PROJECT TITLE: Development of DNA markers for identification of colour varieties and sex in tiger barbs

Executive Summary

Malaysia has a great potential to become a major producer and exporter of ornamental fish. Freshwater fishes constitute the bulk of aquarium fish in Malaysia with its export value for 1993 was RM 17.5 million. Till recently the main supply of aquarium fish came from the natural habitat but due to unfavourable destruction of its habitat, our endemic species are facing critical stresses, which may lead to extinction. Therefor the research to develop novel varieties of indigenous ornamental fish species for new combination of colour patterns and variety has been proposed.

The target species for this research is Puntius spp (Tiger barbs) which is a valuable species of aquarium fishes. This programme will include research projects of this indigenous fish species for enhancing for commercial value using molecular techniques and genetic selection. It can be done through their variation in many morphological, physiological and biochemical features.

A marked proportion of this variation is heredity and this circumstance guarantees of selection work. They have to be selected for desirable genes from existing genetic variation and also from spontaneous mutation that may arose occasionally in their stocks and develop pure lines. The main task in selection program of ornamental fish is in the development of new varieties and strain with attractive pigmentation and body shape. Simple Medelian inheritance (crosses) will be made following the breeding method proposed by Captive breeding Guidelines by Aquatic Conservation Network (1994). The DNA markers are essential to find diagnostic markers between different stocks of the same species, and new technologies for DNA fingerprinting such as RAPD which detects base substitutions will be utilized.

Further to these, isolation, sequencing and cloning of useful genes like those, which control colouration and morphological aberrancies, will be carried out which eventually will entail application to gene transfer. To elucidate the gene control of the colour pattern of the fish will be carried out by screening and carefully picking out spontaneous mutants which may arise in the stocks and subsequently by fixing the gene and building up the new line over several generations or by hybridization between the varieties to achieve new combination of colour patterns. It is important to understand the gene control of the colour phenotypes of different varieties of fish in order to improve the existing varieties and to systematically breed for new one. This can be achieved again through Mandelian Crosses and intense selection. The DNA fingerprinting will be employed to detect how much inbreeding has occurred.

Once the gene such as colour patterns is selected and fixed in a line, the fish can be produced on commercial scale. The direct beneficiaries of the projects include the commercial breeders for they will have propriety tags of their cultured strains. It will allow fish seed producers to monitor the levels of genetic variability's present in their broodstocks and avoid deleterious effects of inbreeding. Selective breeding and crosses made between genetically suitable stocks with new colour patterns and variety whenever necessary to maintain and improve the quality of the broodstocks both in it's aesthetic and price aspects.

This is an important stage fir commercial fish production as well as for conservation and management of Malaysia's rich genetic resources, which is in line with The Rio Declaration. In the initial stage of the projects, outputs will be disseminated by publications in scientific journals and presentation at seminars. The activities of the project will be as an economic contribution as well as a mean to create and establish the Centre of Excellence for Production of Novel Varieties of Ornamental Fish.

Objective Achivements

• Recognition of several genetic markers with diferent fish colours thus allowing for the formulation of breeding programs.
• Five different patterns of fish were obtained
• there are publications both in proceedings and journals as the output from the project
• On the whole the project objectives were achived in both DNAmarkers and selective breeding studies except in the production of Transgenic fish.

Benefits of the Project

• The availability of genetic markers to identify fish stock as well as individual fishes will enable commercial breeders to have propriety tags their cultural strains. It will allow fish seed producers to monitor the levels of genetic variability present in their broodstocks and avoid the deleteriou effects inbreeding. Selective breeding and crosses can be made between genetically suitable stock with new colour patterns and variety whenever necessary to maintain and improve the quality of broodstocks. The production of transgenic fish will boost the aquarium fish industry as new products in the market. It will catch good market price and bring attention to all aquarium fish hobbyist. At the same time availability of genetic markers to identify fish stock as well as individual fishes will enable commercial breeders to have propriety tags of their cultural starins.