Term Project

Mentor-Mr.Rakesh Gupta
Name-Ritu Devi Registration No-11008837 Roll No-RB2051A03 Section-B2051 Programe-M.Sc. Botany

Evaluating cytogenetic aberrations taking place in the weeds inhabiting in metal polluted areas

Contents
1. Objectives 2. Introduction a) pollution b) industrial pollution c) tannery pollution d) phytoremediation e) cytogenetic abberation 3. Review of literature 4. Field demarcation 5. Materials and methods a) Material required b) Procedure 6. Current status 7. Future plans 8. References

Objectives
1. Morphological and phytodistribution studies of the weeds. 2. Cytogenetic and meiotic abberations taking place in the flowerbuds of weeds.

3.

Screening of the weeds with lower frequency of cytogenetic abberations.

Introduction a) Pollution
Pollution is the introduction of contaminants into a natural environment that causes instability, disorder, harm or discomfort to the ecosystem i.e. physical systems or living organisms. Pollution can take the form of chemical substances or energy, such as noise, heat or light. Pollutants, the elements of pollution, can be either foreign substances/energies or naturally occurring contaminants. Pollution is often classed as point source or nonpoint source pollution. Pollution became a popular issue after World War II, due to radioactive fallout from atomic warfare and testing. Then a non-nuclear event, The Great Smog of 1952 in London, killed at least 4000 people. This prompted some of the first major modern environmental legislation, The Clean Air Act of 1956. Pollution began to draw major public attention in the United States between the mid-1950s and early 1970s, when Congress passed the Noise Control Act, the Clean Air Act, the Clean Water Act and the National Environmental Policy Act.

Types of pollution
Pollution is of many types, some important types are mentioned below: 1. Air Pollution 2. Water Pollution 3. Soil Pollution 4. Noise Pollution 5. Thermal Pollution

b) Industrial pollution

is pollution which can be directly linked with industry, in contrast to other pollution sources. This form of pollution is one of the leading causes of pollution worldwide; in the United States, for example, the Environmental Protective Agency estimates that up to 50% of the nation's pollution is caused by industry. Because of its size and scope, industrial pollution is a serious problem for the entire planet, especially in nations which are rapidly industrializing, like China. This form of pollution dates back to antiquity, but widespread industrial pollution accelerated rapidly in the 1800s, with the start of the Industrial Revolution. The Industrial Revolution mechanized means of production, allowing for a much greater volume of production, and generating a corresponding increase in pollution. The problem was compounded by the use of fuels like coal, which is notoriously unclean, and a poor understanding of the causes and consequences of pollution. There are a number of forms of industrial pollution. One of the most common is water pollution, caused by dumping of industrial waste into waterways, or improper containment of waste, which causes leakage into groundwater and waterways. Industrial pollution can also impact air quality, and it can enter the soil, causing widespread environmental problems. Because of the nature of the global environment, industrial pollution is never limited to industrial nations. Samples of ice cores from Antarctica and the Arctic both show high levels of industrial pollutants, illustrating the immense distances which pollutants can travel, and traces of industrial pollutants have been identified in isolated human, animal, and plant populations as well. Industrial pollution hurts the environment in a range of ways, and it has a negative impact on human lives and health. Pollutants can kill animals and plants, imbalance ecosystems, degrade air quality radically, damage buildings, and generally degrade quality of life. Factory workers in areas with uncontrolled industrial pollution are especially vulnerable. A growing awareness of factory pollution and its consequences has led to tighter restrictions on pollution all over the world, with nations recognizing that they have an obligation to protect themselves and their neighbors from pollution. However, industrial pollution also highlights a growing issue: the desire of developing nations to achieve first world standards of living and production. As these countries industrialize, they add to the global burden of industrial pollution, triggering serious discussions and arguments about environmental responsibility and a desire to reach a global agreement on pollution issues

c) TANNERY POLLUTION:Leather industry contributes to one of the major

industrial pollution problems facing the country, and the pollution causing chemicals eg:cadmium, nickel, zinc, arsenic, selenium, and copper,arise mainly from the pre-tanning processes of leather processing.

D) Phytoremediation
Phytoremediation (from the Ancient Greek (phyto, plant), and Latin remedium (restoring balance or remediation) describes the treatment of environmental problems (bioremediation) through the use of plants that mitigate the environmental problem without the need to excavate the contaminant material and dispose of it elsewhere. Phytoremediation consists of mitigating pollutant concentrations in contaminated soils, water, or air, with plants able to contain, degrade, or eliminate metals, pesticides, solvents, explosives, crude oil and its derivatives, and various other contaminants from the media that contain them.

Applications
Phytoremediation may be applied wherever the soil or static water environment has become polluted or is suffering ongoing chronic pollution. Examples where phytoremediation has been used successfully include the restoration of abandoned metal-mine workings, reducing the impact of sites where polychlorinated biphenyls have been dumped during manufacture and mitigation of on-going coal mine discharges. Phytoremediation refers to the natural ability of certain plants called hyperaccumulators to bioaccumulate, degrade,or render harmless contaminants in soils, water, or air. Contaminants such as metals, pesticides, solvents, explosives, and crude oil and its derivatives, have been mitigated in phytoremediation projects worldwide. Many plants such as mustard plants, alpine pennycress and pigweed have proven to be successful at hyperaccumulating contaminants at toxic waste sites. Phytoremediation is considered a clean, cost-effective and non-environmentally disruptive technology, as opposed to mechanical cleanup methods such as soil excavation or pumping polluted groundwater. Over the past 20 years, this technology has become increasingly popular and has been employed at sites with soils contaminated with lead, uranium, and arsenic. However, one major disadvantage of phytoremediation is that it requires a long-term

commitment, as the process is dependent on plant growth, tolerance to toxicity, and bioaccumulation capacity.

How Does Phytoremediation Work?

Phytoremediation is actually a genneric term for several ways in which plants can be used to clean up contaminated soils and water. Plants may break down or degrade organic pollutants, or remove and stabilize metal contaminants. The methods used to phytoremediate metal contaminants are slightly different to those used to remediate sites polluted with organic contaminants.

Metal

Organic

Phytoextraction

Phytodegradation

Rhizofiltration

Rhizodegradation

Phytostabilisation

Phytovolatilisation

Methods of Phytoremediation A. Phytoremediation of metal contaminated sites 1. Phytoextraction (Phytoaccumulation)

Phytoextraction is the name given to the process where plant roots uptake metal contaminants from the soil and translocate them to their above soil tissues. As different plant have different abilities to uptake and withstand high levels of pollutants many different plants may be used. This is of particular importance on sites that have been polluted with more than one type of metal contaminant. Hyperaccumulator plant species (species which absorb higher amounts of pollutants than most other species) are used on may sites due to their tolerance of relatively extreme levels of pollution. Once the plants have grown and absorbed the metal pollutants they are harvested and disposed of safely. This process is repeated several times to reduce contamination to acceptable levels. In some cases it is possible to recycle the metals through a process known as phytomining, though this is usually reserved for use with precious metals. Metal compounds that have been successfully phytoextracted include zinc, copper, and nickel, but there is promising research being completed on lead and chromium absorbing plants.

2. Rhizofiltration
Rhizofiltration is similar in concept to Phytoextraction but is concerned with the remediation of contaminated groundwater rather than the remediation of polluted soils. The contaminants are either adsorbed onto the root surface or are absorbed by the plant roots. Plants used for rhizoliltration are not planted directly in situ but are acclimated to the pollutant first. Plants are hydroponically grown in clean water rather than soil, until a large root system has developed. Once a large root system is in place the water supply is substituted for a polluted water supply to acclimatise the plant. Afer the plants become acclimatised they are planted in the polluted area where the roots uptake the polluted water and the contaminants along with it. As the roots become saturated they are harvested and disposed of safely. Repeated treatments of the site can reduce pollution to suitable levels as was exemplified in Chernobyl where sunflowers were grown in radioactively contaminated pools.

3. Phytostabilisation
Phytostabilisation is the use of certain plants to immobilise soil and water contaminants. Contaminant are absorbed and accumulated by roots, adsorbed onto the roots, or precipitated in the rhizosphere. This reduces or even prevents the mobility of the contaminants preventing migration into the groundwater or air, and also reduces the bioavailibility of the contaminant thus preventing spread through the food chain. This technique can alos be used to re-establish a plant community on sites that have been denuded due to the high levels of metal contamination. Once a community of tolerant species has been established the potential for wind erosion (and thus spread of the pollutant) is reduced and leaching of the soil contaminants is also reduced. (B).

Phytoremediation of organic polluted sites

1. Phytodegradation (Phytotransformation)
Phytodegradation is the degradation or breakdown of organic contaminants by internal and external metabolic processes driven by the plant. Ex planta metabolic processes hydrolyse organic compounds into smaller units that can be absorbed by the plant. Some contaminants can be absorbed by the plant and are then broken down by plant enzymes. These smaller pollutant molecules may then be used as metabolites by the plant as it grows, thus becoming incorporated into the plant tissues. Plant enzymes have been identified that breakdown ammunition wastes, chlorinated solvents such as TCE (Trichloroethane), and others which degrade organic herbicides.

2. Rhizodegradation
Rhizodegradation (also called enhanced rhizosphere biodegradation, phytostimulation, and plant assisted bioremediation) is the breakdown of organic contaminants in the soil by soil dwelling microbes which is enhanced by the rhizosphere's presence. Certain soil dwelling microbes digest organic pollutants such as fuels and solvents, producing harmless pproducts

through a process known as Bioremediation. Plant root exudates such as sugars, alcohols, and organic acids act as carbohydrate sources for the soil microflora and enhance microbial growth and activity. Some of these compound may also act as chemotactic signals for certain microbes. The plant roots also loosen the soil and transport water to the rhizosphere thus additionaly enhancing microbial activity.

3.Phytovolatilization
Phytovolatilization is the process where plants uptake contaminaints which are water soluble and release them into the atmosphere as they transpire the water. The contaminant may become modified along the way, as the water travels along the plant's vascular system from the roots to the leaves, whereby the contaminants evaporate or volatilize into the air surrounding the plant. There are varying degrees of success with plants as phytovolatilizers with one study showing poplar trees to volatilize up to 90% of the TCE they absorb.

Cytogenetic abnormality
An irregularity in the number or structure of chromosomes, usually in the form of a gain (duplication) , loss (deletion), exchange (translocation) or alteration in sequence (inversion) of genetic material.

Review articles
1. Cytogenetical Studies on the effect of omnacortil on root tip cells of Alleum cepa Auti,S., Pagar,R., Ahire,D and Sawale,V. Received: June20,2010 ; Revised: July29,2010 ; Accepted:August7,2010.

Abstract
Omnacortil is steroid, an allopathic drug commonly used as analgesic and suppressant in various diseases. In the present work, onion root tips were subjected with a series of omnacortil concentrations, ranging from 0.2 g/ml, 0.4 g/ml, 0.6 g/ml, 0.8 g/ml and 1.0 g/ml for 24 hours for studying its effect on root mitosis in Allium cepa L. The roots were examined in permanent root tip squash preparations stained by the aceto-orcein. This work has confirmed that omnacortil have various effects on chromosomes and induced different mitotic abnormalities and structural aberration of chromosomes. Mitotic index (MI), relative division rate (RDR), relative abnormality rate (RAR) of treated material has been determined. Obtained results showed that all the concentrations of omnacortil showed higher MI and RDR, lower concentrations such as 0.2 g and0.4 g showed higher values over rest of the concentrations and control. Among the subjected concentrations 0.8 g concentration is most effective and showed highest RAR. Low concentrations of steroids act as stimulants while higher concentrations induced various chromosomal aberrations such as clumping and stickiness, fragmentation, C-mitotic effect, anaphase bridge, chromosomal condensation and contraction, dissolution of chromosome, which clearly showed the mutagenic, condensation and contraction, dissolution of chromosome, which clearly showed the mutagenic, clastogenic , antimitotic and cytotoxic effect of omnacortil.

2. Cytogenetical studirs in wheat XIX. Locationand linkage studies on gene Yr27 for resistance to stripe(yellow) rust. D.B.McDonald, R.A.McIntosh,C.R.Wellings,R.P.Singh and J.C.Nelson. Received: May27,2003 ; Accepted: January23,2004.

Abstract
The stripe (yellow) rust resistance gene Yr27 was located in wheat (Triticum aestivum ) chromosome 2B and shown to be closely linked to the leaf (brown) rust resistance genes Lr13 and Lr23 in the proximal region of the short arm. Gene Yr27 was genetically independent of Lr16, which is distally located in the same arm. While Yr27 was often difficult to score in segregating seedling populations, it is apparently quite effective in conferring resistance to avirulent cultures under field conditions. The occurrence of Yr27 in Mexican wheat germplasm and the current over-dependence on Yr27 for crop protection in Asia are discussed.

Field demarcatrion
I have marked the field for collection of weeds, will use in the term project. The marked area is around about 20 kilometer of the most polluted kala Singhia Drainage which is regularly polluted by the waste released from the leather industry, causing Tannery pollution.The area has divided into four parts as, P1, P2, P3 and P4, where P indicates for polluted area.

Materials and methods a) Material required

Young flower buds, acetocarmine solution, forecep, needle, slide, cover slips, lamp, watch glass, compound microscop, camera microscope.

b) Procedure
1. First we will take the young flower bud and put it on clean glass slide. 2. Now by adding few acetocarmin drops on flower bud on slide, will crushed with the help of glass rod. 3. A slight heat will give to the slide and compressed with thumb. 4. Then we will observe the slide under compound microscope and take the pictures with the help of camera microscope.

Current Status
At now i have only marked out the field for collection of weeds, where i will go after MTEs for collection and then i will start my project work. I have finalise my objectives.

Future Plans
After complete result analysis a better protocol can be prepared for the screening of weeds which show least variation due to metal accumulation and a better reproduction potential that will give an out look for the future potential phytoremediator

References
1. Antiochia,R., Campanella, L., Ghezzi, P., andMovassaghi,K. 2007. The use of vetiver for remediation of heavy metal soil contamination. Anal. Biochem., 2. ANZECC&ARMCANZ. 1999. Australian and New Zealand Guidelines for Fresh and MarineWater Quality. Australian and New Zealand Environment and Conservation Council & Agriculture and Resource Management Council of Australia and New Zealand, Canberra, Australia.

3. Baker, A. J. M.; Brooks, R. R. (1989), "Terrestrial higher plants which hyperaccumulate metallic elements A review of their distribution, ecology and phytochemistry", Biorecovery1 (2): 81126

4. Blaylock, M.J., Salt, D.E., Dushenkov, S., Zakharova, O., Gussman, C., Kapulnik, Y., Ensley, B.D., and Raskin, I. 1997. Enhanced accumulation of Pb in Indian mustard by soilapplied chelating agents. Environ. Sci. Technol.,

5. Bowen, H.J.M. (Ed.) 1979. Plants and the Chemical Elements, London, Academic Press. 6. ^ Burken, J.G. (2004). "2. Uptake and Metabolism of Organic Compounds: Green-Liver Model". In McCutcheon, S.C.; Schnoor, J.L.. Phytoremediation: Transformation and Control of Contaminants. Hoboken, NJ: John Wiley. doi:10.1002/047127304X.ch2 7. ^ Hannink, N.; Rosser, S. J.; French, C. E.; Basran, A.; Murray, J. A.; Nicklin, S.; Bruce, N. C. (2001), "Phytodetoxification of TNT by transgenic plants expressing a bacterial nitroreductase", Nature Biotechnology 19 (12): 116872, doi:10.1038/nbt12011168,PMID 11731787.

8. ^ Subramanian, Murali; Oliver, David J. & Shanks, Jacqueline V. (2006), "TNT Phytotransformation Pathway Characteristics in Arabidopsis: Role of Aromatic Hydroxylamines", Biotechnol. Prog. 22 (1): 208