Analysis Of Mycelia

There are five stages to this experiment: pre-incubation, incubation, heat shock, recovery and growth, and selection. First, two micro test tubes were obtained and labeled “+” and “-”. These labels were representative of transformed (+) and untransformed (-) Escherichia coli (E. coli). 250 microliters of transformation solution (calcium chloride) was added to the test tubes and placed on ice. During the pre-incubation period a single colony was placed into the test tube labeled “+” and was completely dispersed in the solution. This was repeated for the tube labeled “-“ as well. Then the solution was returned to the ice. In the incubation period, the pGLO DNA solution was added to the tube labeled “+” but was not added to the tube labeled “-“. The tube was then returned to the ice and incubated for 10 minutes. While this was undergoing the process, four agar plated were labeled according to what they had been treated with. The plates were treated accordingly: plate 1 contained lysogenic broth (LB)/ ampicillin (amp)/ plasmids with a green fluorescent protein (pGLO); plate 2 contained LB/amp/arabinose sugar with pGLO; plate 3 contained LB/amp without pGLO; and plate 4 contained LB without pGLO. The next step was heat chock. Both test tubes were placed in a water bath that was set at 42°C for 50 seconds. After 50 seconds the test tubes were rapidly transferred back to the ice were they

Chitin is a nitrogen-containing structural polysaccharide present in the cell wall of fungi and exoskeleton of crustaceans, insects and other arthropods. The exoskeleton serves are protection and structural support for the arthropods (Pechenik, 2009). Furthermore, in crustaceans, the chitinous substances is added with calcium carbonate for added protection (Berg, Martin, & Solomon, 2006). Chitin has several derivatives and one of them is chitosan. Chitosan is obtained through deacetylation of chitin under alkaline conditions the acetyl group present in chitin is removed and thus the substance is converted into chitosan (Rinaudo & Younes, 2015). Chitosan is a copolymer composed of N- acetyl glucosamine and glucosamine and possesses various properties such as antimicrobial activity, biocompatibility and biodegradability (Teli

No cell lives in isolation. Cells need to communicate between each other in order for all biological live on Earth to exist. There are several types of cellular signalling (neural, endocrine, exocrine, paracrine, autocrine). In most cases signals are released by signalling cells. These signalling molecules recognise and bind to specific receptors on the target cell. In order for the signal to be generated in most cases there has to be an environmental change.

Each 1.5 ml microcentrifude tube containing 50 µg myceliums, 50 µl of Lysozyme, 450 µl of Lysis solution and 3 µl of RNase A was kept for hour at 37 0C on water bath, vortex several times in duration. Then, addition 250 µl of 2% SDS, gently invert before addtion 250 µl of mixing Phenol- Chloroform- Isoamylclcohol ratio is 25:24:1, vortex and centrifuge at 12,000 rpm in 10 minutes then transfer supernatant on up layer to a new 1.5 ml microcentrifuge and repeat that step again before the next step to completely remove unexpected components. Subsequently, 0.1X of 3M Sodium acetate pH 4.8 was supplemented,

M. oryzae is an ascomycete belonging to a relatively new family, the Magnaporthaceae, which also includes pathogens of other cereals. Only the anamorphic1 state has been found in rice fields, but the teleomorph2 can be produced in culture with a pair of isolates having opposing mating types. The fungus most often presents as diamond-shaped lesions on leaves but is also found on stems, peduncles, panicles, seeds, and has even been detected on roots (TeBeest et al 2007). These diamond-shaped lesions appear with a white to gray center and brown to reddish brown borders (2016). Retrieved from http://ipm.ucanr.edu/PMG/r682100611.html.

Spores were sown in the week of September 10th and no observations were made. First observation was made in the week of September 17th, Friday 3:08 pm, the control displayed some growth with flagella-like hair, however, no germination was observed. The treatment did not display any signs of growth, only air-like bubbles were observed. Second observation was made in the week of September 24th, Friday 3:10 pm, the control displayed germination, gametophytes were visible under the microscope, however, hermaphrodites were not significantly observed. The treatment still does not display signs of germination, only small green spores were observed under the microscope. Last observation was made in the week of October 1st, Friday 3:05 pm, the control displayed significant signs of growth, it appeared that fertilization took place and an embryo was in development. The treatment only displayed minimal germination, however, some spores appeared to be infested with fungus.

The purpose of this lab was to introduce bacterial growth undergoing transformation and to understand the process of transformation.

Genomic DNA was extracted from the fungal mat of Sclerotium rolfsii. Thirty mg of freeze-dried mycelium was ground to a fine powder in an Eppendorf tube in liquid nitrogen. The ground mycelium was resuspended and lysed in 500 µl of lysis buffer (40 mM Tris-acetate, 20 mM sodium acetate,1 mM EDTA, 1% w/v SDS pH 8) (Lerner and Model1981). RNase A (2 µl of 10 mg/ml; Sigma USA) was added and the mixture was incubated for 5 min at 37 °C. To facilitate the precipitation of most polysaccharides, protein and cell debris, 165 ml of 5 mol/l NaCl solution was added and the components mixed by inverting the tube several times. The suspension was centrifuged at 6700 x g for 20 min at 4 °C, the supernatant was immediately transferred to a fresh tube and

Initially the team decided to use genitalium as the donor bacteria but later they decided to switch to M. mycoides as M. genitalium has very slow growth rate. Initially the attempt to extract genome from mycoides and transplant it to capricolum failed because they found that the genome from the donor was methylated while the genomes in recipient yeast are unmethylated. So they methylated the donor with methylases for the process.

Colonies were then selected randomly and the streak plating method employed to purify the stains. The strains were kept in 2 conditions including at 4°C (for MRS and M17 plates ) and at -20°C (for M17 and MRS broth) withh by 20% glycerol.

Two types of pattern were obtained, the relative ability of each gene to cleave coniferin was assessed. Clones containing Agrobacterium tumefaciens B3/73 DNA rapidly and completely hydrolyzed coniferin to coniferyl alcohol. Over the same period, type 2 clones were completely inactive. The different substrate specificities of clones were also evident from their ability to grow on cellobiose Agrobacterium tumefaciens B3/73 was able to use cellobiose as the sole carbon source. Escherichia coli DH5α and type 1 clones were not able to grow on cellobiose. Other clones were able to utilize cellobiose, but grew very slowly (Linda et al., 1992).

fixation. Pea is liable to be attack by many bacterial, fungal, viral, nematode diseases in addition to physiological disorder. However, fungal diseases, especially rust is considered one of the major destructive diseases affecting the crop yield (Hagedron, 1984 and Kraft, and Pfleger, 2001), especially in the north and middle parts of the Delta in Egypt and several countries in the world (Abada et al., 1997; Gupta and Shayam, 1998 and Parilli et al., 2015). The fungus Uromyces pisi is a heteroecious rust pathogen, completing its life cycle on two host plant species. The sexual stages are completed on Euphorbia cyparissias (cypress spurge), while the asexual lifecycle stages are completed on leguminous crop hosts such as Lathyrus, Orobus,

Mycoproteins are derived from the fungus ‘Fusarium venenatum’. This fungus is grown in a fermentation vat filled with glucose syrup and minerals and vitamins. To maintain growth oxygen is pumped in, carbon dioxide is removed and the vat is kept at a temperature between 350C to 420C. To form the all important protein, nitrogen is consistently added to the fungus. The result of all this optimization is that the fungus can double its mass every five hours.

The first thing to be done is to isolate the bacteria with the goal of obtaining a pure culture. This is done by streaking Trypticase Soy Agar (TSA) plate, Dnase plate as well as Mackonkey using the streak method and

aPlant Molecular Biology Laboratory, Department of Botany, Faculty of Science, Dayalbagh Educational Institute (Deemed University), Dayalbagh, Agra-282005, India, Tel: 0562-2801545, Fax: 0562-2801226