Development of an Improved Automatic Chromosome Manipulation System and Optimization of Related Parameters for Pacific Oyster, Crassostrea gigas

牡蠣染色體操作改良型自動化系統及其相關條件之研發

趙乃賢(Nai-Hsien Chao) ； 朱元南(Yuan-Nan Chu) ； 林佳慧(Chia-Hui Lin)

《臺灣水產學會刊》 32卷3期 (2005/09) Pp. 217-227

https://doi.org/10.29822/JFST.200509.0003

摘要

本研究的目的為探討牡蠣大量人工受精和染色體操作自動化之可行性及研發一套又進一步改良之自動化系統。發現牡蠣卵在未受精前分別能耐受暴露於海水和空氣中30和90分鐘。受精卵在濃縮用輪鼓轉速100, 200, 300rpm下配合馬達抽水率2.4, 5.8, 7.6liter/min時轉動20及40分鐘，其分項實驗中各小組孵化率並無差異。但發現以轉速300rpm和抽水率2.4liter/min運轉20和40分鐘，二小組之平均孵化率與它組有顯著性差異；至於抽水率7.6liter/min運轉20分鐘時，輪鼓轉速100, 200, 300rpm三小組之平均孵化率亦然，應予避免。整體改良式染色體操作系統的綜合設計符合牡蠣使用。至於觸控面板及可手動亦可自動化程式之電子控制系統甚具改良效果，有助染色體操作之各項階段性處理得以確實依序準時且高效完成。
貝類染色體操作等較傳統的生物技術因未涉及基因之改變，生產出之貝類可及早獲得認可與上市的機會，應予開發企業化生產。以自動化設施供染色體操作流程應用是現代趨勢，可省力，免失誤，有助提高正確結果的再生率並可開發國際行銷之潛力。

關鍵字

牡蠣 ； 大量人工受精 ； 染色體操作 ； 改良式自動系統

並列摘要

This paper examines two major works, (1) improvement in the design of automatic manipulation system for the complicated chromosomal manipulation of shellfish, and (2) investigation of the optimum parameters needed for automatic chromosome manipulation system in oyster based on species-specific characteristics.
High quality experimental material for ideal fertilization and hatching rates of eggs are highly desired for follow-up trials on chromosome manipulation. The experiments to optimize parameters exclude the use of functional chemicals for induction of triploid or tetraploid with the hope that the automation procedure will not cause any unnecessary damage. Only when high hatching rates of eggs is achieved using the automatic manipulator, that this improved system can be used for follow-up treatments on chromosome manipulation.
Results indicate that fertilization should be performed within 30 minutes after the oyster eggs were stripped and kept submerged in sea water. Any longer delay should be avoided. On the other hand, air exposure of unfertilized eggs within 90 min was considered to be acceptable in the design of the automatic manipulation system.
Hatching rates in individual subgroups using drum speeds of 100, 200, or 300 rpm combined with pump flows of 2.4, 5.8 or 7.6 liter/min for 20 min or 40 min were not significantly different. It was not recommended to adopt a drum speed of 300 rpm for manipulating oyster fertilized eggs if a pump flow of 2.4 liter/min operating for 20 and 40 min were desired because an averaged hatching rate at a drum speed of 300 rpm was significantly lower than that at 200 rpm. Similarly, a pump flow of 7.6 liter/min should be avoided because an averaged hatching rate from three drum speeds at a pump flow of 7.6 liter/min was significantly lower than that at 5.8 liter/min.
Chromosome manipulated shellfish are not related with gene modification and thus are accepted by the market. Therefore it is reasonable to encourage the related enterprise so that value-added product of shellfish can be continuously produced. Improved automation in any manipulation protocol has been further enhanced for the sake of saving energy, saving time, avoiding man-made mistakes, and raising the replication of anticipated results. It possesses the potential of international marketing.

並列關鍵字

oyster ； mass propagation ； chromosome manipulation ； improved automatic manipulation system