Nepal Academy of Science and Technology (NAST)

OVERVIEW OF MOLECULAR MARKERS

Protein-based - eg. Isozymes, Allozymes, Seed proteins

DNA-based or DNA –fingerprinting techniques –

	Non-PCR-based eg. RFLP and DNA-Hybridization (Southern blotting)
	PCR-based eg. RAPDs, microsatellites (SSRs), AFLPs, ISSRs, DNA sequencing and PCR-RFLP

APPLICATION OF MOLECULAR MARKERS

	Medicine – Human disease diagnosis, gene therapy, human genomics
	Agriculture and Forestry – Plant and animal (veterinary) disease diagnosis, germ plasm characterization and diagnosis of difficult to diagnose species, sub-species and plant cultivars, study of genetic diversity and relationship among and within species and populations (Population Genetics), molecular taxonomy or molecular systematics, conservation genetics, marker assisted selection (MAS) and molecular breeding, identification of molecular markers linked to important agronomic traits eg. disease resistance, stress tolerance, salt tolerance, herbicide resistance genes etc. of agricultural crops, identification and isolation of important candidate genes for the study of DNA-sequence variation, gene cloning and sequencing, genetic engineering (Production of GMOs).
	Forensic – Paternity analysis, identification of criminals, identification of illegal plant and animal parts or their products in trade.

CURRENT MOLECULAR PROJECTS AT NAST

Project 1 - Molecular Characterization of Important Medicinal Plants of Nepal for their authentication, sustainable utilization and conservation.

Objectives:

	Study of genetic diversity and systematic relationship of highly important medicinal plants of Nepal (eg. Swertia spp., Taxus wallichiana Zucc. etc.) using PCR and DNA sequencing-based molecular techniques.
	Generate high value medicinal plant species-specific or population specific molecular markers and DNA fingerprints for authentication of Nepalese species and cultivars.

Research Team: Dr. Sangita Shrestha, Ms. Neesha Rana, Ms. Jaishree Sijapati, Ms. Prabina Rana and Ms. Anjana Giri.

Project 2 – Molecular characterization of Bacillus thuringiensis (Bt) strains isolated from various geoclimatic regions of Nepal for their authentication and effective utilization.

Objectives:

	Characterize Nepalese strains of Bt using arbitrarily primed PCR (RAPDs).
	Characterization of Cry genes in Nepalese Bt isolates using specifically primed PCR.
	Study of sequence variation in Cry genes of Nepalese isolates in future for authentication of Nepalese strains.

Research Team: Dr. Sangita Shrestha, Ms. Jaishree sijapati, Ms. Prabina Rana, Ms. Neesha Rana and Ms. Anjana giri

Project 3 – Molecular Characterization of Tea [Camellia sinensis (L) O. Kuntze] germplasm of Eastern Nepal for authentication and sustainable utilization.

Objectives:

	Characterize Nepalese cultivars of Tea using arbitrarily primed PCR.
	Study of genetic diversity and relationship among cultivars based on molecular data.
	Generate cultivar specific DNA fingerprints or specific PCR-based or PCR-RFLP based diagnostic for the Nepalese cultivars for their authentication.

Research team: Dr. Kayo Devi Yami, Dr. Sangita Shrestha and Mr. Iswor Prasad Khanal

Project 4 – Polymerase Chain Reaction (PCR) Based Diagnosis of Citrus Huanglongbing (HLB) Disease in Nepal .

Objectives:

	Reliable diagnosis of citrus HLB disease using two specific PCR assays viz. one based on 16S rDNA and another based on ribosomal protein genes from b operon (rpl KAJL-rpoBC) of HLB organism.
	Study of incidence and prevalence of HLB infestation in various citrus growing region of Nepal based on PCR-based diagnostic result.
	To provide analytical service to farmers and nurserymen for the molecular diagnosis of HLB disease.

Research Team: Dr. Sangita Shrestha, Dr. Chiranjivi Regmi, Ms. Neesha Rana, Ms. Prabina Rana, Ms. Anjana Giri and Ms. Jaishree sijapati.

Note: All above-mentioned projects are seeking funds from National and International organizations for collaboration. Interested NGOs and INGOs are requested to contact RONAST.