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Transport of Minerals and Materials In and Out of the PlantJohn Benedict R. Palo, Kyra Christine A. Rios, Rochelle Anne R. Tan, Nathea Agatha C. Valdez

Submitted to: Maam Elena RagragioSubmitted: Spetember 20, 2012ABSTRACT This exercise is a compilation of different experiments that individually focus on diffusion, osmosis, imbibition, guttation and transpiration in plants. The acquired results supported majority of the groups initial hypotheses. Diffusion rate is affected by temperature and density; high temperature and low density cause faster rate of diffusion. Salt causes a turgid plant cell to shrink through the process of osmosis. Integrity of cell membranes is affected by temperature and acidity of solvent/s. High acid content and high temperature in solvent weakens/diminishes the integrity of the cell membrane. The process of germination is affected by imbibition. Water movement is affected by the presence of the plants xylem. The cuticle and stomatal apparatus of the plant aid in transpiration and guttation provides an alternate process to resolve root pressure.

2Molecular Phylogeny of the Philippine endemic genus Antherostele Bremek. (Rubiaceae) inferred from ITS data (nrDNA) and its conservation statusTransportation of Water and Minerals In and Out of the Plant

Copyright 2012 SciRes. AJPSIntroduction What factors affect the rate of diffusion? The initial hypothesis was that high temperature and low density of solvent cause faster rate of diffusion of the plant pigments. The hypothesis was tested through experimenting on atsuete seeds as they were submerged in different solvents.

Osmosis is the tendency of the solvents molecules to move through a semi-permeable membrane, from a less concentrated to a high concentrated solution. With this definition, the hypothesis formulated was that a turgid cell would decrease in size if it is exposed to a highly concentrated environment.

Cell membranes are mostly composed of lipids, leading to a hypothesis of having the cell membrane become weak if exposed to high temperature and acidic environment. Cold temperature would cause the oils in the cell membranes to somewhat solidify, intensifying its rigidity.

Imbibition is the plants absorption of water through hydrophilic colloids. The hypothesis was that plant imbibition would be much less when exposed to such solvents like kerosene rather than water.

Transpiration is the loss of water vapor in the parts of a plant. The hypothesis tests whether a certain leaf of a plant transpires through the cuticle or the stomatal apparatus.

MethodologyDiffusion of Selected Plant PigmentsFour test tubes were filled with one gram of atsuete seeds each. Test tubes were numbered 1-4. Test tube 1 had a gram of atsuete seeds submerged in 10ml of distilled water. Test tube 2 had a gram of atsuete seeds submerged in 10ml of distilled water and was later boiled in water bath for 5 minutes. Test tube 3 had a gram of atsuete seeds submerged in 10ml of vegetable oil. Test tube 4 had a gram of atsuete seeds submerged in 10ml of vegetable oil which was already heated for 5 minutes earlier. Take note that the vegetable oil for test tube 4 was heated for 5 minutes before the atsuete seeds were put. After 30 minutes, the tubes were shook sideways and color intensities between the four set-ups were compared.OsmosisSections of the lower epidermis of the Bangka-bangkaan were stripped off. Take note that the lower epidermis is the side which is violet in color. The sections were mounted on glass slides wet and cover slips were put. The cells were examined under the microscope using LPO. This was then sketched. Without having to move the slide, the water was drawn off with a piece of paper towel and replaced with a 5% salt solution. The cells were again viewed under the LPO and sketched.

2.3 Factors Affecting the Integrity of Cell Membranes

A sharp blade was used to obtain 7 sections of sugar beet peelings. Take note there were evident intact cells in each section made. Three sections were transferred to test tubes A, B and C. 10ml of distilled water was added to test tubes A-C. Tube A was then placed under room temperature, tube B inside a refrigerator and tube C in a water bat with a 60 degree Celsius temperature. They were occasionally stirred within 30 minutes. After the 30-minute observation and stirring, the peelings were removed and mounted wet. The 3 sections were examined under the microscope, comparing the color intensity of each. The remaining four sections were mounted in different slides, labeled D, E, F and G. A drop of 50% chloroform was added to tube D, a drop of 50% acetone to tube E, a drop of 0.1M NaOH to tube F, and 0.1M of HCl to tube G. The sections were examined immediately after 15 and 30 minutes. Observations were then recorded.

2.4 Imbibition

2 pieces of wood and rubber were prepared and weighed for use in the experiment. 2 sets of 10g of corn seeds were also weighed and prepared. The said materials were separated and put into 2 separate beakers. The first beaker was labeled Beaker 1 and contained 1 piece of wood, rubber and 10g of seeds immersed in water. The other beaker was labeled Beaker 2 and contained 1 piece of wood, rubber and 10g of seeds immersed in kerosene. After 90 minutes, a spatula was used to take out the materials in both beakers and blot dried. The final weights of all the materials were recorded.

2.5 Movement of Water through the Stem

Pechay leaves were first secured for use in the experiment. Then 1cm from the base of the petiole was cut off. After that, the leaves were immersed in a bottle containing 10ml of 0.01% eosin dye solution. Immediately after 10-15 minutes, a leaf was taken and split longitudinally at the stalk. Measurements were taken of the length covered b the dye. Another leaf was taken and a cross-section was cut from it. The tissues stained were observed under the LPO of a compound microscope. A sketch of the cross-section was made showing the stained parts and labeling the identifiable features.

2.6 Comparison of Cuticular and Stomatal Transpiration by Four Leaves Method

4 identical leaves were secured and labeled A, B, C, and D. Leaf A was the control leaf. Leaf B was greased with vasellin in the upper surface, Leaf C was greased on the lower surface, and Leaf D was greased on both surfaces. Then, the leaves were hanged on a thread with both surfaces exposed to the air. After 3 days, the results were recorded.Results3.1. Diffusion of Selected Plant PigmentsTable 1. SpecimenDescriptionColor Intensity

Test Tube 1Atsuete+10ml distilled water+

Test Tube 2Atsuete+10mp distilled water placed in boiling water bath+++

Test Tube 3Atsuete+10ml vegetable oil++

Test Tube 4Atsuete+10ml heated vegetable oil++++

3.2. OsmosisThe wet mounted set-up had turgid epidermal cells. After replacing water with 5% salt solution, the cells plasmolysed and shrunk.3.3. Factors Affecting the Integrity of Cell MembranesTable 2. Sugar beet peeling sections inside test tubes with 10 ml distilled waterSpecimenDescriptionColor Intensity

Test Tube AUnder room temperature++

Test Tube BInside refrigerator+++

Test Tube CIn a water bath with 60oC temperature+

Table 3. Sugar beet peeling sections mounted to slides with different solventsSpecimenDescriptionColor Intensity

Test Tube DWith a drop of 50% chloroform++

Test Tube EWith a dropt of 50% acetone+

Test Tube FWith a drop of 0.1M NaOH++++

Test Tube GWith a drop of 0.1M HCl+++

3.4 Imbibition

MaterialsOriginal WeightImmersed in:Final Weight

Beaker 11 piece of wood1piece of rubber10g of corn seeds 0.5g0.4g10gWater0.6g0.3g0.1g

Beaker 21 piece of wood1piece of rubber10g of corn seeds0.3g0.4g10gKerosene0.4g0.4g11.1g

3.5 Movement of Water through the Stem

Results show that in the longitudinal splitting of the stalk, the dye reached the end of the stalk after a 15-minute time span. Also, in the cross-section, the stains represent the path the water travelled. The stained parts represent the xylem which is the pathway of water through the stem.

3.6 Comparison of Cuticular and Stomatal Transpiration by Four Leaves Method

The leaves that were greased with vasellin wilted much slower comparable to the control experiment. Leaf A wilted easily, while the others so little to no change depending in the surfaces greased.

DiscussionDiffusion of Selected Plant PigmentsHigh temperature and low density cause faster diffusion rate. Oil is less dense compared to water. These concepts resulted to the pigments of atsuete seeds in tube 1 to diffuse the slowest, then tube 3, tube 2 and tube 4 diffusing the fastest. The color intensity translates the diffusion rate; the darker the color, the faster the diffusion rate. Heated set-ups had faster diffusion rates and set-ups containing vegetable oil solvent diffused faster than those with distilled water.OsmosisThe first set-up (wet mount) was turgid because of water abundance in and out of the cells. After replacing water with 5% salt solution, the cell membranes shrunk because the water molecules inside the cell move to a higher concentrated solution (outside the cell). Plasmolysis occurred through osmosis.Factors Affecting the integrity of Cell MembranesThe sugar beet peelings have oil as one of its main components, since cell membranes are composed mainly of lipids and oils. When oil is heated, it decreases in volume; but when oil is cooled it solidifies (but not entirely) into a waxy, butter-like object. This explains why the cooled set-up had the darkest color intensity, and the heated becoming the lightest. Sugar beet peelings in the other half were affected by pH levels and organic solvents. The set-up having the lightest color intensity was the one with a drop of 50% acetone. Acid concentration might attribute to deterioration of plastids and loss of pigments leading to light color intensity. Among the four solvents namely, chloroform, acetone, NaOH and HCl, the 50% acetone has the highest acid concentration. Next was the solvent with 50% chloroform. NaOH had the lowest acid concentration allowing the peelings to preserve its color intensity. ImbibitionThe process of absorption of water by a seed that results in swelling at the beginning of germination is imbibition. The objects grew in weight because they cannot be dissolved in water, instead, they aided in the uptake of water. The materials immersed in kerosene had the same effect because they could not be dissolved.In seeds, the dry seeds uptake water because they cannot be dissolved. This happens just before germination to aid osmosis in growing plant cells.Movement of Water through the StemThe xylem vessels present in the steam of the plant serve as the pathway of water towards the leaves and other parts of the plant. From the roots which absorb water, the water passes through the stem by means of the xylem, which is shown in the experiment. This happens in a small amount of time because the plant constantly needs water to aid in its processes.Comparison of Cuticular and Stomatal Transpiration by Four Leaves MethodThe experiment clearly showed how much transpiration affects the plant. As one can see, the leaves that werent greased in the lower surface wilted and died easily because the water escaped by means of the stomatal apparatus. The ones with the vasellin on the upper surface wilted quite slower because the water that was about to transpire was trapped in the leaf.References[1] Committee on Biology 21, Biology 21 General Botany Laboratory Manual Vol. 2008-2009, Exercise 11: Transport of Water and Minerals In and Out of the Plant[2] James D. Mauseth, Botany: An Introduction to Plant Biology, Jones and Bartlett, Fourth Edition, pp. 267-269

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