KARAKTER VEGETATIF DAN MOLEKULER DUA AKSESI TANAMAN JARAK PAGAR (Jatropha curcas L.) DENGAN PERLAKUAN MUTAGEN COLCHICINE

NOVENTHI SUBANDI, ESTHERINA (2009) KARAKTER VEGETATIF DAN MOLEKULER DUA AKSESI TANAMAN JARAK PAGAR (Jatropha curcas L.) DENGAN PERLAKUAN MUTAGEN COLCHICINE. Other thesis, University of Muhammadiyah Malang.

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Abstract

Effort to use new energy source was an important part in energy diversification caused by the reduction of energy stock and increasing of fuel needs. The utilization of physic nut (Jatropha curcas L.) oil as bio-diesel fuel is an alternative to anticipate the increasing fuel needs. Major variety could be found by plant utilization, like mutation and transgenic. One of Mutation technique used in plant chemical mutation using colchicine compound. The research aimed to discuss the difference of vegetative and molecular character from two accession of physic nut (Jatropha curcas. L) plant caused by several colchicine mutagen concentrate treatment. The research was done in Forestry Experiment Lab, Agronomy Lab Faculty of Farming and Molecular Lab, Center of Biotechnology Development University of Muhammadiyah Malang. The research was done from March 2008 to February 2009. Tools used were: seedbox, polybag, sieve, durran bottle, drop pipette, analytic weighing, microscope, plastic gloves, aluminium foil, camera, gloves, scissors, micropipette (1-10 µL;10-100 µL;100-1000 µl), tube 1,5 ml, tip (1-10 µl,10-100 µl,100-1000 µl), mortar and hammer, spatulae, tissue, beaker glass, measurement glass, analytic weighing, sentrifuse, vortex, magnetik stirrer, pH meter, microwave, centrifuge, vacum frezer, autoclave, electroforesis tool, UV-transiluminator, Thermocycler (Biometra), timer, PCR and writing tool. Material used were: physic nut seed plant Asembagus-Situbondo local and Mukhtiarjo-Kediri local, atonic, alcohol 96%, aquadest, colchicine powder/crystal, sand, fertilizer, and compost, physic nut local Asembagus-Situbondo and local Mukhtiarjo-Kediri leaves which treated with colchicine, extract buffer (EDTA (Etylene Diamine Tetra Acetat) pH 8), Tris HCL pH 8, NaCL,SDS), CI 24:1 (chloroform: isoamyl alkohol), ethanol absolute, ethanol 70%, buffer TE, TBE 1x, gel agarose, Loading dye 6x, EtBr (Ethidium Bromida),dH2O, lambda DNA 100 bp, nuclease free water, PCR mix, primer 10 basa, those were OPA 15 and OPA 20. The research was done by factorial random group consisted of 8 combination of treatment and repeated 4 times consisted of two factors. Factor 1 physic nut plant accession A1 = accession IP-1A, A2 = accession IP-1M. Factor 2: concentration colchicine, that was K0 = without colchicine drop, K1 = dripping colchicine 0,05% concentration; K2 = dripping colchicine 0,10% concentration; K3 = dripping colchicine 0,15% concentration. Next phase was molecular analysis consisted of : (1) DNA isolation, (2) electroforesis / running DNA, (3) PCR (Polymerase Chain Reaction) process consisted of several phases, they were: pre-denaturation in temperature 94ºC in 5 minutes, denaturation in temperature 94ºC for 1 minute, sticking in temperature 36ºC for 1 minute, and elongation in temperature 72ºC for 2 minute, (4) electroforesis/running process of PCR result. From the research, there could be concluded: 1. interaction betwee jarak pagar plant accession IP-1A and IP-1M with colchicine concentration 0,05%; 0,10%, and 0,15% treatment had real influence to the amount of 28 HSP age leaves, leaf width per plant (cm²), and leaf stomata width (µm), but had no real influence to plant height parameter (cm), trunk diameter (cm), leaf stomata length (µm), and leaf clorophil (µg/cm²). in general, accession IP-1M with colchicine 0,15% concentration had higher leaf width per plant (cm²), and leaf stomata widht (µm) than the other treatment combination. 2. in separate treatment, IP-1M had real influence to plant height (cm), but the influence was not real in trunk diameter (cm), leaf amount, leaf width per plant (cm²), leaf stomata length (µm), and leaf chlorophyl contain (µg/cm²). Accession IP-1M had higher plant measurement than accession IP-1A. 3. colchicine concentration had real influence to leaf per plant width (cm²) and leaf somata length (µm), but no real influence to plant height parameter (cm), trunk diameter (cm), leaves amount, and leaf chlorophyl contain (µg/cm²). colchicine concentration 0,15% had leaf width per plant (cm²) and stomata length (µm) larger than colchicine 0,05% and 0,10% concentration. 4. there were difference in DNA ribbon resulted, they were: (a) DNA ribbons with dimension 200 bp, 300 bp, 400 bp, dan and bp at accession IP-1A with colchicine concentration 0,05%; 0,10%; and 0,15% and IP-1A without treatment (control), also accession IP-1M with concentration 0,05% and 0,10%; (b) 5 ribbons with dimension 200 bp, 300 bp, 400 bp, 600 bp, and 700 bp in accession IP-1M with colchicine concentration 0,15%; (c) 7 DNA ribbons with dimension 200 bp, 300 bp, 400 bp, 600 bp, 700 bp, 900 bp, and 1000 bp in accession IP-1M without treatment (control); (d) while DNA ribbon with measurement 200bp and 300bp were thicker in treatment with concentration 0,05%; 0,10%; and 0,15% compared with control. There needed further research to discuss concentration and exact colchicine treatment to get ploidi plant. Colchicine used should be added its concentration, and sprinkling method exchanged by sinking or both combination, sprinkling and sinking.

Item Type: Thesis (Other)
Subjects: S Agriculture > SB Plant culture
Divisions: Faculty of Agriculture & Animal Husbandry > Department of Agronomy
Depositing User: Anggit Aldila
Date Deposited: 21 Jun 2012 03:03
Last Modified: 21 Jun 2012 03:03
URI: http://eprints.umm.ac.id/id/eprint/8846

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