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Vol. 9(1), 2016 ISSN 1675 – 5650 Vol. 9(1), 2016 ISSN 1675 – 5650 MALAYSIAN COCOA JOURNAL ------------------------------------------------------------ Advisor Datin Norhaini Udin Vice Advisor Dr. Ramle Kasin Editor Dr. Rosmin Kasran Editorial Committee Dr. Alias Awang Dr. Ahmad Kamil Mohd Jaaffar Dr. Douglas Furtek Dr. Fisal Ahmad Dr. Hii Ching Lik Dr. Rozita Osman Dr. Samuel Yap Kian Chee Dr. Tan Chia Lock Dr. Zainal Baharum En. Mohamed Yusof Ishak Hjh. Winoryantie Sulaiman Pn. Harnie Harun Pn. Suzannah Sharif Published by MALAYSIAN COCOA BOARD 5-7th Floor, Wisma SEDCO Lorong Plaza Wawasan, Off Coastal Highway 88999 Kota Kinabalu, Sabah, Malaysia ©All rights reserved. No part of this publication may be reproduced in any form or by any means without permission in writing from Malaysian Cocoa Board CONTENTS ASSESSMENT OF SOMATIC EMBRYOGENESIS POTENTIAL OF ELITE MALAYSIAN Theobroma cacao CLONES 1 Siti Norhana, M. A., Haikal Eman, A., Ahmad Kamil, M. J. and Kasran, R. THE EFFECT OF ASCORBIC ACID IN THE MATURATION OF COCOA SOMATIC EMBRYOS 5 Norasekin, T. and Ducos, J.P. IMPROVEMENTS OF Theobroma cacao ACCLIMATIZATION TO EX VITRO ENVIRONMENT 11 Nik Aziz, N. I., Jasli, M., Misak, E., Abdul Asui, N. and Kasran, R. DEVELOPMENT OF SNP GENOTYPING ASSAYS IN COCOA POD BORER USING REAL-TIME PCR Kasran, R., Roslina, M.S., David, A., Nuraziawati, M.Y., Navies, M.., Mellisa, G., Fahmi, W. and 19 Larry, C. IDENTIFICATION OF THE POTENTIAL RESISTANCE GENES IN Theobroma cacao TO COCOA POD BORER INSECT (Conopomorpha cramerella) IN RELATION TO HOST- PATHOGEN INTERACTION USING RNA-SEQ TECHNOLOGY 40 Roslina, M.S., David, A., Anisah, S., Navies, M., Kasran, R., K., Mellisa, G.,Fahmi, W. and Larry, C. DNA FINGERPRINTING OF THE MALAYSIAN COCOA GERMPLASM COLLECTION USING MICROSATELLITE MARKERS 50 Lea, J. and Haya, R. EVALUATION OF CLONAL MATERIALS IN MALAYSIAN COCOA BOARD, CRDC MADAI SABAH 59 Aizat, J., Nuraziawati, M.Y., Azlan, S. and Haya, R. ANALYSIS OF COCOA YIELD COMPONENTS IN RELATION TO ITS YIELD Tan, C.L. and Ling, A.S.C. 64 PHYSIOLOGICAL RESPONSES OF CACAO SEEDLINGS SUPPLEMENTED WITH MINERALS USING FOLIAR APPLICATION 71 Tee, Y.K., Bailey, B.A., Ali, S.S., Strem, M.D., Elson, M. and Baligar, V.C. THE EFFICIENCY OF CACAO SPECIFIC COMPOUND FERTILIZER ON CACAO PRODUCTIONS 79 Helmi, S., Rozita.O.., Haya. R., Ling, A.S.C.and Halim, H. UNRAVELLING THE INTERACTION BETWEEN LEPIDOPTERAN SODIUM CHANNEL AND NEUROTOXIN PEPTIDE: A STRUCTURAL INSIGHT 86 Rahman, M. Z. A. , Jainuddin, H. and Kasran, R. TOXICITY STUDIES OF Bacillus thuringiensis TO COCOA POD BORER LARVA AND ADULT MOTH 97 Tan, C.L., Navies, M. and Anisah, S. CAFFEINE TOXICITY TEST ON COCOA POD BORER ADULT MOTH Anisah, S. 103 EVALUATION OF BEST MANAGEMENT PRACTICES FOR COCOA POD BORER IN COCOA ECOSYSTEM 108 Saripah, B. and Alias, A. ESTIMATING BLACK POD DISEASE PROGRESS RATE USING THE AREA UNDER DISEASE PROGRESS CURVE (AUDPC) FOR DIFFERENT COCOA CLONES IN LABORATORY CONDITION 121 Ling, A.S.C., Ahmad Kamil, M.J., Chong, K.P. and Ho, C.M. PLANT GROWTH HORMONES PRODUCED BY ENDOPHYTIC Bacillus subtilis STRAIN LKM-BK ISOLATED FROM COCOA Ishak, Z., Mohd Iswadi, M.K., Russman Nizam, A.H., Ahmad Kamil, M.J., Ernie Eileen, R.R., Wan 127 Syaidatul, A. and Ainon, H. EFFECTIVENESS OF SYSTEMIC FUNGICIDE (TEBUCONAZOLE) IN CONTROLLING VASCULAR STREAK DIEBACK (VSD) ON COCOA SEEDLINGS 134 Suhaida, S., Shari Fuddin, S. and Alias, A. INHIBITION ABILITY OF COCOA POD EXTRACT ON TYROSINASE ACTIVITY Azila, A.K., Nur Azilah, A. and Azrina, A. 139 THE EFFECTS OF PHYSIOCHEMICAL ACTIVATION ON ADSORPTION CAPACITY, POROSITY AND SURFACE FUNCTIONAL GROUPS OF COCOA (Theobroma cacao) NIB – BASED ACTIVATED CARBON 151 Khairul Adli, N., Fisal, A., Mohd Azmier, A.and Mohd Sukri, H. HUMAN CHEEK SKIN TEXTURE IMPROVEMENT BY COCOA BEAN EXTRACT INCORPORATED IN A COSMETIC FORMULATION 160 Norliza, A.W., Russly, A.R., Puziah, H., Amin, I. and Shuhaimi, M. BIOACTIVITY-GUIDED FRACTIONATION OF POTENT ANTI-CANCER PROPERTIES FROM NON-EDIBLE TISSUES OF Theobroma cacao 170 Zainal, B., Abdah M. A., Taufiq Yap Y. H., Roslida A. H., Mohd Redzuan, S. and Kasran, R. DISINTEGRATION OF COCOA POWDER TABLET Samuel, Y.K.C., Chin, H.H., Sarini, H. and Arief Huzaimi, M.Y. 181 SHELF-LIFE STUDY OF Lactobacillus fermentum IN PROBIOTIC CHOCOLATE Rosmawati, M.S., Abdul Rahim, A.M. and Kasran, R. 187 MALAYSIAN RENAISSANCE COCOA: QUANTIFICATION OF FLAVANOLS AND METHYLXANTINES IN SELECTED MALAYSIAN COCOA CLONES 193 Azhar, M. and Kasran, R. Malaysian Cocoa Journal 2016, Vol. 9 No. 1 ASSESSMENT OF SOMATIC EMBRYOGENESIS POTENTIAL OF ELITE MALAYSIAN Theobroma cacao L. CLONES Siti Norhana, M. A.1, Haikal Eman, A.1, Ahmad Kamil, M. J.2 and Kasran, R.1 1Malaysian Cocoa Board, Cocoa Biotechnology Research Centre, Kota Kinabalu Industrial Park, 88460 Kota Kinabalu, Sabah, Malaysia 2 Cocoa Research & Development Centre Kota Samarahan, Malaysian Cocoa Board, Lot 248, Block 14, Daerah Muara Tuang, Bahagian Samarahan, Locked Bag 3131, 93450, Kuching, Sarawak. Malaysian Cocoa Journal 9(1): 1-4 (2016) ABSTRACT - Somatic embryogenesis provides an efficient in vitro clonal propagation method to propagate elite clones rapidly for research and production. Due to the high heterozygosity nature of most cocoa clones, adjustment on the concentration of nutrients in the culture medium is necessary to increase the efficiency of embryogenic response of the local cocoa clones in Malaysia. Screening of 16 elite Malaysian cocoa clones from Class I and II using both patented and modified Nestle induction medium was carried out. Induction of somatic embryogenesis from floral staminodes of cocoa was studied with respect to primary somatic embryogenesis potential. The result showed five clones that were previously less responsive started to produce primary somatic embryos consistently between four to eight weeks after culture initiation on modified induction medium supplemented with 1mg/L Myo-inositol and 6.25mg/L L- Glutamine. Compared to the patented medium, the use of much lower concentration of Myo-inositol and L- Glutamine (100X and 40X lower, respectively) in the induction medium could reduce the time for somatic embryo production by half and able to stimulate higher frequency of embryogenesis in selected clones. The percentage of primary somatic embryos observed after eight weeks on the modified medium range from 5% to 43.3%. The study is currently concentrating on establishing highly embryonic calli stock and mass propagating secondary somatic embryos for embryo conversion trials. Keywords: Somatic embryogenesis, Primary somatic embryos, Theobroma cacao, Myo-inositol, Glutamine INTRODUCTION of two compounds in the induction medium, myo-inositol and L-glutamine, with respect to Somatic embryogenesis as an alternative for mass primary somatic embryogenesis potential were propagation of elite Theobroma cacao clones has investigated. been well studied for years. It provides an efficient in vitro clonal propagation method to This study involved 16 Malaysian cocoa propagate elite clones rapidly for research and clones from Class I and Class II. These cocoa production. The main advantages of this method clones were previously classified into four classes include the possibility of rapidly generating - Class I, II, II and IV - based on their asexually propagated uniform plants of high adaptability to a wide range of Malaysian agro- genetic value, and the clonal production of climatic conditions, yield potential, pod and bean orthotropic plants with normal dimorphic characteristics, tolerant level to major pest and architecture and taproot formation (Maximova et diseases and possession of high butter content al., 2002). and good flavor (MCB, 2012). Class I clones are generally suitable for planting throughout Due to the high heterozygosity nature of Malaysia with good yield potential (>2.5 most cocoa clones, their degrees of response ton/ha/year) and tolerant to major pest and towards culture media treatments varied. In order diseases, while Class II clones are suitable for to increase the efficiency of embryogenic planting in selective areas (such as inland soil) response of each clone, it is necessary to modify with good yield potential, moderately tolerant to the concentration of the nutrients in the culture. major diseases and possess good pod and bean In this study, the effects of various concentrations quality (MCB, 2012). 1 Malaysian Cocoa Journal 2016, Vol. 9 No. 1 This study aimed to assess the somatic medium contained macro-nutrients DKW, micro- embryogenesis potential of elite Malaysian cocoa nutrients DKW, vitamins DKW, glucose 1 g/l, clones from Class I and II, using both Nestle sucrose 30 g/l, Phytagel (Sigma), pH5.7. patented induction medium as well as modified version of the same medium. Induction of The experiment was carried out in somatic embryogenesis was studied with respect triplicate. 20 explants were cultured per clone per to primary somatic embryogenesis potential. replicate. The average percentage of somatic embryos produced, representing the total number of somatic embryos per total number of explant, MATERIALS AND METHODS was determined eight weeks after the culture initiation. The earliest time taken to produce Staminodes from unopened cocoa flower buds primary somatic embryo from initial culture for (age 2-4 weeks) were used as explants. Samples each clone was also recorded. However, due to were obtained from bud grafted, field and seasonal availability of flowers, the experiments glasshouse-grown clonal plants at Centre for carried out on the patented and modified medium Cocoa Biotechnology Research, Kota Kinabalu were not able to be carried out side by side, hence Industrial Park, Sabah, Malaysia. done on different dates. Unopened cocoa flower buds were washed with tap water and surface-sterilized RESULTS AND DISCUSSIONS using 10% commercial bleach (5.25% (v/v) sodium hypochlorite) for 20 min followed by Using the patented Nestle induction media, the three rinses for five min each in sterile reverse mean percentage of somatic embryos produced osmosis water. Following dissection, staminodes ranged from 8.33 to 18.33 (Table 1).
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