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It is widely accepted that the normal pH condition for successful plant growth is between 5.0 and 6.5. There are natural mechanisms, such as the buffering capacity of soil, that keep pH within this range. However, there are certain occurrences in nature that can raise or lower the pH of a plantÕs environment. Because water is such an important resource for plant growth, the quality of the water is directly related to the health of the plant. Our study seeks to answer the question, ÒHow does the pH of a water source affect the growth patterns of green beans?Ó This study has been undertaken to investigate how acidic pH levels of water sources can affect plant growth. This study examines the effects of an acidic environment on green beans, a crop plant. Many aspects of plant health are included, both at the external level (height, leaf size, weight, etc.) and the internal level (cellular structure, cell wall and membrane functionality, and chloroplast abundance).

Plant growth can be severely affected by the acidity of the surrounding media. While the sources of acidity, such as acid rain, do not directly destroy the plant, low pH levels affect many of the processes necessary for life, such as nutrient uptake and symbiotic relationships [2]. Research concerning soil and water pH and its relationship to plant health can impact many issues that our present in our environment. For example, areas with low soil pH levels tend to have lower species diversity [3]. Also, the nutritional value of crop plants can be degraded if the crops are grown in an acidic environment [9]. This condition has a direct impact on agricultural systems and practices. These effects of low pH environments need to be taken into account when considering the problems of acid rain and acidic soils.

In previous experiments, it has been observed that plants that are exposed to acidic conditions lower than normal range are not able to grow as well [2, 7, 10]. Therefore, we believe our plants will exhibit similar external signs of poor growth as they are subjected to more acidic conditions. More specifically, low pH conditions will decrease the overall health of the plants. Our definition of overall health considers attributes such as plant height, stem width, length between secondary stems, color, root mass, plants mass, overall mass, and reproductive structures. Many of these experiments do not test growth at a pH much lower than 4.0, however, and our experiment examines growth at a pH of 3.0. The pH levels of the water sources used in our experiment bracket the levels of pH normally found in the environment. Furthermore, our research examines the internal effects of a low pH environment on individual plant cells.

Background Information

What is pH?

The pH is most commonly known as the measure of the acidity or alkalinity of a solution, numerically equal to 7 for neutral solutions, increasing with increasing alkalinity and decreasing with increasing acidity. The pH scale ranges from 0 to 14, most acidic to most basic [15]. In chemical terms, the pH scale refers to the concentration of hydrogen ions in a substance, and each whole unit represents a factor of 10 i.e. pH5 is 10 times more acidic than pH6 [16]. Basically a pH scale measures the concentration of H+ and OH-.
The internal pH of most living cells is close to 7. When there is even a slight change in the pH, this can be extremely harmful because the chemical processes of the cell are sensitive to the concentration of hydrogen and hydroxide ions. Biological fluids can resist change to their own pH when acids and bases are introduced because of the presence of buffers. Buffers in human blood for example maintain the blood pH very close to 7.4. A person cannot survive if the pH of their blood drops to 7 or rises to 7.8. Under normal circumstances the buffering capacity of the blood prevents such swings in the pH level. An acid adds hydrogen ions to a solution, but it also removes hydroxide ions because of the tendency of H+ to combine with OH- to form water. The base has the opposite effect with an increasing OH- concentration but also reducing the H+ concentration by the formation of water. Each pH unit represents a tenfold difference of the H+ and OH- concentration. It is this mathematical feature that makes the pH scale so compact. For example a solution of pH 2 is not twice as acidic as a solution of pH 4, but a hundred times more acidic. So when the pH of a solution changes slightly, it actually changes the concentrations of H+ and OH- substantially. H+ ion concentration and pH relate inversely. On the other hand, OH- ion concentration and pH relate directly. Increasing pH means the H+ ions are decreasing. Decreasing pH means H+ ions are increasing. Increasing pH means OH- ions are increasing. Decreasing pH means OH- ions are decreasing [17].
Plant Growth
Water is essential for plant processes such as cell division and elongation, imbibitions and germination, photosynthesis, and nutrient uptake. For plant growth, favorable pH of water is between 5.0Ð 6.5. The pH of a soil can directly impact how each type of plant will best grow; a strongly acidic soil will not best utilize nutrients such as nitrogen, phosphorus and potassium, and will not sustain many plants. For example, a soil pH as low as 5 is ideal for potatoes, whereas a pH nearer 7 will be better for asparagus. The pH of plants cells is maintained in a narrow range. Without it, considerable structure changes could occur affecting the function and viability of the structures. Water is truly vital for growth. All plant tissues consist of cells, including the cell membrane. Water and other small molecules can move through the membrane. There are proteins in the membrane that can quickly move salt ions, usually potassium, into the cell. Other proteins move sugars in and out of the cell. This allows the cell to be saltier and sweeter than the water around it. Water moves from a solution of low concentration into a higher concentration solution to dilute it. Since the cell can actively move salts into the cell, water moves into the cell as some water leaks out, and a balance between inflow and outflow occurs. Plants can make virtually everything they need from water and air with a few nutrients that the roots absorb from the soil. The plant uses sunlight to split water into hydrogen and oxygen. It discards the oxygen as a waste product. The plant uses the hydrogen to make sugar from carbon dioxide in the air. Both plants and people use oxygen in the air to burn sugar and make energy to live. Take a moment to consider the structure of a leaf. It is wide and flat, which it can catch a lot of light but the large surface area means water loss is a problem. It has a waxy cuticle to reduce water loss. The leaves get water from the roots. Air enters the leaves through small holes in the leaves called stomata. When the stomata are open they let air in but they also lose water through evaporation. This is an important point to remember because if those stomata close to conserve water, photosynthesis and sugar production stops. Water is the single most restrictive factor in plant growth. Plants grow in two ways, cell division, which creates more cells, and cell expansion, which increases cell size. Cells grow by taking up water. Young cells expand and produce new cell wall at the same time. Eventually, the cell wall becomes so thick that the cell can no longer expand and growth stops. If water is reduced during growth, final cell size is reduced, which means fewer, smaller leaves, smaller fruit, shorter, thinner stems and fewer roots. Drought stress results in smaller, weaker plants. Most of the water is taken up at the root tips. The roots primary function is to take up water. At the root tip, there is a zone of active cell division where new cells are formed. Directly behind the tip is a region known as the zone of elongation where the new cells grow by increasing in length. This elongation pushes the root into the soil. There are no barriers to water movement in these young tissues and water can move freely into and out of the root. Behind the zone of elongation is the zone of differentiation, where the cells develop into different cell types. In the center of the root is woody xylem tissue, which carries water from the roots to the stems. Around the xylem is a layer of cells called the endodermis. These cells have a thick waxy barrier between them called the 'Casparian Strip', which prevents water loss. Water has to enter the plant cells of the Casparian Strip in order to get into the roots. This barrier is not only effective in keeping water in the root but also prevents its easy entry into the plant. As long as we maintain soil moisture throughout the growing season the roots will be able to maintain an adequate flow of water to the leaves to maintain growth. In total, water takes a continuous path through the soil and the plant then out the leaves into the air [18].

How pH can Affect Plants

At the cellular stage, a change in the pH level can severely damage structures within the cell. In plant cells, the pH is maintained in a generally narrow range that allows for cellular processes to occur. Large quantities of protons are used and produced in these processes. Changes in pH levels are caused by an increase of the concentration of these protons in the cytoplasm. However, there are substances within cells that naturally maintain the necessary pH range, such as amino acids, organic acids, and phosphate groups [5]. There is also variability in the pH levels of substances within the cytoplasm, allowing for normal cellular functions to occur. If this pH range is not maintained, however, structural deformities could occur, leading to the failure of processes and functions within the cell [5].

The pH of the growing media (usually soil) surrounding the plant can have a strong impact on plant growth. This is largely due to the reliance of plants on the soil for several nutrients and minerals. Acidic pH can also be detrimental to microorganisms in the soil that are beneficial to the plant. Most soils have a natural buffering capacity that keeps the pH at a level suitable for plant growth. However, not all soils are capable of maintaining a suitable pH level. Thinner soils, such as those in Northeastern regions of the U.S. have lower buffering capacities than thicker soils found in the Midwest [10].

Low pH levels in soils can have a severe effect on the microorganisms that form symbiotic relationships with plants. A study examining the effects of soil pH on cowpeas showed that poor growth of plants can sometimes be attributed to poor microorganism activity [3]. Microorganisms in the soil aid plant growth by forming symbiotic relationships with roots, aiding in nutrient absorption. These relationships can be affected by low soil pH. In legumes, nodules are not produced at low pH levels lower than 5.0. Also, low soil pH can reduce the diversity of microorganisms and the numbers of rhizobia in the soil available for symbiotic relationships [5].

A high concentration of H+ ions (low pH) in the soil can also lead to nutrient deficiencies. Acidic conditions can lower the levels of phosphorus, calcium magnesium and molybdenum, nutrients that are essential to plant growth [2].
Low pH levels also create mineral levels in the soil that can be toxic, thus harming plants. Low pH can increase the solubility of minerals such as aluminum. Plant growth can be repressed in soils that have severely high levels of aluminum or manganese [2].

Direct exposure to acidic water sources can be harmful to plants as well. For example, in the Mountainous regions of the eastern U.S., acid rain and acidic fog are common [10]. While exposure to acid rain does not directly kill plants, the effects of low pH conditions (leaf damage, nutrient deficiencies and mineral toxicity) are detrimental to the plants. Furthermore, acidic water liquefies essential nutrients in the soil and washes them away before they can be absorbed by plants [10].

Plants that are affected by low pH levels exhibit certain distinct characteristics. Trees affected by acidic soil conditions exhibit retarded growth Also, the leaves of plants turn yellow or brown, and most plants eventually die because of overexposure to low pH [10]. Root health can also be compromised, as mycorrhiza and nodules cannot form at low pH levels.

The pH of soil can also affect plant species diversity and richness. A study done in the Blue Ridge region of the U.S., where highly acidic soils are prominent, concluded that sites with higher pH have more species richness. Also, the average density of species was twice as high in regions with high pH, as compared to regions with low pH. This condition was attributed to the more encouraging growing conditions associated with higher pH levels [3]. The study also shows that species diversity was lower in regions with acidic soil. It was noted that this condition was probably due to the fact that plants must be highly specialized to survive in acidic conditions [3].

The nutritional value of plants can be adversely affected by low pH levels. There is a direct correlation between the nutrients contained in plants and the mineral content of the soil in which they grew, which is affected by pH [9]. However, foodcrops are generally not affected; there are many ways to balance the pH of soil. Low pH can be remedied by adding crushed limestone or acid-tolerant rhizobia species to the soil, or by directly applying lime to seeds [9].

Causes of Soil and Water Acidity

ÒAcid rainÓ is a general term used to describe acids falling from the atmosphere. Because acid falls in solid particles and wet solutions it can be referred to as acid deposition. Dry deposition defines the accumulation of solid acid forms on various surfaces such as trees and buildings. Wet deposition describes the acid falling in the form of precipitation. Wet deposition, when combined with watershed from dry deposition covered surfaces, can cause detrimental effects to plant-life. The acidity in the atmosphere is directly connected to sulfur dioxide and nitrogen oxides produced as by-products from industry. About 2/3 of all sulfur dioxide (SO2) and ¹ of all nitrogen oxides (NOx) come from electric power generated by the burning of fossil fuels. Other causes of NOx and SO2 pollution are automobile and airline industries. These industries are the main contributors to the formation of acid rain. (EPA Acid Rain)

Acid rain doesnÕt directly kill trees and plants; instead it affects the nutrient uptake and starves the plant life. The amount of acidity the soil can neutralize in an efficient way is referred to as Òbuffering capacityÓ. If the buffering capacity is low, the soil will be depleted of the calcium and magnesium needed to sustain plant growth. Also, trees are weakened by an accumulation of acid damaging the leaves and needles; which restricts photosynthesis. (Acid Rain, Air Pollution, and Forest Decline)

Forests with high soil buffering capacity are still at risk, especially in mountainous regions. Forests in northeast United States are at high risk due to elevation. Plant-life, near acidic fog and clouds, has a greater exposure to acid because acidic levels are denser before precipitation. Also, these regions receive more rain because of the geographical structure. Winds can carry acidity from industrialized regions to farmland; however, food crops arenÕt noticeably affected because fertilizers replace nutrients needed for sustained plant growth. Without these supplementing fertilizers the food crops sustain substantial damage. (EPA Acid Rain)

Aquatic regions are also affected by acid rain. Although lakes and streams are slightly acidic, those located in watershed areas are at greater risk because the soil surrounding them cannot filter or neutralize the water. In areas with low buffering capacity in the soil acid rain releases aluminum into the water, which is toxic to many aquatic species. (EPA Acid Rain)

Larger Scopes of Soil and Water Acidity

The affects of acid rain on the health of environment can cause serious problems. Because of the links between fish, plants, and other species, one change can affect the whole ecosystem. Changes in acidic levels can cause toxic aluminum to be released into the lakes and streams altering the biodiversity of an environment. The nitrogen in acid rain gathers along water surfaces in estuaries depleting the water of oxygen. The lack of oxygen results in lower plant production, which is used as food and cover for fish and other organisms. The lack of oxygen also directly affects the health of fish, shellfish, other seafood, and coral reefs. Unhealthy coastal systems produce economic problems for fishermen and those who cater to tourism.

Acid rain can cause problems to plant-life other than trees. Farmland and food crops can be affected by acid rain it usually isnÕt serious because of the fertilizers used to replace nutrients lost in the soil.

Human health is not directly affected by acid rain but rather the fine particles of NOx and SO2 that can be inhaled. These particles are directly related to increased illness and premature death from heart and lung disorders.

Due to the many problems of pollutants causing acid rain or rather acid deposition the EPA, state governments, and academic studies have worked together to reduce and eliminate acid rain problems. The cheapest forms of energy are nuclear, hydropower, and coal, but other forms of alternative power sources, aside from fossil fuels, are being investigated. Because of recent increases in acidic pollutants, the Bush Administration has pledged to Òprotect the air we breatheÓ; in doing so, they proposed a bill to amend the Clean Air Act, which passed in April of 1990. (Acid Rain, Air Pollution, and Forest Decline)
The Acid Rain Program was set up to achieve significant environmental and public health benefits in the United States. The three main objectives for the Acid Rain Program are to 1) reduce the amount of NOx and SO2 in the atmosphere, 2) facilitate active trading allowances and use of other compliance options to minimize compliance costs, maximize economic efficiency, and promote strong economic growth, and 3) promote pollution prevention and energy efficient strategies and technologies. In an attempt to clean the air while not causing economic burdens, the Acid Rain Program has offered companies many compliance options to choose from which will allow companies to operate while reducing polluting emissions. (EPA Acid Rain)

References

1. Correa, O. S., A. Aranda and A.J. Barniex. ÒEffects of pH on Growth and Nodulation of Two Forest Legumes.Ó Journal of Plant Nutrition, Sept 2001: 1367-1375.: Documents the effect of acid pH on two species of forest legumes. Suggests that mycorrhizae are a major factor in determining a plantÕs tolerance to pH.
2. Rohyadi, A., et. al. ÒEffects of pH on mycorrhizal colonization and nutrient uptake in cowpeaÉÓ Plant & Soil, March 2004: 283-290: Documents how fungi affect cowpea plants when they are growing in low pH environments, and the relationship between fungi growth and pH.
3. Peet, Robert K. Jason D. Fridley and Joel M. Gramling. ÒVariation in Species Richness and Species Pool Size Across a pH GradientÉÓ Folia Geobotanica, Dec 2003: 391-401: Studies the association with plant species in acidic soils in the Blue Ridge Mountain Range
4. Hall, J.M., E. Patterson, K. Killham. ÒThe effect of elevated CO2 concentration and soil pH on the relationship between plant growth and rhizoshpere dentrification potential.Ó Global Change Biology, Feb 1998: 209-216: Ryegrass was subjected to different levels of CO2 concentration. Investigates the relationship between plant growth and rhizosphere dentrification potential and soil pH.
5. Rengel, Zdenko. Handbook of Plant Growth: pH as the Master Variable. New York: Marcel Dekker 2002: Outlines the role of pH in plant growth. Describes the biological and chemical effects, as well as interaction of the plants with their encironments
6. Lane, Carter N. Acid Rain: overview and abstracts. Hauppauge, NY: Nova Science Publishers, 2003: Outlines the causes of acid rain, and describes its effects on ecosystems.
7. Abrahamsen, Guunear, Arne O. Stuanes and Bjorn Tviete. Long-Term Experiments with Acid Rain in Norwegian Forest Ecosystems: Describes the methods and results of an experiment that took place in Southern Norway. Shows the long-term effects of acid rain on forest ecosystems from the 1950Õs through the 1980Õs.
8. Styer, Roger C. and David K. Koranski. Plug & transplant production: a growerÕs guide. Batavia, Ill: Ball Publishers, 1997: Describes methods of plant propagation and what factors should be considered.
9. Forbes J.C., and R.D. Watson, Plants in Agriculture. New York, Cambridge University Press, 1992: Describes plant growth in detail, outlining the specific factors that influence growth and how the environment affects parts of the plant.
10. http://www.epa.gov/airmarkets/acidrain/effects/forests.html
11. http://www.necc.mass.edu/mrvis/Mr1_6/start.htm
12. http://www.enviroliteracy.org/article.php/2.html
13. http://www.epa.gov/acidrain/index.html
14. http://www.dpi.qld.gov.au/horticulture/17695.html
15. http://dictionary.reference.com/search?q=pH
16. http://www.gesource.ac.uk/ph-scale.html
17. http://www.elmhurst.edu/~chm/vchembook/184ph.html
18. http://www.canr.msu.edu/vanburen/watergrw.htm

Experiment Design

We will observe100 green bean plants in our experiment. We will have 5 groups of 20 plants, each subjected to a different water source pH Ð 7.0, 6.0, 5.0, 4.0 and 3.0. The solutions were created by mixing sulfuric acid with water. Using pH testing strips, we were able to determine that the pH of the solution was at the desired level. The solutions were contained in glass container, each labeled with the pH level of the contained solution. The plants will be ÒwateredÓ with the solution three times a week. All 100 plants will be watered with the same volume of water at the same time. We will measure a variety of characteristics of the plant to determine the effects of the water source pH on the plants.

Materials and Methods

On Thursday, October 20, 2005, the bean seeds were planted and placed in a stable environment (Boyd Hall greenhouse). Each bean seed was planted in a separate container, each with the same amount of Metromix 360 soil. 20 seeds were planted in each of 5 different flats, separated according to the solution they will be exposed to. The seeds were labeled according to their group. Variables such as amount of sunlight and temperature will be the same for all plants. The only variable in our experiment will be the pH of the water-based solution applied to the plants. On Tuesday, October 25, 2005, using a soil testing kit, the pH of the soil will be tested before the watering process begins. During and after our experiment, we will continue to test the soil pH, to determine what affect the pH solutions have on the soil and to determine if the soil pH affects plant growth. The plants will be subjected to the varying pH levels of water beginning on Tuesday, October 25, 2005. The experiment will continue until November 18, 2005. On this date, the plants will be removed from the soil and will be used in further testing.

During our experiment, many observations will be taken. Observations will be recorded five days a week (Monday through Friday). Photographs taken with a digital camera will be used to more completely document the experiment. The soil pH will be tested, and many characteristics of the plants will be documented. Using metric rulers, height, primary stem width, and the length of internodes will be measured and recorded. Color will be observed and documented through photographs. When the plants start to flower, the number of buds, flowers, pods, beans in the pods, and stem splits will be documented, if these structures exist. On November 18, the plants will be dug out of the soil. Before drying out, the root mass, stem mass and overall mass will be recorded for each plant. These measurements will be taken again after the plants have completely dried. Five plants from every group will then be dissected. Using a microscope, we will observe cellular characteristics, such as membrane and cell wall quality and chloroplast abundance to determine if these structures are affected by pH variability. All of the quantities measured during and after the experiment will be organized into data sheets. We have included a sample data sheet in this packet.

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• great idea (11/9/05) [ Contributor: Katie Bosworth ]
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