CHAPTER THREE

1.0 DESCRIPTION OF WORK DONE

I had my SIWES attachment programme at Forestry Research Institute of Nigeria (FRIN)

Ibadan, Oyo state. The institute has six (6) large specialized Departments. During my 6 months
attachment, I was posted to two departments that are relevant to my field. These departments are;
Sustainable Forest Management (SFM) and Forest Conservation and Protection (FC&P).

1.1 SUSTAINABLE FOREST MANAGEMENT DEPARTMENT

Sustainable Forest Management (SFM) is the heart beat of the institute which Headed by Dr
(Mrs) Adio. In a welcoming lecture delivered by Mr. Iroko, the department has six (6) Sections,
these are;
 Biotechnology section
 Seed section
 Tree improvement section
 Agroforestry / Silviculture section
 Nursery sections
 Tree breeding section

1.1.1 SEED SECTION

At the section, the head of the section (Mr. Kareem) in his lecture gave the definition of seed
as “an embryonic plant enclosed in a protective outer covering called the seed coat, usually with
some stored food’’. Seed is the product of the ripened ovule which occurs after fertilization and
some growth within the mother plant. The formation of the seed completes the process and
production in seed plants (started with the development of flower & pollination), with the
embryo developed from the zygote and the seed coat from the integument of the ovule.
A typical seed consist;
 embryo(rudimentary plant),

1
 endosperm(stored food),
 seed coat(protective surface layer of embryo),
 hypocotyl(shoot of germinating seed)

a. seed coat
b. endosperm
c. cotyledon
d. hypocotyl
Fig 1:A typical diagram of a seed

The Head of the department further explain the seed mandates in a sequecial order. These are;
 Seed Collection / Procurement
 Seed Extraction / Handling
 Seed Storage
 Seed Testing
 Seed Distribution
 Seed Record
 Seed Distribution
 Seed Research

SEED COLLECTION
Seed collection is process of collecting seed from the wild for research or project work
through local collection (within the country) or external (outside the country).
Different steps in seed collection include;
Step 1: Recognizance survey: this is done by visiting the site of collection before going for
collection tour
Step 2: What to collect: collect seed that are free from any infection or insect attack and avoid
picking seeds from those on the ground which might have been infected.

2
Step 3: How to collect: seed are collected from the parent plant tree using sickle. Tapoline is
spread over those seeds that already on the ground to avoid mixing up with new ones.
Always tag or label newly collected seeds which are collected from different locations; even they
are of the same species.
Any seeds collected both locally or externally are passed through Quarantine officer to
check may be they are free of disease.

SEED EXTRACTION
Each species of seed have different method of extraction based on their feacture. The major
measure in seed extraction method is the Moisture content present in the species to extract. The
moisture content present in Gmelina arboreal seed is different from the one present in Tectona
grandis seed.

SEED STORAGE
Seed storage may be defined as the preservation of viable seeds from the time of collection
until they are require for sowing.
When storing seeds for the future, different types of seed should be stored under different
conditions. The ability of a seed to produce plants depend on the time of seed harvest and the
condition of storage.

There are basically two types of seed based on their method of storage, these are;
 Orthodox seed
 Recalciltrant seed
ORTHODOX SEEDS
These are seeds that are able to store for long period of time at low temperature and reduced
humidity, such as
RECALCILTRANT SEEDS
These are seeds that readily loosed their viability when stored for long period of time at low
temperature and reduced humidity, such Irvingia gabonesis, Chrysophylum albidum. They do
not survive drying and freezing.

3
SEED TESTING
Before storage of seeds, seed viability test are done to determine if stored seeds are still
viable. This is done by taking few of them, count them and place them in a flat tray or pot filled
with potting mix. Watered the seeds well and give them plenty of light. Keep the potting mix
moist, and see how many seeds sprout, if more than half to three quarter of the seeds sprout, then
the seeds has a good chance of germinating in garden.
Routing check of the seed stored is necessary.

SEED RECORD
A seed recording is the process of recording the relevant information and data on the seeds
such as collection date, seed weight, location, color of the seed when collected.

SEED DISTRIBUTION
This is the distribution of seeds to those in need of it for research and project.

SEED RESEARCH
Different research is done on seeds to check for their viability, rate of germination, seed
dormancy and dormancy control.

GERMINATION OF SEED.
The life cycle of plants and trees starts with the germination of seeds. Plants and trees produce
seeds and seeds develop into new plants. The cycle thus continues keeping the species alive and
continuing.
Germination is defined as sprouting of seeds, a sprout, a spore, or additional reproductive
body, regularly done after a period of dormancy. Availability of water, health of the seeds, time,
temperature and heat, presence of oxygen, and exposures to light play an important role in the
germination process.
Seed germination can be defined as the resumption of growth of the embryo of the mature
seed; it depends on the same environmental conditions as vegetative growth does. Water and

4
oxygen must be available, the temperature must be suitable, and there must be no inhibitory
substances present.
However, in many cases a viable (living) seed will not germinate even though all the necessary
environmental conditions for growth are satisfied, this phenomenon is termed seed dormancy.

Stages in Germination
 Imbibition:
The initial step in the development of seed germination is to absorb water or imbibition. The
seed absorbs water and as a result it swells. The swelling of seed causes the seed coat to
rupture and allows the radicles to move downward forming the root.
 Respiration:
When the seed absorbs water, it also respires . While in the beginning the respiration is
anaerobic and later it becomes aerobic.

Fig 2: Germination process of bean seed

Factors Affecting Germination Process of Seeds:

 Temperature: Extremely low or cold temperature is not favorable for seed germination.
They prefer higher temperatures. The germination rate of seed is directly proportional to
the rise in temperature.

5
  Moisture or water: Dry seeds do not germinate. Water is an essential factor to trigger off
the process of seed germination.

 Soil: During growth, seeds require mineral elements for further growth which is obtained
from the soil.

 Light: For seed germination light is not essential in the early stages of germination but
plays a main role in the later stages of the life cycle of plants.

 Viability of the seeds: After the seeds are formed, they remain viable up to certain
period which may vary from plant to plant or seed to seed. Many sees die or incapable of
supporting growth after a certain period of time.

 Dormancy period: Many seeds do not germinate abruptly after they are produced.
Certain seeds undergo a resting time through which they stay dormant and germinate
when conditions are favorable. Presence of growth inhibitors like abscisic acid induces
dormancy in seeds.

 Thinness or thickness of seed coat: Different seeds have varying degrees of thickness to
enable the seeds to remain feasible. Seeds with a thin seed coat tend to germinate faster
than those with thicker seed coats.

Changes during Germination Process

Seeds germination is seen when the seed starts growing its shoots and roots . The seed
encloses a baby plant with its cotyledons. The cotyledon store food for the baby plant inside the
seed and provides nutrients till the seed coat ruptures and the plant is capable enough of drawing
nutrients from the soil and the surroundings. The first thing to come out is the main root .The
skin or seed coat starts to split and the tiny shoot straightens, carrying the cotyledons with it.
This is called radicle or the main root which grows bigger. Root hairs appear as the growth takes
place. Gradually, first leaves called plumule starts growing which forms the shoot in initial
stages. To grow, the seed’s growing conditions usually have to be damp, warm, and dark, with

6
moderate soil. A dry seed will stay dormant unless the conditions are favorable. Later, seeds
germinate supporting a new life.

SEED DORMANCY
Seed dormancy is defined as a seed failing to germinate under environmental conditions
optimal for germination, normally when the environment is at a suitable temperature with proper
soil moisture.
Seed dormancy has two main functions, the first is synchronizing germination with the optimal
condition for survival of the resulting seedlings; the second is spreading germination of a batch
of seed overtime.

TYPES OF SEED DORMANCY

There are two types of seed dormancy. These are;
 Coat-imposed dormancy
 Embryo dormancy

COAT IMPOSED DORMANCY: is a dormancy imposed on the embryo by the seed coat and
other enclosing tissues, tissues such as endosperm, pericarp. The embryo of such seed will
germinate readily in the presence of water and oxygen once the seed coat and other surrounding
tissues are removed or damage. Seed coat dormancy is common in California lilac (Ceanothus
spp), Acacia spp
EMBRYO DORMANCY: refers to dormancy that inherent in the embryo and is not due to any
influence of the seed coat or other surrounding tissues. Examples include Corylus avellana
(European hazel).

CONTROL / TREATMENT OF SEED DORMANCY

Various methods have been used by seed scientist and technologists the dormancy of seed.
Simple and widely used methods are,
1. MECHANICAL SCARIFICATION: Mechanical scarification is a technique for overcoming
the effect of an impermeable seed coat. Mechanical scarification can be done by rubbing seeds
between two pieces of sandpaper or using a file, a pin, or a knife to rupture the seed coat.

7
 Care must be taken not to injure the embryo. It may be necessary to open a couple of seeds to
see where the embryo is located in relation to the micropyle, the former point of attachment to the
fruit. Large seeds like those of the bush lupine (Lupinus) are easily scarified with a knife. Examples
are some legume family

2. HOT WATER TREAMENT: For small to medium-sized seeds or large quantities of seeds, the hot
water treatment is more practical than scarification. For this treatment seeds should be dropped into
about six times their volume of 180º-200ºF pre-heated water (rain water is desirable if it is near
neutral in pH). They should be left to cool and soak
In the water for 12 to 24 hours, after which they are ready for sowing. The container used for this
treatment should not be made of aluminum as it may be
Toxic to the seeds.

3. CHEMICAL TREATMENT: Acid treatments are often used to break down especially thick
impermeable seed coats. Since seeds placed in concentrated sulfuric acid (H2SO4) will become
charcoal in time, the temperature of the acid and the length of time the seeds are soaked are very
important. The acid should be used at room temperature for a period of a few minutes to several
hours depending on the species. The seeds should be immersed in acid in a glass, or earthenware
container, and should be stirred occasionally with a glass rod; however, too much stirring will cause
the
acid to heat undesirably.
The seeds must be removed from the acid just before any acid penetrates the seed coats. When the
allotted time is finished, the seeds should be removed promptly and washed thoroughly in several
changes of water to neutralize completely all remaining acid.

8
EXPERIMENT ON THE COMPARATIVE STUDY OF 3 ACACIA SPECIES
At the section, I did practical experiment on the comparative study of 3 different Acacia species;
Acacia nilotica, Acacia auriculiformis, Acacia mangium.

TITLE OF THE EXPERIMENT: COMPARATIVE STUDY OF 3 ACACIA SPECIES.

AIM: The experiment was carried out to check the rate of Germination of the 3 Acacia species.

MATERIAL USED: Seeds of Acacia nilotica, Acacia mangium, Acacia auriculiformis, water,
Copenhagen germination tank, filter paper, beaker, razor blade or scalpel, propagator, polythene pot
filled with top soil.

PROCEDURE: Three different Acacia species (Acacia mangium, A. nilotica, A. auriculiformis)

were picked and scarified using razor blade to cut the hilum in other to allow easy entry of water into
the seed embryo, 30 seeds were scarified and 10 unscarified for each Acacia species. A Copenhagen
germination tank was filled with water to a certain level, then filter papers were put on the glasses
slide in the germination tank and a paper was cut and attached with the filter paper and suspended
into the water in the germination tank which serves to absorb water and keep the seeds on the filter
paper moist. 10 seeds of the Acacia species were distributed on each filter paper making 3 replicates
and 1 control for each Acacia species and the filter papers were labeled with each species name. The
3 Acacia species were subjected to the same treatment (light, temperature and water).
The following day the experiment was observed and the species with emergence of radicle and
result were recorded, it was also observed on the second day and each result was recorded. At the
end of the experiment, the best species was transferred to polythene pots with top soil in the
Propagator.

RESULT OF THE EXPERIMENT ON THE 2 DAYS:

1. Acacia nilotica
DAY 1 DAY2
Replicate No of seeds with emergence of Replicate No of seeds with emergence of
radicle out of 10 radicle out of 10

9
Rep 1 5 Rep 1 Nil
Rep 2 6 Rep 2 Nil
Rep 3 5 Rep 3 Nil
Control No radicle emerged Control No radicle emerged
2. Acacia mangium
DAY 1 DAY2
Replicate No of seeds with emergence of Replicate No of seeds with emergence of
radicle out of 10 radicle out of 10
Rep 1 1 Rep 1 9
Rep 2 1 Rep 2 7
Rep 3 4 Rep 3 5
Control No radicle emerged Control No radicle emerged

3. Acacia auriculiformis
DAY 1 DAY2
Replicate No of seeds with emergence of Replicate No of seeds with emergence of
radicle out of 10 radicle out of 10
Rep 1 1 Rep 1 3
Rep 2 1 Rep 2 3
Rep 3 1 Rep 3 4
Control No radicle emerged Control No radicle emerged

CONCLUSION: At the end of the experiment it was observed that Acacia mangium has the
highest no of peak value (emergence of radicle) out of all the ACACIA SPECIES and the control
seeds without SCARIFICATION did no peak at all(germinate).
 This shows that Scarification of ACACIA SPECIES is necessary to control their dormancy
before planting.
 Since Acacia mangium has the highest peak value, it is recommended for anybody that wants
to plant any ACACIA SPECIES due to its rate of germination.

10
Fig3: Emergence of radicle on the day1. Fig4: Copenhagen germination tank

3.1.2 BIOTECHNOLOGY SECTION

At this section, in a welcoming lecture delivered by Mr. Olomola, he explained details on
what Biotechnology is and their relevant relating to Agriculture, Medicine, and Food Production.
BIOTECHNOLOGY: Is the uses of biological system and living organisms modify or make
useful product for specific uses
In relation to Agriculture: Agriculture has been theorized to have become the dominant way
of producing food in the past. Through early Biotechnology, the earliest Farmers selected and
bred the best suited crops, having the highest yields, to produce enough food to support a
growing population. As crops and fields became increasingly large and difficult to maintain, it
was discovered that specific organisms and their by-product could effectively fertilize, restore
nitrogen and control pests. Through the help of modern biotechnology many crops have been
modify and breed them with other plants to produce new and useful products.
The Biotechnology section of FRIN majorly deals with Tissue Culture.

TISSUE CULTURE
Tissue culture is a technique of growing plant cells, tissues, organs, seed, or other plants part in
an aseptic (sterile) environment on a nutrient medium. The only principle behind tissue culture is
Totipotency (ability of undifferentiated plant to regenerate itself).

11
TERMINOLOGY USED IN TISSUE CULTURE
i. Explant: any portion taken from a plant that will be used to initiate a tissue culture e.g.
seeds, leaves, shoot, node, meristem, embryo etc.
ii. In vitro: To be grown in glass. Propagation of plants in a controlled, artificial
environment using plastic or glass culture vessels, aseptic techniques, and a defined
growing medium.
iii. Plantlets: young or small plant grown from tissue culture.
iv. Aseptic/sterile: Free of microorganisms.
v. Medium (media): A nutritive solution, solid or liquid, for culturing cells.
vi. Micro propagation: In vitro Clonal propagation of plants from shoot tips or nodal
explants, usually with an accelerated proliferation of shoots during subcultures.
vii. Subculture: This is the process by which the tissue or explant is first subdivide, and then
transferred into fresh culture medium.
viii. Acclimatization: Process by which cultured plants (plantlet) are taken out of test
tubes/flasks and then transferred to pots containing autoclaved sterilized mixture of soils
in the Green house.
ix. Inoculation: Inoculation is the process of transferring disinfected / sterilized explant in to
prepared culture medium

MATERIAL NEEDED FOR TISSUE CULTURE

Some of the materials used for tissue culture include;
i. 1 Vial of Murashige and Skoog (MS) media.
ii. 1 L of sterile distilled water.
iii. 10 g of agar/L and 30 g sucrose/L.
iv. 1.5 L or 2 L container in which to prepare the growth medium.
v. Small amounts of 1M NaOH and 1M HCl to adjust the pH of the media.
vi. 60 flat bottom culture tubes with closures, Adhesive / paraffin tape.
vii. 10% Bleach in a spray bottle and 70% alcohol in a spray bottle.
viii. Forceps or tweezers, Gloves, Cutting equipment such as a scalpel blade or razor blade.
ix. Weighing balance, Microwave and autoclave machine
x. Explant and Beaker or jar in which to wash the plant material.

12
xi. Detergent-water mixture - 1ml detergent per litre of water, Bleach sterilizing solution.

STEPS IN TISSUE CULTURE

 Preparation of culture medium
 Collection and disinfection of Explant
 Inoculation
 Transferring to Growth room
 Sub culturing and acclimatization.

1. MEDIA PREPARATION (Murashige and Skoog )

Different types of media use in tissue culture include;
 Murashige and Skoog culture medium
 Preece culture medium
 Woody plant medium (WPM)
 Basal medium
At this section I worked with Murashige and Skoog.
A medium represent the plant food in which Explants are grown. One of the most important
factors governing the growth and morphogenesis of plant tissues in culture is the composition of
the culture medium. The basic nutrient requirements of cultured plant cells are very similar to
those of whole plants.

Plant tissue culture media are generally made up of some or all of the following components:
i. Macronutrients ii. Micronutrients, iii. Vitamins, iv. Amino acids or other nitrogen supplements
v. sugar vi. Solidifying agents and growth regulators.

Murashige and Skoog media are made up of 3 stocks, these are;

Stock 1 (Basically macronutrient): KNO3, NH4NO3, MgSO4.7H2O, KH2PO4
Stock 2 (Basically micronutrient): CuSO4.5H2O, KI, H3BO3, MnSO4. H2O
Stock 3(Vitamins): Pyridoxine HCI, Nicotinic acid, Myo- inositol, Glycine, Thiamine HCL.

Macronutrients

13
The macronutrients provide the six major elements-nitrogen (N), phosphorus (P), potassium (K),
calcium (Ca), magnesium (Mg), and sulphur (S)-required for plant cell or tissue growth. The
optimum concentration of each nutrient for achieving maximum growth rates varies considerably
among species.
Culture media should contain at least 25-60 mM of inorganic nitrogen for adequate plant cell
growth. Plant cells may grow on nitrates alone, but considerably better results are obtained when
the medium contains both a nitrate and ammonium nitrogen source.

Micronutrients
The essential micronutrients for plant tissue culture include iron (Fe), manganese (Mn), zinc
(Zn), boron (B), copper (Cu), and molybdenum (Mo). Chelated forms of iron and zinc are
commonly used in preparing culture media. Iron may be the most critical of all the
micronutrients. Iron citrate and tartrate may be used in culture media, but these compounds are
difficult to dissolve and frequently precipitate after media are prepared. Murashige and Skoog
used an ethylene diaminetetraacetic acid (EDTA)-iron chelate to bypass this problem. Sodium
(Na) and chlorine (Cl) are also used in some media but are not essential for cell growth.

Solidifying Agents or Support Systems

Agar is the most commonly used gelling agent for preparing semisolid and solid plant tissue
culture media. Agar has several advantages over other gelling agents. First, when agar is mixed
with water, it forms a gel that melts at approximately 60°-100° C and solidifies at approximately
45°C; thus, agar gels are stable at all feasible incubation temperatures. Additionally, agar gels do
not react with media constituents and are not digested by plant enzymes. The firmness of an agar
gel is controlled by the concentration and brand of agar used in the culture medium and the pH of
the medium.

Plant growth regulator

Plant growth regulators are the critical media components in determining the developmental
pathway of the plant cells. The plant growth regulators used most commonly are plant hormones
or their synthetic analogues.

14
There are 3 main classes of plant growth regulator used in plant tissue culture, namely:
(1) Auxins: Auxins promote both cell division and cell growth, e.g. (IAA Indole-3-acetic acid)
(IBA Indole-3-butyric acid)
(2) Cytokinins: Cytokinins promote cell division. Naturally occurring cytokinins are a large
group of structurally related (they are purine derivatives) compounds. (3) Gibberellins: They are
involved in regulating cell elongation, and are agronomically important in determining plant
height and fruit-set.
Auxins and cytokinins are the most widely used plant growth regulators in plant tissue culture
and are usually used together, the ratio of the auxin to the cytokinin determining the type of
culture established or regenerated. A high Auxin to Cytokinin ratio generally favours root
formation, whereas a high cytokinin to auxin ratio favours shoot formation. An intermediate ratio
favours callus production.

PROCEDURE:
This step was for 1 L of growth medium which is enough to prepare about 65 growing tubes.
800ml of distilled water was measured using measuring cylinder into a beaker. A stirring bar
was placed inside the beaker containing the distilled water. I measured the stock solutions
(macronutrients, micronutrient, and vitamins). The pH of the solution was adjusted to 5.7 using
1M NaOH or 1M HCl as necessary while gently stirring. Distilled water was added to make the
total volume up to 1L. 7 grams of agar was weighed out and added to the Murashige and Skoog
solution. The solution was homogenized in a microwave (i.e. heat the solution until all the agar
has dissolved) and allows cooling. I dispensed the media into culture tubes. The media was
sterilized at 1210C for 15 minutes in an Autoclave machine. The autoclave machine was Switch
off and allow cooling then unloaded the tubes. The media was left to solidify before used.

2. COLLECTION AND DISSIFECTION OF EXPLANT

The explant is that pieces of plant tissue placed on the culture medium. The choice of explant
depends on the aim and objectives of a planned tissue culture work. If a haploid plantlet is
targeted to be regenerated, the explant to be collected for culturing would be from flower tissue
like anther, pollen. If the intention is to regenerate a virus-tested (virus free) plantlet, the target
organ would be shoot tip or root tip.

15
 Types of explant include;
i. Seed ii. Node iii. Meristem iv. Embryo v. Leaves vi. Root vii. Anther vii.Protoplast

EXPLANT DISINFECTION
Plant tissues inherently have various bacteria and fungi on their surfaces. It is important
that the explant be devoid of any surface contaminants prior to tissue culture since contaminants
can grow in the culture medium, rendering the culture nonsterile. In addition, they compete with
the plant tissue for nutrition, thus depriving the plant tissue of nutrients. Bacteria and especially
fungi can rapidly overtake plant tissues and kill them. Disinfection of the explant is necessary in
other to get contamination/disease free tissue culture.
Disinfection of explant is done in two ways;
(i) Single disinfection
(ii) Double disinfection
SINGLE DISINFECTION: This is usually done for explants such as seeds, leaves, meristem,
embryo etc.

Steps:
i. The disinfection process was done in the air laminar flow hood, the laminar flow was
sterilized using UV light and spraying with ethanol
ii. The collected explant (seed) was placed in a container containing water to prevent withering
due to loss of water
iii. The water was decanted by pouring away the water on the explant in the container.
iv. 70% of ethanol was added to the explant for 3-5 minutes and then decant
v. The explant was rinsed with sterilized distilled water to bring out the effect of the ethanol
vi. 10% of Hypochlorite (NaOCl) and 1-2 drops of tween 20(detergent) was added to the explant
for15-20 minutes to remove dust particle and then decant
vii. The explant was rinsed 3 times in distilled water and then decant twice(i.e. leave the explant
in the third rinse sterile water until its ready to use )
viii. Then Inoculation of the explant into the medium using sterilized forceps was followed

16
DOUBLE DINSIFECTION: This is usually done for explants such as roots and crawling plants
which usually get contaminated due to contact with soil.
Steps:
i. The water on the explant in the container is decanted
ii. 70% of alcohol is added for 3-5 minutes and decant and then rinsed with distilled water
iii. 5% of Hypochloride (NaOCl) and and 1-2 drops of tween 20(detergent) is added to the
explant for 15-20 minutes and then rinsed 3 times with distilled water for proper
disinfection
iv. 10% of Hypochloride and 1-2 drops of tween 20 (detergent) is added for 10-15 minutes
and the rinsed with distilled water 2 times
v. The explant is then inoculated in to the prepared medium

3. INOCULATION:
Inoculation is the process of transferring disinfected / sterilized explant in to prepared culture
medium. Inoculation of explant is done sterilized forceps in the laminar air flow hood to prevent
the entry of Microbes. After inoculation, the test tube was cork with lids and then silled with
paraffin tape to prevent motile organisms. The test tube was labeled with the inoculation date and
medium used.

There are three sources of contamination:

(i). Through utensils and culture medium
(ii).Through explants material
(iii).Through inoculation (transfer of explant to culture medium)

4. INCUBATION
Incubation is the process of taking the inoculated medium in to the growth room for
culturing. The corked test tubes are arrange in the test tube rack, and then placed in the shelf at
the growth room. The growth should be kept cool and with a source of light. When a trace of
yellow is noticed in the leaves, the plantlets have exhausted the nutrient in the medium, and
transfer to acclimatization room or subculture into new medium.

17
 5. ACCLIMATIZATION
Acclimatization is the process of transferring the plantlet from the growth medium to the pots in
the Green House and then to the field.
Once plants are generated by tissue culture, they have to be transferred to the green- house or
field. This requires that the plants be hardened-off before transfer to the field. During this
acclimation process, plants are first transferred to a growth chamber or greenhouse and covered
by domes to minimize the loss of water. In addition, the plants must be “weaned” off the rich
media so they can grow as normal plants in soil. Once the plants are acclimatized under
greenhouse conditions, they are ready for transfer to the field. Acclimation is a very important
step in tissue culture since it is possible to lose plants if they are not properly hardened-off.

CONCLUSIONS
Plant tissue culture is an essential tool in plant biotechnology that has enabled mass clonal
propagation, production of secondary metabolites, preservation of germplasm, and production of
virus-free plants. Moreover, it serves as an indispensable tool for regenerating transgenic plants.
All this has been possible by manipulating plant tissues and various kinds of media developed by
plant tissue culturists and by the use of plant hormones. It has been one of the very exciting
discoveries for plant biologists and will continue to be most useful in the coming years.

Fig 5:pH metre Fig 6: Microwave Fig 7: purified agar

18
Fig8: Acclimitization room Fig9: Growth room Fig10: medium Salt

Fig11: In vitro plantlet Fig12: Nitrogen tank Fig13:Taking of medium pH

3.1.3 NURSERY SECTION

FRIN has 2 Nursery units, these are;
1. Ornamental Nursery
2. Central Nursery

ORNAMENTAL NURSERY
The head of the Unit, Mr Onilearo, in his lecture defined Nursery as a place where plants are
propagated and grown to usable size.
The Aim of this unit is to carry out propagation and production of flowering plants. At this unit,
I had the opportunity to know different categories of Ornamental plants, preparation and
propagation using cuttings, how to care for nursery, landscaping, and I also participated in a
research work using 3 different plants leaves as Green manure to raise Garcinia kola (Bitter
kola).

19
CATEGORIES OF ORNAMENTAL PLANTS
Ornamental plants are plants that are naturally decorated or adorn which make them relevant
and useful for Landscaping and Beautification. Some ornamental plants are not beautiful or
attractive to eyes but some are planted based on their different characteristics to man and
environment. E.g Scented plants.

Different categories of Ornamental plants include;

1. Hedge plants: These are ornamental plants for making walls or fence. They are planted
at about 20cm, 30cm close to each other in such a way that they will form canopy to each
other.
Examples include; Ixora coccinea, Duranta repens, Duranta erecta, Ficus nitida(yellow
bush) ,Muraya paniculata, Acalypha wikisiena, Eugenia uniflora, Abies spp (Fir) ,
codieum veariegatum.

2. Scented plants: these are plants that are not beautiful to the eyes but have scented
fragrance.
Examples include; Cestrum nocturnum (queen of the night), Muraya paniculata,
Jasminum sambac(Jasmine), Cananga odorata (king of the night), Gardenia jasminoides.

3. Defensive plants: These are planted as defence against any kind of intruders.
Examples include; India forma(west indes), Agave Americana, Agave sisiliana,
Euphorbia milli(Christ thorns), Opuntia macroysis,Cereus peruviana, Petiveria
alleacae(Anti snake), Datura metele(Gegemu), Dactura stratemonium
4. Indoor plants: These are plants grown indoor without Ethiolation. They perform well
without much sunlight.
Examples include; Caladium bicolor (blood of Jesus), Aglaonema commutatum,
Euphorbia splendes, Dieffenbachia picta (Dumbkin), Draceania sanderiana, Sanseviera
trifasetia(Mother in-law tongue).

5. Ornamental palms: Examples include; Arcanthophoenix alexandreae (Kings palm),

Roystonea regia (Royal palm), Ravenala madagascariensis (Traveler palm), Borassus

20
 aethiopium (Fan palm), Caryota mitis (Fish-tail palm), Syagrus romanzoffianum (Queens
palm), Phoenix dactylifera (Date palm).

6. Spot plants: These are ornamental plants grown on a spot. Few of them are needed for
proper ventilation.
Examples include; Thuja orientalis, Lagastromia indica, Tecoma stance, Mussainda
erythrophylla (Queeen of phillipines), Cassia biflora, Boungevillea glabra(variegated
ones).

7. Ornamental trees / Shade trees: they are planted to create partial shade. Examples
include; Poliathia longifolia, Terminalia cattapa, Terminalia mantaly, Eucalyptus
camaldulensis, Hura crepitens, Pinus carabian, Acacia auriculiformis.

LANDSCAPING
At this unit, I had lecture on what Landscaping is, steps to take when landscaping and I
also participated in the landscaping of the nursery entrance using different ornamental
plant such as, Ixora coccinea, Duranta repen.
Landscaping is the art and science that deals with planning, design, and indoor
environment using ornamental and non-ornamental plant materials. It is the changing of
the scenery of natural environment for the purpose of beautification.

TYPES OF LANDSCAPING

 Soft Landscaping: the uses of plant components and materials for beautification.
It entails the use of plants of all categories(shrub, palms, tree, herbaceous plants)
 Hard landscaping: the use of non-plant material for beautification.

STEPS IN LANDSCAPING

As a landscape designer, these are the few steps to take in Landscaping work of building and
environment.

21
  Recognisance survey: This is done by visiting the area to check the Topography (such as
valley, slope, hills), and bush of the site. All these are done to be able to know the type of
design suitable for the site.
 Measurement of the Site: Measurement of the site is taken to know the number of plants
that will occupy the whole site. The distance /area of the site is measure and the area for
planting are lined and pegged. For hedge plants, measurements of 30cm to 30cm between
the plants are required.
 Designing: The sketch of the whole site is done to get the roads, building to landscape
and the best plants that fit each.
 Application of Manure: Application of manure to the hole dig for planting on the site to
enhance the quick growth of the plants.
 Expenses: estimate the cost of the plants, labour, equipment, and miscellaneous.
Addition of workmanship profit should be added. The proposal is then submitted to the
client for evaluation.

CUTTING PREPARATION / PROPAGATION

Cuttings are severed plant pieces with at least one node. Various plants organs can be used for
cuttings: stem, shoot, and root or leaf cuttings. Cuttings are usually placed into a suitable rooting
substrate and kept under high humidity until roots and shoots have formed.
At this unit , I worked on the propagation of Ficus Benjamina using Stem cutting.
Steps:
 The Cuttings were taken early in the morning before the sun from the mother tree.
 The cuttings were made below a node using a sharp secateurs.
 The leaves on the cutting were reduced to 2 leafs to prevent loss of little water in the
cutting through transpiration
 The cuttings were put into a bowl containing water to avoid shock
 Polythene bags were filled with top soil, holes were made on th polythene bag to prevent
water log when watered and were arranged at the nursery terrace
 A hole was made into the soil in the polythene pot using stick
 The cuttings were inserted into the hole and soil was press round it
 The new cutting was watered using Watering Can

22
3.1.4 AGROFORESTRY / SILVICULTURE SECTION

At this section, in a lecture delivered by Mr Odewo, gave the definition of Agroforestry,

categories of Agroforestry and benefits of Agroforestry

Agroforestry system is the deliberate combination of woody perennial plants with

Agricultural crops or /and animals on the same land management in a spacial / seqeuecial form
for Ecological or Economic purpose.

Agroforestry system is an integrated system of rural land resource management based on

combining shrubs and trees with crops and/or livestock, whose interactions generate economic,
environmental and social benefits.

TYPES OF AGROFORESTRY

There are various types of agroforestry systems, some of which are listed as follows:

 Silvo - pastoral: These are mostly trees with pastures and livestock. It is essentially the
practice of animal production along with trees and pastures.
 Agrosilvopastural: This is the deliberate combination trees, pastures and animals on the
same land.
 Aquaculture / Aqua forestry: is the planting of Arable crops and trees at the bank of
easily Dry River, in which the tree later form canopy over the river to prevent easy drain.
 Apiculture: this deals with raising of Trees for the production of honeybee introducing
bees.
 Agri -silvicultural :This is subdivided into;
1. Alley farming: This is a process where an Agricultural crop is grown simultaneously
with a long- term tree crop to provide income while the tree crop matures. Fine
hardwood, like walnut, oaks, ash are the favourable species in Alley farming system
2. Strip farming: planting of trees in rows, and the space in between the trees are used
for Arable crops.

23
 BENEFIT OF AGROFORESTRY SYSTEM
Successful agroforestry practices benefits the farmers in the following ways:
 Consistent restoration of the fertility status of the soil through the recycled litter deposition
and nitrogen fixing mechanism of trees.
 A variety of products, firewood, poles, woodcraft, fodder, medicinal herbs and food for
livestock and man respectively.
 Prevention of wind and water erosion by trees acting as wind break and intercepting the
raindrop impact on the soil respectively.
 Improving the micro-climate effect of the immediate and adjourning environment.
 Restoration of water table to an absorbable level for crops use.
 Increased income opportunities.
 Increased economic stability
 Reduce cost for establishing plantation
 Increased ability to manage for sustained yield.

SILVICULTURE: Silviculture is the science of tending and raising forest crop from Nursery to
Maturity. Forest is the carbon sink of the world. It plays roles in reducing the effect of carbon
monoxide (CO) in the Air from Industries. Which reduce the action of Ozone layer depletion,
global warming, and Greenhouse effect.

Reasons for Plantation Establishment include;

 Wood demand (Timber, pole)

 Pulp and paper production
 Ecological purpose
 Employment
 Firewood and Charcoal

WAY OF ESTABLISHING PLANTATION

 Enrichment plant: A way of enriching / replenishing a land that has been degraded is by
planting trees at random or spot e.g. using Tectona grandis, Gmelina arboreal ,because

24
 the grown very fast (Exotic), but these are not advisable to be planted because of high
demand of it legally and illegally. Kit sis better to use Indigenous trees which is of low
demand and growth.
 Afforestation / Afforestation Method: the planting of new trees where plant exist
before but might have been degraded are cut down. It is the act of renewing trees cover
by establishing young trees, generally after the previous stand or forest has been
removed.

In establishing forest plantation the following should be done;

 Environmental Impact Assessment should be done on the environment, people in the

area.
 Surveying , Map out }recognisance survey
 Purpose of establishing
 Cleaning of the area using Mechanical or Manual cleaning method. During clearing
method, raising of the nursery plants should be done around December to May.
 Burning
 Packing of burnt bush
 Re-burning of debris

3.1.5 PHYSIOLOGY AND TREE BREEDING SECTION

The section is divided into two units, which are Fruit tree unit and Pine tree unit. At this
section I had opportunity to work only at Fruit trees unit, where I learnt different Vegetative
method of propagation used at the unit.

Fruit trees unit have their mandate on the following;

1. Domestication of Indigenous fruit tree species found in this Ecological zone

2. Domestication of undomesticated and semi-domesticated (those that are found around us but
in small quantity than in the wild) and

25
3. Domestication of endangered species of plants for mass production.

The only method of propagation used at this unit is Vegetative method of propagation which
include; Grafting, Cutting, Air layering, Budding.

1. GRAFTING: is the method of Vegetative propagation by which new plant (offspring) is

produced from the joining together of a plant from mother plant (plus tree) called the Scion
and a growing seedling of at least a pencil stock diameter called Root Stock.

Plus Trees: plus tree are plant tree that has all the desirable character that is desire in a particular
species. It must be a tree that is fruiting or fruited before. It must not have any manifestation of
disease or insect infestation.

Types of grafting method

Different types of Grafting include;

i. Top cleft grafting: the root stock is open and the scion is inserted
ii. Side vennier: the root stock and scion is cut slantly into the cambium layer and the scion
is joined with the root stock and tight with rope. The grafting is watered for five week,
after the scion has been stabilized with the root stock; the existing top of the stock is cut
out.

One of the advantages of Grafting is that, when the mother plant is fruiting, no matter how
short and smaller the grafted plant is, it will start fruiting at least 2 years after stabilizing
itself. Therefore, it reduce gestation period of fruit trees.
The failure of grafting could be due to several things, such as:
 Disease attacking the joining (connection)
 Scion grows faster before the joining has been properly united
 Mechanical damage of phloem tissue
 Rootstock does contain enough carbohydrate to be translocated toward scion (uneven
distribution of carbohydrate).
 Family difference
 Different behavior between rootstock and scion.

26
2. CUTTING: Plant cutting may be root cutting, stem cutting and leave cutting. The major
motive of cutting is for mass production of fruit tree species.
Materials needed for cutting include; Plant Propagator, Growth promoter (IBA, IAA, and
NAA), Germination trays, cleaned washed and sterilized River sand, sprayer, and secateurs.
Preparation of Cutting
 The plant cutting (stem, leave and root) is produced from disease free plant.
 The cutting is done using secateurs to cut to avoid rough or wounded cutting and the
cutting must have at least a node (a point on the plant stem where new leafs grows.)
 River sand is packed, washed and sterilized using Autoclave sterilizing machine.
The sands are allowed to cool and then transfer into Germination tray.
 The cutting are dipped into growth promoter(hormone e.g IBA, IAA, GA) to
enhance quick root and shoot initiation
 The cutting is the quickly transfer to the sterilized sand in the Germination tray and
the germination is the taking to propagator to control the temperature, relative
humidity, and Air of the environment.
 When they are stabilized in the propagator, they are then pricked out into polythene
pot containing Top soil under weaning shed for 3/4 months of Acclimitization
before taking to permanent field.

3. AIR LAYERING / MARCOTING: Air layering or marcoting is used for all types of
propagation in which roots are formed while the stem is still attached to the mother plant.
Only after the root formation, the layer is detached and planted as a new plant. Air layering
or marcoting can be done with almost any woody plant and is an excellent method to
propagate small numbers of individual trees.
In principle, marcoting is conducted by removing (peeling) barks of the branch or stem up
to the cambium layer. Afterwards, the peeled area is covered with moist growth medium and
is tightly wrapped with plastic sheet, covered fibre or other available materials. Root
primordial will emerge from the marcotted stem. To stimulate root growth, all peeled area is
polished with growth hormone such as IAA, NAA, and IBA.

27
4. BUDDING: In principle budding is attaching axillary bud to Branch or stem which is desired
from a rootstock. This technique has been developed on Tectona grandis for developing
clone bank. Factors which should be considered are among other things:
 Growth characteristics of bud which is attached
 Physiological status (dormant or active)
 Plant species
 Attack by pest and disease.

ADVANTAGES OF VEGETATIVE PROPAGATION

 Production of species plants with Genetic makeup of the mother tree
 It helps in reducing the gestation period, plant height and fruiting period of fruit trees
species.

3.1.6 TREE IMPROVEMENT SECTION

The mandate of this section is the raising of indigenous species of plants and to work on the
reduction of Gestation period of trees that have late germination using vegetative propagated
method such as, Grafting, cutting, layering, marcotting, budding etc.
At this section, under the supervision of Mrs. Akinnate, in her lecture, she made me to
understand that seedlings are young plant grown from seed or any young plant grown in a
nursery for transplanting.
Seedlings are raised using viable seeds. Raising of seedling using Top soil is less preferable
than River sand. River sand is easily drained, but top soil gets wet and sticky together when
watered. Therefore, the plumule gets difficulty in germination, but easily pick when using River
sand to raise seedlings in the germination tray. The seedling are pricked out of the germination
tray or bed in to the polythene bag under weaning shed, which are later harden -off to the normal
environmental condition at the field.

VEGETATIVE PROPAGATION
I worked on the propagation of Syncepalum dulcificum, using stem cutting.

28
Materials used: River sand, Scalpel, Secateurs, Autoclave Sterilizing Machine, Germination
tray, water
Steps:
1. River sand was packed and it was washed until the water get cleaned
2. The river was allow to drained and dried
3. The sand was packed into Autoclave Sterilizing machine and sterilized for 1hour to kill
all the microorganism in the sand
4. After 1hour, the sand was packed out from the autoclave machine and allow to cool
5. Stem cutting was made using secateurs and scalpel
6. The stem cutting was dipped into growth regulator (hormone) to enhance formation of
root and shoot.
7. The sterilized sand was packed into different germination tray
8. The stem cutting was inserted to the sterilized sand
9. The cutting was watered and placed in the glass house

Fig 14: Cuttings in germination tray Fig15: Autoclave sterilizing machine

Fig16: Potted plants at the section Fig17: Pricked seedlings

29
 3.2 FOREST CONSERVATION AND PROTECTION DEPARTMENT
Forest conservation and protection (FC&P) department is one of the research departments at
FRIN. The department has 5 sections, these are;
 Taxonomy sections
 Wildlife section
 Pathology section
 Entomology section
 Ecology section

3.2.1 TAXONOMY SECTION

The section is headed by Dr (Mrs) EKPO. The section carries out research on plant collection,
entobotanical survey and plant preservation. At the section I had opportunity to work on
Herbarium process / techniques and Botanical Rambling under the supervision of Mr
EKUNDAYO.
In his lecture, He defined taxonomy as a systematic way /method of naming, classifying, and
identification of plant species based on already laid down principle using taxonomic keys
(features of plant, apex shape, leaf blade type, leaf venation, flower shape, leaves arrangement,
petiole length, plant growth forms etc.)
He further defined Herbarium as collection of plant specimens that usually have been dried
and pressed, are arranged in the sequence of an accepted classification and are available for
reference or other scientific study. Forestry research institute of Nigeria (FRIN) herbarium is the
largest Herbarium in the whole West Africa with over 120,000 species of plant. He later
explains herbarium techniques and process procedure.

HERBARIUM TECHNIQUES / PROCESS

In a herbarium, plants that cannot be kept in the fresh state are preserved to serve as a
reference collection for botanical comparison and research. Plant specimens are pressed and
dried between sheets of smooth, heavy paper by means of adhesive glues. Loose bits of plant
material, such as fruits and seeds, are placed in an envelope attached to the herbarium sheet.
Each sheet is carefully labelled with the name of the plant and its habitat, the date and place of
collection, the name of the collector, and other pertinent information.

30
STEPS IN HERBARIUM PROCESS AND TECHNIQUES
These are the following steps to follow in botanical collection and processing of herbarium
specimens;
 Collection of plant specimen
 Note taking (i.e. gathering of information on the collected specimen)
 Pressing and Drying
 Treatment with poison
 Mounting
 Labelling and
 Laying away

 COLLECTION OF PLANT
In the collection of herbarium plant specimens certain items of equipment are indispensable to
plant collection. They include Plant presses, Absorbent paper (flimsy paper), twine or straps,
polythene bags, secateurs, field note book, hand lenses, machete, collecting pick, pen and pencil,
metric ruler.
The specimen intended for herbarium must not be a sterile specimen i.e. it should be of
tolerable quality and should be complete with leaves & flowers and or fruits and full note. Sterile
specimens are rarely of value and are not commonly not identifiable with certainty; they should
be collected and accompanied with leaves, and or flowers. Collection should be done in the
morning and evening, it may not possible to collect as whole plant but it is usually to collect
samples from the plant pieces about the size of herbarium sheet should be taken. The collected
plant should be store temporarily in a vasculum bag until they are ready for pressing.
 NOTE TAKING
Every specimen should be accompanied by comprehensive notes retained in a collecting note
book. These notes may not only aid in identification of the material, but will later be used to
complete the information on the herbarium label. Therefore, in summary, the field note should
give full details of any character which may not remain permanent with plants and are not
indicated by the plant itself.
The note should contain the following;
 State: specific state of collection

31
  Locality: village or town of collection
 Habitat: These include vegetation type, soil type, altitude etc.
 Habit: shrub, tree, herbs etc.
 Stem: bole, slash, buttress, branches
 Leaves: venation, prominent
 Flower: calyx, corolla, stamen, pistil, colour
 Fruit: shape of fruit, colour of fruit during collection

 PRESSING AND DRYING

Different methods of pressing include; Oven drying or Air drying.
The most important thing to do with freshly collected material is to dry it out as fast as possible.
This prevents fungal infections and preserves colour. It is an important aspect of plant collecting
that enough time be left at the end of the day to process the specimens. When plants have to be
left overnight they should be put in a cool place. Sometimes woody specimens can be placed in
water for a day or so to force buds or restore wilting leaves.
The collected specimen are then placed inside the folded sheets of absorbent papers
(newspaper), another flimsy is placed between the two specimens. Two or three double sheets of
absorbent paper should be placed against the wooden frames of the plant press to prevent the
specimen from being damaged by the wood. The plant press wooden frame containing the
specimens is tied firmly with strap/ twine to apply pressure. The plant should be carefully
arranged to show the upper and the lower surface of the leaves. Flowers should also be properly
displayed.
The plant press containing the specimen is then put in away in a cool place, not in sun for
about 24hours for drying. During drying, the absorbent papers (not the one in which the
specimen is placed) must be change every 24hours or more often if they become damp.

 TREATMENT WITH POISON

Treatment with poison (chemical) follows drying to prevent the plant from fungi attacks and
growth, and insect attacks. There are several methods of treating specimens from being attacked
by insect of fungi, these are;

32
 Chemical treatment: This is done by immersion /dipping the specimen into the poison or by
spraying the chemicals on the specimen. Examples of chemicals (poison) used are; Mercury
chloride, Methanol and insecticides like raid, baggon etc.
 Deep freezer treatment: Instead of using highly corrosive chemicals like mercury chloride,
storage of the specimens wrapped with polythene in deep freezer for a few weeks(2-3wks).
By this, all stages of insect development will be destroyed.

 MOUNTING OF SPECIMEN
Mounting is the process of affixing a dried pressed plant and its label to a sheet of heavy
paper. Mounting of the specimen follows treatment with poison. Specimens are attached to the
mounting sheets about 341/2 cm using glue. The mounting sheet is cut using paper cutter to
341/2cm. An efficient and effective way is to coat the surface of the mounting tray with glue
using painters brush. The specimen is then placed on the glue using forceps, and the specimen is
quickly transferred to the mounting sheet. If the glue is not evenly distributed, more may be
applied using a short brush.
All fragment, fruits and seeds should be placed in packets or envelops and these are affixed to
the Herbarium sheet at this time. A heavy board or weight is then placed on the sheet.

 LABELLING
A plant specimen is incomplete without label data. Label data is a form of field data and must
be accurate. Space should be left in the lower right hand corner for the herbarium label. Label
generally is applied using the same glue and put on before the specimen.
The following are important elements:
 Scientific name : genus, species, authority, intraspecific information
 Determiner of the scientific name: the name of the person who identified the plant
 Detailed location: the location is used by researchers on several levels
 Habitat : the type of plant community where the plant is growing and, if known, other
plants growing in association
 Plant Habit: describes the form of the plant (tree, shrub, vine, and herb) and its height.
Examples: tree, ca. 50 ft. tall. sprawling herb

33
  Frequency: is the plant rare, occasional, frequent or common?
 Plant description : describe characteristics of the plant which may be lost upon drying,
such as flower/fruit color and fragrance, leaf orientation and aroma
 Collector name: it is recommended that the collector be consistent and use their full first
name, middle initial (or full name) and full last name.
 Date of collection: a format with the month spelled out or abbreviated and 4 digit year
will prevent confusion. E.g., 22 Feb 2015, not 22/2/15 or 2/22/15.

 LAYING AWAY
Laying away is simply the arrangement of the pressed and mounted specimens into the
herbarium shelf or cabinet in a Phylogenetic order (Genetic relatedness) and alphabetical order.

USES /RELEVANCE OF HERBARIUM

1. To confirm the identity of plants by comparing with documented ones
2. Herbaria are essential for the study of plant taxonomy, the study of geographic
distributions, and the stabilizing of nomenclature.
3. Specimens housed in herbaria may be used to catalogue or identify the flora of an area.
4. Herbaria also preserve a historical record of change in vegetation over time. In some
cases, plants become extinct in one area or may become extinct altogether. In such cases,
specimens preserved in an herbarium can represent the only record of the plant's original
distribution.

 At this section, I also went to the field for botanical rambling(plant identification) of some
trees, herbs and shrubs under the supervision of MR OBA, where i knew some plants names
and their families, such as;

34
Plant name Common name Habit Families
Ficus thongii Igi Odon trees Moraceae
Triplochyton scleroxylon Igi Obese trees Sterculaceae
Tetraplura tetraptera Igi Aidon trees Ephorbiaceae
Vocanga Africana Apocynacea
Newbouldia laevis Biglonaceae
Persia americana Pear trees Lauraceae
Bixa orrelana Bixaceae
Antigonium leptopus Polygaceae
Petivera allaceae Anti snake shrub Phytolaceae
Cnidoscolus acontifoline Iyanapaja Ephorbiaceae
Entandrophragma angonese Igi Gedu tree Miliceae
Venonia amygdalina Ewuro Asteraceae
Costus afar Zingeberaceae
Bligha sapida Ishin trees Sapidaceae
Khaya senegalensis Meliaceae
Eugenia uniflora Mataceae
Pheltophtorum pterocapus Ceasaphinaceae
Citrus auratifolia Rutaceae
Telfaria occidentalis Ugu Cucurbitaceae
Platycerium aleicorne Polypodiaceae
Cassia fistula Ceasaphinaceae

Fig18: Herbarium cabinet Fig19: Plant press

35
Fig20: Mounted specimen Fig21: mounting tray

3.2.2 ENTOMOLOGY SECTION

Entomology is the scientific study of insects. The mandate of this Section is to carry out
research on Forest Entomology (Forest trees related insects), their Economic importance,
harmful effect and their control.
At this section, I learnt about Importance of insects, harmful effect and control, Apiculture,
Sericulture, and other aspect of entomology beside Forest Entomology under the supervision of
Mr Oloruntoba and Mr Gbade
Mr Oloruntoba, in his lecture explains insects as member of ecosystem found in every Habitat.
They play important role in food chain and food web; they help in conservation of ecosystem.
Usefulness of insects includes;
 Some insects are very edible with good proteins, which are under Entomophargy, e.g.
grasshopper, pupa stage of silkworm, cricket, locust etc.
 As nutrient recyclers; they break down large organic molecules which are made available
for new plants e.g. termite
 Some are producer of useful products e.g. Honeybee, silkworm etc.
 As a pollinator e.g. Order Lepidoptera
 As a specimen in the laboratory e.g. Drosophila melanogaster
36
 At this section, I had lectures on Sericulture and Apiculture which is part of the research
work at the section. The lectures were delivered by Mr GBADE.

SERICULTURE
Sericulture is the cultivation of silk through rearing of silkworm (Bombyx mori). It is an agro-
based industry. It involves the raising of food plants for silkworm, rearing of silkworm for
production of cocoons, reeling and spinning of cocoon for production of yarn etc. for value
added benefits such as processing and weaving.

 REARING OF SILKWORM
Silkworm (Bombyx mori), is a larva stage of insects called silkmolt (adult) which produce silk.
It is a monophagous insects i.e. it feed only on Mulberry (Morus alba) leaves. The egg stage
generally last for 10 days, if the diapause is broken. The life cycle is 25-30days. The silkworm
moult 4times to caste off the old skin. The pupal stage which is the formation of cocoon round
itself lasts for 8-10days.
SILK: Silk is a protein fibre produce by the silkworm for spinning a cocoon. There are two
proteins e.g. fibroin and sericin. These proteins are synthesized by the silkworm from the
mulberry leaf it feed on during its larva period.
MULBERRY: Mulberry constitutes the only food of Bombyx mori (Silkworm). Very hardy,
perennial and belonging to the genus Morus under the family of Moraceae. Mulberry is a very
quick growing plant and its leaves can be harvested several times in a year. The lean period is
about 6-8 months.

 MATERIALS NEEDED FOR REARING

These are the materials needed for successful rearing of silkworm.
1). Mulberry plantation: A period of 7-8 month plantation is needed. The mulberry is planted
through cutting. Regular pruning of old / brown foliage is necessary.
2). Rearing room 3). Mountage 4). Cutting knife 5). Formalin 6). Net 7). Cutting board.
8).Rearing tray etc.

37
 STAGE OF PRODUCTION
The stages of production are as follows;
 The rearing room for the rearing should be disinfected from manifestation of disease using
formalin. The disinfectant of materials used is done using formalin. The disinfectant is
sprayed on the materials using sprayer.
 The silk moth lays eggs. When the eggs are freshly laid, they are yellowish in colour, later
turn brown and later turn blue giving signs for hatching. An eggs layed by one female is
about 350-600 eggs.
 After the eggs have hatched, the larvae are spread out on Rearing tray to grow. They are fed
with chopped young mulberry leaves for a month.
 Foam is damp inside water and squeeze to drain. The foam is placed around the larva and
then cover with brown paper to keep the relative humidity of the new environment and to
prevent the mulberry from wildering or rotting.
 The larvae are observed for few days (10days) and the foams are removed. The larva may be
fed with young or old mulberry leaves.
 The insect have 4 larvae stages and they moult 4 times. During moulting the insect do not
feed. After the completion of the Larvae stages, they give sign for pupa stage (they started
raising their head).
 They are transferred from the Rearing tray to Montage; this serves as twig or corner for the
spinning of their silken cocoon, after 19-21days of feeding. The produce Cocoon; comprise
of Silk and Pupa. Good and quality cocoon is gotten when the Larva is feed with good and
quality mulberry.
 After 8days of Pupa inside the cocoon, the pupa secretes protolithic enzymes to break the
cocoon. So the silkworm pupa is render inactive by transferred into Ovum for 8days, after the
8days, they are immersed in hot water. In the hot water, the cocoon become softens; the
SILK are gotten and the pupa gets separated.
 To produce adult for the production of new eggs, the cocoon are preserved.

USES OF SILK
 It is used for sutures on human being in the hospital during surgery

38
  Used for preparing Parachute, because of the silk tensile strength.
 Dress materials like ready-made garments, curtains,stoles, insulating materials for wires
and cable.

Fig22: Preserved Cocoon Fig23: Montage

Fig24: New layed eggs Fig25: Mulberry plant(Bombyx mori)

39
DESTRUCTIVE EFFECT OF INSECTS ON FOREST TREES
Diseases in plants are grouped into those caused by Fungi, Nematodes, Bacteria, and Virus.
In his (Mr. GBADE) lecture, he explain the destructive effect of insect on Forest Trees and
Agricultural product using Iroko gall bug(Phytolyma species), that affect Iroko trees (Milicia
spp), as an example.
Iroko gall bug (Phytolyma species), is a pathogenic insects that attack IROKO TRESS species.
 Phytolyma lata attacks Milicia excelsa (Nigeria Iroko)
 Phytolyma fusca attacks Milicia regia (Ghana Iroko)
Iroko trees (MILICIA SPP), is a hardwood obtained from several Africa trees of the genus
Cholorophora, used for commercial timber.
Iroko gall bugs (Phytolyma species), belong to order Hemiptera. The insect attacks the
meristematic region of the trees seedlings, and bore hole into the trees, causing stunted growth
and lateral branching.

Ways to control / stop these attacks includes;

 The attacks and the infection can be prevented by bringing the MILICIA REGIA (Ghana
iroko) to EXCELSA (Nigeria iroko) for planting, vice versa, because PHYTOLYMA
LATA cannot attack MILICIA REGIA.
 Through biotechnology method(Gene/ DNA sequencing)
 Silvicultural method( i.e. growing of different species of trees together)
 Genetic transformation using Agrobacterium tumefaciens or Escherichia coli.

CONTROL OF INSECTS
Different ways of controlling insects are:
 Botanical control: this involves the using of plant extract to control insect. Some of these
extract are gotten from plants such as, Azadirachta indica, Angeratum connosoides,
cedrela odorata etc.
 Biological control: involve the using of one organism (e.g. insect) to control other insects.
e.g. using USCARIA LARIOPHAGA to control Cowpea beetle(Calobrocus maculatis).
 Chemical control: uses of synthetic chemicals e.g. insecticide
 Physical control: using trap or by hand picking

40
 Integrated pest management: this is the combination of more than one control measure
together with less involvement of chemicals
 Breeding control: development of disease resistant varieties.

COLLECTION OF INSECTS
At this section, I was lectured and worked on the collection and killing of insects.
Material for collection
Different materials used for collection of insects at the section are;
 Aspirator: The Aspirator has two mouth linked by a container, one mouth is placed
closed to the insect and air is drawn from the other side, at the same time the insect is
drawn into the container. They are used for the collection of tiny insects e.g. Aphid.
 Johnson Taylor Suction Trap: The trap is powered at the field using generator. The
fan inside the trap blows with high speed and drawn any insect flying at the
circumference of the trap fan into the container at the base of the trap. They are used
for flying insects that cannot be collected using Sweep net.
 Sweep net: they are used for collecting flying insects by sweeping the net on the
insect at the field.
 Light trap: for the collection of nocturnal insect (insects active at night)

KILLING OF INSECTS
Killing of insects is done by putting the collected insects into kilner jar and 2-3 drops of
ethanol or chloroform is added.

MOUNTING OF INSECTS
Mounting is the laying or setting of insect on a standard setting board using
Entomological pin after killing with relevant data attached to it. The entomological pin is
pierced through the thorax of the insect.

41
 Fig26: Johnson Taylor suction trap Fig27: Mounted specimen

3.2.3 PATHOLOGY SECTION

This section is coordinated by Mrs. Olusola. In her (Mrs Olusola) lecture, she defined Pathology
as a branch of medicine concerned with the study of the nature of diseases and its causes,
processes, development, and its consequences. She further list different Areas of pathology such
as;
 Forensic pathology Plan
 Anatomical pathology
 Histopathology
 Neuropathology
 Veterinary pathology
 Phytopathology(plant pathology)

42
PHYTOPATHOLOGY (PLANT PATHOLOGY)
Plant pathology is the study of plants diseases. A plant disease is any abnormal condition that
alters the appearance or function of a plant. It is a physiological process that affects some or all
plant functions. This Disease may also reduce yield and quality of harvested product.
A few examples of devastating diseases are that affects plants are:
 Potato late blight
 Ergot of Rye
 Black Stem Rust of Wheat
 Necrosis leaf spot disease
 Bacterial Canker on Citrus
All these are infectious diseases caused by plant pathogens such as Fungi, Bacteria, Nematodes,
Viruses and Parasitic Plants.
Control of plant diseases:
 Biological control: uses of organisms to control other organism
 Chemical control: using synthetic chemicals

FUNGI
Fungi are grouped into two;
 Micro fungi e.g. Penicillum spp, Aspergillus spp
 Macro fungi e.g. mushroom
A mushroom is the fleshy, spore-bearing fruiting body of a fungus, typically produced above
ground on soil or on its food source.
This section carry out research work majorly on the cultivation of 3 species edible mushroom.
These are;
 Pleurotus sajor-caju
 Pleurotus florida
 Lentinus squarosullus
At the section, I was taught how to cultivate mushroom and different substrate used.
MUSHROOM CULTIVATION
Materials needed are;

43
  Sawdust (from Tectona grandis, Gmelina arboreal, Triplochyton scleroxylon are good
for the cultivation of mushrooms.)
 Nylon bag, Powdered Lime(CaCo3), Autoclave machine or pressure pot, water,
Aluminium foil, Weighing balance, Wheat bran, Mushroom spawn.
Procedure for the cultivation of Mushroom using Gmelina arboreal sawdust as Substrate
 The Gmelina sawdust was spread to open air, to allow well dried
 1% of lime was dissolved in small quantity of water, it was well mixed with the dried
Sawdust
 Wheat bran was added to the moist Sawdust (Substrate) to improve the nutrient content
and to enhance yield
 The Substrate was packed into nylon bag, it was compressed to avoid space at the bottom
of the nylon bag and the nylon bag was tied with rubber band.
 The substrate was transferred to Autoclave machine for sterilization at 1210C for 2 hours
 After sterilization of the substrate, it was allow to cool
 The Mushroom spawn (mycelia) was Inoculated to the Sterilized Substrate in a sterile
laminar air flow
 After the Inoculation, the Nylon bag was tied and was arranged in Mushroom house
 Wetting of the Mushroom is done on daily basis

Fig28: Drying of Sawdust Fig29: Cultivated mushroom

44
3.2.4 WILDLIFE SECTION
This section is divided into 2 units; GRASSCUTTER UNIT and SNAILERY UNIT.
GRASSCUTTER UNIT
The mandate of this unit is Grasscutter Domestication and Multiplication.
At Grasscutter unit, under the supervision of Mrs. Shoronpe. In her lecture, she made me to
understand that Grasscutter is a large rodent animal, belonging of the order- RODENTA, family-
THRYONONIDEA and of the genus- THRYONOMYS. It is also called Cane rat because they
feed on Grasses and Canes. The section carries out research only on two major species of
grasscutter. These are; A giant grasscutter (Thryonomys swinderianus) and the dwarf grasscutter
(Thryonomys gregonomusterminch).

 GESTATION PERIOD: The gestation period of grasscutter is between 150-156days.

Maximum number of 10 offspring and minimum of 4 offspring.
 COLONY: Family of Grasscutter is called Colony, and a Colony of grasscutter
comprises of 1 Male and 4 Female. The Male grasscutter is called DOE and the Female
grasscutter is called BUCK.
 HOUSING: The grasscutter house is called HUTCH. The 3 major types of Hutch are;
Block hutch, Cage hutch, and Floor hutch. The hutches must b constructed in such a way
that it must prevent cold and predators.
 FEEDING: They are Herbivorous animal and majorly feeds on elephant grass
(Pennisetum purpurenum), Guinea grass. Feeding should be done early in the morning
and later in the evening. The forage should be added with Biovite supplement such as
pineapple, sweet potatoes, maize etc.
 PREDATOR: The common predator that attacks them is soldier ants and Snake. These
are prevented by pouring engine oil around their house and planting of anti- snake plants
such as Petivera alleaceae.
 DISEASES: The common diseases that affect them are PHEUMONIA. This is prevented
by covering their hutch with nylon during cold and removing it during heat for proper
ventilation
 SEX IDENTIFICATION: The male grasscutter have Roundish and Broad head, the
female have Elongated head

45
  REPRODUCTION: Grasscutter reproduce twice in a year. Day1 are called offspring,
day1-8weeks called Weaners. The sub- adult Female is between 8weeks – 6 month, the
sub- adult Male is between 8weeks – 8months. They attained Adult stage between
8months- 12month.

SNAILERY UNIT
The mandate of this unit is domestication and multiplication of Snails.The unit is under the
supervision of Mrs .T. Adeshina. In her lecture, she explained Heliculture as the domestication /
rearing of snails and Snailery as a place where Snails are bred or kept.
The unit work on the domestication of 3 different species of Snails which are; Acharchatina
marginata, Achartina fulica, and Achartina achartina
SNAILS belong to the phylum – Mollusca, and class - Gastropoda
IDENTIFICATION: A. marginata grows very big comparing to the other species. It is the
biggest of the species. A. marginata has round shell and other species having pointed shell. They
are nocturnal animal (active at night)
HOUSING: The housing made for them depends on the rearing plan; small scale e.g. cage and
large scale. The house must have soil (loamy soil), the soil in their house must be wet always.
FEEDING: They feed on Leaves e.g. cocoyam, pawpaw, cassava leafs. Fruits e.g. pawpaw,
tubers (yam and potatoes).Vegetable e.g. Water leaves, Amaranthus. They also feed on
household wastes.
PEST / PREDATOR: There pests and predator include, snake, lizard, rat and soldier ants.
REPRODUCTION: They are all Hermaphrodites. Acharchatina marginata lays bigger eggs than
the other species. It lays minimum of 3 Eggs, and maximum of 10/15 Eggs. When their eggs are
layed, they are buried in the soil for Incubation, and then hatch by itself on the 21days.
ECONOMICS / NUTRITION VALUE
 They are rich in protein, calcium, iron
 They are good for people with Cardiovascular disease
 It cures stroke and hypertension due to low cholesterol
 It serve as source of income

46
Fig30: grasscutter Fig31: Snails feeding

CHAPTER FOUR
4.1 SKILLS DEVELOPED AND TECHNIQUES LEARNT
During my SIWES attachment at Forestry Research Institutes of Nigeria (FRIN), the experience
gained was enlightening and eye opening. Majority of the works I was introduced to were not
new theoretically, but new practically.

At Sustainable Forest Management department;

 I learnt how to prepare tissue culture medium, disinfection of explants and Techniques in
Tissue Culture
 I learnt how to test for Seed Viability
 I learnt how to propagate Ornamental plants
 I learnt how pricked, thinned, hardened off and beating up seedlings
 I learnt how to Scarified seed that have dormancy
 I learnt how to identify plants (Shrubs, Trees, and Herbs) and families they belong during
Botanical Rambling.
 I learnt how to carry out Vegetative Propagation through Cuttings, Marcotting, and Layering.
 I learnt how to operate some laboratory machine and equipment which are not available in
my school.

47
At Forest conservation and Protection department;
 I learnt how to collect, pressed and dried, poison, mounted, labeled and laying away
herbarium specimens
 I learnt how to prepared and cultivate Edible Mushrooms
 I learnt how to multiply Mushroom Spawn
 I learnt how collect, mounted, and finish insects
 I learnt how to reared grass cutters
 I learnt how to prepared botanicals (plant extracts) used to control insect and pests.

4.2 CHALLENGES FACED AT WORK

The major challenge faced at the work was the issue of power failure which delayed many of
the practical and resulted in repetition of some of the analysis. Another challenge encountered
was the issue of inadequate laboratory furniture; most times we remain standing throughout the
day due to insufficient availability of furniture.

4.3 CONTRIBUTION MADE TO THE COMPANY

 I helped in the packing of top soil and river sand which was used to propagate
some seedling and cuttings
 I helped in fetching water for the watering of some propagated plants at the
company nursery unit
 I also gave them advised that the plants at the ornamental should be tagged for
easy identification
4.4 RELEVANCE OF STUDENT INDUSTRIAL WORK EXPERINCE
SHEME(SIWES) TO PLANT SCIENCE AND BIOTECHNOLOGY

48
 CHAPTER FIVE
5.1 GLOSSARY OF WORDS
 Explant: This is any portion taken from a plant that will be used to initiate a tissue
culture e.g. seeds, leaves, shoot, node, meristem, embryo etc.
 Thinning: This is done in the forest to create space by removing some excessive plants
to create space for others
 Pruning: This is the removal of excessive plants part e.g. Root, Branches to alter / reduce
their growth.
 Acclimatization: this is the process of changing the environment of plantlet raised in
growth medium to normal environment
 Laying away: This is the arrangement of mounted and labeling herbarium specimen into
the Herbarium cabinet
 Inoculation: Inoculation is the process of transferring disinfected / sterilized explant in to
prepared culture medium
 Dormancy:
 Totipotency: A cell characteristic in which the potential for forming all the cell types in
the adult organism are retained.
 Subculture: This is the process by which the tissue or explant is first subdivide, and then
transferred into fresh culture medium.
 Sterile Techniques: The practice of working with cultures in an environment free from
microorganisms.
 Agar: this is a polysaccharide powder derived from algae used to gel a medium. Agar is
generally used at a concentration of 6-12 g/liter.
 Hormones: Growth regulators, generally synthetic in occurrence, which strongly affects
growth (i.e. cytokinins, auxins, and gibberellins).
 Terrace : this is an area in the nursery where seedlings are be laid in bed
 Harden –off: The process of gradual exposing of seedlings grown under weaning shed or
green house to the field.
 Etiolation: The process of raising plant without much sunlight i.e under partial shade.
 Domestication: This is the bringing of plants from wild and planting / propagating them
to man living environment.

49
 Pricking – out: This is the transplanting or removal of young seedlings from polythene
pots to the green house when its first true leaves emerge.
 Scarification: This the process of cutting the seed coat or ileum of dormant seeds for
easy imbibed of water.
 Beating –up: The act of replacing dead seedlings of plants with new ones
 Diapause: is a temporary pause in the growth and development of insects due to adverse
environmental condition
 Heliculture: this the rearing or domestication of snail

50
5.2 RECOMMENDATION
The industrial training should be highly encouraged in all institutions, most especially
Universities. As a result of the technical and practical aspect of education being acquired during
this period of training, I wish to suggest that the industrial training fund (ITF) should ensure
proper and thorough supervision of students, not only by the institution-based supervisors but
also the ITF officials. I hereby recommend FRIN to Plants science and Biotechnology students
that have interest in industrial and research work because is well equipped and their research
works deals with plant relatedness with wide range of raw materials and upgrades of indigenous
production techniques.

5.3 CONCLUSION
Students’ Industrial Work Experience Scheme (SIWES) was able to expose me to get acquainted
to work experience and to cope in labor market. It provided an avenue for me to understand the
practical aspects of the theories I was thought in school. It has given me the privilege to handle
various machines and equipment relevant to plant science and biotechnology and be well
grounded in their mode of application and operation.

The industrial training has been highly beneficial in experience, knowledge, innovativeness,
sense of responsibility, and interpersonal relationship with work mate and factory workers. This
training also gave me the opportunity to interact, share knowledge and ideas with other students
from different institutions.

During this period of six months, I have been privileged to personally bridge the gap between
my practical and theoretical knowledge to an extent. Indeed, the SIWES programme has been an
eye opener.

51