Title: DNA-Based Identifications of Tilapia in Hawaii
1DNA-Based Identifications of Tilapia in Hawaii
Jinzeng Yang and Harry Ako
College of Tropical Agriculture and Human
Resources University of Hawaii at Manoa
2Tilapia Aquaculture
- High-quality food fish
- Easy to reproduce, breed and farm
- Farming for freshwater and brackish water
- Excellent species for aquaponics
- A potential growth area for Hawaii with
groundwork having been laid in the marketing
area.
3Challenges of Tilapia Farming in Hawaii
4The Goal and Objectives
- The long-term goal of the project is to have
fast-growing tilapia for farmers in Hawaii. - Main Objectives
- Identification of the tilapia strains and hybrids
existing in the wild and captive populations by
DNA barcoding method - Successfully allow importation of fast growing
strains or develop DNA-based testing tools for
selection of high-growth tilapia by selectively
breeding local tilapia resources.
5- What is DNA and genome?
- Permanent part of the body cells, inheritable
- Genome a whole collection of the DNA of a
species - Different species are made of different genomes
- Strains with one species have slightly different
DNA sequences.
6How Can DNA-based Technology be Helpful for
Tilapia Farming?
7The PCR Song There was a time when to amplify
DNA, You had to grow tons and tons of tiny
cells. Then along came a guy named Dr. Kary
Mullis, Said you can amplify in vitro just as
well. Just mix your template with a buffer and
some primers, Nucleotides and polymerases,
too. Denaturing, annealing, and extending. Well
its amazing what heating and cooling and heating
will do. PCR, when you need to detect
mutations. PCR, when you need to recombine. PCR,
when you need to find out who the daddy mommy
is. PCR, when you need to solve a
crime. www.cnpg.com/video/flatfiles/539/
8PCR and DNA Sequencing
- PCR is a technology used to amplify genes of
interest. - Template DNA, primer, denaturing, annealing and
extending. - Sequencing amplified genes from fin clip.
- Each MBBE lab has about 4 PCR machines.
DNA Sequencing
PCR Amplification
9DNA Barcoding
- It is DNA sequence-based taxonomic method of
species identification. - A 648 base-pair region in the mitochondrial
(maternal) cytochrome c oxidase 1 gene (COI) has
been extensively studied. FISH-BOL database.
Obtain sequences of known species as niloticus,
mossambicus, aureus and so on. - The DNA sequence COI can identify parents
(mothers) of existing species in Hawaii,
including niloticus.
10- Amplify and sequence DNA of available tilapia in
Hawaii. - If we have niloticus here already, DOA may let us
import. - If we have niloticus here already, we can reverse
breed to get back a fast growing strain. - (These methods have been used to identified
mislabeled fillets.)
11Microsatellite DNA Marker
- Simple nucleotide repeat, very short,
tandem-repeat sequences, such as
ACACACACACAC-(AC)6 - Highly polymorphic- most individual animals are
heterozygous at each locus this is because there
is no selective pressure to maintain the
sequences they are not translated. - Genetics analysis DNA identification,
marker-assisted selection, parental assignment.
Useful to distinguish closely related organisms
including siblings of same species. - Can use this method in marker assisted breeding.
12Microsatellite DNA Analysis
CACACACACACACACACACACACACACA
13Plan of Work
14Summary
- Determination of genetic resources in Hawaii
- To allow importation of fast growing species
since they may already be here or - To back engineer to fast growing strains that
originally got in - To use microsatellite markers if there is a
selected breeding program. - Sponsored by CTSA.