<p>Several<br />tropical plant species contain high concentrations of phenolic<br />compounds, which become oxidised when their cells<br />are wounded or when the plant parts become senescences.<br />In tissue culture, the phenolic compounds usually leach into<br />the medium from the cut surfaces of explants. The phenolic<br />compounds caused the culture medium turns to dark brown<br />in colour due to oxidation. This is detrimental to the culture,<br />because it causes the isolated tissue fails to grow. The<br />browning of tissue culture and the medium can often be<br />prevented by one of the several different approaches, such<br />as by removing the phenolic compounds produced, modifying<br />the redox potential, inactivating phenolase enzymes,<br />reducing phenolase activity and substrate availability, as well<br />as pre-treatments by soaking and preconditioning on a basal<br />medium.</p>
a b s t r a c tIncreased production of sugarcane in Indonesia can be done with extensification sugarcane plantations which largely dominated by acidic upland red-yellow podzolic soil. High aluminium (Al) content and low pH of the soil can inhibit plant growth and development. Tolerant sugarcane in acid soil is the most efficient way, but the adaptive variety is still limited. In vitro culture technique can increase genetic variability to assemble new varieties through somaclonal variation combined with mutation using ethyl methane sulphonate (EMS). The new characters was directed by in vitro selection using AlCl 3 .6H 2 O with pH ¼ 4 as a component of selection for resistance to high aluminium. VMC 7616 and PS 862 varieties were used as materials. Mutation induced using EMS at concentrations of 0.1%, 0.3%, and 0.5% for 30, 60 and 120 minutes. Plantlets mutant obtained through callus formation, immersion callus in EMS, in vitro selection, and regeneration of callus. Result of study showed that the long immersion in the EMS solution caused greater damage to the cells, as indicated by the change in callus colour. Callus immersion time in EMS gave greater influence to regeneration compared to concentration of EMS. PS 862 had higher Al tolerance than VMC 7616. Rooting of shoot induced using indole-3-butyric acid (IBA) 3 mg/L.
ABSTRACT Shoot Multiplication and Root Induction on "Ubi Kelapa" (Dioscorea alata L.) and "Gembili" (Dioscorea esculenta L.) Through In Vitro Culture. Sri Hutami, Ragapadmi Purnamaningsih, Ika Mariska, and Surya Diantina. Dioscorea sp. (yam) is one of the minor tuber crops which grows wildly in the forest and only a few of its species are cultivated and used as main or secondary food. Conservation is needed to preserve plant genetic material. The objective of this research was to obtain methods of plantlets propagation of D. alata L. and D. esculenta L. through in vitro culture. The research was conducted at Tissue Culture Laboratory of ICABIOGRAD in 2012. The research consisted of three stages. First, shoot emergence. In this experiment, young shoots were planted in MS basic medium combined with benzyl adenine (BA) (0, 1, 3, and 5 mg/l) and gibberelic acid (GA) (0 and 5 mg/l). Second, shoot multiplication. Shoots of Dioscorea which were planted in the best medium of the first experiment were subcultured in MS medium combined with thidiazuron (0, 0.1, 0.5, 1, 2, and 3 mg/l). Third, root initiation. Shoots of Dioscorea which were planted in the best medium of the second experiment were subcultured in MS medium (½ MS and 1 MS) combined with indole-3-butyric acid (IBA) (0, 1, 3, and 5 mg/l). Result of these experiments showed that shoot emergence of D. alata L. and D. esculenta L. began at 2 weeks after planting in MS medium. More plantlets of D. alata L. and D. esculenta L. were obtained by shoot multiplication in MS media. Root initiation of the Dioscorea began at 4 weeks after planting in MS media. The addition of IBA (3-5 mg/l) on D. esculenta L. could not stimulate rooting but led to the formation of callus at the base of the stem buds.Keywords: Dioscorea alata L., Dioscorea esculenta L., shoots multiplication, root induction, in vitro culture. Eksplan berupa tunas muda ubi kelapa dan gembili ditanam pada media dasar MS dengan penambahan zat pengatur tumbuh benzyl adenine (BA) (0, 1, 3, dan 5 mg/l) dan asam giberelin (GA 3 ) (0 dan 5 mg/l). Kedua, multiplikasi tunas. Tunas Dioscorea yang terbentuk pada media terbaik pada penelitian inisiasi tunas, disubkultur pada media multiplikasi menggunakan media dasar MS ditambah thidiazuron (0, 0,1, 0,5, 1, 2, dan 3 mg/l). Ketiga, inisiasi dan pertumbuhan akar. Tunas Dioscorea yang terbentuk pada media terbaik pada penelitian multiplikasi tunas disubkultur pada media perakaran, yaitu kombinasi dari dua media dasar MS (½ MS dan 1 MS) dengan pemberian indolebutyric acid (IBA) (0, 1, 3, dan 5 mg/l). Hasil penelitian menunjukkan bahwa pertumbuhan/munculnya tunas ubi kelapa dan gembili dimulai pada umur 2 minggu setelah tanam, dengan media terbaik untuk ubi kelapa maupun gembili adalah media MS. Pembentukan planlet ubi kelapa dan gembili juga dapat dilakukan dengan multiplikasi tunas pada media MS. Perakaran ubi kelapa dan gembili diinisiasi pada umur 4 minggu setelah tanam pada media MS. Pemberian IBA (3-5 mg/l) pada gembili tidak dapat memacu perakaran, tetapi menyebabkan ...
Improvement of Plant Genetic Variability through Somaclonal Variations. Sri Hutami, Ika Mariska, and YatiSupriati. High genetic variability's are important factors in the development of new crop varieties. In vitro techniques are applicable for development of crop variability that is not found in the gene pool. One of the in vitro techniques that can be used for this purpose is the somaclonal variation technique. Somaclonal variation may be derived from genetic variations in explants and genetic variations in tissue cultures. Variations in the explant may be obtained from cell mutations or polysomic mutations of a certain tissue. Genetic variations in tissue culture may be caused by ploidy of chromosomes (endomitosis fusion), changes of chromosom structures (crossings), as well as changes of genes and cytoplasms. Changes of genetic characters may be improved if anorganic compound was added into the medium. To improve the plant tolerances to biotic or abiotic factors, selection components may also be added to the medium. Research results showed that somaclonal variation in tissue culture can improve genetic variations in plants. The variation produced in tissue culture provide chances to develop new plant genotipes. Many selection components, such as Gamma-ray irradiation, Al contents and low pH, pure toxin or filtrate, polyethylene glycol (PEG), and plant growth regulators can be used to improve somaclonal variations in many plants to produce new genotipes.
<p>Cell suspension culture could be defined as a<br />process that allows rapidly dividing homogenous suspension<br />of cells to grow in liquid nutrient media. There are two main<br />types of suspension cultures: (1) Batch cultures in which<br />cells are nurtured in a fixed volume of medium until growth<br />ceases and (2) Continuous cultures in which cell growth is<br />maintained by continuous replenishment of sterile nutrient<br />media. Plant cell suspension cultures are mostly used for the<br />biochemical investigation of cell physiology, growth, metabolism,<br />protoplast fusion, transformation and for large scale<br />production of seed by bioreactor and production of secondary<br />metabolites. Contamination is one of the largest problems<br />when dealing with cell cultures. Differences between<br />the products of cell suspension culture and whole plant are<br />frequently observed. These phenomena’s may be resulted<br />from lack of differentiation and organization and cell cultureinduced<br />variation. Utilization of cell suspension culture in<br />Indonesia is still limited, some of them for mass production<br />of plantation seed with bioreactor system and for production<br />of secondary metabolites. The success of this study give the<br />opportunity for mass production of seeds from other plants<br />and also production of secondary metabolites.</p>
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