Plant tissue culture biotechnology in Ethiopia

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ISSN : 0974 - 7532
Volume 7 Issue 7
Research & Reviews in
BioSciences
Trade Science Inc.
Review
RRBS, 7(7), 2013 [249-255]
Plant tissue culture biotechnology in Ethiopia: Challenges and
opportunities
Mohammed Adefa Seid
Arba Minch University, College of Natural Sciences, Department of Biology, P.O.Box 21, Arba Minch, (ETHIOPIA)
E-mail : [email protected]
KEYWORDS
ABSTRACT
Ethiopia is the second most populous nation where 80% of its inhabitants
acquire their livelihood from agriculture. However, the long-lasted
subsistence agricultural mode of production of the country could not
enabled the population to increase agricultural productivity and boost
the national economy. In this regard, plant tissue culture technology is
the likely opportunity for Ethiopian agricultural system towards improving
agricultural yields. Indeed, so far, conventional breeding and selection of
plants and animals played a vital role in agricultural productivity. Yet,
unlike many developed nations, Ethiopian is at initial stages of getting
benefits from modern biotechnological products. But, nowadays, Ethiopian
government is giving due attention towards designing and implementation
of policies and strategies related to biotechnology, particularly in
agricultural sectors. In fact, it remained difficult to use such tools to obtain
solutions for major agricultural problems due to lack of resources and well
skilled personnel. But, state universities and research institutes, and some
private enterprises, are playing a promising role in conducting research
and producing skilled manpower in the area of biotechnology. In this
regard, Ethiopian Institute of Agricultural Research (EIAR) and Addis
Ababa University take the leading role in doing researches related to
plant tissue culture biotechnology in agricultural sectors so as to produce
drought and diseases resistant crop varieties. Though biotechnology and
genetic engineering has a significant contribution by addressing
environmental and food safety concerns with rigorous bio-safety
regulations, it could also cause hazardous problems upon human health
and the ecosystem when it is applied irresponsibly and unsafely.
 2013 Trade Science Inc. - INDIA
INTRODUCTION
Ethiopia is the second most populated nation in
Africa where agricultural sector is the leading national
income supporting more than 80% of the population[1].
Ethiopia;
Tissue culture;
Biotechnology;
Micropropagation;
Agriculture;
Explants;
GMOs.
Ethiopian agricultural system is predominantly characterized by high level of subsistence production and low
improvement of traditional farming practices resulting
in declining of agricultural productivity. In fact, persistent dependency on rainfall and frequent occurrence of
250
Plant tissue culture biotechnology
in Ethiopia
.
RRBS, 7(7) 2013
Review
drought and other natural calamities add up for low
agricultural production.
Hence, application of tissue culture biotechnology
in the field of agriculture seems very crucial so as to
increase agricultural productions for the purposes of
feeding the population with no need of international aids.
Tissue culture is an in vitro culture of cells, tissues,
organs or the whole plant parts under aseptic and controlled nutritional and environmental conditions often to
produce the clone of plants. Nowadays, plant tissue
culture is globally being widely used for large scale plant
propagation and improvement as well as production of
secondary metabolites[11].
In Ethiopia, application of tissue culture is premature. At this time, conventional plant and animal breeding and selection techniques are widely used as major
tools for increasing agricultural productivity[17]. Until
recently, research guidelines related to genetic engineering do not exist in the country discouraging Ethiopian
scientists from conducting projects of genetic engineering and consequently significantly hampers the research
and capacity building process in modern biotechnology
research and development in the country. But, nowadays, the government has approved biosafety law that
encourages genetic engineering and marketing of its
products in the country. In this regard, a wide range of
crop production problems that are either difficult to
address using conventional research techniques are likely
to be solved using crops genetically engineered for specific traits and adapted to local conditions.
but laid down the foundation of tissue culture technology for which he is regarded as the father of plant tissue
culture. After that some of the landmark discoveries
took place in tissue culture are summarized under
TABLE 1 below.
Techniques of plant tissue culture
Micropropagation
Micropropagation starts with the selection of plant
tissues (explant) from a healthy, vigorous mother
plant[14]. Any part of the plant (leaf, apical meristem,
bud and root) can be used as explant.
Preparation of donor plant
Any plant tissue can be introduced in vitro. To enhance the probability of success, the mother plant should
be ex vitro cultivated under optimal conditions to minimize contamination in the in vitro culture[4].
Initiation stage
In this stage an explant is surface sterilized and
transferred into nutrient medium. Generally, the combined application of bactericide and fungicide products is suggested. The selection of products depends
on the type of explant to be introduced. The surface
sterilization of explant in chemical solutions is an important step to remove contaminants with minimal
damage to plant cells[10]. The most commonly used
disinfectants are sodium hypochlorite[13,19], calcium
hypochlorite[9], ethanol[18] and mercuric chloride
(HgCl2)[10]. The cultures are incubated in growth chamHistorical development of tissue culture technol- ber either under light or dark conditions according to
the method of propagation.
ogy
Multiplication stage
The science of plant tissue culture takes its roots
The aim of this phase is to increase the number of
from the discovery of cell followed by setting out the
[16]
propagules
. The number of propagules is multiplied
cell theory. In 1838, Schleiden and Schwann proposed
that cell is the basic unit of all living organisms. They by repeated subcultures until the desired number of plants
hypothesized that cell is capable of autonomy and there- is attained.
fore is capable of regenerating or differentiating into Rooting stage
whole organisms (plants, animals or microorganisms) if
The rooting stage may occur simultaneously in the
given an appropriate environment to grow. Based on same culture media used for multiplication of the exthis premise, in 1902, Gottlieb Haberlandt, a German plants. However, in some cases it is necessary to change
physiologist, attempted to culture isolated single pali- media, including nutritional modification and growth
sade cells from leaves in knop’s salt solution enriched regulator composition to induce rooting and the develwith sucrose for the first time. The cells remained alive opment of strong root growth.
for up to one month, increased in size, accumulated
Acclimatization stage
starch but failed to divide. Though he was unsuccessful
At this stage, the in vitro plants are weaned and
RRBS, 7(7) 2013
Mohammed Adefa Seid
251
Review
TABLE 1 : Summary of historical development of tissue culture
Years
1902
1904
1922
1926
1934
1939
1941
1946
1954
1955
1957
1959
1960
1960
1960
1962
1964
1966
1970
1971
1972
1974
1977
1978
1981
1983
1984
1987
2005
Events in the history of tissue culture
Haberlandt proposed concept of in vitro cell culture
Hannig cultured embryos from several cruciferous species
Kolte and Robbins successfully cultured root and stem tips respectively
Went discovered first plant growth hormone –Indole acetic acid
White introduced vitamin B as growth supplement in tissue culture media for tomato root tip
Gautheret, White and Nobecourt established endless proliferation of callus Cultures
Overbeek was first to add coconut milk for cell division in Datura
Ball raised whole plants of Lupinus by shoot tip culture
Muir was first to break callus tissues into single cells
Skoog and Miller discovered kinetin as cell division hormone
Skoog and Miller gave concept of hormonal control (auxin: cytokinin) of organ Formation
Reinert and Steward regenerated embryos from callus clumps and cell suspension of carrot (Daucus carota)
Cocking was first to isolate protoplast by enzymatic degradation of cell wall
Bergmann filtered cell suspension and isolated single cells by plating
Kanta and Maheshwari developed test tube fertilization technique
Murashige and Skoog developed MS medium with higher salt concentration
Guha and Maheshwari produced first haploid plants from pollen grains of Datura (Anther culture)
Steward demonstrated totipotency by regenerating carrot plants from single cells of tomato
Power et al. successfully achieved protoplast fusion
Takebe et al. regenerated first plants from protoplasts
Carlson produced first interspecific hybrid of Nicotiana tabacum by protoplast Fusion
Reinhard introduced biotransformation in plant tissue cultures
Chilton et al. successfully integrated Ti plasmid DNA from Agrobacterium tumefaciens in plants
Melchers et al. carried out somatic hybridization of tomato and potato resulting in Pomato
Larkin and Scowcroft introduced the term somaclonal variation
Pelletier et al. conducted intergeneric cytoplasmic hybridization in Radish and Grape
Horsh et al. developed transgenic tobacco by transformation with Agrobacterium
Klien et al. developed biolistic gene transfer method for plant transformation
Rice genome sequenced under International Rice Genome Sequencing Project
(adopted from Hussain et al., 2012)
hardened. Hardening is done gradually from high to low
humidity and from low light intensity to high light intensity. The plants are then transferred to an appropriate
substrate (sand, peat, compost etc.) and gradually hardened under greenhouse.
Current status of tissue culture biotechnology in
Ethiopia
Several developed nations in the Western hemisphere and the Asia-Pacific region are already benefiting significantly from modern biotechnology[6]. However, the majority of the developing countries, including
Ethiopia, have very little access to this emerging economic sector.
Ethiopian government development strategy rec-
ognizes the leading role of agriculture in the economy
and stipulates that for the country to record rapid economic prosperity. The strategy identifies information and
communication technology and biotechnology as essential tools for rapid transformation of largely subsistence mode of production to market-oriented production enterprises that ultimately lead to industrialization[1,7].
Modern agricultural biotechnology research and development are recently being developed along with long
established conventional agricultural research in Ethiopia. In this regard, public institutions with significant
activities in the area of biotechnology include the Ethiopian Institute of Agricultural Research (EIAR), Addis
Ababa University, National Veterinary Institute, The
National Animal Health Research Laboratory, Institute
Plant tissue culture biotechnology
in Ethiopia
.
252
RRBS, 7(7) 2013
Review
TABLE 2 : Development status of tissue culture protocols at Ethiopian institute of agricultural research (adopted from
Abraham, 2009)
Plant Name
Banana
Explant Used
Shoot tips
Main Purpose
Micropropagation, Virus cleaning
Status
Completed and being scaled up
Black pepper
Shoot tip
Micropropagation
Ongoing and in good progress
Brassica spp.
Anther
Double haploid line development
Initial stage
Cardomom
Rhizome lateral bud
Micropropagation
Completed and being scaled up
Cassava
Meristem
Micropropagation, Virus cleaning
Initial stage
Citrus
Seed
Micropropagation, virus cleaning
Ongoing and in good progress
Coffee
Leaf
Micropropagation, in vitro disease screening
Completed and being scaled up
Enset
Shoot tip
Micropropagation
Ongoing and in good progress
Garlic
Meristem
Micropropagation, virus cleaning
Initial stage
Geranium
Shoot tip
Micropropagation
Completed
Ginger
Rhizome lateral bud
Micropropagation
Initial stage
Grapevine
Shoot tip
Micropropagation
Completed
Cardamon aframomum
Rhizome lateral bud
Micropropagation
Completed
Niger
Anther
Double haploid line development
Ongoing and in good progress
Pineapple
Shoot tip
Micropropagation
Completed and being scaled up
Potato
Node
Micropropagation, virus cleaning
Completed and being scaled up
Sweet potato
Shoot meristem
Micropropagation
Initial stage
Tef
Floral part
Double haploid line development
Ongoing and in good progress
of Biodiversity Conservation, the International Livestock
Research Institute and the Regional Agricultural Research
Institutes.
In Ethiopia, plant tissue culture has been given the
highest priority in biotechnological research and development. This is because tissue culture provides large
quantities of disease-free plant materials in short time.
Tissue culture activities first started in Ethiopia in 1980’s
at Addis Ababa University with focus on
micropropagation of indigenous forest species like
Podocarpus sp., Cordia africana, Hagenia
abyssinica and Annengeria sp. that are either difficult
to regenerate vegetatively or require long time propagation[8]. This was followed by micropropagation works
carried out on some Ethiopian plant species like
Phytolacca dodecandra[5], Eragrostis tef[2] and Enset
ventricosum[3,15].
In 2000, a more comprehensive and concerted emphasis was given for a protocol of optimization for mass
propagation, disease cleansing and in vitro conservation of economically important crop species. The major achievements from these works include the distribution of large number of tissue culture plants of banana, hybrid coffee, pineapple and potato to farmers in
various parts of the country (see TABLE 2). In addition to micropropagation and virus cleansing, in vitro
techniques are also being used for production of
dihaploid plants to reduce breeding cycle by obtaining
pure line for further improvement in crops like tef and
Niger[1] and for in vitro screening of crops like coffee
for resistance to coffee berry diseases.
Challenges and opportunities of application of tissue culture in Ethiopia
In Ethiopia, application of technologies generated
by conventional research has significantly improved the
country’s agricultural productivity in the past[1]. If biotechnology is properly integrated into these technologies, it would complement these efforts by providing opportunity to speed up such processes giving new
solutions to the old and emerging problems in a more
precise and cost-effective manner. The potential of tissue culture biotechnology in improving crop and livestock productivity is very huge and rapid progress is
being made worldwide on its application. In this regard, Ethiopia has developed favorable policies to facilitate the safe application of biotechnology in agricultural sectors in particular. Yet, nowadays, it remained
difficult to use such tools to acquire solutions to major
agricultural problems owing to lack of resources (infrastructures) and well skilled personnel.
The advantage of tissue culture for rapid and large
RRBS, 7(7) 2013
Mohammed Adefa Seid
253
Review
scale multiplication of plants has been widely recognized in the country in recent years and research efforts
are now being extended to other institutions as well.
For instance, regional agricultural research institutes
namely the Amhara Agricultural Research Institute and
Southern Agricultural Research Institute have recently
developed their capacity and initiated tissue culture work
for micropropagation of selected crops of importance
in the respective geographical area. Furthermore, private enterprises like Mekele Plant Tissue Culture Laboratory have recently engaged in large scale multiplication and propagation of tissue culture of crops including
banana, sugarcane, grapevine and flowers to producers in northern Ethiopia.
Genetic engineering offers several benefits when
used responsibly by addressing the environmental and
food safety concerns with rigorous biosafety regulations. Until recently, guidelines with genetic engineering
research and deployment of genetically modified organisms (GMOs) do not exist in the country. This situation discouraged Ethiopian scientists from initiating
genetic engineering projects and participating in similar
network activities at regional and international level and
consequently significantly hampering the research and
capacity building process in modern biotechnology research and development in the country. Three years
back, the Ethiopian government has approved biosafety
law which is expected to encourage genetic engineering research as well as marketing of its products in the
country. A wide range of crop production problems that
are either difficult or impossible to address using conventional research techniques are likely to be solved
using crops genetically engineered for specific traits and
adapted to local conditions. For some of these constraints, transformation technology is already developed
elsewhere and commercially available and only needs
to be introduced and adopted to local conditions with
minimum value inputs. Recently, the private sector has
expressed keen interest in introducing Bt cotton to boost
its production and thus satisfying the booming textile
industry in the country[1].
It is also possible to use transgenic plants as parents to transgress the desired genes to locally preferred
cotton varieties by conventional breeding. On the other
hand, there is a need to develop local capacity in genetic engineering technologies in terms of infrastructure
and manpower to address constraints on indigenous
crops like Eragrostis tef and Enset ventricosum.
Potential impacts of GMOs
Genetic engineering, if not managed safely and responsibly, could impose some dangerous problems on
the environment and wellbeing of human. For instance,
transgenic plants harm non-target species directly and
indirectly down the food chain. For instance, Bt-cotton
harms bees which are major pollinators, Bt-maize harms
lacewings fed on pests that have eaten Bt-maize, and
Bt-maize pollen also harms larvae of Monarch butterflies. Transgenic potatoes with snowdrop lectin harms
ladybirds fed on aphids that have eaten transgenic potato[12].
Transgenic varieties are unstable, do not breed true,
and do not perform consistently. Herbicide tolerant
transgenic crops are incompatible with sustainable agriculture dependent on mixed cropping and crop rotation. Broad-spectrum herbicides harm earthworms and
microoragnisms that maintain natural soil fertility in organic farming. Transgenic plants with bt-toxin undermine pest control for organic farming and are toxic to
major pollinators and other beneficial insects.
Transgenic lines are even more genetically uniform
than conventional monoculture crops and may hence
be more susceptible to diseases and environmental exigencies. Viral resistant transgenic plants can generate
new, often super infectious viruses. Terminator technologies destroy seed fertility[12].
The hazards are inherent to the hit or miss technology. Random gene insertions give random genetic abnormalities and unexpected effects. New genes, gene
constructs and products from viruses, bacteria and nonfood species are introduced into our food for which no
safety tests exist. Interaction between introduced gene
and host genes increases unexpected effects including
toxins and allergens. The technology enhances horizontal
gene transfer and has the potential to generate new viruses and bacteria that cause diseases and spread drug
and antibiotic resistance.
Horizontal gene transfer and recombination spread
antibiotic resistance genes and have created new pathogens in recent years. Strains of four dangerous bacteria, including the one causing tuberculosis, are resistant
to all antibiotics and hence are untreatable. So far, reports reveled that at least 40 new viruses that cause
disease in human beings have emerged between 1988
Plant tissue culture biotechnology
in Ethiopia
.
254
RRBS, 7(7) 2013
Review
and 1996. Transgenic plants were found to transfer
transgenes and antibiotic resistant marker genes to soil
microorganisms and fungi[12]. Usually, DNA released
from dead or viable cells persists in all environments
and remain infectious.
CONCLUSION
Biotechnology is a scientific technique based on biology, especially used in agriculture, food and medicine, which uses living organism or substance of organism to make or modify product, to improve plants or
animal or to develop microorganism for specific uses.
The genetic modification leading to improvement of organism is necessary for production of certain products.
The most common techniques of biotechnology include
tissue culture, in vitro regeneration, organogenesis, genetic engineering (transformation) and embryogenesis.
Tissue culture biotechnology is the most widely used
tool in agricultural sectors for improving and producing
disease-free plants and animals, and their products. In
this regard, developed nations have significantly benefited from the product of biotechnology in general and
tissue culture technology in particular.
Unfortunately, underdeveloped countries including
Ethiopia missed such opportunities so far as the result of
lack of resources and required knowledge, and lack of
awareness. However, nowadays, the Ethiopian government started planning and implementing policies and strategies related to biotechnology in agricultural sectors.
In Ethiopia, application of plant tissue culture biotechnology is at initial stage. In fact, presently, many government universities and research institutes are taking initiatives towards developing curricula, educating skilled
manpower and conducting problem solving researches
in the area of biotechnology. Despite the fact that biotechnology would solve societal problems, irresponsible
and unsafe application and management of biotechnological products could severely harm the wellbeing of
human, his possession and the environment at large. So,
application of biotechnology and its product should be
handled in a responsible and safe ways.
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