Biotechnology in Nepal

BIOTECHNOLOGY IN NEPAL - A SYNOPSIS

Professor Dr. Sarbajna Man Tuladhar Research Centre for Applied Science and Technology (RECAST) Tribhuvan University, Kirtipur, Kathmandu, Nepal.

1. INTRODUCTION

Biotechnology has been around us for centuries. In industrialized countries, modern biotechnology has become well advanced.

In the last decade of the 20th century, Nepal witnessed several biotechnology-derived products in the market :  Human health care products  Seeds and plants of high yielding variety of agricultural and horticultural crops  Food products and drugs  Improved livestock  Animal health care products

They entered into the country from the neighboring and the developed countries. Modern biotechnology has entered into Nepal like bandwagon and everybody is jumping onto it. In fact biotechnology programs are running without any policies and plans. Modern biotechnology in Nepal is still in a state of infancy.

2. THE CONCERNED MINISTRIES

1. Ministry of Environment, Science and Technology 2. Ministry of Forest and Soil Conservation 3. Ministry of Agriculture and Co-operative 4. Ministry of Health 5. Ministry of Education and Sports 6. Ministry of Industry, Commerce and Supply

3. PRESENT STATUS OF DIFFERENT AREAS OF BIOTECHNOLOGY

3.1 PLANT TISSUE CULTURE

 Plant tissue culture is the major biotechnology activity of public as well as private sectors. 2

 Public laboratories developed protocols for in vitro propagation of different plant species.  Private laboratories produced elite plants commercially using protocols developed by public laboratories.

3.1.1 PUBLIC SECTOR INITIATIONS

Prominent laboratories involved in plant tissue culture are:

1. DEPARTMENT OF PLANT RESOURCES (DPR)

 First tissue culture laboratory established in 1976 at Godavari to initiate and promote plant tissue culture.  Developed protocols for tissue culture propagation of more than 100 plant species including orchids, agricultural and horticultural trees, ornamental plants, and bamboo.  Most important achievement is the development of sand rooting technique – the tissue-cultured micro shoots can be rooted in the non-sterile sand.  Sand rooting technique is easy, cheap, less time consuming, economic, conveniently done in nurseries, can easily be established for field plantation.  Provides tissue culture training for entrepreneurs. Consequently four private plant tissue culture factories have been established.

2. NEPAL AGRICULTURE RESEARCH COUNCIL (NARC)

 Potato Research Program (PRP) with the assistance of Swiss Development Corporation has been producing virus-free pre-basic seeds of potato at its tissue culture laboratories and green house facilities at Khumaltar.  Established tissue culture laboratories at regional agricultural research stations at Dhankuta (Eastern development region) and Lumlay (Mid-western development region) to promote micro propagation.

Division of Agriculture Botany of NARC has initiated some works on  anther culture of rice and wheat,  germ plasm conservation and diagnostic facilities  Polymerase Chain Reaction (PCR) technology.

3. HORTICULTURE DEVELOPMENT PROJECT/ DEPARTMENT OF AGRICULTURE

Its activities were:  Tissue culture propagation of apple rootstocks.  Micro grafting of sweet orange (JUNAR) for the production of virus-free and greening-free plants. However the facilities remain unused at present. 3

4. INSTITUTE OF AGRICULTURE AND ANIMAL SCIENCES (IAAS) / TRIBHUVAN UNIVERSITY.

 Established tissue culture facilities with the grant assistance from Rockefeller Foundation.  Assisting thesis works for M. Sc. Degree.

3.1.2 UNIVERSITY SECTOR ACTIVITIES

1. CENTRAL DEPARTMENT OF BOTANY/ TRIBHUVAN UNIVERSITY

Activities:  Tissue culture propagation of different medicinal plants.  Anther culture.  Rhizobium inoculation in leguminous plants

3.1.3. PRIVATE SECTOR INITIATIONS

1. BOTANICAL ENTERPRISES PVT. LTD.

 In vitro propagation of many species of orchid, potato, fodder, Chrysanthemum, Gerbera, African violet, Lily etc.  Export directly to The Netherlands.

2. NEPAL BIOTECH NURSERY

 Produces banana, orchid, and ornamental plants by tissue culture and non-sterile sand rooting technique.

3. RESEARCH LABORATORY FOR AGRICULTURE BIOTECHNOLOGY AND BIOCHEMISTRY (RLABB)

 Tissue culture propagation for pine, Artocarpus, Brassica.  Anther culture of cold tolerant rice  Modest facilities for DNA work by PCR technology and enzyme analysis.

Facilities are used for research and teaching.

4. MICROPLANTS NEPAL PVT. LTD.

 Commercial propagation of banana, orchid, lily, mums etc.

5. GREEN RESEARCH AND TECHNOLOGY (GREAT)

 Developing virus testing and elimination facilities on horticultural crops such as potato, citrus, banana, cardamom, strawberry, and some ornamental plants using tissue culture techniques.  Has modest screen-house facility for indexing against citrus greening disease. 4

 Cleaned two potato cultivars of Bangladesh namely Petronese and Multa from potato viruses such as PVX and PVY.  Cleaned three leading cultivars of large cardamom (Amomum subutalum) namely Ramshahi, Golshahi, and Dambarshahi from Chhirke and Furke viruses.  Virus-free plants derived from meristem culture are available for further multiplication.

6. HIMALAYAN BOTANICAL RESEARCH CENTRE (HIMBORCE)

 Established infrastructure for micro propagation of medicinal and ornamental plants indigenous to Nepal.  Embarking upon micro propagation of Gladiolus.

3.2 BIOFERTILIZERS

1. ROYAL NEPAL ACADEMY OF SCIENCE AND TECHNOLOGY (RONAST)

 Developed Rhizobium inoculant technology for pulses.  Developed microbial inoculant technology for composting.  Use of ecto and endomycorrhiza in conservation of forest soils.  Quality production of organic fertilizers.

2. DIVISION OF SOIL SCIENCE / NARC

 Production and distribution of legume inoculant packets for more than a decade.  Distributed 2627 Rhizobium inoculant packets to the farmers for important pulse and pasture crops.

3. DIVISION OF PLANT PATHOLOGY / NARC

 Engaged in spawn production technology for mushroom culture using two genera namely Agaricus and Pleurotus.

3. RESEARCH CENTRE FOR APPLIED SCIENCE AND TECHNOLOGY (RECAST) / T. U.

 Improvement of traditional composting technology.  An integrated approach for the utilization of municipality solid wastes.

5. CENTRAL DEPARTMENT OF MICROBIOLOGY / T. U AND 6. CENTRAL DEPARTMENT OF CHEMISTRY / T. U.

 Impact of industrial waste on land and water pollution. 5

3.3 BIOPESTICIDES

 Several isolations of Bacillus thuringiensis (Bt) from Nepalese soils for biological control of Lepidoptera caterpillar in crucifer vegetables.  Field trial of indigenous pesticide plants like Eupatorium adenophorum, Lantana camara, Azadirachta indica etc. in collaboration with Department of Agriculture (Ministry of Agriculture).

2. DIVISION OF ENTOMOLOGY / NARC

 Reported efficacy of Acorus calamus (Root powder), neem oil (Azadiractin, 0.15%), and neem seed extract (Azadiracta monotetala) against storage insect like Sitophilus zeamais.

3. RESEARCH CENTRE FOR APPLIED SCIENCE AND TECHNOLOGY / T. U.

 Survey of plant pesticides for grain storage.

3.4 MEDICAL BIOTECHNOLOGY

1. THE LEPROSY MISSION, NEPAL

 Determination of cytokines by ELISA for blood immune response studies in leprosy patients.  DNA diagnosis of Mycobacterium leprae by PCR.

2. CENTRAL VETERINARY LABORATORY

 Vaccine production against PPR (Pestes des Petites Ruminant) using tissue culture.

3. NEPAL AGRICULTURE RESEARCH COUNCIL (NARC)

 Performing embryo transfer and artificial insemination in cattle.

4. EVEREST BIOTECH PVT. LTD.

 Production of specific antibody required of Human Genome Project, England.

5. CENTRAL DEPARTMENT OF CHEMISTRY/ T.U.

 Antibacterial, antifungal, and cytotoxic activities of medicinal plants.  Study of specific inhibitors for proteolytic enzymes.  Taxol from endophytes of Himalayan conifers. 6

6. CENTRAL DEPARTMENT OF MICROBIOLOGY / T. U.

 Antimicrobial activities of some plant extracts.

7. RESEARCH CENTRE FOR APPLIED SCIENCE AND TECHNOLOGY / T. U.

 Bioprospecting of ethno medicinal plants of Nepal for the conservation of biological and cultural resources.

3.5 DISEASE DIAGNOSTICS

 Diagnosis of Asian citrus greening diseases or Huanglongbing of citrus using PCR based technique.  Diagnosis of Citrus tristeza virus disease using Double Antibody Sandwich Enzyme Linked Imunosorbent Assay (DAS-ELISA) technique.

2. INSTITUTE OF AGRICULTURE AND ANIMAL SCIENCES (IAAS) / T. U.

 Diagnosis of viral and bacterial diseases in rice using PCR technology.

3. GREEN RESEARCH AND TECHNOLOGY (GREAT)

 Use of PCR technology for routine diagnosis of citrus-greening disease (Huanglongbing).  Provides training on virus testing using DAS-ELISA and biological indicator plants for the diagnosis of Huanglongbin on citrus.

3.6 BIOENERGY AND BIOCONVERSION TECHNOLOGY

1. RESEARCH CENTRE FOR APPLIED SCIENCE AND TECHNOLOGY (RECAST) / T. U.

 Improvement of biogas production by using weeds such as Eupatorium adenophorum and water hyacinth.  Bio fuel from the Physic nut (Jatropha curcas L) for running diesel engine.

3.7 MOLECULAR CHARACTERIZATION

1. NEPAL AGRICULTURE AND RESEARCH COUNCIL (NARC)

 Genetic diversity analysis of Fagopyrum spp. (wild sweet and bitter buckwheat) using Randomly Amplified Polymorphic DNA (RAPD).  Genetic diversity analysis of isozymes in indigenous crops like rice, finger millet, barley, pigeon pea, buckwheat, taro, cucumber, sponge gourd, citrus, and Swertia spp. 7

 Initiated research work on molecular characterization of medicinal plants like Swertia chirata, Camellia sinensis etc. and microorganisms such as Bacillus thuringiensis.

2. RESEARCH CENTRE FOR APPLIED SCIENCE AND TECHNOLOGY (RECAST)

 Selection of appropriate genotypes of Mulberry and exploitation of genetic yield potential of common buckwheat ecotypes.

3.8 FERMENTATION

Fermentation is the major traditional biotechnology and national heritage of Nepal.

Significant ancient biotechnologies still surviving in Nepal are confined to cottage industry scale or individual family consumption; they are:

 Fermentation and distillation of characteristic alcoholic drinks from millet (KODO), rice, brown sugar (SAKHAR) by using indigenous yeast.  The traditional process of making yeast is still a closely guarded secret.  Fermentation of leafy vegetables into GUNDRUK, the favourite food specialties of rural Nepalese.  GUNDRUK from the leaves of mustard, radish or cauliflower.  SINKI from radishes cut into pieces.  SINAMANI from the whole radishes without their leaves cut out.  Browning of rice into HAKUWA or HAKUJAKI (literally meaning black rice) by indigenous fermentation.  Production of jute (NALU) from the bark of the hemp through fermentation process.  Production of yogurt (DAHI) and hard cheese (CHHURPI).  Dung used as fertilizers and dung cakes used as fuel for cooking.

Interesting to note that traditional biotechnology is being conserved predominantly by womenfolk.

3.9 PRACTICAL APPLICATION OF BIOTECHNOLOGY PRODUCTS/ PROCESSES

3.9.1 AT COMMERCIAL / SEMI-COMMERCIAL LEVEL

 Tissue culture raised plantlets of sisso tree, orchids, medicinal plants, ornamental plants, and virus free banana and citrus. 8

3.9.2 AT COTTAGE INDUSTRY LEVEL

 Characteristic alcohol from millet (KODO), rice, and brown sugar (SAKHAR).  Fermented vegetables – GUNDRUK, SINKI, SINAMANI.  Yogurt (DAHI) and Hard Cheese (CHHURPI).

3.10 BIOTECHNOLOGY IN ACADEMIA

3.10.1 AT UNDERGRADUATE LEVEL

 Biotechnology is taught only as a small part of the course unit.

3.10.2 AT GRADUATE / POST-GRADUATE LEVEL

 Central Department of Botany, T. U. – M. Sc. Thesis works in tissue culture.  Central Department of Microbiology, T. U. – M. Sc. thesis works in agriculture biotechnology, medical biotechnology, environmental biotechnology, genomics, and proteomics.  Institute of Agriculture and Animal Sciences, T. U. – thesis works in diagnosis of viral and bacterial diseases in rice using PCR technology.  Two year M. Sc. in Biotechnology –opening soon in Tribhuvan University, the oldest and largest university of Nepal.  Kathmandu University – B. Sc. with specialization in Biotechnology.  Purbanchal University – B. Sc. with specialization in Biochemistry (with more emphasis on Biology).  Pokhara University – B. Sc. with specialization in Biochemistry (with more emphasis on Chemistry).

3.11 HUMAN RESOURCES

 Persons specialized in biotechnology are only nominal.  Most biotechnology activities are due to personnel specialized in Agriculture and Botany.

3.12 NATIONAL ORGANISM BANK

 Established in 1985 at Royal Nepal Academy of Science and Technology. Objectives: Isolation, identification, and maintenance of the microorganisms (fungi and bacteria).  Acts as a depository to supply authentic microbial cultures for the use in various research activities of research organizations, universities, and industries. This is an attempt to conserve and utilize microbial genetic resources of the country. 9

3.13 BIOTECHNOLOGY INFORMATION NETWORK

 Biotechnological research findings and activities are communicated through occasional papers and annual reports published by respective organizations or national journals.  Ministry of Environment, Science and Technology plans to develop the information networking in Biotechnology.  A resource centre as the focal point of SAARC Technical Committee on Science and Technology will be developed in due course of time.

3.14 REGULATORY PROCEDURES

 Nepal has signed the Cartagena Protocol on Biosafety on March 2, 2001.  Focal point for the Biosafety Protocol is the Chief of the Environment Division in the Ministry of Forests and Soil Conservation.  Ministry of Forests and Soil Conservation has enforced Biosafety Regulations 2062 BS throughout the Kingdom of Nepal on and from May 25, 2005.

OTHER PERTINENT LEGISLATIONS : - Export Import (Control) Act, 2013 BS. - Food Act, 2023 BS. - Plant Protection Act, 2029 BS - Livestock Feed Products Act, 2033 BS - Medicine / Drug Act, 2035 BS - Seed Act, 2045 BS - Livestock Health and Livestock service act, 2055 BS

3.15 REGULATORY MECHANISM

IMPLEMENTING AGENCY RESPONSIBILITY Ministry of Forests and Soil Conservation National Focal Point for the Biosafety Protocol. Department of Livestock, Animal For enacting The Livestock Health and Quarantine Section and 25 Quarantine Livestock Service Act. check posts / Ministry of Agriculture and Cooperatives. Plant Quarantine Office, Department of For enacting The Plant Protection Act and Agriculture/ Ministry of Agriculture and The Seed Act. Cooperatives Department of Food Research and Quality For checking whether any food products Control are polluted or of quality below the permissible standard for human consumption. Department of Drug Administration / For issuing permission for production, Ministry of Health import, export, and clinical trial of new medicines. 10

Department of Industry / Ministry of For issuing license to establish industry and Industry, Commerce and Supply for export and import of goods. Department of custom / Ministry of For issuing of custom clearance of goods Finance only after seeking recommendation or permission letter from the concerned Departments.

For dealing with the issues of the LMOs / GMOs, capacity of the above-mentioned ministries should be increased. Capacity increment refers to:  Human resources development  Infrastructure (laboratory and equipments) development  Development of administrative mechanism  Networking of concerned ministries  Formulation of legislation

3.16 INTERNATIONAL AGREEMENTS

 Nepal has always expressed its solidarity and commitment toward several international efforts related to the environment conservation. Consequently it has become a signatory to a number of international legal instruments.

LEGAL INSTRUMENT DATE SIGNED Plant Protection Agreement for the South- 27 February 1956 East Asia and Pacific Region. Convention on Wetlands of International 2 February 1971 Importance Especially waterfowl Habitat. Convention for the Protection of the World 23 November 1972 Cultural and Natural Heritage. Convention on International Trade in 3 March 1973 Endangered Species of Wild Fauna and Flora. World Conservation Strategy 1980 World Charter for Nature 1982 International Tropical Timber Agreement. 18 November 1983 Agreement on Network of Aquaculture 8 January 1988 Centres in Asia and the Pacific. Conservation on Biological Diversity. 22 May 1922 The Cartagena Protocol on Biosafety. 2 March 2001 World Trade Organization (WTO) member 23 April 2004 [147th].

4. MAJOR CONSTRAINTS

 Acute lack of national program on biotechnology. 11

 Biotechnology programs are usually dependent on the initiative of a few foreign- trained scholars and enthusiast scientists at home.  National Biotechnology Policy is non-existent as yet. Draft of the Policy prepared by Ministry of Environment, Science and Technology has been years in gestation in the Cabinet.  Lack of qualified and properly trained research and development manpower – the limiting factor both for private and government sectors.  The availability and quality of infrastructures relevant to biotechnology research is negligible.  Lack of public sector-private enterprise links, university-industry interaction, and university-university cooperation.

5. PROSPECTS

Despite serious handicaps, the prospects for growth of biotechnology in Nepal are numerous. Biotechnology potentials worth mentioning are:

AGRICULTURE / FOOD SECTOR

 Plant tissue culture  Biofertilizer  Biopesticide  Animal feeds and supplements from agricultural products

INDUSTRY SECTOR

 Food and industrial enzymes  Fermentation products  Bio energy, bio fuel and bioconversion

HUMAN HEALTH CARE SECTOR

 Development of rapid diagnostic kits and techniques  Production of culture media  Production of monoclonal antibodies  Vaccines against malaria, measles, diarrhoea, jaundice, cholera etc.  Therapeutic agents from local plant resources, for example, anticancer / antitumour drugs, sweeteners, dyes, flavours, fragrances etc.

ANIMAL HEALTH CARE SECTOR

 Veterinary diagnostics and vaccines  Immunology  Embryo transfer 12

MINING SECTOR

 Mineral leaching is particularly important in the context of Nepal having several mineral deposits scattered all over the country in relatively lower volume of mineral deposits.

DNA TECHNOLOGY

 Identification and isolation of genes for use in molecular hybridization.  DNA fingerprinting for forensic, diagnosis of genetic disease, paternity analysis, immigration, and citizenship record.  Studies on polymorphism and molecular mapping of endangered and commercial crops.  Development of diagnostic kits.  Selection in breeding programme.

AQUACULTURE

 Feeds  Diagnostics

HUMAN RESOURCES DEVELOPMENT SECTOR

 Introduction of biotechnology and bioinformatics at the graduate and post graduate levels in universities.  More Ph. D.s at home and abroad.  Academic collaboration with leading universities in the neighbour and abroad.  Develop programs to foster biotechnology entrepreneurship.

ENVIRONMENT AND BIODIVERSITY

 Biological control agents  Urban waste management  Industrial waste treatment  Monitoring tools and biosensors

6. POSSIBLE ROLE OF BINASIA

 A serious constraint is the lack of information on biotechnologies. No institution has developed capability in information collection, retrieval and dissemination. Therefore it relies on industrialized countries for specialized information.  Solution to information shortage is to conduct a thorough literature search on the existing information, which should then be stored in the institutes having the 13

capability to manage information and make it easily accessible to biotechnology researchers. OR to set up a biotechnology information centre and establish a computer link with institutions in the industrialized countries. However this depends on the willingness of the institutions in the developed countries to share their biotechnological information with developing or least developed countries.  Hence pivotal role of BINASIA could be to make avail of publications with full text.

T H A N K Y O U