Year | Section: 12 – STEM B.

Nebres Approved Working Title:

Names: Task / Due:

Leader: Bandiola, Pink Joy D. Research Design, Materials and Setting,
Members: Bacani, Richmaffi B. Experimental Set-up, Analysis of Data
Cajotoc, Joshua M. November 28, 2023
Gabutin, Gino P.
Garcia, Muswenda Veniz D.
PART I. RESEARCH DESIGN

The experiment was carried out by planting a tomato with the utilization of soil induced

with R. solanacearum and has numerous replicates to ensure statistical robustness. To

guarantee that the soil we used was infested by R. solanacearum, it was brought by the

researchers to a testing center to undertake soil testing that indicates its composition, nutrient

content and soil pH. Before being incorporated into the soil contaminated by R. solanacaerum,

crushed eggshells were sanitized to remove any potential pollutants. Treatment comprised

eggshell concentrations that range from 0% (control) to 10% by weight, with 2.5% increments.

These pots of tomato plants are labeled as T1, T2, and T3 to present the differences of each

pot's containment. T1 contains no amount of eggshell powder, T2 has 100 grams of eggshell

powder poured in its soil and T3 has an amount of 200 grams in it. In these pots, the soil was

properly mixed to ensure their consistency. R. solanacaerum was determined by carefully

observing signs such as wilting, yellowing, and necrosis in tomato plants, with information

collected on a regular basis during its growing season (Liewes, 2020).

By analyzing the impact of eggshell amendments on disease incidence, soil pH, and microbial

community composition, statistical analyses were performed using relevant software such as R

or SPSS. Analysis of variance (ANOVA) was used to evaluate whether there are significant

differences between treatment groups, and post-hoc tests was used for pairwise comparison

(Chua, 2019). Correlation analysis was used to investigate the connections amongst eggshell
concentrations and the occurrence of R. solanacaerum. These findings will provide in-depth

comprehension of the quantitative effects of using eggshell as a soil amendment on the soil-

plant system, providing helpful insight into sustainable agriculture practices aimed at

managing R. solanacaerum in tomato crops.

PART II. MATERIALS AND SETTING

To make eggshell powder, researchers began the process by boiling eggshells along with

their membranes in hot water for a duration of 5 to 10 minutes to eliminate pathogens.

Afterwards, researchers allow the eggshells to dry naturally under the air and sunlight for 5

days, followed by additional drying in an oven at 110°C for 15 minutes before finely

powdering them using a blender. The powdered eggshells were then soaked in water (1 liter

for each treatment) overnight. Following this, four holes were made around each plant, and the

water containing the soaked eggshell powder was transferred into these holes. This eggshell

powder was applied twice, 10 to 20 days after transplanting for growing tomato plants.

Moreover, the researchers' innovative method includes incorporating finely crushed eggshells

into the soil of pots, intending to harness the effectiveness of eggshell powder within the

presence of Ralstonia solanacearum.

The pots, each measuring 25 cm (10 inches) in diameter and 20 cm (8 inches) in height,

were prepared a week prior to the random collection of soil from locations prone to bacterial

presence. After the collection of soil samples, they were taken to the Advanced Geotechnical

Engineering Services (AGES) located in East Fairview, Quezon City, on January 22, 2024, to

undergo soil testing for Ralstonia solanacearum. Subsequently, the researchers obtained the
necessary laboratory materials, including a digital light microscope, 3 pipettes, bunsen burner,

and a weighing scale, from the Chemical Laboratory of Batasan Hills National High School to

observe the growth of sample tomato plants.

Figure 1. Sample pot for

the Figure 2. Sample pot for
the
study study

Figure 3. Advanced Geotechnical Figure 4. Google Map View of Advanced

Engineering Services (AGES) Geotechnical Engineering Services (AGES)
 Figure 5. Batasan Hills National Figure 6. Google Map View of Batasan
High School Hills National High School

The digital light microscope served as the primary research instrument for the observation

of the three different tomato plants: Tomato 1, which did not utilize any eggshell powder

treatment; Tomato 2, which was given 100 grams of eggshell powder; and Tomato 3, which

received 200 grams of eggshell powder. The first objective of the experiment was then

accomplished by using a microscope to discern and compare any observable differences in the

occurrence, development, or suppression of Ralstonia solanacearum among the different

tomato plants. Afterwards, soil testing was employed once more to assess the main objective,

which aims to determine the difference in the presence of Ralstonia solanacearum among the

sample tomato plants before and after the experiment.

PART III. EXPERIMENTAL SET-UP

Using an experimental approach, Tomato 1 stands as the control group, serving as the

baseline against which changes are measured. It remains untreated, providing a standard to

compare the effects of the eggshell powder on the other tomatoes. Tomato 2 and Tomato 3, on

the other hand, represent the experimental groups subjected to varying amounts of eggshell

powder, receiving 100 grams and 200 grams, respectively. The study's dependent variables

encompass crucial aspects such as the size of the tomato plants, the number of leaves and

branches, which were meticulously assessed during both the pretest and post-test phases.

Moreover, the water mixed with different amount in grams of eggshell powder was a

deliberate strategy aimed at creating diverse datasets within each experimental setup. This
systematic manipulation allowed for the exploration of potential correlations between the

different concentrations of eggshell powder and the resulting variations observed in the tomato

plant samples, contributing to a more comprehensive understanding of their relationship.

Collection of Materials

In pursuit of a comprehensive and diverse collection of soil samples for the testing of R.

solanacearum, each researcher took to collecting soil samples randomly from

their respective households. Subsequently, the accumulated samples were transported to the

AGES. Moreover, the researchers borrowed multiple pieces of equipment from the laboratory

facility of Batasan Hills National High School, located on IBP Road in Batasan Hills, Quezon

City. Among the borrowed equipment were a digital light microscope, specifically utilized to

examine the soil samples under a low-power objective (LPO) at 10x magnification, a weighing

scale for correctly measuring the mass of the eggshell powder, 3 pipettes, and a set of bunsen

burners for the purpose of boiling eggshells as part of the researchers’ experimental

procedures.

Solanum lycopersicum Treatment

Using the microscope borrowed from the school laboratory of Batasan Hills National

High School, the researchers conducted the pre-test for both the control setup and

experimental set-up of the study. The S. lycopersicum samples underwent initial preparation

involving cutting them into 1x1x1 mm dimensions to ensure they fit within the confines of

glass slides. Light microscopes operate by allowing light to enter the specimen from beneath

and subsequently focusing it for observation by the viewer's eye, as stated by Rodriguez
(2019). This necessity underscores the requirement for the leaf samples to be exceptionally

thin. Ensuring thinness enables light to effectively pass through the sample, facilitating clearer

observations and analysis under the microscope. Afterwards, the eggshell powder was utilized

in T2 (100 grams of eggshell powder) and T3 (200 grams of eggshell powder).

T1. Control Variable T2. Experiment Var. T3. Experiment Var.

Figure 7. Control and Experimental Variables

PART IV. ANALYSIS OF DATA

PART V. PLAGIARISM RATE

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