2022 Fiscal Year Annual Research Report
Basis of clownfish pigmentation plasticity
| Project/Area Number |
22F22768
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| Allocation Type | Single-year Grants |
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| Research Institution | Okinawa Institute of Science and Technology Graduate University |
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| Host Researcher |
LAUDET Vincent 沖縄科学技術大学院大学, 海洋生態進化発生生物学ユニット, 教授 (20898423)
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| Foreign Research Fellow |
MITCHELL LAURIE 沖縄科学技術大学院大学, 海洋生態進化発生生物学ユニット, 外国人特別研究員
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| Project Period (FY) |
2022-11-16 – 2025-03-31
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| Keywords | Pigmentation / Ecology / Coral Reef Fishes / Ontogeny / Cell Death / Color Pattern Change |
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| Outline of Annual Research Achievements |
The purpose of this project is to investigate the ecological and genetic drivers of pigmentation and color pattern diversity in anemonefishes. To achieve this goal, we have determined the ontogenetic transcriptome of the tomato anemonefish (Amphiprion frenatus) during its juvenile color pattern change and linked aspects of ecology with the timing of its pigmentation transition.
To further explore interspecies color pattern differences, we have crossed the (horizontally) striped species A. sandaracinos with the (vertically) barred A. ocellaris to produce a viable first generation of hybrids. The second generation of hybrids will be used to analyze colour patterns and identify key genetic loci that correspond to aspects of color pattern variation.
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| Current Status of Research Progress |
Current Status of Research Progress
4: Progress in research has been delayed.
Reason
Major delays in setting-up our husbandry has delayed the establishment of stable breeding fish couples. Moreover, the presence of marine fish egg parasites in our aquarium system has meant that time has been spent on identifying and testing the treatment of infected eggs. This has meant that the final part of this project in using CRISPR/Cas9 genome editing to test the function of specific genes in white bar formation has also been significantly delayed. Our recent successful treatment of artificially incubated eggs gives us confidence that genome-editing can resume soon, and results will be generated.
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| Strategy for Future Research Activity |
The work is now going in three primary directions:
1° We are conducting behavioural experiments testing the functional importance of bar number after larval settlement. We are also performing immunostaining assays to confirm that the mode of pigment cell death is apoptosis.
2° We are applying CRISPR/Cas9 genome-editing to perform the targeted knockout of specific genes which are strongly suspected to be involved in white bar formation.
3° In collaboration with Acadamia Sinica in Taiwan, we are attempting to raise the first-generation hybrid fish to produce a second generation. This second-generation will be photographed and sampled for DNA to perform a quantitative trait loci analysis to identify key genes contributing to aspects of color pattern.
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