Responses of Selected Fungal Communities in Soil under Plant-based Fertilizer

with Calamansi (Citrus microcarpa) and Lime from Freshwater Perna viridis

(1) PROJECT PROFILE

Project Title: Responses of Selected Fungal Communities in Soil under Plant-

based Fertilizer with Calamansi (Citrus microcarpa) and Lime from Freshwater

Perna viridis

(2) STATEMENT OF THE PROBLEM

This study is to present the plant-based fertilizer with calamansi from

freshwater Perna viridis, the use of this plant-based fertilizer is to promote the

growth of beneficial microorganisms in the soil. It seeks to answer the

following questions:

1.) How does the plant-based fertilizer affect the following:

a. Fungal Count

i.Verticillium dalhiae

ii.Phytophthora

iii.Rhizoctonia

iv.Pythium

b. Microbial Interaction

c. Physical Properties of the Soil (color, texture, structure, porosity, density,

consistence, aggregate stability, and temperature)

(4) HYPOTHESIS

Alternative hypothesis – The use of plant-based fertilizer with calamansi

(Citrus microcarpa) from freshwater Perna viridis significantly alters the

population count of selected fungal communities in soil compared to the

control group without the application of the fertilizer.

Null hypothesis - The use of plant-based fertilizer with calamansi (Citrus

microcarpa) from freshwater Perna viridis does not significantly alter the

population count of selected fungal communities in soil compared to the

control group without the application of the fertilizer.

(3) EXECUTIVE SUMMARY

Agriculture is one of the major issues our country is facing at this

moment.Fertilizers are chemical or organic substances that provide essential

nutrients to plants, such as nitrogen, phosphorus, and potassium. The use of

organic fertilizers, like plant-based fertilizers, is becoming a popular alternative

to chemical fertilizers, which have negative impacts on the environment and

human health. In the Philippines, agriculture is a significant sector of the

economy, but many farmers struggle to earn a sustainable income due to a

range of challenges, including low market demand and access to buyers. As a

result, many harvested crops are left unused or go to waste, leading to

economic losses and environmental degradation . In this study, the researchers

investigate the responses of selected fungal communities to plant fertilizers ,

including Calamansi (Citrus microcarpa) from freshwater Perna virdis. Taken

from the primary study, the researchers have used Lemons along with fresh
from the primary study, the researchers have used Lemons along with fresh

water bivalves as their components. This study will be making use of an item

more accessible to many which is Calamansi and Fresh water Perna Virdis.

Citrus fruits, like calamansi, are rich in essential nutrients and organic matter,

making them valuable natural fertilizers for promoting healthy plant growth.

The study seeks to compare the outcomes of using this plant-based fertilizer

with a normal fertilizer and to determine the impacts of the fertilizer on plant

growth and nutrient absorption. Taking this into account, results suggests the

potential of Calamansi (Citrus microcarpa) from the freshwater Perna virdis as

an alternative organic fertilizer to improve soil fertility and promote sustainable

agriculture.

(5) INTRODUCTION

Fertilizers are substances that supply plants with essential nourishments,

such as phosphorus, nitrogen and potassium, that are crucial for their growth

and development. These nutrients are frequently lacking in soil, and without

them, plants cannot grow or produce an optimal yield. This is why fertilizers

are crucial for modern agriculture, as farmers must produce enough food to

feed a growing global population. Although, their impact on plant health and

quality should also be considered to ensure that we are not only producing

enough food, but also producing healthy, high-quality crops in a sustainable

way.

Synthetic or chemical fertilizer is another type of fertilizer which is

made from refined organic or more often artificial matter, where nutrients have

been extracted and combined with chemical fillers to create a synthetic

solution. Chemical fertilizers, according to Bisht and Chauhan (2021),

solution. Chemical fertilizers, according to Bisht and Chauhan (2021), are

employed in excessive and disproportionate amounts to increase agricultural

yields in order to resist specific situations.

Chemical fertilizers, according to Bisht and Chauhan (2021), are

employed in excessive and disproportionate amounts to increase agricultural

yields in order to resist specific situations. This may be because of numerous

reasons such as its quicker results and less expensive price compared to organic

fertilizer which makes it easier for anyone to afford and provide for their crops.

However, excessive use of chemical fertilizers has led to several issues such as

serious soil degradation, nitrogen leaching, soil compaction, reduction in soil

organic matter, and loss of soil carbon. In addition, the efficacy of chemical

fertilizers on crop yield has been decreasing over time (Lin et al., 2019). To

decrease and remove the negative impacts of synthetic fertilizers on human

health and the environment, new farming approaches, including all so-called

organic agriculture, have been created (Sharma & Chetani, 2020). This includes

the creation of organic fertilizers which can either be plant-based, animal-based

or nitrogen-based.

Plant fertilizers made from organic matter, or what we call organic fertilizers,

like animal manure, compost, and plant residues are seen as a common

alternative to chemical fertilizers. They slowly release essential nutrients to

plants. Plant-based fertilizers also promote the growth of beneficial

microorganisms in the soil.

Many studies have been conducted to prove that citrus fruits can be used as

organic fertilizers for plants. Citrus fruits such as calamansi (Citrus microcarpa)

contain high levels of essential nutrients such as nitrogen, phosphorus, and

potassium, which are necessary for healthy plant growth. Nitrogen is essential

for the production of chlorophyll, which is necessary for photosynthesis, the

process by which plants convert sunlight into energy. Phosphorus is necessary

for root development, and it plays a crucial role in the transfer of energy within

the plant.

Potassium helps to regulate water balance within the plant and is

important for the production of strong stems and leaves. In addition to these

essential nutrients, citrus fruits also contain other beneficial nutrients such as

calcium, magnesium, and sulfur, which can help improve soil fertility and

support plant growth. It also contains nutrients such as vitamins A, E, and B

(thiamine, riboflavin, and niacin), minerals, and antioxidants such as

flavonoids, vitamin C, phenolic compounds, and carotenoids. Overall, the high

nutrient content and organic matter found in citrus fruits make them a valuable

natural fertilizer for promoting healthy plant growth, making calamansi a

potential fertilizer for plants ( Uthman, A. & Garba, Y., 2023).

Green mussel, which is also known as Perna Viridis, can also be used as

fertilizer due to its high nutrient content. Mussels are filter feeders that absorb

nutrients and minerals from the water in which they live, including nitrogen,

phosphorus, and calcium. As a result, the shells and meat of the mussels are

rich in these nutrients, making them a valuable source of organic fertilizer. This

information is also supported by a study that investigated the use of green

mussel shells as fertilizer for rice fields. According to Chandran et al. (2011),

the shells of green mussels release

the shells of green mussels release nutrients such as nitrogen, phosphorus, and

calcium into the soil and improve soil fertility and crop growth. Thus, green

mussels, or Perna viridis, can also be a good source of organic fertilizer for

plants.

The application of organic fertilizer shifts the composition and

abundance of fungal communities, and may increase fungal diversity (Semenov

et. al., 2022). Fungal communities refer to the populations of fungi that exist

within a particular ecological environment. Fungal communities that exist in

forest soils are extremely complex and diverse, with different species having

different functions such as decomposing dead organic matter, living within

plant tissues, causing diseases, and forming beneficial relationships with plant

roots. These fungal communities are distributed differently over time and

space. These also play important roles in plant community dynamics,

subsurface trophic interactions, and biogeochemical cycles in forest ecosystems

(Izquierdo et al., 2021).

Assessing fungal biodiversity in soil can provide insights into their

functions in affecting soil quality and plant health (Frąc et al., 2015). Different

types of organic manure can influence soil health indirectly through

physicochemical characteristics and directly by affecting soil fungal

communities.
(6) RATIONALE

Bisht and Chauhan (2021) stated that chemical fertilizers have been

used excessively by many farmers and agriculturists to raise more crops

quickly. However, excessive use of chemical fertilizers also poses negative

effects on our environment. Chemical fertilizers are more resistant to the

environment, reduce soil fertility, and actually cause a lot of degradation of soil

and land (Liu et al., 2009). Given this, there is still a need to seek out more

studies about the creation of organic fertilizers to reduce and eliminate the

negative impacts of these chemical fertilizers on our environment.

Fruit peel residues are part of the waste that accumulates in large

quantities every day. This is a serious problem that needs to be addressed in

order to keep the environment free from pollution. Fruit peels are very rich in

macro and micro elements that have a beneficial effect on plant growth. By

using calamansi peeling as fertilizer, we can reduce waste and get more

benefits than inorganic fertilizers. Organic fertilizers decompose relatively

quickly and provide the lawn and garden with a wide range of nutrients, which

promote plant growth and soil regeneration.

The pH of the maximum growth rate is called the optimal growth pH.

Based on the optimal pH for growth, microbes can be divided into three groups:

acidophiles grow best at pH <; 5: Neutrophils grow optimally between pH 5

and 9 and alkaliphiles grow fastest above pH 9 (Horikoshi, 1999; Baker-Austin

and Dopson, 2007). pH can also affect microbial metabolism and hence

microbial community structures by modulating the thermodynamics and

kinetics of redox reactions. Microbial respiration catalyzes redox reactions to

kinetics of redox reactions. Microbial respiration catalyzes redox reactions to

synthesize ATP.

In this study, the researchers will investigate the responses of

selected soil fungal communities to plant-based fertilizers, including Calamansi

(Citrus microcarpa) from the freshwater Perna viridis, which is a citrus fruit

widely used in the Philippines for its medicinal and culinary properties.

Freshwater Perna viridis, also known as the Asian green clam, is an

economically important mollusk that is abundant in Philippine waters.

By conducting this study, the researchers will be able to

determine whether calamansi (Citrus microcarpa) from the freshwater Perna

Viridis may be used as a plant-based fertilizer. This will also assist the

researchers in controlling fertilization and regulating soil microorganisms. The

purpose of this study is to evaluate the effects of plant fertilizers containing

Calamansi from freshwater Perna viridis on the abundance and diversity of

selected fungal communities in soil.

The results of this study may provide insight into the potential of

Calamansi (Citrus microcarpa) from the freshwater Perna Viridis as an

alternative organic fertilizer to improve soil fertility and promote sustainable

agriculture. It may also mitigate or solve the negative impacts of chemical

fertilizers in our environment.

By conducting this study, the researchers will be able to determine

whether calamansi (Citrus microcarpa) from the freshwater Perna Viridis may

be used as a plant-based fertilizer. This will also assist the researchers in

controlling fertilization and reducing soil microorganisms. The purpose of this

controlling fertilization and reducing soil microorganisms. The purpose of this

study is to evaluate the effects of plant fertilizers containing Calamansi from

freshwater Perna viridis on the abundance and diversity of selected fungal

communities in soil.

The results of this study may provide insight into the potential of

Calamansi (Citrus microcarpa) from the freshwater Perna Viridis as an

alternative organic fertilizer to improve soil fertility and promote sustainable

agriculture. It may also mitigate or solve the negative impacts of chemical

fertilizers in our environment.

(7) SCIENTIFIC BASIS INVOLVED

According to the research of Ryan Christian Balce, fertile soil was an

essential requirement for mass agricultural production. In the statement of

frontiers, plants are factors of our living, they provide us oxygen and food. It is

said that in their ecosystem, plants interact or co-exist with various types of

bacteria.

Numerous plants have been detrimental anticipated by certain bacterias

spreading throughout its body. Given this finding, the researchers have

established a proposal to lessen certain bacterias to travel inside the plants. Soil

and substrates were thoroughly mixed until an average homogeneous mixture

was obtained. The samples were then subjected to maintenance of the reference

temperature in the laboratory.

In a 2022 study by Semenov MV, Krasnov GS, and Van Bruggen A.,

fungi represent a diverse group of organisms that play key roles in maintaining
fungi represent a diverse group of organisms that play key roles in maintaining

soil health and ecosystem function. Long-term use of fertilizer provides the soil

with nutrients that can offset the effects of plants on fungal communities.

Organic fertilizers significantly increased microbial biomass and fungal

abundance and suppressed fungal pathogens from plants. This shows that

organic fertilizers can then be used to manipulate fungal communities in soil.

In addition, study says that some bacteria's can be harmful to plants. For

example, cause them to rot or possibly make them sick. Certain bacteria's

cause harm to a plants growth and health. The aimed results of the researchers

are to lessen the spread of bacteria's and help with the growth of plants all over

our surroundings.

(8) REVIEW OF RELATED LITERATURE

A. CITRUS FRUITS

Calamansi peels have been shown to have a beneficial effect on plant

growth, as it contains nutrients such as nitrogen, phosphorus, and potassium.

One of the available nutrients, phosphorus, is an essential macronutrient for

plant growth, development, and production. It plays an important role in signal

transduction and photosynthesis in plants and has a decisive impact on stress

resistance, plant dependency on vesicular-arbuscular mycohrrhizal for P uptake

in order to produce its maximum growth, yield and quality of crop.

Leghari, S. (2016) also stated that nitrogen is a nutrient which greatly

contributes in agriculture and is considered the most imperative element for

proper growth and development of plants which significantly increases and

proper growth and development of plants which significantly increases and

enhances the yield and its quality by playing a vital role in biochemical and

physiological functions of plant.

According to the study of Xin, YH., et al., evaluation of the phenotypic

diversity of Calamansi (Citrus microcarpa) germplasm in Hainan Island In Sci

Rep 12, 371 (2022), calamansi (Citrus microcarpa), or Philippine lime, is an

important local economic crop in Hainan, China. It originated in Southeast

Asia, mainly growing in Southeast Asia and tropical regions of China, and it

has a long history of cultivation on Hainan Island.

Calamansi fruit is rich in vitamins C, aromatic oils, carotenoids, and

other natural substances that have lots of health benefits for humans, such as

beneficial effects for human eyes, being good for treating cough, asthma, high

blood pressure, preventing arteriosclerosis, etc. However, the commercially

cultivated Calamansi were mostly seedling trees, and their genetic diversity and

improvement had not been studied, which caused a series of problems such as

no stable commodity supply period and uneven fruit quality. Hainan Island is

the main growing area of Calamansi in China. The investigations and

evaluations of the germplasm of Calamansi on Hainan Island hold great

significance for Calamansi genetic improvement and fruit quality.

According to the study of Schipani, S. (2019), citrus peels can be used

to add acidity to the soil and provide a nutrient boost for plants that thrive in

acidic conditions, such as nasturtium, blueberries, and radishes. By grinding

dry peels into a powder and sprinkling it over these acid-loving plants, you can

enhance their growth. Additionally, contrary to the myth that citrus rinds deter
enhance their growth. Additionally, contrary to the myth that citrus rinds deter

beneficial insects in compost, they can actually help keep scavenging rats and

birds away from the compost pile. To facilitate quicker decomposition, tear the

citrus peels into smaller pieces before adding them to the compost.

The natural environment relies heavily on the pH of the soil, which has

an impact on numerous biogeochemical processes in the soil. According to the

study of Neiana, D. (2019), it is viewed as the “master soil variable” because

of its broad effect on soil properties and cycles that influence plant

development and biomass yield. The effects of soil pH on substance

availability, mobility, and biological processes in the soil are the subject of this

abstract. It also investigates the potential applications of soil pH in nutrient

cycling, plant nutrition, and soil remediation (both bioremediation and

physicochemical remediation). Additionally, it investigates how living

organisms regulate soil pH. In general, this paper emphasizes the significance

of soil pH in managing and comprehending soil biogeochemistry 

        The effect of pH on phosphate adsorption varied between soils. In

unfertilized soils, adsorption decreases as the pH increases to approximately

5.5. A further increase in pH resulted in a further decrease in adsorption in one

soil and an increase in adsorption in the other three soils. For fertile soils, the

measured adsorption amount increased with pH. When sodium chloride was

used instead of calcium chloride, the trend of decreasing adsorption with

increasing pH was more pronounced. 

Differences between soils can be attributed to differences in their

properties. One of them was the rate of decrease of the adsorption surface
properties. One of them was the rate of decrease of the adsorption surface

electrostatic potential with increasing pH. Only small differences in the rate of

change of potential were required to reproduce the differences observed

between soils. The electrostatic potential decreases faster in sodium salt

solutions than in calcium salt solutions, which explains the differences

observed between these media. Another soil property affecting the observed

sorption was the release of phosphate from the soil. The amount released was

the greatest.

Acidity does not directly limit plant growth, but it negatively impacts

biological processes that are beneficial for plants. It decreases the availability

of important nutrients like phosphorus and molybdenum while increasing the

availability of toxic elements like aluminum and manganese. It also harms soil

organisms and can inhibit the survival of useful bacteria like rhizobia.

Additionally, acidity can cause essential plant nutrients to leach below the

rooting zone. (Agriculture Victoria, 2020)

B. Perna Viridis

According to the study of Álvarez, E. et al. (2012), large

quantities of mussel shells (66 000–94 000 t year−1), an alkaline material that

can be used as a soil amendment, are generated as waste in Galicia, NW Spain.

A field trial was carried out by planting different pasture species in a Haplic

Umbrisol using a randomized block design with four blocks and six treatments

(not amended control or soil amended with lime, finely ground shell, coarsely

ground shell, finely ground calcined shell, or coarsely ground calcined shell) to

compare the effects of lime and mussel shell additions on a soil with a low
compare the effects of lime and mussel shell additions on a soil with a low

cation exchange capacity and high Al saturation. The trial was established in

March 2007, and samples of plants and soil were collected when plots were

harvested in summer 2008 (separating bulk and rhizosphere soil). The soils

were analyzed for pH, total C, total N, available P, exchangeable cations,

effective cation exchange capacity, and available micronutrients. Dry matter

yield was measured in all plots, and plants were analyzed for nutrients.

Application of mussel shells and commercial lime resulted in an increase in pH

and exchangeable Ca and a decrease in exchangeable Al and Al saturation. The

stability of pH over time was high. These effects were most noticeable in the

rhizosphere. The amendment also had a positive effect on dry matter yield and

the concentration of Ca in the plant.

C. Plant-Based Fertilizer

According to the study of Ma, X., et al. (2022), Fertilization is an

important part of citrus crop management. However, limited details are

available about the fertilization approach to citrus plant development. In this

study, a pot experiment for fertilization approaches and fertigation levels was

conducted. Four fertilization approaches, namely, drip fertigation (DF),

broadcast fertilization (CK), hole fertilization (HF), and pour fertilization (PF),

were tested. The fertigation level treatment included 100% (DF-337.5), 80%

(DF-270), 60% (DF-202.5), and 40% (DF-135) fertilizer supply with DF, and

the 100% fertilizer supply with broadcast fertilization served as control (CK).

The results showed that DF not only increased the absorptions of nitrogen (N),

phosphorus (P), and potassium (K) but also promoted citrus plant height, stem

diameter, and dry weight. In terms of fruit quality, DF had the highest fruit
fruit total soluble solid (TSS) and titratable acidity (TA) contents. For fertilizer

loss, DF had the lowest N and K leaching losses of 9.26% and 4.05%,

respectively, and the lowest N and K runoff losses among the approaches.

Isotopic tracing with 15N indicated that DF had the highest fertilizer use

efficiency. Based on the analysis of fertigation levels, a DF approach with a

60% fertilizer reduction could improve citrus plant development. Therefore,

DF promoted citrus plant growth and fruit quality by accelerating fertilizer

utilization and impairing fertilizer loss. The fertilizer amount in citrus

production could be reduced significantly using DF.

D. Microbial interaction

According to the study of Microbiol B. (2016). These interactions involve

changes in the environment, exchange and conversion of metabolites, genetic

exchange, and signaling. Species diversity plays a role in the establishment of

microorganisms in an environment, and these associations result from a co-

evolution process that allows for adaptation and specialization. Molecular

mechanisms such as secondary metabolites, quorum sensing, and gene

expression are involved in these interactions. The article focuses on molecular

mechanisms involved in microbial interactions in the environment and not just

in microbial-host interactions.

The application of fertilization resulted in a notable increase in the

dominant fungi phylum Ascomycota, along with changes in fungal diversity

and richness in the soil surrounding cassava roots (rhizospheric soil). Different

fertilization methods also influenced the structure of the fungal community in

the rhizospheric soil of cassava. These alterations led to an increase in

the rhizospheric soil of cassava. These alterations led to an increase in

potentially beneficial fungi and a decrease in potentially harmful fungi,

creating a favorable environment for cassava growth and enhancing both its

yield and quality. Moreover, the availability of phosphorus (P), potassium (K),

and organic matter (OM) exhibited the greatest influence on the microbial

community. There was a positive correlation observed between soil nutrition

and the structure of the fungal community following fertilization. Therefore, it

is crucial to prioritize the maintenance of a stable micro-ecology in cassava’s

rhizospheric soil during actual production. In the future, the use of specific

microbial fertilizers for cassava planting and production may emerge as a new

trend.(Cai, J.et al., 2021)

E. Selected Fungal Communities

Due to their ability to produce a wide variety of extracellular

enzymes, they are able to break down all kinds of organic matter, decomposing

soil components and thereby regulating the balance of carbon and nutrients

Zifčáková et al., 2016.

The diversity and activity of fungi is regulated by various biotic (plants

and other organisms) and abiotic (soil pH, moisture, salinity, structure, and

temperature) factors (López-Bucio et al., 2015; Rouphael et al., 2015). Fungi

can be found in almost every environment and can live in wide range of pH

and temperature (Frac et al., 2015).

According to Semenov M. (2022), fungi are a varied collection of

organisms that play an important role in soil health and ecosystem functioning.
Fungi play a significant role in the decomposition and recycling of refractory or

labile organic compounds. Although they frequently form symbiotic

relationships with plant roots, they can also be soil-borne plant diseases.

Verticillium wilt, one of the soil-borne fungal diseases in plants, has

always been a serious disease of many economically important agricultural and

horticultural crops. It affects approximately 400 plant species, including trees

and shrubs, vines, flowers, and vegetables (Song H. 2021). Verticillium dahliae

and Verticillium albo-atrum are the two forms of soil-borne fungus. These

fungi dwell in the plant xylem, a network of small tubes that carry water and

nutrients throughout the plant. They effectively cause a clog in the plant’s

vascular system, preventing it from receiving adequate water. They enter

damaged plant tissue through the roots and multiply. As a result, some diseased

plants may wilt. Toxins produced by Verticillium are also harmful to host

plants.

However, there are several methods for preventing or managing this

condition. If verticillium is present in the soil, start by picking plant varieties

that have been developed to be resistant or immune to verticillium wilt.

Furthermore, keeping healthy plants and using the right fertilizer can prevent or

stop the fungi from spreading. Additionally, in a study by Fisher S. in 2022,

Verticillium wilt can be characterized by these symptoms:

a) Yellowing, withering, or shriveling leaves appear first on older

growth, branches dieback.   

b) Fruit crop output is lowered due to stunted or deformed growth. 

 c) In hot weather, sudden wilting, occasionally followed by rebound

when circumstances cool and damp.   

d) Cutting through an infected stem or branch exposes black tissue

discoloration.

In addition to this, high N dosages, N/K imbalances, and frequent

watering are all known to encourage Verticillium wilt. The effect of fertilizer

and its relationship with irrigation frequency on the development of

Verticillium wilt of olive (VWO) has been studied. A split-split-plot design was

developed in microplots with two naturally infected soils of varied texture to

examine three fertilization treatments (NO3Ca, NPK, and no fertilization).

Regardless of soil texture, the final disease incidence in plants treated to

NO3Ca daily treatment was (100%), N-P-K-daily treatment (61.1%) and no

fertilization (9.6%). Hence treatment 1 did the best out of the 3 (Perez-

Rodriguez, 2022).

As stated by Semenov, M. V. et al., (2022), they found that the type of

fertilizer led to distinct fungal communities in the rhizosphere, overriding the

effects of plant species. Organic fertilizers increased the abundance of certain

fungal groups, while mineral fertilizers increased others. Organic fertilizers

also increased fungal abundance but decreased diversity. The proportion of

various kinds of fungal phytopathogens increased, but there was only a minimal

impact on diversity from mineral fertilizers. The study suggests that long-term

fertilization can influence the abundance and diversity of rhizosphere fungal

communities, and fertilization management could be used to manipulate these

communities and control soilborne pathogens.

(9) METHODOLOGY

This chapter discusses methods and procedures used in the study. The

focus of this research study is to determine the responses of selected fungal

communities in soil under plant-based fertilizer with calamansi (citrus

microcarpa) from freshwater perna viridis. The following information provides

details on the research design, research informants, research tools, source of

information, data gathering procedure, and statistical analysis the researchers

will use in the study.

A. Research Design

Experimental design of research will be used in this study. The

experimental design is used to study the interactions between independent and

dependent variables, which can be used to determine cause and effect. The

purpose of an experimental research design is to determine the effect that a

factor or independent variable has on the dependent variable. This type of

research design aims to help readers understand how the data was obtained and

thus help them to properly analyze the results. For this reason, this type of

research is the most appropriate for this study since this involves an assessment

that is suitable for the given condition.

B. Research Locale

This study will be conducted at Vinzons Pilot High School. Vinzons

Pilot High School will be the site of this investigation. Due to its ideal climatic

conditions and importance in supporting various fields of agriculture, this

location was chosen. Warm temperatures and abundant rainfall are

location was chosen. Warm temperatures and abundant rainfall are

characteristics of Vinzon's tropical climate, which creates an environment that

is favorable to soil fertility. The fact that crops are grown in Region V suggests

a wide variety of fungal communities exist there. As a result, Vinzons is the

perfect place to study how particular fungal communities react to plant-based

fertilizers. Vinzons Pilot High School makes for an excellent practical research

site by giving the researchers access to crucial resources like laboratories and

equipment. Potential opportunities for collaborative research and educational

engagement related to these topics are provided by the school's collaboration

with other teachers and student researchers. Collaboration with local farmers,

governmental organizations, and agricultural groups can yield useful insights

and important information.

C. Materials and Instruments

Calamansi

Also known as citrus microcarpa, it is a fruit native to the Philippines.

Citrus microcarpa is rich in vitamin C, essential minerals, and a variety of

antioxidant compounds. It has also been shown to have some benefits for the

immune system, cardiovascular system, gut health, and skin health.

Perna Viridis

The green mussel, or Perna viridis, is a species that is remarkable

enough to be able to colonize artificial marine habitats, reach very high

biomass levels, withstand environmental changes, concentrate a variety of

organic and inorganic environmental pollutants, and invade new geographic

areas.