Present status of biotechnology in Nepal

In recent years, there has been a growing awareness about the scope and potentiality of genetic engineering. The plant tissue cultured was started in Nepal in 1976 at the National Herbarium and plant Research Laboratory in Godawari. This laboratory basically has been doing clonol propagation of hard wood trees, horticulture crops, medicinal plants, ornamental plants and over 30 kinds of orchids. They have been involved in some anther culture as well as some karyological work.
The biggest laboratory in private sector is the Research Laboratory for Agriculture Biotechnology and Biochemistry (RLABB) which was established in 1986. Research Laboratory for Agriculture Biotechnology and Biochemistry initiated for the first time various molecular works in the field of agriculture and forestry. This lab is highly sophisticated having various biotechnological apparatus. This lab has developed many biochemical techniques to create cold tolerant rice plant, sex-determination of dioecious plant and micropropagration of many plants like pinus, Autocarpous, Rice, orchids, etc. Anther culture of rice was initiated in
Research Laboratory for Agriculture Biotechnology and Biochemistry for the first time.
Other private enterprises are Botanical Enterprise, Nepal Biotech Nursery, Himalayan Floratech, Microplant Nepal, etc. They are commercializing the tissue culture grown plants and their products.
Potato Development centre at Khumaltar has also included some tissue culture activities. This main objective of this centre is to create and multiply the virus free seeds potatoes to meet the need of farmers. They have been successful in creating potato varieties of virus.
Basic research on tissue culture is also initiated at Central Department of Botany, Kritipur mainly of medicinal activities. RECAST, Kirtipur has also the facilities available for plant tissue culture.
A new biotechnology laboratory Green Research Technology has been established with the aim of creating virus free plants of Citrus, Cardamom and potato plants using meristem culture and make them available to the Nepalese farmers.
To carryout the molecular work which includes ELISA test. Laboratory for Agriculture Biotechnology and Biochemistry, Sanepa, Potato Development Centre at NARC, Institute of Agriculture and animal Science, Rampur and Teaching Hospital are developing some basic facilities with international collabration.

Plant tissue culture

plant cells have the potential to grow as a plant. This potential of the plant cell is called to be totipotency. Due to that potential any plant part can develop a plant as a whole. These plant tissue culture is a technique of growing plant cells, tissues or organs in an artificially prepared nutrient medium under an ascptic condition in the laboratory.

Historical background
Haberlandt (1896) was the first person to culture Bolated vegetative calls of higher plants in simple nutrient solutions. Although Haverlandt was able to maintained the cells in the nutrient media the cell division was not recorded until much later.
Embryos were the first plant tissue to be successfully cultured in vitro on artificial medium. The first result of successfully in vitro culture was by Hanning (1904) who grew plants under aspetic conditions relatively mature embryos of Raphanus, R. landra, R. sativus and cochleasia danica (Brassicacesae) on Tollens medium with sugars, amino acids and plants extracts and obtained transplantable seedings.
The first major breakthrough in the field of plant tissue culture was made in 1922 when Robbins (USA) and Kotte (a student achieved success in growing isolated roots tips. white (1935) grew cultures of tomato root tips for a longer period ( more than 20 weeks) using sucrose, inorganic salts and yeast extract.
Further development in tissue culture of bio technology was made in 1934 by Gantheret of France who culture cambium cells of some tree species (populus higra) on knop's solution. skoog (1944) and skoog and tusi (1951) reported that in tobacco pith tissue cultures the addition adenine and high level of phosphate increased callus growth and bud formation in the presence of IAA. Musrashighe (1961) demonstrarted the technique in vitro culture of propagation of various plant species. Gaha and Maheshwari (1964) first of all successfully reported the anther culture of Datura innoxia

Application of biotechnology

Biotechnology has wide applications. Here its applications in the field of agriculture and medicine have been discussed.
In the field of agriculture – biotechnology has been applied for the improvement of agricultural, horticultural and forestry crops. Following are the application of biotechnology in the are of agriculture:
1. Micropropagation – using plant tissue culture, rapid vegetative multiplication of the plant material for agriculture, horticulture and forestry is possible. Micropropagation is beneficial because by this technique, large number of off springs or plantlets can be obtained in short period of time.
2. Production of disease – free plants – meristem tip culture is a technique of plant tissue culture in which meristem is used as explant. Using this method, virus free plants may be obtained.
3. Androgenic haploids – by anther culture androgenic haploid embryos or plantlets are obtained from the microspores or pollens. Haploid plants are significant because mutation ( spontaneous or induced ) can express themselves and can be easily detected. It is very important for research point of view.
4. Development of resistant plants – plant tissue culture can also be useful for creating varieties tolerence to pests, diseases or stresses.
5. Protoplast culture and fusion – protoplast can be used for genetic manupultion and somatic cells fusion in plant cells. This provides a chance to get a hybrid variety between non-related species.

Branches of biotechnology

1 plant tissue culture – It is a technique of growing plants cells, tissues or organs in an artificially prepared nutrient medium under aspectic condition. In this technique, a growing tissue is excised from plant and kept in a callus inducing artificial medium to produce calli. Calli are the undifferentiated and unorganized cells. These calli can regenerate green plants after putting them into
a regeneration medium.
Tissue ? callus ? plant regeneration
Environmental conditions like light, temperature, hormones( auxins and cytokinins), and nutrients can have important role in achieving plant tissue in vitro.
This technique has become extemely significant in agriculture.
Following are the benefits of plants tissue culture (PTC):
• Rapid clonal propagation of economically important plants in large number,
• Maintaining pathogen free plants,
• Possibility of wide hybridization,
• Increase in plant productivity,
• Improvement of variation among plants,
2 Protoplast
Protoplast is a cell without cell wall. Protoplast can be obtained, cultured and fused. A complete plant can be obtained from a single protoplast culture. It is useful in-
? Obtaining germ free plants,
? Introduction of foreign genes, and
? Fusion of two incompatible plants.
3 Gene transfer
Gene is a part of a DNA segment with a particular character. ( DNA segment that expresses a character is called gene). Many useful genes like Nif gene, kanamycin resistant gene, luciferease gene, etc. have been known and isolated. In this technique, a foreign gene of interest can be inserted into other plants by the following methods:
• Direct gene transfer – It is the process of transfer of a gene interest directly into plants by conventional breeding, protoplast fusion, etc.
• Agrobaterium mediated DNA transfer technology – it is the process of transfer of gene from a plant to other plants through Agrobacterium tumefaciens.

4 Hybridoma technology – this technique is used to fuse a normal antibody-producing lymphocytes (B cells or plasma cells) with myeloma cell (a kind of tumor cells) giving a hybridoma. Hybridoma has the potential to grow indefinitely in culture and hence can be source of unending supply of an antibody of choice. Since the antibody produced by a hybridoma is biochemically pure, it is called monoclonal antibody. Monoclonal antibodies are used for detection of animals and plants diseases.
Using this technique, various types of antibodies, vaccines, hormones, enzymes, kits for early diagnosis of diseases, etc. have been developed.
Genetic engineering – it is also known as recombinant DNA technology. In this technique, a piece of foreign DNA is inserted into DNA of a vector (foreign DNA + vector DNA) is multiplied and/or expressed by introducing the vector into a bactrium. This approach is used for DNA sequencing and commercial production of proteins.

Bio technology

Biotechnology is a new discipline of science in which latest technology has been applied to biological organisms for the welfare of society. Biotechnology is the application of scientific and engineering principles to the processing of materials using biological agents to provides goods and services. It is defined as “any technique that uses living organisms to make or modify products to improve plants and animals or to develop micro-organisms for specific uses”.
Biotechnology is as old as our civilization. The old biotechnology is like alcohol fermentation using yeast to make wine and beer.
The modern biotechnology stands on the understing of molecular basic of biological cell function and alter the cell function to make it produce products required by society.
The major new techniques available with modern biotechnology are plant tissue culture, recombinant DNA technology and hybridoma technology.
It is a completely genetic engineering. It involves the knowledge of biology, microbiology, biochemistry, immunology, molecular biology, genetic engineering etc.
Biotechnology holds potentials for developing products and processess in various sectors of agriculture, animals husbandry, health, energy and environment protection.

Historical perspectives
Biotechnology processes are actually in use since prehistoric times. The primitive people were all acquired with alcoholic fermentation and vinegar fermentation from sugary substances. The Sumerians were experts in brewing different types of beer.
Ethanol was first obtained 1150 by distillation of alcoholic beverages. Since than it is being manufactured throughout the world. In 14th century, first vinegar manufacturing industry was established in France. Artificial cultivation of mushroom was initiated in 1650 in France. 1680 Leeuwenhoek first observed yeast cells. Erxleben (1818) discovered the fermentative properties of yeast. In 18 79 Louis Pasteur highlighted the latic acid fermentation by microbs. Hansen (1897) reported the enzymes extracted from yeast could convert sugar into alcohol.
Towards the end of the 19th century the large-scale sewage purification system employing microbs was established in Germany and France. During the same period, three improtant industrial chemicals ( acetone, butanol and glycerine) were obtained from bacteria. Penicilline was discovered by Alexender Flemming in 1928. The large-scale manufacture of penicilline started in 1944. From 1950 several antiboties were dicovered and added to the list of “ wonder drugs”.
In 1953 Watson and Crick reveated the structure of DNA which actually laid the foundation of genetic engineering. First successful genetic engineering experiment was done in 1973. In 1982 genetically engineering insulin was approved for use. In the middle and late eighties several new antiboties and vaccines were developed. Interferous are being widely employed for the treatment of some diseases and even certain viral induced cancers

bio-technology

Bio-technology is a discipline of biology, food science and medicine. latest use of the of the term state forward to genetic engineering and to cell biology and also tissue culture technologes.However, the concept encompasses a large range and history of procedures for the manupulating living organisms according to human purposes, as we move back to cultivation of plants and improvement of plants and domastication of animals by breeding good quality of animals. the employment can be done by artificial selection and hybridization. Comparing to biotechnology bioengineering is usually througt of as related field with its emphasis some what more on mechanical and upper class systems approaches to interfacing with and exploiting living things.United Nations convention on Biological Diversity defines Biotechnology.

"Technology with its application that uses biological systems, living organisms or its derivatives there fo to make or modify goods or processes for specific use".

Biotechnology draws on the pure biological sciences, ( microbiology, genetics, animal cell culture, molecular biology, biochemistry, embroyology, cell biology) and in large number of instanes is also dependent on knowledge and its process from outside the sphere of biology

(chemicaengineering, bioprocess engineering, information technology, biorobotics).

concquently latest biological sciences (including even concepts such as molecular ecology) are intimately entwined and dependent on the methods developed through biotechnology and what is commonly thought of as the life sciences industry.