Assessments of genetic structure and population connectivity of Hawaiian flame wrasse may provide a model for gene dispersal in Hawaiian reef fishes and potentially be applied to other economically, culturally, and biologically important reef fish species. Combining tools such as genetic analysis with aquaculture techniques is considered a valuable strategy in the conservation of Hawaiian reef fishes. However, minimal funding and lack of genetics expertise often act as barriers in including genetics in conservation programs. Here, we outline a collaborative initiative among University of Hawai‘i at Hilo’s Department of Biology undergraduate genetics class, Tropical Conservation Biology and Environmental Science graduate program (TCBES), and Pacific Aquaculture and Coastal Resources Center (PACRC). This initiative aims to generate preliminary genetic data investigating genetic structure in endemic Hawaiian flame wrasse (Cirrhilabrus jordani), by involving undergraduate student coursework. Genetic comparisons of wild populations coupled with larval rearing will indicate the duration of the larval cycle and allow us to determine if this species is a large metapopulation or separate island populations. Undergraduate students are working with flame wrasse samples collected from Oahu, Maui, and Hawaiʻi Island obtained through the PACRC larval rearing program. During the undergraduate’s teaching lab, fish are genotyped to assess cytochrome oxidase I (COI) genetic variation. The COI gene is often used in genetic structure analysis, however future directions include conducting a genome-wide SNP analysis. This project will be used to increase undergraduate involvement in hands-on research with local relevance, as well as increase interdepartmental collaborations at UHH.
- 15 specimens of Hawaiian Flame Wrasse from Oahu sequenced for the COI gene
- Comparisons between Oahu Hawaiian Flame Wrasse sequences and preexisting GenBank sequence
- Examine the % similarities between island populations
- Determine if the amount of similarity is enough to delineate genetic structure between islands
- Currently, all exports of Hawaiian reef fishes for the marine ornamental fish trade are wild-captured (Ogawa & Brown 2001)
- Understanding the population structure of Hawaiian reef fishes has important conservation implications
- Minimal funding and lack of genetics expertise often act as barriers in including genetics in conservation programs
- Genetic connectivity of Hawaiian Flame Wrasse (Cirrhilabrus jordani) between the main Hawaiian Islands has not yet been assessed
- The aim is to generate preliminary genetic data investigating genetic structure this endemic species through a collaborative initiative among University of Hawai‘i at Hilo’s Department of Biology undergraduate genetics class, TCBES graduate program, and PACRC
H0: There is no genetic distinctiveness among Hawaiian Flame Wrasse (Cirrhilabrus jordani) populations between the main Hawaiian Islands.
HA: There is genetic distinctiveness among Hawaiian Flame Wrasse (Cirrhilabrus jordani) populations between the main Hawaiian Islands.
- Increased undergraduate involvement in hands-on research and interdepartmental collaborations at UHH
- Comparisons will be indicative of the possible variation within the COI gene among Hawaiian Flame Wrasse populations from neighboring islands
- This study may provide insight into whether this species is grouped genetically as one metapopulation or contains distinct island populations
- If there is a population that experiences a decline on one island, loss of genetic diversity
- Neighboring islands have the possibility of replenishing genetic diversity (Salles et al. 2016)
- Hawaiian Flame Wrasse study may be serve as a model for other species with similar life history
- Samples collected from the island of Oahu
- Chelex 100 DNA Extractions
- FISH F2 and R2 (Ward et al. 2005) targeting cytochrome oxidase I gene (COI)
-Sequences will be aligned and compared using the software Geneious
Chelex 100 DNA Extractions:
1. 600uL of 5% Chelex
2. Small amount of tissue and 5uL of 20mg/mL pro K
3. Digest at 55C in rocker, check after 1 hour, then overnight
4. Spin down at 12000rpm for 1 min
5. Put aqueous in fresh tube
6. Precipitate using 20uL of 5M LiCl and cold 100% EtOH, gently invert
7. Spin tube for 8000rpm for 1 min to pellet DNA, loop with fresh tip
8. Wash with 70% EtOH, drain and air dry to remove EtOH
9. Resuspend in 100uL of H2O
Use Q5 Hot Start (Denature: 98C for 30s, Gradient from 61-69C over 35 cycles 98C for 8s, 53C Ta for 20s, 72C for 20s, Final Extensiom: 72C for 2 min)
- Collect samples from the islands of Maui, Hawaii, Kauai, and Northwestern Hawaiian Islands for further comparisons
- Genetic comparisons of wild populations coupled with larval rearing will indicate the duration of the larval cycle and allow us to determine if this species is a large metapopulation or separate island populations
- Genome-wide SNP analysis
Ogawa, T., & Brown, C. L. (2001). Ornamental reef fish aquaculture and collection in Hawaii. Aquarium Sciences and Conservation, 3(1-3), 151-169.
Salles, O. C., Pujol, B., Maynard, J. A., Almany, G. R., Berumen, M. L., Jones, G. P., ... & Planes, S. (2016). First genealogy for a wild marine fish population reveals multigenerational philopatry. Proceedings of the National Academy of Sciences, 113(46), 13245-13250.
Schultz, J. K., Pyle, R. L., DeMartini, E., & Bowen, B. W. (2007). Genetic connectivity among color morphs and Pacific archipelagos for the flame angelfish, Centropyge loriculus. Marine Biology, 151(1), 167-175.
Ward, R. D., Zemlak, T. S., Innes, B. H., Last, P. R., & Hebert, P. D. (2005). DNA barcoding Australia's fish species. Philosophical Transactions of the Royal Society of London B: Biological Sciences, 360(1462), 1847-1857.