Corpuz Student ID DTN1454290095 Thesis Title Seed Health Testing of Selected Forest Tree Seeds and Their Control Using Sodium Hypochlorite and Capsicum Extract Soap Supervisors Marceli
INTRODUCTION
Rationale
Seeds are the fertilized and ripened ovules of flowering plants, containing an embryo that can germinate to produce new plants For successful reproduction of healthy plants, seeds must be genetically and physically pure, possess high vigor and germination potential, maintain the right moisture content, and be free from pests and diseases Contamination and seed-borne pathogens can lead to infections that hinder plant growth, resulting in decreased productivity in both quantity and quality.
Seed health testing is essential for assessing the overall condition of seeds, including their germination capacity and disease tolerance This procedure identifies the presence or absence of seed-borne pathogens in a seed lot, ensuring that only healthy seeds are used for planting.
Seed technology ensures the genetic nutritive value, high yield, and overall health of seeds The Seed Health Test (SHT) is a crucial procedure for identifying seed-borne pathogens, pests, and contaminants that may originate from the mother tree during harvesting, drying, and storage These pests, including insects and microorganisms, can significantly reduce plant germination and lead to poor seedling growth or stunting The International Seed Testing Association (ISTA) has established various methods applicable to both agricultural and forestry seed species in tropical and temperate conditions.
This study aims to assess the health status of selected forest tree seed species and to disinfect the seeds by eliminating microorganisms using Sodium hypochlorite and Capsicum extract soap solution This approach is intended to reduce microbial infections, thereby enhancing both the quantity and quality of crop production and the germination capacity of the seeds The use of disinfectants instead of fungicides prioritizes a lesser chemical impact on both users and the environment (Sauer & Burroughs, 1986).
Objectives
To determine the different types of seed pathogen of four (4) selected forestry seed species and their control/ fungus contamination reduction using Sodium hypochlorite and Capsicum Extract Soap Solution
To determine the microorganism associated with four (4) forestry seed species through seed health test/ blotter method
To determine different types of fungal pathogens and/or storage fungi that cause seed rots on or in the test seed species
To determine the effects of seed pre-treatment on the selected four (4) forestry species
To know the level of effectiveness of Sodium Hypochlorite and Capsicum Extract Soap Solution.
Content of Work
Petri dish sterilization in hot air oven at 80 degree Celsius for 2 hours Distilled water sterilization and filter paper sterilization in automatic autoclave at
212 degree Celsius, 120 Pressure per square inch (psi) for one (1) hour
To create a Capsicum Extract Soap Solution, combine 25 pieces of hot chili, which have been sieved through gauze, with one teaspoon of liquid soap and one liter of distilled water The mixture should then be steam sterilized at 212 degrees Celsius and 120 psi for one hour.
Prepare four (4) kinds of forestry seeds that will be tested;
Plant the seed onto Petri dish with three layers of filter paper that can hold 5ml distilled water
Incubate the plated seeds under height (12 inches space between) day light tubes Petri dish for 7- 10 days under alternating light (12 hours) and 12 hours of darkness
Seven days post-incubation, the seeds were initially inspected with the naked eye and then analyzed under a binocular microscope at 10-40x magnification Seeds exhibiting microbial growth were subsequently examined using a compound microscope at magnifications ranging from 100 to 400x, allowing for the identification of certain fungi producing spores, potentially up to the species level.
Other data collected were bacterial contamination and rate of seed germination
Data were tabulated and statistically analyzed
Profile of the Cooperating Agency
The Ecosystem Research and Development Bureau (ERDB) serves as the primary research division of the Department of Environmental and Natural Resources (DENR) in the Philippines Its research and development, along with extension activities, concentrate on five key ecosystems: forests, upland farms, grasslands and degraded areas, coastal zones, and freshwater and urban environments.
Established in June 1987 under Executive Order (EO) 192, the Ecosystems Research and Development Bureau (ERDB) combines the functions of the former Forest Research Institute (FORI) and the National Mangrove Committee Its headquarters is situated at the University of the Philippines Los Baños, within the College of Forestry and Natural Resources (UPLB-CFNR) campus, located approximately 65 kilometers south of Manila in the province of Laguna.
Relevant research, development and extension toward a healthy environment and sustainable natural resources for improved quality of life
The goal is to deliver suitable technology and information through research, development, and extension services aimed at improving the productivity and sustainability of natural resources This initiative also focuses on environmental protection to enhance the quality of life for Filipinos.
Formulate, implements, coordinates and evaluated integrated research, development and extension agenda/ programs on Environment and Natural
Resources (ENR) and other related cross-cutting concern to address the need of the various sector in support of the thrusts and program of the Department
Conduct basic research and develop standard, protocol and science- based technologies to address the need of the different DENR Bureaus, end-user and stakeholder
Provide technical assistance by developing and maintaining network and linkages with DENR Bureaus and office, local, and international institution, other stakeholder and civil society; and
Perform other functions that may be assigned by higher authorities
The article outlines the formulation, implementation, monitoring, and evaluation of action-oriented Environment and Natural Resources Research, Development, and Extension Programs (ENR-RDE-PAPs) These programs focus on various critical areas, including watersheds, water resources, land management, agroforestry, upland farming systems, biodiversity, coastal and wetland ecosystems, ecotourism, mining, degraded areas, forest and timber resources, as well as the management of toxic and hazardous wastes.
Pilot test and verify the standards and protocol developed by ERDB-MO;
Provide the latter feedback towards their improvement;
Conduct coordination and networking services with RDE stakeholder; and
Provide technical assistance and extension services on ENR technologies
1.4.5 ERDB’S Research, Development and Extension Program
Forest Ecosystem Resiliency and Sustainability
Sustainability of Laboratory and Experimental/ Demonstration Service in Support to RDE Program, Activities and Projects of ERDB Main Office and Research Centers
Client- Based Technology Transfer and Extension Service of ENR Management
Addressing Hazard in the Watershed for Water Supply Sufficiency and Resources Availability
Land Management and Sustainability and Agroforestry and Upland Farming System
Enhancing Resilience of Wetlands and Coastal Areas to Support Biodiversity Conservation and Ecotourism Development
Conservation and Sustainable Management of Natural and Plantation Forest
Action Program to Stabilize Mined- Out and Degraded Areas through Rehabilitation Technologies
Pollution Abatement though Integrated Management of Toxic Substances, Hazardous Wastes in Highly Urbanized Areas
Vulnerability Assessment of Priority Watershed
National Greening Program- Production of Quality Planting Materials (NGP- PQPM)
Enhanced- NGP (Bamboo and Mangrove)
An Integrated Science Based Approch Rehabilitation of /mined- out and Wasted Dump areas, in Bagacay, Hinabangan, Western, Samar
Development of Micro- Propagation Protocol for Four Economically Important Bamboo Species in the Philippines
Rubber, Coffee and Cacao: Building Site Matching Function for Improved Farm and Agroforestry Development
Mrs Marcelina V Pacho holds a Bachelor of Science in Agriculture with a major in Microbiology from the University of the Philippines at Los Baños, Laguna She furthered her education at the Graduate School of UP Los Baños, earning a degree in Forestry with a major in Forest Biological Science and a minor in Plant Pathology Additionally, she received specialized training in Seed Pathology at the Institute of Seed Pathology in Hellerup, Denmark, and in Tree Surgery at the College of Forestry and Natural Resources, UP Los Baños.
Description of the Activity
This study focused on assessing the health of four forestry seed species: Lanete, Taluto, Ipil-ipil, and Molave The primary objective was to identify the pathogens, pests, and microorganisms associated with these seeds, as well as to evaluate the effectiveness of two treatments—Sodium hypochlorite and Capsicum Extract Soap Solution—in disinfecting the seeds Under the guidance of the instructor, students were trained to recognize the various organisms linked to the test seeds.
Timeline
The internship training started on March 5 to July 4, 2018; committed to work 8 hours per day and 5 days per week at the ERDB Los Baňos Laguna
Philippines Log book was be used for recording of work time and working days/ hours were completed Field work activities were counted as a working day or
An Official Business (OB) trip is an essential component of a project or order, enabling employees to leave the office and perform necessary tasks outside of their usual work environment.
Table 1: Timeline of Working Days of On-The-Job Training
Month March April May June
REVIEW OF RELATED LITERATURE
Seedborne Pathognes
Seeds are essential for crop production, but they are vulnerable to pathogens that can compromise their viability Ensuring high-quality seeds, characterized by uniform size, appropriate moisture content, and overall health, is crucial for successful plant reproduction and harvests Pathogen-free seeds are vital, as infections from pathogens or contamination by storage fungi can hinder germination and affect crop yields.
1941 & Epner, 1964) because of infection to seedlings and growing plants Healthy seeds are considered as an important factor for successful crop production
Seed health testing is essential for detecting and controlling seed pathogens to prevent infections and diseases Since 1924, the International Seed Health Testing Association (ISTA) has focused on seed gene purity and germination Seed testing adheres to the International Rules for Seed Testing (ISTA, 2015), encompassing procedures such as sampling, moisture content determination, purity analysis, and seed weight assessment The germination testing standards are based on protocols developed at the Petawawa National Forestry Institute, the British Columbia Tree Seed Centre, and the Alberta Tree Improvement and Seed Center (ATISC), which have demonstrated optimal performance at ATISC Additionally, the procedures for determining seed equilibrium and relative humidity are aligned with the Genebank Standards for Plant Genetic Resources and Food & Agriculture (FAO).
2013) and research at the Royal Botanic Gardens, Kew (Smith et al., 2003, Gold
Seed Treatments
Seed treatments in the United States are essential for controlling seed-borne pathogens, utilizing chemical, physical, and mechanical methods These pathogens can significantly damage seeds, making effective treatment crucial By disinfecting contaminated seeds, seed treatments enhance germination rates and promote healthier crop growth.
In the United States, chemical seed treatment is employed to combat seed-borne pathogens, primarily using thiram and seed disinfectants Thiram serves dual purposes as a fungicide and a repellent for birds and animals, particularly in nurseries This treatment has proven effective against F oxysporum in ponderosa pine and Douglas-fir seeds, as well as against Fusarium spp in longleaf pine seeds.
Thiram has been shown to have a toxic effect on the seeds of many conifer species, particularly at high dosages, which can hinder germination under operational conditions (Abbott, 1958; Nolte & Barnett, 2000) While other fungicides not specifically labeled for tree seed use have been tested with some positive outcomes, they have not demonstrated significantly better control of fungal infestations or improved germination rates compared to disinfectants (Barnett & McGilvray, 2002; Barnett & Varela, 2003).
Disinfectants like sodium hypochlorite, hydrogen peroxide, and hydrogen dioxide (Zero Tol®) are effective in reducing fungal infections and enhancing seed germination Hydrogen peroxide, particularly at a 30% concentration, can eliminate seed-borne mycoflora and significantly stimulate the germination of conifer seeds, while also effectively removing seed coat contamination from longleaf pine seeds caused by F circinantum and other Fusarium species Additionally, a 3% concentration of hydrogen peroxide has proven effective in reducing seed-borne contamination of Fusarium spp and maintaining high germination rates Sodium hypochlorite also contributes to reducing Fusarium contamination and improving germination rates in various western conifers.
& Dumroese, 1987) Pretreatment of seeds with ethyl alcohol can increase the efficacy of sodium hypochlorite in agricultural crops (Sauer & Burroughs, 1986)
Physical seed treatment methods, such as water rinses, can help reduce seed-borne pathogens and enhance germination (Riffle & Springfield, 1968) While stratification may increase fungal presence in certain seed lots, running water can effectively decrease surface contamination (Alxelrood et al., 1995) However, it's important to note that water rinses alone cannot completely eliminate fungal contamination from seeds (Riffle & Springfield, 1968) Therefore, seed lots suspected of harboring seed-borne pathogens should also undergo treatment with disinfectants or fungicides (Campbell & Landis, 1990).
Heat treatment effectively manages seed-borne pathogens while preserving seed viability Techniques such as hot water, aerated steam, and microwave radiation have been extensively studied for agricultural crops (Agarwal & Sinclair, 1997) Notably, microwave hot water treatment has successfully controlled Fusarium spp in Douglas-fir seeds without significantly impacting germination rates (James, Gilligan, Dumroese, & Wenny, 1988) However, further research is needed to enhance understanding of these methods.
Mechanical seed treatment methods effectively remove dead and fungus-damaged seeds to enhance germination rates and minimize seed-borne pathogens Techniques such as the gravity table have proven successful in separating damaged seeds from healthy ones (Karrfalt, 1983) Additionally, the Incubation, Drying, Separation (IDS) system has been utilized to distinguish viable seeds from filled-dead seeds, demonstrating effectiveness in removing damaged seeds from seed lots (Simak, 1984; Donald, 1985; Downie & Wang, 1992), particularly in species like Plantanus x acerifolia (Ait.) Willd (Falleri & Pacella, 1997).
Effective control of seed-borne fungi typically relies on chemical fungicides; however, their application to grains is limited due to pesticide toxicity (Ferrer, 1991; Harris, 2001; Dukic, 2004) This necessitates the exploration of alternative, cost-effective, and eco-friendly methods for safe seed storage Research has shown that organic soap solutions can help reduce seed contamination According to a National Academy of Science report (1986), fungicides present a higher carcinogenic risk compared to insecticides and herbicides, highlighting the urgent need for safer alternatives Over the years, significant efforts have focused on discovering new antifungal agents from natural sources for food preservation The current study aims to evaluate the antifungal activity of aqueous Capsicum frutescens (hot chili pepper) extract against key seed-borne pathogens Capsicum frutescens has demonstrated effectiveness in reducing A flavus and A niger at relatively low concentrations However, MVP (1985) noted that neither chemical nor organic treatments consistently eliminate contamination that can damage seeds, and factors such as the hardness of the seed coat or prolonged storage periods may hinder treatment efficacy, potentially leading to seed death.
Methodology
The standard blotter method, as outlined by the International Rules for Seed Testing Science and Technology (1996), involved washing seeds with 0.1% Sodium Hypochlorite for 10 minutes Subsequently, 10 seeds per Petri dish were placed on three layers of moistened blotter, with the incubation occurring in a B.O.D incubator at 25± 2˚C for 7-10 days under a 12-hour alternating light and dark cycle On the 8th day, seeds were examined for seed-borne fungi using a stereo binocular microscope, and pathogens were identified with a compound microscope The percentage of seed infection and germination was calculated, and isolated fungi were identified using key monographs and relevant literature.
(Fennell, 1965) (Booth, 1971) (Ellis, 1971) (Hunter, 1972)the untreated seeds were used as control All experiments were carried out in triplicate However
In 1982, research on Shtsjorea robusta seeds identified 21 fungal species using the blotter method, while only 5 species were found on potato dextrose agar (PDA) Subsequently, Mittal (1983) reported 26 fungal species through the blotter method, with 10 species detected on the seeds.
Cedrus deodara in PDA method
The health of seeds is crucial for preventing the survival and transmission of pathogens, which can be present as contaminants on the seed surface or within the seed lot itself, potentially introducing diseases to previously clean areas (Henning, 2005) Fungi represent the largest group of pathogenic agents associated with seeds and grains, as noted by Souza et al (2007), who recommended using sodium hypochlorite (2.0-2.5% active chlorine) for effective treatment.
Sodium hypochlorite (5.0% NaOCl) effectively inhibited the in vitro mycelial growth of Cercospora longissima Additionally, previous studies have assessed the allelopathic effects of plant extracts on lettuce (MAGIERO, 2009) The results indicated that there was no significant reduction in germination time at any of the tested concentrations.
A study on the fungi associated with the seeds of Dalbergia sissoo, Azadirachta indica, Paulownia fortunei, Albizia procera, Strychnos nux-vomica, and Plantago ovate in India identified 22 fungal species across 15 genera The findings revealed significant variability in the seed mycoflora between stored and freshly collected seeds, with stored seeds exhibiting a higher number of fungi Additionally, seed treatments with fungicides such as Thiram, Dithane M-45 (mancozeb), Bavistin (carbendazim), and Emisan (2-methoxyethylmercury chloride) effectively controlled the seed mycoflora, with varying efficiency among the different fungicides.
2.4.Seed Borne Fungi of Some Forest and Their Control
The research conducted by Singh and Khann (1999) focused on the mycoflora of seeds from Prosopis chinensis, Albizia procera, and Michelia champaca, which were sourced from a forest in Dehra Dun, Uttar Pradesh.
In a study conducted in India, a total of 16 fungi from 8 genera were identified, with moulds being the most prevalent The highest diversity of fungi, totaling 8 species, was found on the seeds of Paulownia fortunea, while the lowest, with only 4 species, was observed on Prosopis chilensis Aspergillus spp were detected on the seeds of all tree species except Albizia procera Additionally, Penicillium citrium was found on M champaca, Albizia procera, Paulownia fortunea, and B purperea, but was absent from the other tree species.
Three species of Phoma were detected on P.fortunei only, while Trichohecium roseum was present on seeds of Albrizia procera only Fusarium avenaceum
[Gibberella avenacea] was present on seeds of Albizia procera, Accia nilotica, Acia catechu and B purrurea whereas it was absent in M champaca and Paulownia firtunei Application of Emisan [2-methoxythylmercury chlorite],
Topsin (thiophanate-methyl) and Bavistin (carbendazim) were effective at a dosage of 2g/kg, while Dithane M-45 (mancozeb), copper ox chlorite, and Thiram showed efficacy at 4g/kg In the case of M champaca, Albizia procera, and Acacia nilotica, Emisan was followed by Thiram, whereas Dithane M-45 was effective for Acacia catechu Topsin and copper ox also demonstrated significant results.
2.5 Impact of Seed Mycoflora and Forest Seed on See Quality anf Their Management
Seed ( Mamatha et.al, 2000) of Dendrocalamus strictus Phyllanthus emblica
The study investigated the mycoflora incidence in Hardwickia binata and Dalbergia latifolia, revealing the presence of both field and storage fungi The dominant mycoflora were isolated, and their impact on seed quality, particularly germination and vigor, was assessed Seeds inoculated with these dominant fungi exhibited a significant reduction in germination rates and seedling vigor Additionally, four different pre-treatments were applied to the seed samples, with treatments involving Trichoderma spp proving most effective in decreasing mycoflora incidence while enhancing both germination and vigor.
This study aims to highlight the research priorities in seed pathology, emphasizing the critical need for seed testing to prevent catastrophic losses from seed-borne diseases Given the historical neglect of forestry studies on seed-borne transmission, it is essential to implement protective measures for forestry seed species Seed health testing can identify seed-borne pathogens, necessitating treatments such as Sodium hypochlorite and plant extracts like Capsicum sp These pre-treatments have proven effective in reducing seed contamination and enhancing germination rates The research focuses on identifying seed pathogens and evaluating the effectiveness of these treatments in minimizing infections in forestry species seeds.
STATUS OF THE CONSIDERED ISSUE AT THE TRAINING
Title
Seed Health Testing of Selected Forestry Seeds and Their Control Using Sodium Hypochlorite and Capsicum Extract Soap Solution
Time and Place
The experiment took place at the ERDB Seed Center and the Seed Pathology Laboratory of the Environmental Research and Development Bureau (ERDB) in Los Baños, Laguna, Philippines, from April 2018 to June 2017 The experimental area maintained normal humidity levels After 7 to 10 days, the seeds in the incubator were checked for the presence of microorganisms.
To determine different kinds of seed pathogens/other microorganism contaminating or infecting four (4) kinds of forestry seeds
To determine the level/percent seed pest or microbial infection
To determine the effectiveness of sodium hypochlorite and capsicum extract -soap solution as seed disinfectant.
Statement of the Problem
What are the seed borne pathogens found in four selected forestry seed species?
What are the level/percentage of seed pests or microbial infection?
Which is more effective treatment, Sodium Hypochlorite or Capsicum Extract-Soap Solution?
Hypothesis
Percentage fungal seed infection/contamination is low
The treatments are not effective as fungal control
Percentage fungal seed infection/ contamination is high
The treatments are effective as fungal control.
Scope and Limitation
The research faced several limitations, including insufficient time to assess the long-term growth and performance of seeds in the nursery, as the experiment was conducted solely in a laboratory setting Additionally, the ERDB seed center offered a limited variety of forestry species, restricting the number of desired experiments The study primarily aimed to evaluate fungal diversity and contamination in seeds, while bacterial infections were noted but not characterized due to a lack of necessary facilities and reagents for physiological analysis Germination was the only parameter examined in relation to microorganism infection, with factors such as seed moisture, weight, size, and color excluded from the study.
Materials
All the materials and equipment are provided by Ecosystem Research Development Bureau, University of the Philippines, Los Banos Laguna Philippnes
3.7.1 Four selected forestry seeds species
Lanete seeds (100 seeds per treatment)
Taluto seeds (100 seeds per treatment)
Ipil-ipil seeds (100 seeds per treatment)
Molave seeds (100 seeds per treatment)
Shaped like cylinder they are usally made of glass or plastic That Scientist uses it to grow cell from fungus and diseases of the seeds
Heated container used for chemical reaction and other processes using high pressure and temperature e.g., steam sterilization
Optical instrument for viewing sample at high magnification has three, four or five objective lenses that range in power from 4x too 100x
Is an electrical device which use dry heat to sterilize Petri dish and othe equipment was use in the experiment
Ditilled water is one type of purified water This is used for watering the seeds and for the application of the treatments
Three layers of filter paper on the Petri plates that can hold 5 ml of distilled water
Are a handheld, hinged instrument used for grasping and holding object This is used to make sure that seeds are protected for other microorganisms
3.7.9 De- natured Alcohol and Alcohol lamp
Is a piece of laboratory equipment used to produce an open flame This is used for sterilization of the disease
3.7.10 Glass slides and Glass cover
This is used to hold objects for examination under a microscope Typically the object is mounted on the slide, and then both are inserted together in the microscope for viewing.
Methods
Petri dish sterilization in hot air oven at 80 degree Celsius for 2 hours Distilled water sterilization and filter paper sterilization in automatic autoclave at
212 degree Celsius, 120 Pressure per square inch (psi) for one (1) hour
To create a Capsicum Extract Soap Solution, combine 25 pieces of hot chili, sieved through gauze, with one teaspoon of liquid soap and one liter of distilled water The mixture should then be steam sterilized at 212 degrees Celsius and 120 psi for one hour.
Prepare four (4) selected forestry seeds that will be tested;
Plant the seed onto Petri dish with three layers of filter paper that can hold
Incubate the plated seeds under (12 inches space between) day light tubes for 7- 10 days under alternating light (12 hours) and 12 hours of darkness
Seven days post-incubation, visually inspect the seeds, first with the naked eye and then using a binocular microscope at 4-10x magnification Seeds exhibiting microbial growth should be further analyzed under a compound microscope at 100 to 400x magnification, allowing for the identification or suspicion of certain fungi species based on spore production.
Other data collected were bacterial contamination and rate of seed germination
Data were tabulated and statistically analyzed
Research Approach
Research Design
The research design that was used is Complete Randomized Design (CRD) as layout in the table below
Treatment 3 (T3) - Capsicum Extract Soap Solution (CESS)
Data Gathering
Data collection will occur after seven days of incubation (baseline data) and again seven days post-treatment The study will evaluate seeds based on parameters such as the number of infected seeds, germinated seeds, and those exhibiting fungal growth or bacterial contamination Fungi will be identified using various references, while bacterial presence will be noted without specific identification Physiological tests, however, cannot be conducted in the laboratory due to a lack of available reagents and other necessary materials.
Statistical Analysis
Statistical analyses were conducted using the Multiple Comparison of Turkeys within the Statistical Analysis System (SAS) to assess the percentage of infected seeds among four selected forestry species subjected to various disinfection treatments for fungal infections This analysis focused on growth parameters related to fungal and bacterial infections, as well as germination rates.
This chapter presents a summary of the data analysis results, utilizing tables to display information on infected seeds, germination rates, bacterial presence, and fungal counts The study employed specific labels to denote various treatment levels, including T1, which represents the control group with no treatment applied.
T3- Capsicum Extract Soap Solution Soap/CESS
Table 2: List of Seedborne Pathogens/Contaminants
Forest Tree Seed Species Type of Seed Borne Pathogens
Apocynacea wrightia pubescens ( Lanete) Aspergillus flavus, Verticillium sp Pterocymbium tinctorium (Taluto) Fusarium spp
Vitex parviflora (Molave) Aspergillus niger, Chaetomnium sp.Verticillium sp Leucaena leucocephala (Ipil- Ipil) Fusarium spp, Chaetomnium sp
Seed-borne pathogens were identified through their distinct characteristics, utilizing references such as the "Monograph on Fusarium: Illustrated Genera of Fungi Imperfecti" and recent studies on seed-borne fungi affecting forestry species.
Table 3: Percentage of infection of Fungi on four slected forestry
Seeds and Effect of Pre-treatment
Seeds Treatments Percentage Std Err Trurkeys
The results presented in Table 3 indicate that the highest fungal infection in Lanete seeds occurred in the seeds treated with CESS at 21% (with 100 seeds per treatment replicated three times), followed by the control group with a mean infection rate of 19% The lowest infection rate was observed in seeds treated with Sodium hypochlorite, which recorded only 7% This suggests that treatment 3 is the most effective against fungal infection in Apocynacea wrighita pubescens R.Br (Lanete), while treatments 1 and 2 showed no significant difference in effectiveness The P-value of