pin code, e-mail etc at the bottom of first page. Below the author (s') name (s), provide an abstract emphasizing the key aspects of results but without reference to the body of the text of the full articles. After the abstract, provide a list of the Keywords: (4-6). The introduction should have a brief statement of the problem and explain the aim or the objectives of the investigation. The materials and methods should be very clear with all details of experimental design, treatments, location, period of study, methods adopted etc. Methods should be clearly written so that the reader of your article should be able to use it in pursuing her/his studies elsewhere. The results and discussions should provide data organized into tables, figures and photographs, suitably compared and discussed with earlier published findings. All data must be presented in metric units. For abbreviations, consult the B.S. (British Standards) on "Letter symbols, signs, and abbreviations". Abbreviations are alike in singular and plural. The placing of a dot (stop) after a standard abbreviations should be avoided unless the abbreviation form is likely to be taken for another word, e.g., in for inch, where a full stop would be necessary. In the text, references should be quoted according to the Harvard system, in which the author's name and date are given in parenthesis, e.g. (Magat, 2003) except when the author's name is part of a sentence, e.g. Magat (2003) reported …. When there are two authors, both names should be given, e.g. (Magat and Margate, 1990). If there are more than two authors, give the first author's name only followed by the words et al. e.g. Magat et al. (1989). The table should carry appropriate titles, which should be typed in bold letters. Each table should be self-explanatory, numbered consecutively. Try to avoid presenting table that is too large to print across the page. Use - (dash) when no observation was taken and 0 for zero reading. Express values less than unity as 0.25 and not .25. The figures should be restricted to the display of results when large number of values can be comprehended more easily; tables and figures should not reproduce the same data. Numbering and lettering should be kept to an absolute minimum. The photos and plates should be of high quality and be able to make a definite contribution to the value of the paper. The acknowledgment should follow the section on discussion and immediately precede references. Only references quoted in the text should be listed. References in the text should be arranged chronologically. Reference to publications in periodicals should be cited in the following order surname of author, followed by the initials, year of publication, title of paper, name of journal (italics), volume (in bold Arabic letters), and number in bracket and after a colon the first and last pages of the reference (without the prefix "pp.") e.g. Morin, J.P, Sudharto, P.S., Purba, R, Desmier de Chenon, Kakul, T., Laup, S., Beaudoin-Ollivier, L. and Rochat, D. 2001. A new type of trap for capturing Oryctes rhinoceros (Scarabaeidae: Dynastinae), the main pest in young oil palm and coconut plantings. Cord 17(2): 13-21. Reference to books and monographs should include author and/or editor's name full title, number of pages, edition, the publisher's name and place of publication. The title should be in italics. A list of references should be given at the end of the text and be arranged alphabetically on authors' name. If an author's name in the list is also mentioned with co-authors the following order should be used: publications of the single author, arranged according to publications dates, publications of the same author with one co-author and publications of the author with more than one co-author. If there are several references by the same author in one year, they are distinguished by the addition of small letters a, b, c, etc., e.g. Magat, 2003a, Magat, 2003b. Abbreviations of journal titles should be according to the "World list of Scientific Periodicals.” Note: The author(s) of selected paper will receive a token honorarium, as well as one-year free subscription of Cord for senior author if there is more than one author. Articles not accepted will not be sent back to the author. For each published article, only the senior author will receive one complimentary copy of the journal. For more copies, the cost of the journal has to be borne by the authors. Papers published in the Cord may be reprinted or published with prior permission from the APCC Secretariat.