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水產種原庫多功能建置及科技產業化應用

  • 日期:107-02-20
  • 計畫編號:107農科-1.3.5-水-A1
  • 年度:2018
  • 領域:農業科技管理及產業化
  • 主持人:吳豐成
  • 研究人員:楊順德、曾福生、謝恆毅、蔡惠萍、杜金蓮、陳鏗元、張格銓、王敏儒、江姿吟

(一)優質九孔之選育與技術整合應用(水產養殖組) 為了解105年建立的九孔(Haliotis diversicolor)精子的冷凍保存精子的實用性 ,使用該精子在相同解凍條件下冷凍保存180天,在23-25 ° C受精,胚胎畸形率和 沒有凍存處理的對照組分別為46%和28%,比較精液的濃度為102 / mL和103 / mL,結果 冷凍保存180天之孵化情況分別為67.16個(0.00015%)和114.28個(0.00026%)孵化相 當低,顯示冷凍保存的時間對後續的發育影響非常大。 (二)臺灣重要經濟性養殖魚貝類精液冷凍保存-利用冷凍精液進行雜交育種(東部海 洋生物研究中心) 以點帶石斑 Epinephelus coioides 及條石鯛 Oplegnathus fasciatus 為實驗材料 ,對精子稀釋液、抗凍劑種類和適宜濃度之冷凍保存液進行篩選。結果顯示利用 Hank’s稀釋液、海水魚生理食鹽水及EM1-2稀釋液均適合於點帶石斑精子冷凍保存 ,分別以500 μl冷凍麥管 (Cryo-straw) )為容器並將之凍結保存於液態氮中,解 凍後精子活力可達73.00%±2.00、71.67%±1.53及68.00%±3.00。另利用Hank’s稀釋 液為基礎稀釋液對抗凍劑保護劑進行篩選試驗,針對二甲基乙砜(DMSO)、甘油 (Glycerin)、、丙二醇(Propylene Glycol)、甲醇(Methanol) 4種抗凍劑進行篩選 試驗,結果顯示12%和18%的DMSO及Propylene Glycol 冷凍保存後精子活力與新鮮精 液無顯著差異(P>0.05),其中以12%DMSO冷凍保存精子效果最佳,解凍後精子活力可 達91.00%±2.00。而條石鯛冷凍精液試驗結果以Hank’s稀釋液冷凍保存精子其活力 較佳,同樣以500 μl冷凍麥管為容器並將之凍結保存於液態氮中,解凍後精子活力 為54.33%±7.37。利用Hank’s稀釋液為基礎稀釋液對抗凍劑保護劑進行篩選試驗 ,結果顯示12% Glycerin 冷凍保存後精子活力較佳且與新鮮精液無顯著差異 (P>0.05),其冷凍保存精子解凍後精子活力可達85.67%±4.51。另利用於106年度冷 凍保存的龍膽石斑精液與點帶石斑卵進行受精,其受精率分別可達97.72%±1.64及 96.72%±1.09,與新鮮精液95.56%±1.75無顯著差異(P>0.05);其孵化率分為 79.22%±2.88及78.27%±2.21,略低於新鮮精液90.82%±0.48。本試驗已初步完建立點 帶石斑及條石鯛的精液冷凍保存技術,為石斑魚及條石鯛的人工繁殖及雜交育種提 供基礎方法。 (三)吳郭魚種原之產業應用(淡水繁養殖研究中心) 本計畫之主軸為吳郭魚種原之產業應用,本年度之研究與應用包含下列3個項目 (1)持續技轉遺傳雄性尼羅吳郭魚技術 (2)推廣快速成長尼羅吳郭魚苗50萬尾 (3)進 行多雌成長遺傳研究,了解多雌成長遺傳特性對不同成長期的吳郭魚造成的影響。 本年技轉部分未有完成件數,明年仍須努力,推廣魚苗的部份則完成約23萬尾。另 外,本研究進一步分析多雌遺傳之成長相關特性,結果顯示多雌遺傳搭配P1型的特 與單純帶遺傳雄性P1型之成長沒有顯著差異。另外,多雌成長遺傳的性別決定力僅 次於遺傳雄性P1型,顯示其有更多的研究價值。 (四) 長硨磲蛤共生藻離體培育及復合(澎湖海洋生物研究中心) 利用人工繁殖長硨磲蛤之不同外套膜顏色之個體進行外套膜樣本採集、離體培養、 以及利用分子工具針對共生藻品系進行確認,結果顯示不同外套模樣態的長硨磲蛤體內所含有之共生藻品系皆為A品系,顯示外套膜不同的樣態並非因為具有不同共生 藻品系所致。本年度亦已利用離體培育共生藻進行人工繁殖幼生復合。體長1公分左 右之稚貝,足絲再生時間可在24個小時內完成、而3-6公分的個體,則是5-7天可完 成。2017年野外移植殼長6公分左右的個體,一年後野外的存活率介於0-20%(東吉 0%、七美5%、嶼坪20%),籠具內3公分個體存活率96.3%。

研究報告摘要(英)


1. In order to understand practicablity cryopreserved sperm of small abalone (Haliotis diversicolor )sperm, which have established in 2016, using this sperm that has been cryopreserved in liquid nitrogen (LN) for 180 days under the same thawing conditions fertilized test at concentration of 102 / mL to 103 / mL at 23-25 ° C, comparing the deformity of embryo stage and the control group, there were respectively 46% and 28%, and the hatch-out was respectively 67 (0.00015%) and 114 (0.00026%). The results showed that the length of time for cryopreservation in LN has a great impact on subsequent development. 2. In study the cryopreservation of Epinephelus coioides and Oplegnathus fasciatus sperm , mature E. coioides semen and O. fasciatus semen were used as experimental material, and concentration of various semen diluents, and concentration of cryoprotectant were screened.We used liquid nitrogen and 0.5 ml cryo-straws to preserve sperms by step cooling procedure. The result showed the cryopreservation of  E. coioides semen that Hank’s solution、Marine Fish Ringer and EM1-2 semen duluents was better , which preserved 73.00%±2.00、71.67%±1.53及68.00%±3.00 sperm motility after unfrozen. Using Hank’s solution as the liquid foundation to prepare cryoprotectant, no significant difference was observed in sperm motility by 12% and 18% DMSO or Propylene Glycol cryopreservation (P>0.05) . The best of all is 12% DMSO, which preserved 91.00% ± 2.00 sperm motility respectively after unfrozen. The cryopreservation of O. fasciatus semen that Hank’s solution semen diluents was better in sperm motility, which preserved 54.33% ± 7.37% sperm motility after unfrozen. Using Hank’s solution as the liquid foundation to prepare cryoprotectant, no significant difference was observed in sperm motility by 12% Glycerin cryopreservation, which preserved 85.67% ±4.51 sperm motility respectively after unfrozen.Using cryopreserved E.lanceolatus semen to fertilize E. coioides  eggs, the rates of fertilization were 97.72%±1.64 and 96.72%±1.09 without difference with fresh sperms, the rates of hatching were 79.22%±2.88 and 78.27%±2.21. We established a sperm cryopreservation protocol for E. coioides and O. fasciatus on this study, and it will provide a basis for the artificial breeding and cross breeding. 3. The purpose of this program is to provide the tilapia industry with new techniques that were developed by the Germplasm Bank of Freshwater Fish, Fisheries Research Institute. These techniques will be promoted to the industry with following goals to enhance the competitive strength.The technology transfer of genetically male tilapia.The supply of 500,000 fries of Nile tilapia with the feature of high growth rate.The application of molecular protocols to analysis the reason of low maleratio of tilapia fry.Although the FRI could provide the aquaculture industry with a high-growth tilapia to commercialization but could not match and through licensing agreements in this year. In addition, we try to analyze the sex-determining markers of genetically both female and male ,try to realize the growth characters. The results show that the final weight in the group C (genetic female and male) with group D (genetic male only) have no significant difference. Otherwise, the genetic female marker could be applied on genetic breeding in the future. 4.Symbiotic alga of Tridacna maxima were isolated and cultivated from artificially produced juveniles with different mantle colorations. Molecular genetic data verified all symbiotic alga from different hosts are the same clade (A). It means different mantle colorations of T. maxima are not determined by symbiotic alga. The cultivated alga were also prepared for infection trail for artificially produced larvae. Byssus of juveniles with shell length of 1 cm will regenerate within 24 hours whereas those of 3-6 cm will do in 5-7 days. One-year survival rates of transplanted juveniles in 2017 are between 0-20%, while those protected in cage are 96.3%.