Genetics characterization, nutritional and phytochemicals potential of gedi leaves (Abelmoschus manihot (L.) Medik) growing in the North Sulawesi of Indonesia as a candidate of poultry feed

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Mandey et al., 2014 INTRODUCTION Abelmoschus manihot (L.) Medik is a native plant which is 1.2 – 1.8 m height and is widely distributed in the tropical regions. This plant has various local names such as aibika. It was hypothesized that the origin of this plant from the survey of literature the local names of Abelmoschus manihot (L.) Medik varies and the data available were largely derived from studies carried out in the polynesian-pacific regions (Preston, 1998). In North Sulawesi of Indonesia this plant is called “gedi” and its leaves provide essential ingredient for cooking porridge as a special gourmet food among the North Sulawesi cuisine. According to Jain and Bari (2010), gedi leaves contain polysaccharides and protein containing mucilage (gum) that enables the porridge to have a special viscosity. Morphologically, gedi plants vary in shape, color and other properties regardless of geographical differences suggesting some genetic variation may occur after a long period of adaptation. Gedi plants have been reported to posses medicinal properties that may benefit to human health. Puel et al., (2005) reported that the female wistar rat which feeding 15 % of gedi leaves prevent osteopenia that was attributable to the calcium content of gedi leaves. Other authors, Jain et al., (2009) reported that woody stem of gedi plant contain stigmasterol and γ-sitosterol, and also contain isoquercitrin, hyperoside, hibifolin, quercetin and isohamnetin that have anti consulvant and anti depressant-like activity (Guo et al., 2011; Wang et al., 1981; Wang et al., 2004). Gedi leaves have active pharmacological properties against analgesic effect (Jain et al., 2011). Sarwar et al., (2011) stated that Abelmoschus manihot has a profound anti-inflammatory and anti-diabetic effect. From these reports it can be concluded that gedi plants posses herbal medicine properties that can be used to manipulate the human and animal health. In spite of its phytopharmaceutical benefits there is paucity in information dealing with genetic diversity of gedi plant in Indonesia. Most information of Abelmoschus manihot derived from the studies carried out in the polynesian pacific regions (Preston, 1998). Gedi as a culinary herb and medicinal herb may have beneficial effects in animals. The phytochemical and nutritional compounds of leaf material may affect to poultry health and productivity. Cross et al., (2007) reported that culinary herbs in diets affect chick performance, gut health and endogenous secretions. Al-Sultan and Gameel (2004) suggests that feeding Curcuma longa (turmeric) to chicken through diet can induce hepatic changes and that these changes are not dose or time dependent. Windisch et al., (2008) cited several research, i.e. that phytogenic product also reduced activities of intestinal and fecal urease enzyme in broilers. Ashayerizadeh et al., (2009) reported that garlic powder and turmeric powder in diet significantly reduced abdominal fat percent, LDL and VLDL concentration in serum of broiler. Moreover, Yang et al., (2003) reported that green tea by product affect the reduction of body weight gain and meat cholesterol in broilers. Khatun et al., (2010) observed using in vitro model that viscous water-soluble portion of the fruit of Abelmoschus esculentus (L.) Moench has significant capacity to reduce the glucose diffusion form the dietary fiberglucose systems. The study was undertaken to investigate the compositional characterization of gedi. The samples were an alysed for the molecular characterization and identification, the proximate composition of the leaf part, energy content and the phytochemical composition, in order to get some useful information to be used in the preparation of poultry feed. Because there are no major reports in the literature, this would be an information for the detailed utilization of gedi to poultry feed. 1277 Journal of Research in Biology (2014) 4(2): 1276-1286
Mandey et al., 2014 MATERIAL AND METHODS Plant Identification Eight accessions of gedi (Abelmoschus manihot) collected from Manado, the North Sulawesi, Indonesia were used for this study. Herbarium specimens were identified for plant species at the Research Center for Biology, Indonesian Institute of Sciences, Bogor, Indonesia. DNA extraction, quantification, and sequencing DNA was extracted from 80-100 mg of fresh leaf tissue from each of the 5 randomly selected samples using a protocol of AxyPrep Multisource Genomic DNA M i n i p r e p K i t ( A x y g e n B i o s c i e n c e s , www.axygenbio.com). Three samples were scored as missing because of unable to isolate the mucilage. The final DNA supernatant were diluted for DNA quantifications with PCR technique. PCR analysis were performed using a Thermocycler machine, and in 50 µl reaction mixture containing 2 µl template of DNA, 2 x master Mix Vivantis 25 µl (Vi Buffer A 1 x; Taq Polimerase 1,25 unit), Primer F1 (10 pmol/µl) 1 µl (0,2 mM), Primer R1318 (10 pmol/µl) 1 µl (0,2 mM), MgCl 2 (50 mM) 1,5 µl (3 mM dNTPs 0,4 mM), H 2 O 20,5 µl, sample 1 µl.Initial trial was run on 5 samples and Taq quantity was Taq Polimerase 1,25 unit. Two primers were initially screened for amplification in PCR, they are Primer ndhF-F1 with product description 5’-GAA-TAT- GCA-TGG-ATC-ATA-CC-3’ (length 20) dan primer ndhF-R1318 with product description 5’-CGA-AAC- ATA-TAA-AAT-GCR-GTT-AAT-CC-3’ (length 26). PCR conditions were pre-hot 94°C (5 minutes), denaturation 94°C (45 seconds), annealing 54°C (45 seconds), primerization 72°C (1 minute 30 seconds) in 35 cycles and hold at 72°C (5 minutes). All PCR products were separated by electrophoresis in 1% agarose gel in 1 x TBE ran for 2 hours followed by ethidium bromide staining (5 µg ethidium bromide/ml). The gel was then stained and rinsed in water for about 10 minutes, and after that visualized under UV-light in trans -illuminator. All PCR products were sequenced. Sequence data were identified at First Base Laboratories Sdn, Bhd (1 st base), Taman Serdang Perdana, Selangor, Malaysia. Sequences were aligned using BLAST programme, and the building of a phylogenetic tree was established by Bioedit 7.19 and Mega 5 programme (http:// megasoftware.net). Phytochemical Screening Chemical tests were carried out to evaluate the presence of the phytochemicals such as alkaloids, Table 1: Identification/Determination of Gedi Leaves from Manado, North Sulawesi No Place of Collection Species Tribe 1 (1) (GH4) Abelmoschus manihot (L.) Medik Malvaceae 2 (2) (GH5) Abelmoschus manihot (L.) Medik Malvaceae 3 (3) (GH2) Abelmoschus manihot (L.) Medik Malvaceae 4 (4) (GM2) Abelmoschus manihot (L.) Medik Malvaceae 5 (6) (GH3) Abelmoschus manihot (L.) Medik Malvaceae 6 (8) (GM1) Abelmoschus manihot (L.) Medik Malvaceae 7 (9) (GH1) Abelmoschus manihot (L.) Medik Malvaceae 8 (11) (GH6) Abelmoschus manihot (L.) Medik Malvaceae Notes: GH = green leaf; GM = reddish green leaf; GH1= Bumi Nyiur; GH2 = Wanea; GH3 = Bumi Beringin; GH4 = Teling; GH5 = Bahu; GH6 = Kleak; GM1 = Tingkulu; GM2 = Wanea. Journal of Research in Biology (2014) 4(2): 1276-1286 1278
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Genetics characterization, nutritional and phytochemicals potential of gedi leaves (Abelmoschus manihot (L.) Medik) growing in the North Sulawesi of Indonesia as a candidate of poultry feed

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