The Effect of Soil pH on the Stomatal Density of Tradescantia spathacea

Annelyn Ruth Lanoche Sucayan

Group 4 Section 3L

December 5, 2019

A scientific paper in partial fulfilment of the requirement in Bio 11.1 Laboratory under Professor
Kristine Abenis, 1st Semester, 2019-2020.
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ABSTRACT

The buds of Tradescantia spathacea plant of the same age were planted on nine
pots with equal amount of loam soil mixed with sand and pebbles. Three treatments
(soil watered with tap water, vinegar solution and baking soda solution) were
obtained with three replicates. The plants were watered 20 mL twice a day of the
corresponding treatments prepared for a span of four days. Leaves with the same
size and location on the plants were collected. Imprinting method was used to
determine the number of stomates and epidermal cells in order to compute for the
stomatal density of treatments. The pH level of each treatments was measured using
the digital pH meter to distinguish whether the treatments were acidic, neutral or
basic. The results of the experiment showed that soil watered with tap water has
greater stomatal density of 0.27 with a pH level of 6.1 followed by soil watered with
vinegar solution having a stomatal density of 0.23 with a pH level of 5.9 and soil
watered with baking soda solution has a stomatal density of 0.21 with pH level of
7.3. The pH level of the soil affects the stomatal density of the leaves of
Tradescantia spathacea such that the different treatments used varied from each
other. Since the pH level of soil affects the plant growth and plant growth varies
with the nutrients present in a plant which affects the stomatal density of a leaf.
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INTRODUCTION

Plants are living organisms living on either land or water. Plants can be classified

by their different forms such as trees, herbs and others are bushy in form. They provide

products which are essential to humans. On the same manner, plants help maintain balance

in oxygen which is the most important gas enabling us to breath. While animals emit carbon

dioxide, which is essential to plants, by the process of inhaling the oxygen (Sarma, 2017).

Tradescantia spathacea is a succulent introduced to South Asia and many Pacific

Islands. The plant is very invasive on some areas especially in Florida wherein it invades

native plant communities and it was almost declared as pest. Tradescantia spathacea

prevents native plants from germinating by creating a dense groundcover on the forest

floor. Tradescantia spathacea has diverse reproductive methods and grows in areas other

plants cannot. These two characteristics make this plant a potential danger to many areas.

Monitoring is recommended wherever this species is present (Global Invasive species

database, 2019). The plant is also considered as a landscape plant for it is commonly found

and used on gardens.

Just like other plants, Tradescantia spathacea exhibits natural plant processes such

as transpiration and respiration. Transpiration is a process wherein evaporation of water

vapor takes place, while respiration is the process of releasing oxygen due to another

process called photosynthesis. These processes occur at the tiny pores found usually and

abundantly found on the abaxial side of a leaf called stomates or stomata. With this,

transpiration and respiration rely on the number of stomates on a leaf. The number of

stomata per unit area in a leaf is called stomatal density. Another element that contributes
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in these processes as well as to the plant growth is the soil. The soil serves as the foundation

of a plant wherein the roots of a plant anchors to supply oxygen to the environment and

deliver enough water and minerals. However, the soil has different pH levels that affects

the plants. Soils that are highly acidic or basic results to a poor soil. Given this, the best

soil to grow a plant is said to be at pH level between 6.5 to 7.0.

Therefore, the pH level of the soil affects the growth of the plants. Thus, the pH

level of the soil may affect the stomatal density of leaves. This study aimed to determine

the effect of the pH level of the soil on the stomatal density of the leaves of Tradescantia

spathacea. The main objective is to determine the effect of soil pH on the stomatal density

of Tradescantia spathacea.

The experiment was conducted by group four of 3L on November 20, 2019 during

BIO 11.1 class at the Institute of Biological Sciences Laboratory, room C-117, University

of the Philippines Los Baños, Laguna, Philippines administered by Professor Kristine

Abenis.

MATERIALS AND METHODS

Buds of Tradescantia spathacea plant of the same age were planted on nine pots

with equal amount of loam soil mixed with sand and pebbles. There were three treatments

in the experiment, first is the soil watered with tap water which served as the negative

control from the set-ups. Then the soil watered with vinegar solution which was obtained

by mixing two tablespoons of Silver Swan vinegar with two liters of tap water. It was the

same for the third treatment which is the soil watered with baking soda solution which was
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obtained through mixing two tablespoons of baking soda with two liters of tap water. Every

treatment had three replicates to ensure validity of the obtained results. The plants were

watered 20 mL twice a day of the corresponding treatments prepared (soil watered with tap

water, vinegar solution and baking soda solution) for a span of four days. Afterwards,

leaves, with the same size and location on the plant, on each replicates of every treatments

were collected. Followed by the imprinting method, both sides of a sample leaf were

painted with a clear nail polish to determine whether the adaxial or abaxial side has a

greater number of stomates.

After determining that the abaxial side of leaves has a comparative number of

stomates, leaves of each treatment were painted with clear nail polish on an area in abaxial

side that contains few veins and away from the midrib. The leaves were set aside and

allowed the nail polish to dry completely. After a few minutes, a transparent tape was used

to remove the nail polish from the surface of the leaves. Then, the tapes with the nail polish
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were placed on a clean glass slide and was properly labeled ready to be observed on the

microscope under HPO.

Thereafter, imprints of stomates and epidermal cells were counted, then the

stomatal density of the samples was computed with the use of the formula: 𝑆. 𝐷. =

𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑠𝑡𝑜𝑚𝑎𝑡𝑒𝑠
. The pH level of the treatments used in watering was
𝑡𝑜𝑡𝑎𝑙 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑒𝑝𝑖𝑑𝑒𝑟𝑚𝑎𝑙 𝑐𝑒𝑙𝑙𝑠

measured using the digital pH meter to distinguish whether the treatments were acidic,

neutral or basic. Also, soils with the treatments were diluted with the same amount of dH2O

in a petri dish to measure the pH level of the soils.

RESULTS AND DISCUSSION

Stomata are crucial for a plant’s life function for it allows carbon dioxide gas that

contains carbon to enter the plant tissues. The carbon atoms that enters are used by plants

to create sugars, proteins, and other materials essential for life. Without the carbon dioxide

reaching the plant tissues, plants could not create organic molecules which is needed to

survive and grow. Most plants open their stomata at day to maximize carbon dioxide intake

to harvest energy from the sunlight. But there are plants that are mostly found in desert

environments that open their stomata at night which is cooler. In this way, they lose less

water rather than opening their stomata at the scorching desert sun ad they would

eventually store the carbon dioxide and use it for photosynthesis when daylight appears

(Biology Dictionary, 2017).

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Table 1. Number of stomates and epidermal cells on each replicate of the three treatments.

REPLICATE 1 REPLICATE 2 REPLICATE 3

TREATMENTS number number of number of number of
(soil watered of epidermal number of epidermal number of epidermal
with) stomates cells stomates cells stomates cells
TAP WATER 4 15 3 11 3 11
VINEGAR
SOLUTION 3 14 4 15 2 10
BAKING
SODA
SOLUTION 3 17 2 9 2 9

There were three treatments, each had three replicates. On the first treatment which

is the soil watered with tap water, results showed that on the first replicate there were four

stomates and 15 epidermal cells found while the other two replicates had three stomates

and 11 epidermal cells. The second treatment is the soil watered with a vinegar solution, it

has three stomates and 14 epidermal cells on the first replicate, four stomates and 15

epidermal cells on the second replicate and two stomates and 10 epidermal cells were on

the third replicate. Lastly, for the soil watered with baking soda solution, there were three

stomates and 17 epidermal cells on the first replicate, two stomates and nine epidermal

cells on both second and third replicates.

Table 2. Computed stomatal density of the three replicates of each treatments.

STOMATAL DENSITY
TREATMENTS
(soil watered with) REPLICATE 1 REPLICATE 2 REPLICATE 3
TAP WATER 0.266666666 0.272727272 0.277272727
VINEGAR
SOLUTION 0.214285714 0.266666666 0.2
BAKING SODA
SOLUTION 0.176470588 0.222222222 0.222222222
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The first treatment which is the soil watered with tap water had a stomatal density

of 0.266666666 for the first replicate and 0.277272727 for the other replicates that clearly

shows how close the results were. On the other hand, the soil watered with a vinegar

solution had a stomatal density of 0.214285714 for the first replicate, 0.266666666 for the

second replicate and 0.2 for the third replicate which shows that the data gathered were not

corelated. While on the soil watered with baking soda solution had a stomatal density of

0.176470588 or the first replicate and 0.222222222 for the other replicates which shows

how close the obtained results compared to the second treatment.

Table 3. Average Stomatal density of the treatments.

Treatments Tap water Vinegar solution Baking Soda solution

Average Stomatal Density 0.27 0.23 0.21

The average stomatal density for the soil watered with tap water is 0.27 while for

the soil watered with vinegar solution is 0.23 and for the soil watered with baking soda

solution is 0.21. The data gathered clearly shows that the tap water treatment has the highest

stomatal density followed by the vinegar solution treatment and lastly the baking soda

solution treatment.
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Table 4. Soil pH of each treatment.

Soil Treatment

Tap water Vinegar solution Baking soda solution

Soil pH 6.1 5.9 7.3

The measured soil pH of the soil watered with tap water is 6.1 which is slightly

acidic and according to Horiba (2015) soils with pH level of 6.0-7.5 is acceptable to plants

for their growth. On the other hand, the soil watered with vinegar solution is 5.9 which is

in the range of pH level 5.5- 6.0 that reduces soil microbial activity (Horiba, 2015). Soil

microbial activity may influence global warming because the soil can either be a source or

acceptor of CO2 (Maier, 2015). Thus, according to Maclachlan and Defeo (2018), the

microbial activity is a steep chemical gradients resulted from anaerobic decomposition of

organic matter following the catabolic pathway using molecular oxygen, nitrate or nitrite,

manganese, iron, to sulfate reduction and methanogenesis which plays an important role in

the environment . While, the soil watered with baking soda solution has a pH of 7.3 which

is alkaline and iron availability in this pH level becomes a problem.

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0.3
0.25

Average Stomatal Density 0.2

0.15
0.1
0.05
0
Tap water Vinegar solution Baking Soda solution

Treatments

Figure 4. Bar graph showing the comparison between the stomatal density of the

treatments.

The graph shows the difference between the stomatal density of the treatments. The

tap water has the highest average stomatal density followed by vinegar solution and the

smallest stomatal density is the baking soda solution.

The pH of the soil is measured in order to know whether the soil is acidic, alkaline

or neutral. A pH scale of 7. Is considered neutral. Below 7.0 is acidic and above 7.0 is

alkaline or basic. Highly acidic soil is more toxic due to the availability of minerals such

as aluminum and manganese while the minerals such as calcium, phosphorus and

magnesium are less available to the plant. In highly alkaline soil, lesser availability of

phosphorus and other micronutrients. The pH of the soil greatly affects the nutrients

available for plant growth and processes such as transpiration and respiration (Horiba,

2015). Also, according to Desai (1937), loss of water and gaseous exchange is an effect

caused by stomatal behavior that remarkably influence plant behavior and there are factors
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that affects the behavior of stomates. The importance of the mineral nutrient supply is

considered significant in plant growth and processes (transpiration and respiration).

SUMMARY AND CONCLUSION

The physical traits of leaves vary across environmental gradients, influence leaf

functions. Stomata influences leaf function by its changes in density and size that causes

potential water loss, carbon dioxide uptake and leaf cooling (Hill et al., 2014).

Buds of Tradescantia spathacea plant of the same age were planted on nine pots

with equal amount of loam soil mixed with sand and pebbles. The treatments were soil

watered with tap water, soil watered with vinegar solution and soil watered with baking

soda solution each had three replicates to ensure validity of the obtained results. The plants

were watered 20 mL twice a day of the corresponding treatments prepared for a span of

four days. Leaves with the same size and location on the plants were collected. Imprinting

method was used to determine the number of stomates and epidermal cells in order to

compute for the stomatal density of treatments. The samples were observed on the

microscope under HPO. Also, pH level of the treatments used in watering and the soil

were measured using the digital pH meter to distinguish whether the treatments were

acidic, neutral or basic.

The experiment proved that the pH level of the soil affects the stomatal density of

the leaves of Tradescantia spathacea such that the different treatments used varied from

each other. The results were obtained using imprinting method. The results showed that

soil watered with tap water has greater stomatal density of 0.27 with a pH level of 6.1
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followed by soil watered with vinegar solution having a stomatal density of 0.23 with a pH

level of 5.9 and soil watered with baking soda solution has a stomatal density of 0.21 with

pH level of 7.3. Even though tap water had the greatest stomatal density, it was proven

that the pH levels of both vinegar solution and baking soda solution are not harmful to the

plants since according to Horiba (2015), only highly alkaline and acidic soil can harm the

plant due to the lesser availability of certain mineral nutrients which is essential for the

stomates. Due this, the pH level of soil affects the plant growth and plant growth varies

with the nutrients present in a plant which can affect the stomatal density of a leaf.

It is recommended for future researchers to try other landscape plants and other materials

to alter the soil’s pH in order to compare the stomatal density of other plants.

LITERATURE CITED

Biology Dictionary. (24 June, 2017). Stoma. Retrieved from

https://biologydictionary.net/stoma/?fbclid=IwAR0J7SIDMEdnY-

eprjYEtgu0U1cGFpjZRMurKwZ1rpBqAflmigyhKf9SjMw. Accessed December 3, 2019.

Global Invasive Species Database (2019) Species profile: Tradescantia spathacea. Downloaded

from http://www.iucngisd.org/gisd/species.php?sc=493 on 28-11-2019.

Hill, K. E., Gruerin, G. R., Hill, R. S., & Watling, J. R. (2015). Temperature influences

stomatal density and maximum potential water loss through stomata of Dodonaea

viscosa subsp. angustissima along a latitude gradient in southern Australia.

Australian Journal of Botany, 657–665. doi: https://doi.org/10.1071/BT14204

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Horiba. (22 September, 2015). Soil pH and Nutrient Availability. Retrieved from

https://www.horiba.com/en_en/applications/food-and-beverage/agriculture-crop-

science/soil-ph-and-nutrient-

availability/?fbclid=IwAR1aBffcFhqVRn8oaau7E0Foo_HsRyDTXx22ua14hhvD

LsqPCoMUDhotD84. Accessed December 4, 2019.

Maclachlan, A. and Defeo, O. (2018). The Interstitial Environment. Academic Press. The Ecology

of Sandy Shores (Third Edition). pp. 37-62. Retrieved from

https://www.sciencedirect.com/science/article/pii/B9780128094679000035?via%3Dihub.

Accessed December 4, 2019.

Maier, R. (2015). Biogeochemical Cycling. Environmental Microbiology (Third Edition).

Retrieved from https://www.sciencedirect.com/topics/earth-and-planetary-sciences/soil-

microbial-activity. Accessed December 4, 2019.

Sarma, S. (2017). Importance of plants in our life. Science India. Retrieved from

http://www.scienceindia.in/home/view_article/59. Accessed December 3, 2019.