Improvement of Cold-Shock Treatment Conditions for Triploidy Induction in Pacific Abalone Haliotis discus hannai
Choul Ji Park, Min-Su Kwon, Eun Jeong Kim and Yoon Kwon Nam
Genetics & Breeding Research Center, National Institute of Fisheries Science, Geoje, Korea Department of Fisheries Biology, Pukyong National University, Busan 48513, Korea
Optimized protocols of cold shock treatment for the induction of triploidy were developed in Pacific abalone Haliotis discus hannai. Time windows of polar body extrusions were analyzed and various technical parameters including initial treatment times, treatment temperatures (-1ºC, 1ºC and 3ºC) and treatment durations (10 to 24 min) were tested in order to develop the best combination(s) among parameters enabling the efficient blocking the polar bodies. Based on a series of tests regarding hatchability of embryos, normality of larvae and induction efficiency of triploidy, the most appropriate protocol to block the first polar body was proven to be the cold shock treatment at 1ºC for 14 min with an initial treatment at 11 min post insemination. On the other hand, the most efficient condition for blocking the second polar body was the cold shock at 1ºC for 14 min with an initial treatment at 25-27 min post insemination. Under these treatment conditions, larval yields of cold-shock treated groups relative to non-treated control groups were up to 70% with a fairly consistent triploidy rate up to near 100%. Efficacy and reproducibility of triploid induction were further validated under scaled-up conditions; however, the yield of triploid larvae was highly influenced depending on the gamete quality used. Results from this study could be a useful basis for developing the practical protocol for mass production of triploid seedlings in this abalone species.
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