Study area:
Present study is based on laboratory work; however the plant material for experimental purpose will be collected from experimental farm of CSIR-IIIM (32.73N and 74.87E). Objectives:
1. Full length cloning of squalene synthase, squalene epoxidase, β- amyrin synthase CYP88D6, CYP172A54, the five known genes involved in glycyrrhizin biosynthesis from Glycyrrhiza glabra.
2. Real time expression analyses of the targeted genes in various in-vitro regenerated plant organs in relation to organ specific glycyrrhizin accumulation.
3. Phytochemical analysis of in vitro regenerated plants.
4. Functional and biochemical characterization of at least one protein expressed in heterologous plant host.
REVIEW OF LITERATURE
The
…show more content…
Biosynthesis of glycyrrhizin is not fully elucidated at gene level. The early stages of triterpenoid saponin biosynthesis via isoprenoid pathway involve the dimerization of two molecules of farnesyl diphosphate catalyzed by squalene synthase, epoxidase–mediated oxidation then produces 2,3-oxidosqualene. The first diversifying step in triterpenoid biosynthesis is the cyclization of 2,3-oxidosqualene a common substrate of oxidosqualene cyclases (OSCs) and a precursor of both triterpenes and sterols [Abe et al., 1993]. The biosynthesis of glycyrrhizin involves the initial cyclization of 2, 3-oxidosqualene by a specific OSC, β-amyrin synthase (bAS), to the primarily oleanane triterpene β-amyrin which is one of the most commonly occurring triterpenes in plants. The subsequent steps involve a series of oxidative reactions at positions C-11 (two-step oxidation) and C-30 (three-step oxidation), followed by glycosyl transfers to the C-3 hydroxyl group.
Genes involved in the early stages of glycyrrhizin biosynthesis, namely, squalene synthase and bAS, have been functionally isolated from G. glabra [Hayashi et al., 1999, 2001]. The bAS gene has also been functionally isolated from several other plants including Arabidopsis thaliana [Shibuya et al., 2009], Avena strigosa [Qi et al., 2004], Lotus japonicus [Sawai et al., 2006], and Medicago truncatula [Suzuki et al., 2002]; however, most of the steps in the modification of the
In this study, rice will be exposed to osmotic stress condition, whereby it will show: a higher accumulation of proline and soluble sugars, reduced levels of MDA and minimized water loss rate compared to wild types of plant. Still, the stress-responsive gene OsHsfC1b will exhibit a significantly higher expression levels in rice than in transgenic plants under similar environmental conditions. Although OsHsfC1b has been evaluated purposely for its significance in drought
Nearly 50 years after the first lysozyme crystal structure had been published, the missing piece, the SN2 reaction of the glycosyl-enzyme intermediate, was found. The first lysozyme structure was that of a hen egg white, and it provided a deeper understanding of the mechanism of enzyme reactions. Lysozyme enables the transfer of a glycosyl group to water to occur more quickly. The cleavage of the C-O bond in glycosides without the lysozyme occurs rather slowly.
Overall, the results suggest that, this was somewhat of an effective method in isolation the enzyme GAPDH from yeast
Discussion Temperature and photoperiods are important for the life cycle of insects. The temperature of an environment provides the insect with an optimal living condition so it may develop into adults to then start mating to make the next generation. The temperature can then allow them to either go into diapause as adults to wait to start reproducing or start reproducing immediately after becoming adults. Photoperiods are also important to insects because the day/night cycle provides the insect with information about the change in weather for the upcoming months. The change in photoperiods tells insects to start going into diapause for the winter or for months that will not be optimal for development and reproduction.
Gibberellin Acid is a plant hormone that can affect plant growth by manipulating the cell division, stem elongation, and even mobilizes food resources within the endosperm to increase seed germination (Wiathrop, 1998). This experiment took place to test the factor of whether or not gibberellin could allow a seed to germinate and grow in the absence of light. Plants were distributed five drops of gibberellin and then placed in the cabinet for a total of three weeks. Each week, we recorded the growth of each plant. As a result of the three weeks, only one of the two hormone-induced plants successfully sprouted to a total of 16.2 centimeters. The other hormone-induced seed showed germination however, resulted to zero vertical growth,
Similar to thioredoxins, glutaredoxins (Grx) are small antioxidative proteins that use glutathione and NADPH for their regeneration. In addition to this, recent studies have shown that some classes of Grx can bind iron in Fe-S clusters and thus reducing the availability of iron in the Fenton and Fenton-like reactions [108]. Although Grx and Trx are very similar in their structure and mode of action, only Grx possess dehydroascorbate-reductase activity (DHAR) and can take part in the regeneration of an important cellular antioxidants - vitamin C [108]. Since Grx expression was opposite to Trx expression in O. nubilalis (Figure 18), this indicates spatial adjustments of the antioxidative defense systems in this species, in order to optimize the
On Plate I (LB) and Plate II (LB + Ampicilin), E.coli growth was expected to occur. Since the environment was not selective in Plate I, growth was expected to be found as lawn. This prediction was confirmed by the experiment. On Plate II, it was expected that either lawn or colonies would be found. The observed growth occured as lawn. On Plate III (YPD) growth was predicted, given the presence of all the nutrients that the auxotrophic yeasts cannot synthetize. Due to the usual non-selectivity of the medium, lawn was the expected type of growth. However, growth actually manifested as red colonies, an occurrence which will be explained in the Discussion section. On Plate IV (YNB, -AA) and Plate V (YNB, -AA, +5% AS) no growth was expected to happen given the absence of amino acids, which the auxotrophic yeast could not synthetize. The results were as predicted. On Plate VI (YNB, +AA-Arg, -AS ), it was expected to obtain colonies, due to the selectivity of the environment. This indeed was observed, in the form of white colonies.
This report will be analysing the effectiveness of Gibberellic Acid on different plants to see if it helps them germinate faster. This experiment will be looking at the plant types, Raphanus sativus (Radish), and Echinocactus grusonii (Cactus). It aims to see if the Gibberellic Acid increases the rate at which seeds germinate. Then, if the Gibberellic Acid does, this experiment will look at what ppm ration of GA (Gibberellic acid) results in the fastest germination time. The hypothesis being explored, states, “The seeds containing a 500ppm of Gibberellic Acid will germinate faster than one with more or less.”
The free hydroxyl group of galactose at 6th carbon is essential for galectin binding49.Another very significant structural feature of ST6Gal-I is that it contains six cysteine residues which are also involving disulfide linkages, Cys142-Cys406, Cys184-Cys335, and Cys353-Cys364. These disulfide bonds give important features such as hydrophobicity and rigidity. The other major structural feature of this enzyme is containing a large glycan (with ten monosaccharide units) covalently bonded to Asn149. This specific glycan is also the acceptor for the reaction catalyze by ST6Gal-I. Therefore, The ST6Gal-I is acting as an enzyme as well as a
Engineered s. cerevisiae with a reconstituted 7-gene pathway is feasible for the synthesis and production of codeine and morphine from (R)-reticuline (Fossati, et.al, 2015). S. cerevisiae cells fed “(R)-reticuline, salutaridine or codeine as substrates showed that all enzymes were functionally co-expressed in yeast and that activity of salutaridine reductase and codeine-O-demethylase likely limit flux to morphine synthesis” (Fossati, et.al, 2015). This study describes a significant advance for S. cerevisiae and paves the way for complete synthesis of substances not naturally produced by microbes. Morphine alkaloids are narcotic analgesics and the most powerful naturally produced alkaloids used to treat severe and chronic pain.
Thioredoxins (TRX) are small proteins (around 12 KDa) which are present in all organisms. In addition to these ‘classical’ thioredoxins, numerous proteins also exhibit thioredoxin like domains or multiple thioredoxin domains (P., 2001). Thioredoxin is a general name for small proteins and conserved WC(G/P)PC motif to catalyze thiol disulfide interchanges (Meyer, Buchanan, Vignols, & Reichheld, 2009). Trx proteins function in disulfide reductases to reduce H2 O2 to non-reactive H2O and to scavenge free radicals, This protects cells against oxidative stress and apoptosis (Wang et al., 2013). A large number of Trx proteins have been identified in plants(Meyer, Reichheld, & Vignols, 2005).There are six different types (m, f, x,
Genetically modified crops are produced using a process called ‘genetic engineering’. Using the process of genetic engineering enables to add a different gene into the target plants’ cells. It then alternates the plants’ genetic content to make new characteristics and give additional abilities such as adding more proteins and vitamins. The first method of genetically modifying a crop involves identifying and copying the gene of the donor organism with a desired trait, this procedure is called mapping. The second method involves transferring the gene into the target plant and when it is injected into the target plants’ cell; it then becomes part of the cells genetic characteristic. To be able to transfer the gene into the target plant, two
To date, FragariaCyc is the only resource that provides a comprehensive knowledgebase of strawberry metabolism and allows users to upload and analyze data of their choice within the framework of cellular-level metabolic network supported by extensive manual curation. FragariaCyc is based on the sequenced genome of F. vesca. version 1.0 (Shulaev et al., 2011) and shares functionalities with other BioCyc pathway databases including several species-specific plant pathway databases (Mueller et al., 2003;Urbanczyk-Wochniak and Sumner, 2007;Dharmawardhana et al., 2013;Monaco et al., 2013;Naithani et al., 2014) available from http://pathways.cgrb.oregonstate.edu.
Once the plants go through abiotic stresses like drought, temperature, and salinity, ABA is produced to rescue the plants from these harsh conditions. ABA evokes signal transduction pathway leading to the activation of the members of SnRK2s, namely SnRK2.2, SnRK2.3 and SnRk2.6 (Fujita et al., 2013). The SnRK2s leads the activation of downstream AREBs/ABFs. SnRK2s are responsible for ABA-dependent phosphorylation of AREBs/ABFs in multiple phosphorylation sites (RXXS/T) in protein sequence (Figure 2) (Nicolas et al., 2014). AREB1/ABF2, AREB2/ABF4, ABF3, and ABF1 are the major TFs which are downstream of ABA-activated SnRK2s in ABA signalling pathway during vegetative growth.
The evolution of ACC also brings about the production of 5¢-methylthioadenosine-nine. Increase in the rate of respiration of the fruit gives ATP (Adenosine triphosphate) need for the methionine cycle and can lead to induced ethylene production without high levels of intracellular methionine. SAM is a crucial methyl giver and relates to numerous aspects of cellular metabolism. Thus, the two steps involved in the synthesis of ethylene is the formation of ACC and its conversion into ethylene. The genes encoding ACS and ACO have therefore been studied more deeply than other enzymes in the pathway, although there is proof that a few other genes concerned with methionine synthesis and methionine salvage pathway are differentially demonstrated throughout ripening and in response to ethylene (Alba and others 2005; Zegzouti and others 1999). ACS and ACO are encoded by multigene families in higher plants, with tomato having at most nine ACS (LEACS1A, LEACS1B, and LEACS2-8) and five ACO (LEACO1-5) genes (Barry and others 1996; Nakatsuka and others 1998; Oetiker and others 1997; Van-der-Hoeven and others 2002; Zarembinski and Theologis 1994). Expression survey has disclosed that at least four ACS (LEACS1A, LEACS2, LEACS4, LEACS6) and three ACO (LEACO1, LEACO3, LEACO4) genes are differentially shown in tomato fruit (Barry and others 1996, 2000; Nakatsuka and others 1998; Rottmann and others 1991). LEACO1, LEACO3, andLEACO4 are shown at low levels in green fruits that are in the first