DEBRE TABOR UNIVERSITY

COLLEGE OF AGRICULTURE AND ENVIRONMENTAL SCIENCE

DEPARTMENT OF HORTICULTURE
For Practical Attachment Course (HORT392)
Title:- micro clonal propagation of bamboo

By:- Fanaye Kelile

ID no: - 0649
SUBMITTED TO DEPARTMENT OF HORTICUITURE
ACADAMIC YEAR 2016EC
SEMISTER II
DEBRE TABOR,ETHIOPIA
P.O.BOX 27
TABLE CONTENTS

Acknowledgment
Executive Summary
CHAPTER ONE.................……........................
……….................................1
1 INTRODUCTION............................................
……………………................1
1.1 Historical Background of EFD and Establishmen......
……………………..1
1.2 Main Function or Core Activity of
EFD..................................................2
1.3 Vision and Mission Statement of the
Institution....................................2
1.4 Organization Structure of the
Site.........................................................2
CHAPTER TWO
2.1 Staff establishment of the department in the term of number of
employees and roe and responsibilities
....................................................................
2.2. Main activities….....................................................
……........................
2.2.1defination of Tissue
cculture ............................................................
2.2.3
advantage ................................................................... ..................
..
2.2.4 micro clonal propagation of lowland
bamboo .....................................
2.3.1 source of Experimental
material........................................................
2.3.2 componet of culture
media..................................................................
2.3.3 culture media
preparation...................................................................
2.3.4 explant surface
disinfection........................................... ...................
2.3.5. Establishment of culture
shoot………………………...................... ......
Chapter three
3 Evaluation of the attachment
period......................... .................................
3.1 Benefit you gained from the
attachment.....................................................
3.2 descriptions of the strength and weakness of assignment that
you were handling..………………………………………….
3.3 description of the threats and opportunities of organization that
you doing your professional practices...........................................
3.4 possible problms that you have noticed respect specific
task.................
3.5 conclusion...............................
………………..............................
4 Recemendation.......................
……................ ...... ..................................
5
Reference ........................................................................................
......
 Acknowlegment

First of all I would like to thank you my god to I went to

appeciata to Ethiopian forest development research
center supervisor Dr, mehari for his follow up and parently
approach at every stage during for leering in that site’s am
also great foul to Mr. Adugnwu adimas lab techinicial of
EFD for given me to conduct my practical attachment in
the site and thank you for all community of in that worker
for help me.

I
I
 Executive summary

Ethiopian forest development is a critical component of

the country's environmental conservation and sustainable
development efforts. The government of Ethiopia, in
collaboration with various stakeholders, has been
implementing initiatives to address deforestation, promote
afforestation and reforestation, and ensure the sustainable
management of its forest resource.Key activities in
Ethiopian forest development include the planting of native
tree species, protection of existing forests, promotion of
sustainable land-use practices, and engagement of local
communities in forest conservation. These efforts aim to
enhance biodiversity, mitigate climate change, improve soil
health, and provide livelihood opportunities for rural
communities dependent on forest resources. Challenges
such as deforestation, land degradation, climate change,
and unsustainable land-use practices continue to threaten
Ethiopia's forests. To address these challenges, the
government has developed policies and regulations to guide
forest management practices and promote sustainable
development.

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II
 CHAPTER ONE
1. Introduction
Bamboo is a perennial grass belonging to the Poaceae
(Gramineae) family and Bambuseae subfamily. Bamboos
have a long history as an exceptionally versatile and widely
used resource in the world. Nowadays, it is becoming so
increasingly important in the world’s forest economy,
because it is: a superior wood substitute, cheap and
efficient to produce and utilize, environmentally friendly
and the world forest is shrinking hence requiring
alternative sources.
There are two indigenous bamboo species in Ethiopia
namely lowland bamboo (Oxytenanthera abyssinica A.
Rich. Munro) and Ethiopian highland bamboo (Arundinaria
alpina K. Schumach) (Phillips, 1995). These two species are
found restricted in limited agro ecological regions, i.e.
highland bamboo in highland areas of altitude 2200-3500
ma.s.l. and lowland bamboo from 500-1800 ma.s.l
(Yigardu, 2016). Ethiopian lowland bamboo is used for
housing, handicrafts, pulp and paper industries, energy
source, and food; it fits ISO standards for the production of
an array of lumber-based and stick-based products. It has
also high value in carbon sequestration (EFCCC and
INBAR, 2020). Medical use of O. abyssinica is documented
in different countries including Ethiopia [4]. O. abyssinica

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has also important phytochemicals with a resultant
antioxidant property (Bartholomew and Maxwell, 2013

1.1 Historical Background Of Ethiopian Forstry

Development

The Ethiopian Forestry Development is an Autonomous Federal

institution established by The Federal Government of Ethiopia
Council of minister EFD was resulted by merging together
( The Ethiopian Enviromental and Forest Research Institition
(EEFRI)
Research Center(FRC) and The then wood Utilization Research
Center (WURC) in 1975 and 1979 respectively under the
forestry and Wildlife conservation Development Authority
(FaWCDA) EFD headequarter is located in addis ababa. Yeka sub
city.wereda 9 (Gurd shola)
behind athletics federation and Ethiopian construction works
corporaton building

1.2 Main Function or Core Activity of Ethiopian Forestry

Development

Ethiopia's forests serve key sectors of the economy with

immense potential to contribute to social and economic
development through maximizing a range of environmental,
ecological, economic, and social services.
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1.3 Vision and Mission Statement Of Institution
Mission
promote and ensure the protection and conservation of the
environment as a valuable asset for the people of Ethiopia.
To protect our people and the environment from harmful
effects of Pollution, Climate change and Biodiversity loss.”
Vision
All Ethiopian Society and sectors of development value and
undertake sound environmental management to clean,
healthy, safe, and protected environment that supports the
national aspiration to be African bacon of prosperity by
2030.

1.4 Organizational Structure Of Site

EFD is the federal autonomous governmental institution

organized under the Ministry of Agriculture and led by
DIRECTOR GENERAL. Here is a complete organizational
hierarchy.

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 Objective
To create awarnes in The fild and in the office
To get different skill about horticulture
To get adoption in different work about horticulture
To get practical knowledge about horticulture

CHAPTER TWO

2. HOST ATACHEMENT DEPARTMENT ORGANIZATION

2.1 Staff establishment of the department in the term

of number of employees and role and activity

4
There are 501-1000 employees in the institution in
different role and responsibility picket to manager.
Dr. Wubalem Tadesse……………….manager

2.2 Main Activities

2.2.1 Defination of Tissue culture
Tissue culture is a technique in which fragments of
plants are cultured and grown in a laboratory. Many times
the organs are also used for tissue culture. The media used
for the growth of the culture is broth and agar.
This technique is also known as micropropagation. It has
proved beneficial for the production of disease-free plants
and increase plant yield in developing countries. It only
requires a sterile workplace, greenhouse, trained
manpower, and a nursery.
Oil palm, banana, eggplant, pineapple, rubber tree, tomato,
sweet potato have been produced by tissue culture in the
developing countries.

2.2.3 Advantages of Tissue Culture

Following are the various advantages of tissue culture

technique:
The plantlets are obtained in a very short time with a small
amount of plant tissue.

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The new plants produced are disease-free.
The plants can be grown throughout the year, irrespective
of the season
A large space is not required to grow plants by tissue
culture technique.
The production of new varieties in the market place speeds
up.
This technique is being used for the production of
ornamental plants such as dahlia, chrysanthemum,
orchids, etc.

2.2.4 Micro Clonal Propagation Of Lowland bamboo

Material and Method

2.3.1 Source of Exipermental material

The seeds for this study were obtained from Assosa,
Healthy seeds were selected carefully and they were stored
in plastic bag at +4°C in refrigerator.
Culture Media Preparation
Sterilisation of material in Autoclave at 121oC FOR 15
minutes

2.3.2 Culture Medium components

One of the most important factors governing the growth
and morphogenesis of the bamboo tissues in culture is the
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composition of the culture medium. The basic nutrient
requirements of cultured plant cells are very similar to
those of whole plants. The basic requirements of mineral
elements required for the growth of plant tissues are
fulfilled by providing their common salts in the medium.
 Macronutrients.
 Micronutrients
 Vitamins
 Amino acids or other nitrogen supplements
 Carbohydrates or sugars
 Solidifying agents or supporting systems and
 Growth regulators (plant hormones)

2.3.3 Culture media preparation

Componet Element 1000mg/L 500mg/L

MS media

Macro Potassium 1900.00 9.5

nutrients calicum 440.00 2.2
maginsum 180.00 1.85
ammonum 1650 8.25

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Micro Zinc 2.23 1.115
nutrients sulphate 0.86 0.43
copper 0.62
sulphaet 0.3
0.246
maganes 0.123
sulphat
molybdi acid
Ironestok Iron sulphat 1.8 0.9
Iron chlorid 0.14 0.07
Vitamin Thiamine 0.04 0.02
pyridoxine 0.04 0.02
nicotinic 0.1 0.05
Potassium 0.08 0.4
iodide

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The medium consists of inorganic salts and vitamins of MS
medium, 30 g/L sucrose, 100

mg/L myo-inositol and 8 g/L agar Using:

1. powdered MS medium (salts and vitamins) 4.4 g/

2. Sucrose
3. myo-inositol stock solution (10 mg/ml)

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4. Agar
The pH is 5.8
In 250 ml clean beaker, put 100 ml distilled water and
dissolve:
1.1 g powdered MS medium
7.5 g sucrose
2.5 ml myo-inositol stock solution (10 mg/ml)

Up to 225 ml with distilled water

Adjust pH to 5.8

Up to 250 ml with distilled water

Add 2 g Agar and boil with continuous stirring till

disappearance of Agar

Divide into 5 jars

Autoclave for 20 minutes at 121 o

Preparing Stock Solution
for each cultutre media to pick 10ml in the Baker
Then prepared stock solution for growth of bamboo
Measure pH value for the growth PH 5.2-5.7
Add agar to soldify solution
Use maginetic sterile to dissolve agar

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Give heat in oven dry

2.3 .4 Explants Surface Disinfection

➤ Selected healthy seeds were sterilized to get ride –off all

micro –organisms.
➤ Also, the seeds were washed with tape water to remove
debris.
➤ Then, to get clean seeds it soaked in distilled water for 2
hrs by shaking and washed by double distilled water
(DDW) with liquid soap with 2-3 drops of Tween −20 for 20
minutes.
➤ Then, treated by antifungal of mancozine 20 g/l for 20
minute and washed the seeds with DDw three times.
➤ By following those procedure, seeds were treated by 2%
of NaOCl for 20 minute and washed the seeds three times
by 2-3 drop of Tween-20 for five minute.
➤ After pre-treatment, the seeds were treated with 1%
NaOCl and washed three times by DDw.

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➤ Finally, it treated with 70% ethanol for 30 seconds
under laminar air flow cabinet. After sterilization of the MS
medium,
➤ for shoot initiation three jars for each treatments (0.003,
0.004 and 0.005 g/L BAP) five seeds were placed randomly
in completely randomized design (CRD) arrangement.

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Leveled jar

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2.3 .5 Establishment of Culture Shoots

Disinfected seeds were cultured in 9 jars that contained the

above mentioned nutrient 50 ml of MS medium with BAP
and 3 jars PGRs free medium for shoot initiation study.

. Shoot Multiplication
To avoid the carry over effect of shoot initiation media
during shoot multiplication, initiated propagules consisting
of three shoots each were subculture on PGRs free MS
medium for two weeks. Each propagule was placed
vertically and lightly pressed into the culture medium
supplemented with of 0.003-0.005 g/litre of BAP with each
activated charkol for inhibition of oxidants of the cells
mostly for phenol exudation. MS medium without PGRs
was used as control. 12 jars each with three propagules
were used and kept under light conditions. Then, after two
weeks multiplication of new leaf were best at 0.004 g/litre
of BAP as it showed in Figure 1
.
. Rooting of Shoots
The in vitro regenerated three shoots in bunch were used
for rooting studies after sub-cultured on PGRs free MS
medium for 2 weeks. The rooting response of these shoots
was studied on different concentrations of IBA (0.004,
0.005 and 0.006 g/L) and with for each treatments used
0.1 g/L activated charkol for inhibition of oxidants of the

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cells mostly for phenol exudation on MS medium and
without hormone was used as control. For each treatment
three jars, each with three clumps were used. All shoots
were incubated on rooting medium for 4 weeks and kept
under light conditions in culture room in CRD
arrangement. Among all treatment, 0.005 g/L of IBA
solution experiment were best for root formation as it
shown in Figure 2 .

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 Chapter three

Evaluation of the attachment period

3.1 benefit you gain from the attachment

Better knowledge retention

To get understand plant growth in Laboratory
To develop Interpersonal skill
Improve skill set
Something that need to be experience to be understoode

3.2 descriptions of the strength and weakness of

assignment that you were handling

3.2.1 Strong side of institute

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Favorable enviromental condition
Time managment
The expert use standard Instrument
Full filling material

3.2.2 Weak side of institute

Lack of electricity power

3.3 description of the threats and opportunities of

organization that you doing your professional
practices

To opportunity media preparation by Itself

To supply of tissue culture material
Threat by lab expert

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3.4 possible problems that you have noticed respect
specific task

Lack of transportation

3.5. Challenges encountered by the student during

the attachment period.

3.6. How the challenges were solved

4 Conclusion
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5 Recommendations

5.1 To the university

The university should improve the practical attachment in

the winter time and for longer time.

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 5.2. To the Research center

The research centers should be schedule to show us for

practical
Should have skillful human resources with numerous

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References

Arya S, Sharma S, Kaur R, Arya ID. (1999).

Micropropagation of Dendrocalamus asper by shoot
proliferation using seeds. Plant Cell Report. 18:879–882.

Arya ID, Kaur B, Arya S. (2012). Rapid and mass

propagation of economically important bamboo
Dendrocalamus hamiltonii. Indian Journal of Energy/
1(1):11–16.

Banik RL (1995). Selection criteria and population

enhancement of priority bamboos. INBAR Technical Report
N°5 Genetic enhancement of bamboo and rattan INBAR
New Delhi.

ereket H. (2008). Study on establishment of bamboo

processing plants in Amhara Regional State. M.Sc. Thesis
Addis Ababa University, Addis Ababa, Ethiopia

Demelash A, Zebene T, Yared K. (2012). Effect of storage

media and storage time on germination and field
emergence of Oxytenanthera abyssinica
seeds. International Journal of Basic and Applied Sciences,
1(3):218–226.

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