Transport of Water and Minerals In and Out of the Plant

Anya Pena, Christian Solis, Jerick Bolintiam, Justine Alain Sy, Noemi Nunez, Venus Banaag
Cell activities, such as diffusion, osmosis, imbibition, and guttation, involve the transport of water and materials in and out of the cell membrane- which is essential for a plant to maintain equilibrium in an unstable environment. The transport of the dissolved substances in different environments affects the plant cell in different ways. To further understand the cell activities and the environment¶s effect on the plant cell, seven different tests were conducted. Diffusion of selected plant pigments was observed in the first experiment where Bixa orellana seeds were placed in different test tubes containing different substances, namely: distilled water, boiled water, vegetable oil, and heated vegetable oil. Osmosis was observed in the second experiment where small strips of the lower epidermis of the Rhoeo spathodea was put two different slides- one with water and the other with salt solution. The factors affecting the integrity of the cell membrane was determined in the third experiment using Pyrus Malus peelings which was placed in different temperatures and substances. Imbibition was observed in the fourth experiment where wood, rubber, and corn seeds were put in two beakers- one with water and the other with kerosene. It was also noticed that each of the material had an increase in weight when placed in kerosene. The movement of water through the stem was observed in the pechay stalk of the the fifth experiment. Transpiration was observed in the sixth experiment which also compared four identical leaves with different applications of vasellin. Finally, guttation was observed in rice seedlings which were covered with a wide mouth jar. Keywords: diffusion, osmosis, cell membrane, imbibition, active transport, transpiration, guttation

The cell membrane is the biological membrane separating the interior of a cell from the outside environment. It is a semipermeable membrane surrounding all cells, which controls the movement of substances in an out of the cells; therefore, responsible for maintaining a steady and stable living condition²or a state of equilibrium²even in the midst of a transient environment. It is

involved in a wide variety of cellular processes, such as growth, absorption, and respiration, and serves as the attachment point for the intracellular cytoskeleton and, sometimes, the extra-cellular cell wall. There are two ways in which transport can occur across a membrane, either by passive or active, depending on the energy required during the process. Simple diffusion, a kind

of passive transport, moves water from regions of high water concentration (low solute concentration) to regions of low water concentration (high solute concentration). This process however, is only possible for solutes that are readily permeable such as nonpolar and small polar molecules. Other processes include: bulk flow, osmosis, imbibition, and active transport. Through this lab exercise, the student should have been able: (1) To determine some factors that affect the diffusion process; (2) To differentiate between

After 30 minutes, these test tubes were shook and the color intensities in each was compared by using +, ++, and +++.

Osmosis ± Thin sections of the lower epidermal side of Bangka-bangkaan

(Tradescantia spathacea) were cut by Group number 3 and these were placed into a wet mount. Using this examination under the LPO of the microscope, a sketch of a turgid cell was made. Next, without moving the slide, water was drawn off using a paper towel and was replaced with a 5% salt solution. A sketch showing the change in the cells was then made based on the

diffusion and imbibition; (3) To determine some factors that affect the permeability of cell membranes; and (4) To demonstrate the various processes by which materials are transported and transpired.

observations under a microscope.

Factors Affecting the Integrity of Cell Membranes ± Apple peels were acquired by

METHODOLOGY Diffusion of Selected Plant Pigments ± Atsuete (Bixa orellana) seeds were weighed by Group number 1 and placed 1g of seed was placed into 4 test tubes. These test tubes were labelled 1-4. In test tube number 1, 10ml of distilled water was placed. In test tube number 2, the group put 10ml of distilled water and then placed it in a boiling water bath. In test tube number 3, they put 10ml of vegetable oil. In test tube number 4, 10ml of pre-heated vegetable oil was placed.

our group by using a sharp blade. 7 sections of peels were placed in a beaker filled with distilled water. The first three sections were transferred into three test tubes each with 10ml of distilled water and labelled A, B and C. Test tube A was placed under room temperature (250C) while Test tube B was placed inside a refrigerator (100C) and Test tube C was placed in a water bath (600 C). After observing for 30 minutes, each of the three sections were placed into wet mounts and viewed under the microscope. Color

intensity in each was compared by using +, ++, and +++. For the remaining 4 sections, they were also placed in wet mounts and labelled D-G. A drop of pure chloroform was added to D. A drop of 50% acetone was added to E. 0.1 M of NaOH and 0.1 M of HCl were added to F and G respectively. These four were observed under the microscope after 15 minutes and after 30 minutes. recorded. The observations were then

leaves were immersed in a bottle filled with 10ml of 0.01% eosin dye solution. After 15 minutes, a leaf was removed and the stalk was split longitudinally. The length covered by the dye was measured. From another leaf, a thin cross section of the stalk was cut and viewed under the LPO of the microscope. The stained tissues were then identified through being viewed under the microscope.





Stomatal Transpiration by Four Leaves Imbibition ± The weights of 2 pieces of wood and 2 pieces of rubber were weighed by Group number 4. 2 sets of 10g corn seed were also weighed. In one beaker, one piece of wood, one piece of rubber and 10g of corn seeds was placed. Water was added until each of the materials was completely immersed. In the other beaker, one piece of wood, one piece of rubber and 10g of corn seeds was and immersed with kerosene. After 90 minutes, all the materials from the two beakers were taken out and dried gently. The final weights of these materials were then measured. Guttation ± 5 rice grains were planted on a container by Group number 1. The lower portion of the container was immersed Movement of Water Through the Stem ± Pechay leaves with intact petiole were gathered by Group number 3. 1 cm was cut off from the base of the petiole. The with water. When the seedlings are 2-5cm long, they were covered with a transparent bell jar (a wide-mouthed bottle can also be used). This set-up was observed and the Method ± 4 identical leaves were gathered by Group number 4. These were then labelled A, B, C, and D. Leaf A was the control. Using vasellin, each of the three remaining leaves were greased. The upper surface of B was greased while Leaf C had its lower surface greased. Both sides of D were greased. These leaves were then hanged by a thread to expose both sides to air. The set-ups were observed after one meeting.

droplets that formed on the leaf surfaces were noted.

Results Diffusion of Selected Plant Pigments ± The diffusion of the pigments of the seeds of Bixa orellana is relatively faster and greater in the setups exposed to higher
Fig. 1 Tradescantia spathacea cells in a water solution

temperatures. Also, the diffusion was greater in oil than in water, heated or not. Substance Test Tube 1 (Distilled Water) Test Tube 2 (Hot Distilled Water) Test Tube 3 (Vegetable Oil) Test Tube 4 (Heated Vegetable Oil) Observation + +++ ++ ++++
Fig. 2 Tradescantia spathacea cells in a 5% salt solution

Factors affecting integrity of cell membrane ± The results produced from this experiment can be divided into three parts: temperature, pH solvents. With temperature, the peeling of Pyrus malus that was exposed to the lowest temperature exhibited more damage to the cell membrane while the one that was put in a water bath exhibited less damage. For organic solvents, the experiment that pure chloroform is much more effects, and organic

Table 1. Shows the color intensity of the different setups, + being the lightest and ++++ being the darkest

Osmosis ± To observe how the cell changes in plasmolysis, wet mounts of Tradescantia spathacea were made and the water was later on replaced with a 5% salt solution. The cells were larger and turgid when it was exposed in water (Fig. 1). When the water was replaced with the 5% salt solution, the cells became flaccid (Fig. 2).

damaging than 50% acetone. As for pH effect, the one that was subjected to NaOH, a base, had less damage than the one subjected to HCl, an acid.

Fig 3.1 Pyrus malus soaked in water with a temperature of 10°C

Fig 4.2 Pyrus malus subjected to pure chloroform after 30 minutes

Fig. 3.2 Pyrus malus soaked in water that has a temperature of 25°C

Fig 5.1 Pyrus malus subjected to 0.1M NaOH solution after 30 minutes

Fig. 3.3 Pyrus malus soaked in water that has a temperature of 60°C

Fig 5.2 Pyrus malus subjected to 0.1 HCl solution after 30 minutes

Imbibition ± In this experiment, the affinity of wood, rubber, and seeds to different solvents, kerosene and water, were compared. Wood and rubber makes good .
Fig 4.1 Pyrus malus subjected to 50% acetone solution after 30 minutes

imbibant of water as they showed greater significant change in weight, while rubber showed greater change in weight when it imbibed kerosene.

Initial Weight Final Weight % in Weight (Wi) (Wf) {(Wf-Wi)/Wi} * 100 Water rubber 0.5g 0.7g 40% wood 19.95g 21.6g 8.27% seeds 10g 12.1g 21% Kerosene rubber 0.4g 0.7g 75% wood 16.9g 17.9g 5.92% seeds 10g 10.6g 6% Table 2. Compares the affinity of certain plant materials to different solvents Movement of water through the stem ± The 10mL of 0.01% of eosin dye solution went up through the stem in a straight line manner through the xylem tissues. leaf C smeared with grease on the lower surface, and leaf D smeared with grease on the both sides, a week of observations were made. Leaf A was the most desiccated one and curled on both sides. Leaf B and C were both half-dry with leaf B curling outward and leaf C curling inward. Leaf D remained fresh, waxy, and moist, with no occurrence of desiccation and curling.



Fig. 6.1 Cross-section of pechay stalk

Guttation ± The appearance of xylem saps happened at the tip of the leaf blades after the plant was covered with a wide mouth jar.

DISCUSSION Diffusion of selected plant pigments ±
Fig. 6.2 External view of stained pechay

The greater the concentration gradient between the outside and the inside of the

Comparison of cuticular and stomatal transpiration by four leaves method ± Leaf A being the control, leaf B smeared with grease and vaselin on the upper surface,

membrane, the greater the diffusion. If the concentration of the pigments outside and inside the membrane were greater, then it would diffuse more quickly. The opposite is

also true. Another factor affecting the rate of diffusion is the size of the particles. The smaller the size of the particle, the faster it would be diffused. For the Bixa orellana seeds, the carotenoids diffused more quickly when it was submerged in the heated vegetable oil solution. Generally, increases in temperature speeds up the movement of molecules and faster movements of molecules means faster rates of diffusion. This is why the heated distilled water and the heated vegetablle oil had the fastest rates of diffusion. As for the competition between oil and water, faster rates of diffusion happened in oil because the pigments of the Bixa orellana seeds are insoluble in water.

solution on the other side of the membrane. In a plasmolyzed cell, the water would continually move out of the cell until the concetration of the impermeable solutes equals to that of the hypertonic solution.

Factors affecting the integrity of cell membrane ± In this experiment, the apple peelings exhibited diffecrent intensities of the color of their pigments. The darker ones indicate more damage and stress to the cell membrane while the lighter ones indicate less stress and damage. The ones with the damaged cell membranes exhibited darker colors because when the cell membrane gets destroyed, the pigments from inside the cell leak out. Temperature ± 3 test tubes containing

Osmosis ± When the Trandescantia spathacea was immeresed in a hypotonic solution, the cells became turgid. The water was moving from a smaller concentration of solutes than the solution on the other side of the membrane. In a turgid cell, the water will continually move into the cell until the concetration of the impermeable solutes equals to that of the hypotonic solution. When the water was replaced with a 5% salt solution, the water bacame

immersed apple peelings in water were subjected to different temperatures. In this experiment, we have arrived with a

conclusion that lower temperatures inflict more damage to the cell membrane and that normal and high temperatures did not do much damage. pH effects ± Wet mounts of 0.1M NaOH solution and 0.1M HCl solution were observed for 30 minutes. The results show that the more acidic a solution is, the more damage it can inflict to the cell membrane. The more basic it is, the less damage.

hypertonic. The water then moved from a larger concentration of solutes than the

Organic solvent ± Chloroform inflicted more damage in the membrane than the acetone. In this one, the pH again got involved. Chloroform is more acidic than acetone.

because imbibition in water can burst the seed coat, which would signal the start of germination.

Movement of water through the stem ± The 0.01% of eosin dye solution rose up

Imbibition ± Woods and seeds imbibed better in water while rubber imbibed better in kerosene. Water is composed of 2 molecules of hydrogen and a molecule of oxygen, hence H20. Kerosene is an oil distillate commonly used as a fuel or solvent. It is a thin, clear liquid consisting of a mixture of hydrocarbons and is primarily derived from refined petroleum. Imbibition occurs with or without the help of living cells. According to Ferdinand Sachs¶ imbibition theory, water moves in tubes in the walls of plants without the cooperation of living cells and not within the cell cavities. There are no living cells in the veneer. Veneer comes from the peeling of tree trunks and these are secondary xylem, which are dead cells. Living cells are involved in the

through the stem into the leaves. This only shows that water moves through the xylem elements in the stem to spread the water. The eosin dye stain reached up to the cross section of the leaf which means that the water is diffused all throughout the plant through the stem.

Comparison of the cuticular and stomatal transpiration ± Too much

transpiration can cause the dessication of the leaves. Cuticle helps in preventing this. The control set-up, leaf A, where nothing was applied, desiccated while leaf D which was smeared with grease on both sides remained fresh and waxy without any occurrence of desiccation.

Guttation ± Since the jar was closed, there was an excess of moisture in the environment of the rice seedlings, hence, it could not undergo transpiration, which why guttation was induced. Guttation is the release of water from plants the hydathodes, which is induced by

imbibition of the seed. This is because the entry of water takes place in the testa, which may be actively dividing. The swelling effect of imbibition in seeds is important in seed germination

root pressure, mainly because of the high moisture content of the soil while title=Imbibition&oldid=295094135 LYCOPENE. (2009, September 21). In Wikipedia, The Free Encyclopedia. Retrieved 10:35, September 21, from

transpiration is the process of water vapor loss from the internal atmosphere of the plant. Another difference is that in


transpiration, water vapor is released while in guttation, xylem saps are released. title=Lycopene&oldid=315268969 STERN, K. R. 1997. Introductory Plant

LITERATURE CITED ANTHOCYANIN. (2009, September 23). In Wikipedia, The Free Encyclopedia. Retrieved 14:25, September 23,

Biology TONICITY. (2009, September 23). In Wikipedia, The Free Encyclopedia. Retrieved 2009, 12:58, September 23, from

2009, from w/index.php?title=Anthocyanin&old id=315714581 CAROTENOID. (2009, September 18). In Wikipedia, The Free Encyclopedia. Retrieved 2009, 18:59, September 18, from title=Tonicity&oldid=315700881 TRANSPIRATION. (2009, September 16). In Wikipedia, The Free 10:18,



September 16, 2009, from http:// Transpiration&oldid=314314913 title=Carotenoid&oldid=31476936 GUTTATION. (2009, May 15). In

Wikipedia, The Free Encyclopedia. Retrieved 16:21, May 15, 2009, from title=Guttation&oldid=290118315 IMBIBITION. (2009, June 8). In Wikipedia, The Free Encyclopedia. Retrieved 02:19, June 8, 2009, from

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