Each year, there are enormous amount of waste products and hazardous chemical by-products which are being realized to the environment. It is estimated that there are millions of chemical compounds already being produced industrially, which release almost half a billion kilograms of toxic substances, annually. This is an indication of the extent to which the environment suffers from pollutants. As a result of the effects caused by the pollutants to the environment, the reduction or elimination of such contaminants has become a matter of great concerns. Accordingly, a lot of efforts from different scientific fields are put to curb or overcome the negative environmental effects of the pollutants. In the recent past, ecologists, environmentalists, microbiologist among other individuals from other fields related to biological sciences have shown interest of studying, understanding and finding an everlasting solution, through the establishment of means to eliminate the contaminants from the environment (Glick and Pasternak,1998, 59).
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In the endeavor of establishing a clean environment, scientists have employed different methods and techniques to clear the already deposited harmful products in the environment. The most commonly used methods include use of thermal processes which entails excavation and transportation of contaminated materials for further treatments. Other commonly used traditional techniques those involving use of chemical and physical processes that have been used for years despite the fact that they are a bit costly/expensive and ineffective.
The continued used of these traditional methods which are considered as inefficient and obsolete has almost been brought to a halt through the establishing of new functions of plants, particularly the purposes of cleaning of the polluted environments. The current world has slowly been diverting from these old physical and chemical techniques, and embracing bioremediation techniques, which are now considered as the modern ones. This has been seen as an additional function of the plants that would help man in maintaining a healthy environment for both economic productions and for his own good health. The bioremediation procedures are believed to be most effective that in the past decades have proved to be useful way of destroying hazardous substances/ contaminants in the environment at low costs and without addition of further environmental damages as it is with most other techniques. The Arabidopsis species are some of the plant materials which have been proved to be good instruments for use in the bioremediation work. It is an exotic species found grown in the Australian regions. The biggest question which arises concerning this is whether it is possible to establish an indigenous species that has similar abilities of cleaning the environments as it is with Arabidopsis species. The application of the discovery has however been limited mainly because of the lack instruments to identify or establish those areas which are polluted in ease and quick. It is our believed genetic studies would also serve as of great help in setting such a situation which would allow identification of those areas which are polluted and then enable the subsequent use of the plant species which have the power of performing to rhizoremediation process to be extensively used in those areas where it can survive (Lamp et al, 1990,49).
Though the discovery of the bioremediation techniques has been a crucial one in the advancement of scientific work as well as in the creation of a better environment for the living of man, this has only been possible with certain type of plant species that have shown ability to survive in certain environmental conditions. As well known, plant survival in any given area is depended upon plants external features. These external features are however physical expression of the plants internal features which are closely linked to genetic materials, particularly the DNA. The structure of the DNA dictates the external features the plant express which have widely been used to determine the kind of a plant environment in which they fit. The DNA has plays crucial part in the accomplishment of many internal metabolic functions, especially physiological functions that utilizes enzymatic substances. While many of the previous scientific research work has concentrated on the physical external and internal features to establish an indigenous plant that can perform bioremediation functions in a pervasive environmental conditions like the one in Australia, we believe that they is none who has done so through with the study of the plants genetic materials. That I why this study endeavors to establish whether it is possible to have a plant of a perverse category to be used in rhizoremediation purpose, together with identifying whether G3pDH genes could be used as indicators of plants with the potential of performing rhizoremediation functions.
Materials & Methods
DNA and Genome Extractions
Three Australian grasses were studied. This included 2 isolated grass species namely: Themeda triandra, commonly known as the kangaroo grass, and the second one was microlaena stipoides which is commonly known as the weeping grass. These two together with another DNA sample sequence product of the plants species known as Arabidopsis athaliana were provided by the department of Biotechnology of Flinders University of South Australia School of biological sciences, as primary grass species found in Australia (Wrigley and Fagg, 2003, 36).. All the two plant materials of Themeda triandra and microlaena stipoides were placed under process of DNA homogenization and protein denature using DNAses (papain) and lysing detergents. Interface formations: – the extracted mixture was heated to accelerate protein degradation processes, cooled and cold alcohol of ethanol added leading to formation of an interface at the center of the two profiles/layers. Precipitation of DNA: – the DNA mixture was let to undisturbed to precipitate the DNAs and then centrifuged for about 2 min. using micro-centrifuge tubes. Separation: – the centrifuged tubes were spun and the alcohol portion, the supernatant solution was poured off, and then the DNAs made to re-solubilize using a buffer. The extracted DNAs would then be compared with the DNA sample of the Arabidopsis athaliana.
PCR Processes: – the extracted DNAs were then run on a PCR- procedure to estimate the size of the DNA. This was done by observing the lanes formed in a PCR-machine that gave graphs of the different DNAs (McPherson and Møller, 2006, 68).
DNA sequence analysis of G3pDH gene locus. The reporting of study G3pDH loci in the plant species incorporated the partial and full analysis of the DNA sequence of the G3pDH locus. This entailed the downstream and upstream scanning of the G3pDH locus. It was observed for the DNA locus to evaluate whether it contained a DNA and produced the gene substances/products. The segment of the DNA was important in determining the cells’ ability to produce the materials needed in the conversion of hydrocarbons to useful plant products. The same Arabidopsis athaliana DNA sample library was used to make comparison of the DNA sequences of the other two species (Brown, 2007, 93).
Expected Results and discussion
According to the general knowledge on genetics, it was our view that the expected result of the gene sequences would be different for each of the various species. These kinds of speculated results were assumed because after a consideration of several factors which determine the plant species feature. The frequencies of the loci of the genes were expected to be almost the same for genetic materials which were of close relationship. However, we believed that wide difference of the frequencies of gene loci for the G3pDH gene would have been different for plant cells that were quite different in there functioning.
The analysis of the G3pDH loci for each of the two species would have shown similar structures if the species functioning of the substances produced by it has the purposes to the plant. Otherwise, any structural difference of the revealed gene locus for the gene responsible in the production of enzymatic materials, G3P responsible for the conversion of petroleum hydrocarbons would be altered and different. The locus closer in resemblance to that of Arabidopsis athaliana, would generally give some indication of having similar materials coded from this area being almost the same as those produced by the locus of the same kind of gene from Arabidopsis athaliana (Primrose and Twyman, 2006, 105).
Depending on the theory information, themeda triandra and microlaena stipoides would display little similarities in their genetic structural data. Despite the fact that they belong to the same family of poaceae, much differences are expected in the assessment of the DNA strands of the two species. This is because the genetic make up must have been the contributing factors that led them have the disparities in their physical features as causing them to be placed in two difference genera. It was seen that through the analysis of sequences of the DNA materials, certain loci would either be added or missing for the expression of dissimilar features on both species. It was also established that these two plants establishes and grows in a variety of habitats (Metzenberg, 2007, 35). According to the soil and shade needs, the microlaena stipoides survives well in shady and not well drained soils, while themeda triandra habitats sandy to clay soils. Such external features are useful in predicting the internal structures not only on the basis of tissues and organs but at in-depth level of genetic make up of the plant species.
Under the consideration of the DNA structure, the total amount of DNA, that is the genome may so form the basis of the ability of a plant to carry out bioremediation activities. In this case, the analysis of the PCR giving the size of DNA was also important in acting as a correlation between the plant potential to rhizo-remediate. The first part was therefore a means to establish the genome of the plants species and see whether they were of comparable quantities as to those of the Arabidopsis athaliana. To the understanding on the knowledge of genetic, one would obvious say that there should have been a less difference between each of the two species because they all belonged to the same family.
While the Arabidopsis athaliana acted as the control in the experiment, the species genomes for the both species of A.athalaina and T. triandra were expected to be a bit different from that of the control species. There several reasons which could have led to these differences. First and foremost, the species which were being used in the experiment were both belonging to angiosperms but the control species was a dicot while the others were monocots. This implies their gene make up must also have been different. As revealed, the microlaena stipoides are prone to frequent genetic changes thus they are undergoing some sort of micro-evolutionary changes, while the themeda triandra are a bit stable but likely to be affected by mutations.
The studying of plant species is of crucial importance in the endeavor of finding an everlasting solution to the nations. However, through the exploration of the physical features of the plants has proven unfruitful in combating with the prevailing condition of pollutions under various habitats. From the studying A.athaliana and M. stipoides, it can be concluded from the hypothetical work, they are enormous challenges surrounding the establishing of an indigenous plant to use in the cleaning of contaminants in the aquatic regions and marshy areas. The microleana and tremeda species which appeared to promising species for such areas shows great disparities in their genetic structure, hence making it impossible to be used as cleaning instruments for these areas.
Brown,.A. 2007. Genomes 3. New York: Garland Science.
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Glick, R., and Pasternak, J. 1998. Molecular Biotechnology. Washington: ASM Press.
Lamp,.A. Forbes, J. and Cade, W. 1990. Grasses of Temperate Australia. Melbourne Inkata Press.
McPherson, M., and Møller, S. 2006. PCR. New York: Taylor & Francis.
Metzenberg, S. 2007. Working with DNA. New York: Taylor & Francis Group.
Primrose, B., and Twyman, M. 2006. Principles of Gene Manipulation and Genomics, Malden: Blackwell Publishing.
Wrigley, W. and Fagg, M. 2003. Australian Native Plants. Reed New Holland.