presentation pollutan bender

8/12/2019 Presentation Pollutan Bender

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Effects of copper and cadmium onheavy metal polluted waterbody

restoration by duckweed (Lemna minor)

Presented by:

1. Whicliffe Feirnaleonardo Pasedan NIM: 1104015035

2. Yoana Sakramentia NIM: 1104015036

3. Saepuddin NIM: 1104015043

Chemistry Forest ProductFaculty of Forestry

Mulawarman University


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OUTLINE

INTRODUCTION

MATERIAL AND METHODS

RESULT AND DISCUSSION

CONCLUSION


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Over the last 2 decades,pollutan, especially heavy

metal has been

increased.

Seriousproblems inorganism

Cu & Cd

ManyTechnologieshave beenapplied to

prevent and treatwater polutan.

Some technoloiges canbe expensive, especially

in large volume.

Developingplant method toreduce heavy

metal pollutan.

Lemna

minor


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Material and methods

A. Plant material and growth condition1. Lemna minor form lake Taihu (China), maintained in glass

aquarium.2. Condition room : 26 2 oC.3. Light intensity : 72mol m-2s-1 in LD cycle of 16h : 10h for 3

months.4. The medium of pre-treatment was 1/10 Hutner medium, with a

pump to keep circulation.5. The medium was replaced every 1,5 months.


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B. Experiment1. Lemna fronds were disinfected by immersing them in 1 %

(v/v) NaClO for 3 5 min, then rinsed with distiled water 3times.

2. 2 g fronds were incubated in 500 ml beakers containing 300ml medium.

3. The incubation conditions is as follows: temperature 28 oC inlight and 26 oC in dark ; light intensity 36 mol ; LD cycle 16h: 8h ; humidity 60 %

4. Medium for incubation is the modified Steinberg.

5. Lemna fronds are with Cu and CD of concentrations 0, 0.05,0.5, 5, 10, 20 mg l-1in modified steinburg.

6. After 4 days exposure to heavy metal (FW), soluble protein,photosynthtic pigments, POD, CAT, SOD, and MDA of frondswere determined. A minimun of 3 replicates were peformed ineach experiment.


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C. Photosynthetic pigments and soluble proteindetermination1. 150 g FW lemna fronds without root were homogenized on ice

with mortar and pestle in 3 ml of g mM phosphate buffer pH 7.2with 10 mM KCl, then extracted with cold acetone (80 %)

2. The absorbance of pigment extract was measured at wavelengthog 470, 626, 645, 663, and 730 nm.

3. The contents of Chl a, Chl b and carotenoid were calculated inaccordance with experimental equation as described byLichtenthaler.


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D. Enzyme extraction and assay1. 500 g FW of lemna fronds was homogenized in 5 ml cold

potassium phosphate buffer (0.1 M, pH 7.8). The homogenate

was centrifuged at x G (4 oC) for 15 min.2. All the work for preparation of enzyme extract was carries out at

4 oC.3. POD activity was determined spectrophotometrically by

measuring the increase in absorbance at 470 nm after 20 minincubation at room temperature. The reaction mixture contained

potassium phosphate buffer (50 mM, pH 7.0, 1 ml), H2O2(0.2%,v/v, 2 ml), guaiacol (0.2%, v/v, 0.95 ml) and enzyme extract (50ml). The reaction started by adding H2O2.

4. CAT activity was evaluated spectrophotometrically by measuringthe consumption of H2O2 at 240 nm where the testing mediumcontained in final volume of potassium phosphate buffer (50 mM,pH 7.5, 750 ml), H2O2 (200 mM, 100 ml), and enzyme extract(150 ml) in a final volume of 1 ml.


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5. MDA activity was determined to indicate the level of lipidperoxidation of fronds as described by Zhao. Enzyme extract (1.5ml) and thiobarbituric acid (0.5%, v/v, 2.5 ml) were boiled for 20min and then centrifuged at 10,000 _ g for 5 min. The supernatantwas measured spectrophotometrically at wavelength of 532 nm, 600nm and 450 nm.

E. StatisticAll data presented in the paper are the means of at least threereplicates. Significance of differences of samples was calculated byStudents t-test. Results of testing were consid-ered significant ifcalculated p-values were _0.05.


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Result


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Conclusion

1. Of 2 heavy metals, cadmium was more toxic than copper. The

antioxidant system had become disordered when lemna frondswere exposed 0.5 mg l-1of cadmium in grown medium.2. At the lowest concentration (0.005 mg l-1), copper did little harm

to duckweed. When the concentration reached up to 10 mg l-1 ,the antioxidant system of plants began to break down .

experiments showed that Lemna minor could tolerate low levelheavy metal stress (Cu2

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