The use of plant cell and tissue culture techniques have been widely used for producing secondary metabolites, both on a small scale (research) and industry with a large enough capacity, especially the use of bioreactors. Studies that have been carried out include the use of plant cells that are useful to produce useful chemical products and how to control the metabolism of the cells so as to obtain the target secondary metabolites. Plant cell cultures is done through callus proliferation, so should be able to synthesize, particularly accumulate some kind of secondary metabolites in particular compounds such medicinal alkaloid compound saponin, kardenolid, anthraquinone, poly phenols, and terpenes.
For decades research related to natural products chemistry is growing by leaps and bounds. Cases of degenerative diseases and anti-aging activity of free radicals caused mainly by stress and pollutants encourages various studies regarding the use of antioxidant compounds.
What are antioxidants?
Antioxidants are compounds that can prevent or delay the oxidation reaction on the substrate in lower concentrations when compared to the oxidized substrate. Reaction overhaul of oxygen reactive / reactive oxygen species (ROS) are memungkinkankita understand the reaction mechanism of antioxidants. Through the binding reaction with the aid of a metal or inhibition of enzymes capable of preventing the increase in free radical reactions. Actually there is a cell in the cell endogenous antioxidant, which is an antioxidant that is produced by cells such as ascorbic acid, ureat acid, glutathione, tokoperol, and others. The antioxidant activity of the product of secondary metabolites have been widely known through the technique of in vitro and some have been known reaction mechanisms.
Why should the in vitro?
Cracked natural materials have been used as raw material antioxidants have been widely used as agricultural and horticultural products (corn, carrot, tomato apple tea leaves damlain etc.) or medicinal plants such as pine, Curcuma, suren etc. Even the industry of liquor in several countries in Europe and olive oil have been using waste products (waste product) as an industrial raw material antioxidant compounds.
Then why should the in vitro? Currently biotechnology science and engineering is growing rapidly, especially in producing secondary metabolites to be more economical and able to overcome the problems in the conventional cultivation. When compared with conventional farming techniques, biotechnology does have some advantages, namely extraction and purification easier, some compounds not found in nature, does not depend on the weather and climate, it is easier to control the process of biosynthesis, and the last exploration through genetic engineering is more likely performed compared with when we are introducing GMOs into the environment.
What methods are used?
Of the few studies of tissue culture, the isolation of secondary metabolites, antioxidant activity is rarely noticed by researchers. Knowledge of the antioxidant properties of a compound often come from research on the theme ethnobotany medicinal plants, especially in terms of conservation. Testing antioxidant compounds depends on the form of the compound that we examine. The simple method is done by using a calorimetry color test which detects changes in metal reduction, test the free radical synthesis (2,2 & prime-azinobis- (3-3thilbenzothiazolene-6-sulfonate) and DPHH (1,1-diphenyl-2-picrylhydrazyl free radical ).
Jasmonate can increase the expression of genes involved in plant resistance. Generally jasmonate contained in approximately 160 species of angiosperms and gymnosperms on. JA induction by wounding and pathogens can push fatty acids produced by the destruction of the cell membrane which is produced through the activity lipoxigenase JA (LOX3) which mediates the oxygenation of alpha-linolenic acid to form Cyclopentanone thereby increasing production jasmonate that cause reinforcement signal that stimulates resistance mechanisms more effectively . This enzyme has been identified in chloroplasts.
Allene Oxide Syntase (AOS) is an enzyme involved in the biosynthesis of JA, evidenced by the close relationship between the AOS with JA content during the process of wounding in Arabidopsis plants. AOS gene expression in tomato activated by wounding, the addition of 12–oxophytodienoic acid and table. Phospolinase D (PLD) activity shows that have promoted the formation of linoleic acid and stimulates the biosynthesis of JA.
Jasmonate studies in response to resistance has been carried out in the Arabidopsis mutant plants coil1 more susceptible to pathogenic fungi and bacteria that cause soft rot. Even mutant jar1 (insensitive Jasmonic acid) and fad13, fad7, 8-deficient diets linolenic acid required in the biosynthesis of JA) susceptible to non-pathogenic bacteria and insect pests.
PLD activity associated with the occurrence of stress which play a role in the destruction of the membrane signal transduction. Treatment jasmonat exogenously has done much to reduce the effect of antioxidants, and photosynthesis. Besides the addition of the plant jasmonat given a fairly high salinity treatment led to a balance of endogenous hormones such as ABA.
Some genes are induced due to the presence of jasmonate like seed storage protein gene does not play a role in plant resistance, but an important signal in the control of gene expression during growth. Even as wounding induced gene may not do in plant resistance but only helped change the metabolism of plants under stress. Generally jasmonate concentration in plants ranged between 10-50 ng / g wet weight of plant tissue, a concentration sufficient to affect the physiology of plants.
Increased endogenous JA for the elicitation process increases the synthesis of certain secondary metabolites. Alkaloids in cell culture can be increased after elicitation by yeast (yeast). Most of the gene encoding the enzyme for the biosynthesis of nicotine transcription factors regulated by JA. Besides anthocyanins, glucosinolates and morphine were able to accumulate because of the influence of JA.