Adama Science And Technology University

Plant Tissue Culture Challenges in Ethiopia and Alternative

Options for Low-Cost Media

Seminar II Submitted To
Department of Applied Biology
School of Applied Natural Science
By
Kasahun Amare
Supervisor: Geleta Dugassa (Ph.D.)

June 2022
Adama, Ethiopia
1
 Presentation outlines
1. Introduction
2. PTCs in Ethiopia and current status
3. The major challenges of PTC
4. Low-cost PTC options
5. Conclusion

2
 1. Introduction
• The growth of plant organs, tissues, cells, or protoplasts in
specially formulated nutritional media under sterile and well-
controlled conditions is known as plant tissue culture(PTC).
• With the increasing demand for agricultural products, PTC
market was valued at US$382.305 million in 2020 and
• It is also projected to grow to US$895.006 million by 2030,
with annual growth rate of 8.5% from 2021-2030.
• It is estimated that the current market of tissue cultured
products is 500 million to 1 billion plants annually.
• Due to labor cost in the developed countries, the PTC
system has been challenged by high production cost and
low gross profit.

3
 Cont.…
• To meet market demand and provide planting material, production must be
plentiful.

• However, one of the major limitations of PTC is

• The high cost of culture media, facilities, energy, and labor
• Inadequate funding and
• Poor mgt practices, and
• Staff turnover are serious bottlenecks.
• Due to these small and medium-sized labs with limited resources cannot
take advantage of PTC.
• These difficulties have been solved by
• the development of reliable, and inexpensive methods
• Alternative materials such as solidifying agents
• isabugol, sago, cassava starch, barley starch, and bulla can be
substituted for agar
4
 Cont.…
• Carbon sources such as
• table sugar, potatoes, cane juice, sugar cubes, and
brown sugar
• Other options
• liquid media, reusable glass beads and bioreactors, can
reduce production costs
• The intensive labor along with the production cost there is
need to develop scale-up systems and automation.

• In this case, this paper aims to review the current

challenges of PTC in the development of various low-cost
alternatives in Ethiopia.

5
 2. PTCs in Ethiopia and Current Status
• The scope of biotechnology in Ethiopia includes
• Several institutions with a total of 24 branches
• Dealing with
• Tissue culture
• molecular markers,
• Embryo transfer,
• Immunology, vaccines,
• Diagnostic kit development, epidemiology, and genetic
diversity.
• The most widely used technique in agricultural
biotechnology is PTC.

6
 Cont.….
• In Ethiopia, training in biotechnology has been initiated in
AAU, dating back to the 1980s and 1990s.

• Following AAU, other institutions of higher learning in the

country have started graduate and undergraduate programs.

• However, a more comprehensive and concerted PTC

research program was rolled out by EIAR.

• Regional agricultural research institutes such as

ARARI(Bahir),
SNNPRARI (Areka),
ORARI (Adami Tulu), and
TRARI (Mekelle)
7
 Cont.…
• In addition, PTC in private companies; for example,
• VCI was active until 2017,
• Narus Biotechnology, ceased operations.
• Waginos Biotech operational focused on ornamental
plants for local market and small-scale seed potato
plants.
• Tigray Biotechnology Center,
• Dessie Tissue Culture Center, and
• Bahir Dar Tissue Culture Enterprise
• Play a role in the commercialization and dissemination of
research outputs.

8
 Cont.…
• Currently, there is a fertile ground to exploit advantage from
biotechnology, particularly from PTC.

• Efforts came to results and commenced to solve the

prevailing shortage of planting materials.

• Tigray Biotechnology Center alone had a production

capacity of 40 million, and

• Dessie Tissue Culture Center has 20 million.

9
 3. The Major Challenges of PTC
• Ethiopia, cannot benefit from modern biotechnological.

• Thus, emerging subject both as

• academic subject
• industrial application.
• Few research and academic institutes in Ethiopia are
engaged in basic, advanced and research in biotechnology
with limitations in

• tools, techniques, and management as well as financial

constraints.

10
 Cont.…
funding and supplies in government institutions,
 low management autonomy in maintenance,
technical and administrative problems,
 limited capacities, improper networking
• In addition,
• Finding skilled workers.
• Prices for media and PGR, equipment, and energy
sources remain a challenge

11
 Cont.…
• To realize the potential of PTC in Ethiopia, developing
solutions to these limitations is necessary.
• The government established a national biotechnology
roadmap, which will
• Design, implement policies and strategies, prioritizing the
agricultural sector.
• Public universities, research institutes, and some private
companies play a role in producing a skilled labor forces
• Alternative options for culture media,
• Carbon source
• Macro and micro nutrients
• Galling agents are researched
12
 3.2. Inadequacy of Funding
• The early emergence of biotechnology in industrialized
countries was the strong background of budgets

• Various countries pay special attention to the development

of biotechnology and allocate sufficient funds to R&D.

• For example,

• Lithuania (29%), Austria (3.3%), Germany (2.8%), and the Russian

Federation (1%) spent by the fiscal year of 2020 on the research
and development of biotechnology companies.

• R & D by biotechnology companies as a proportion of the

indicator for the country's research focuses on modern
biotechnology.

13
 Cont.…
• In FY2019-20, 0.16 billion Birr for the Ethiopian Institute of
Biotechnology was funded,
• However, most public and private labs are foreign fund
source dependent .
• Some of the labs are for tissue culture, and others are for
molecular biology.
• Therefore, the fund is not as sufficient to undergo plant
biotechnology tasks as needed.
• Due to high costs associated with this,
• long-term planning is required when allocating resources.
• Medium-term, limited operating budgets poor performance

14
 Cont.…
• Therefore, 4 separate steps must be recognized in the
funding process
• Funding to establish the physical laboratory
• Funding to purchase the critical mass of equipment
needed for molecular work.
• Funding to cover operational costs of expensive
reagents, renewable consumables, and technical
support.
• Funding to specific research projects for the period
necessary to obtain results.

15
 3.3. Skilled Workforce
• Labor is a major expense at PTC in developed countries.
• On the other hand, labor in developing countries is
generally cheaper.
• The typical cost profile of a tissue culture facility production
system shows that
• 40% of the costs are labor,
• 10% are materials,
• 20% are overhead, and
• 30% are sales, general and administrative operations.
• However, in the EU, labor can account for 60-70% of the
cost of plants produced in vitro.

16
 Cont.….
• Another bottleneck is
• supportive staff for repair and maintenance of
equipment.
• To overcome the scarcity of manpower, there should be a
system that encourages
 Students to attend universities and research centers
Others technology transfer and alliances, are needed.
• To reduce staff turnover,
Achieved by providing significant incentives.
 Incentives encouraging employees to be more loyal and
productive.
17
 3.4. Poor Facilities
• The physical components of PTC include
equipment and
buildings with a operational rooms,
soil growing area (greenhouses, plastic tunnels),
chemical storage area, containers and extras.
• The size of the physical components of PTC depends on its
functional requirements.
• In Ethiopian, PTC labs are characterized by:
 inadequate infrastructure, and
 unreliable utility services (electricity and water supply).
• Therefore, finding a reliable solution to such problems
requires adequate
funding and maintain facilities to operate.
18
 4. Low-cost PTC Options
• cost reductions are achieved by
• improving process efficiency and better use of
resources.
• is a high priority in agriculture, horticulture, forestry, and
floriculture
• Several low-cost alternatives can be used to simplify various
operations and reduce costs. For example,
• cost of electricity, culture vessels, media, chemicals,
devices, and instruments for micropropagation.

19
 4.1. Low-Cost Options for Sterilization
• The success of micropropagation can be influenced by
• effectiveness of surface sterilization,
• type of culture medium, and
• mgt of browning symptoms.
• Various reasons have been proposed for contamination. one
reason is
• method used to sterilize the explants, tools, and
equipment.
• Another concern is the external contamination
• explants from contaminated tools, equipment, and
workers in media preparation and culturing.

20
 Cont.…
• Surface sterilization of explants is one of the critical steps in PTC.
• Commonly used sterilants are
sodium hypochlorite (NaOCl),
calcium hypochlorite Ca(ClO)2,
mercuric chloride (HgCl2),
silver nitrate (AgNO3), and
hydrogen peroxide (H2O2).
• Accordingly, the use of locally available materials like
• Berekina 5% chlorine for 25 min
• JIC 3.5% NaOCl for 20-45 min cowpea, rice, and sorghum explants
• JIK at 40% v/v containing 1.5% NaOCl for 20 min cane shoot tips
• Pressure cooker
21
 Cont...

Figure 1: Substitution of preparation room equipment

with low cost options has been successful

22
 4.2. Low Cost Options for Culture Vessels
• Several types of culture vessels
are used in PTC labs, such as
• lab-specific culture jars,
• Erlenmeyer flasks,
• magenta bottles, and
• Petri dishes
• To reduce the costs, locally
available glass jars that are used
for food canning can be used in
place of traditional vessels.
• Hence, at least 85% of the cost
could be reduced.
Figure 2: Food canning culture vessels
may be used in PTC (45, 60, and 100 mL
23
left to right)
 Cont.…

Figure 3: Costly, delicate pipettes and Cylinders Replaced

by cheap, hardy syringes

24
 4.3. Bioreactor-Based Low-Cost Option for PTC
• Bioreactors are vessels designed for large-scale cell, tissue,
or organ culture in liquid media.
• An increasing number of plants have demonstrated several
important advantages over
• traditional semisolid micropropagation,
• multiple increases in propagation rates and a
• reduction in space, energy, and labor.
• The major advantages of bioreactor
• scaling up in a shorter time limit,
• reduction in the production cost and automated control
of physical and chemical environments

25
 4.4. Low Cost Options for Light
• Artificial light generates heat that must
be dissipated by cooling, and the fan
further contributes to the electrical load.
• Light is a very expensive and ineffective
method in tissue culture technology.
• In developing countries, electricity costs
can account for up to 60% of production
costs.
• While the need for electrical power is
essential, adopting low-cost options can
reduce production costs.
• the use of light-emitting diodes in
micropropagation can reduce electricity
prices by 50 to 75%
• This is done by replacing artificial light
with natural light .
Figure 4:Natural light
26
 4.5. Low-Cost Options for Distilled Water
• Double distilled water is the main component of all tissue
culture media and is considered free of ions and impurities.
• However, distilled water produced through electrical
distillation is expensive and adds to the cost of tissue
culture;
• Cheaper water sources that could reduce the cost of the
media without altering the composition are needed.
• Tap water (free of heavy metals and contaminants) can be
substituted for distilled water
• Reverse osmosis water can be used for stock solutions
and hormone preparations,

27
 4.6. Low-Cost Options for Media Components
• For semisolid and solid media, a typical PTC medium
includes a base solution containing
• major and minor minerals,
• carbon source (sucrose),
• vitamins, growth regulators, and a gelling agent
• However, there is an ongoing search for alternatives
• The major goal is to substitute nutritive solutions derived
from less expensive sources for macro- and micronutrients,
sugars, and gelling agents.

28
 4.6.1. Low-Cost Options for Gelling Agent
• Agar is widely used most expensive component of PTC
usually applied at 0.6-0.8% w/v.
• Various inexpensive sources,
• potato flour, rice, barley, wheat, bulla, and corn flour, have been
used.
• Combination of laundry starch, potato starch, and semolina
reduces the cost by more than 70%
Bulla: was first used for the micropropagation of pineapple
• The production costs were reduced up to 76%
• 60 - 100 g/L bulla reduced the cost of vanilla by 50 - 72.5%

29
 Cont.…
Isubgol:
• The price of "Isubgol husk" is cheaper than agar
• Replacing agar 0.8% with Isbugol (1.5%) produced a much
similar response to agar for callus formation in sugarcane
• Isubgol at 30 g/L was cost-effective for best growth of
shoots, leaves, and roots of orchids
Potato, Barley, and Corn Starch:
• Starch is cheapest and its use may reduce costs.
• corn, potato, and barley starches at 50 g/L as a
solidifying substitution for the micropropagation of potato.
• The highest cost reduction was 61-66% observed.

30
 Cont.…
• Sago starch: is a complex polysaccharide that serves as a
storage product in a variety of plants.
• It contains a small number of sugars, fiber, protein, calcium,
and other minerals.
• Sago, as a gelling agent, had a significant influence on
shoot proliferation and in vitro rooting of
• ginger, turmeric, and banana.
• Sago at 70 g/L gave proper solidification and normal growth
in ginger and turmeric.
• Sago at 80 g/L for banana reduced the media cost by
69.69%

31
 4.6.2. Low-Cost Options for Macro and Micro Nutrients

• Elements in large quantities,

N, P, Ca, P, Mg, and S.
• Elements required in small amounts
Fe, Cu, Zn, B, Mo, Mn, and Cl
• Low-cost options.
• macronutrient (monopotassium phosphate, potassium fertilizer,
Epsom salt, and ammonium quarry salt) and
• micronutrients (stanes iodised microfood) .
• Cost Reductions of 87.8% to 68.6%, for macronutrients,
and micronutrients, respectively.
• Easygro vegetative fertilizer reduced the cost by 96.2%,

32
 Cont.…
• Hydro Agri’s fertilizer is also another alternative option to
replace MS media for micropropagation of cassava,
• resulted in a cost reduction of 93.1%

• In addition, palm oil mill effluent is a free and

nontoxic material that contains valuable
macronutrients required for plant growth.

33
 4.6.3. Low-Cost Options for Carbon Source
• In plant cell culture media, sucrose is mainly used as an
energy source.
• However, fructose and glucose may be used.
• Generally, 20-40 grams of sucrose is added to each liter of
liquid medium.
• The unavailability and the prohibitive cost of lab-grade
sucrose are among the major constraints of PTC labs found
in developing countries.
• Alternatives such as: Table sugar, cane molasses, banana
extract, and coconut water are good for reducing costs.

34
 Cont.…
• The cost of potatoes TC reduced by 34 to 51% utilizing table
sugar

• Table sugar at 30 g/L was able to support the growth of the

banana plantlets.

• Low-cost alternative gelling and carbon sources will

significantly reduce production costs by 90-97%.

• Vermicompost 50 g/L+ table sugar 30 g/L, wheat flour, and

coconut water reduces production costs by 58 to 97 percent

35
 5. CONCLUSION
• PTC is considered to be a mass propagator of agriculturally
important crops within a finite period regardless of the
season.
• However, its application is hampered by costs required.
• One solution for developing countries to use PTC
applications is to
• replace low-cost media alternatives, gelling agents,
locally available culture vessels, natural light, tap water,
table sugar as a carbon source, and others.
• Furthermore, the government should pay attention to
• funding, training, provide incentives, and focus on
expanding laboratory facilities
36
 The end

Thank you

37