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水產種原庫多功能建置及科技產業化應用

  • 日期:106-02-21
  • 計畫編號:106農科-1.2.3-水-A1
  • 年度:2017
  • 領域:農業科技產業化領域
  • 主持人:許晉榮
  • 研究人員:曾福生、謝恆毅、蔡惠萍、張格銓、杜金蓮、陳鏗元、王 敏儒

(一)優質九孔之選育與技術整合應用(水產養殖組) 為了解2016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天和1天,精液的濃度為10 2 / mL~10 3 / mL在23-25 ° C受精,比較兩個細胞階段的胚胎的發生率高低,作為精子液氮儲存 質量評估的參考。結果表明,受精率為4.07±4.09%和20.74±5.48%。另外,利用公佈 的與鮑肌肉萎縮症病毒(Abalone shriveling syndrome associated virus, AbSV)相關的基因作為精子篩查的遺傳標記,排除疑感染病原體的樣本。

(二)臺灣重要經濟性養殖魚貝類精液冷凍保存-利用石斑冷凍精液進行雜交育種(東 部海洋生物研究中心) 以龍膽石斑(鞍帶石斑)Epinephelus lanceolatus 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稀釋液及海水魚 生理食鹽水較適合於龍膽石斑精子冷凍保存,以2 ml冷凍小管為容器並將之凍結保 存於液態氮中,解凍後精子活力可達90.33%±0.58%及91.00% ±1.00%,較優於MPRS稀 釋液。利用Hank’s為基礎稀釋液對抗凍劑保護劑進行篩選試驗,結果顯示12%和 18%的二甲基乙砜(DMSO)冷凍保存後精子活力無顯著差異(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74.67%±6.51%。利用冷凍保存 的龍膽石斑精液與分別與雲紋石斑卵進行受精,其受精率分別可達90.37%±8.71,與 新鮮精液無顯著差異(P>0.05)。本試驗顯示利用Hank’s稀釋液配制12% DMSO可用於 冷凍保存龍膽石斑精液。 本試驗已初步完建立龍膽石斑的精液冷凍保存技術,為石 斑魚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) 本年度之研究與應用包含下列3個項目 (1)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)進行多雌遺傳研究,解析多雌遺傳特性降低魚苗的 雄性比例原因。本年度已公開技轉,但至目前為止仍未媒合成功,為順利完成技轉 ,已評估將技轉內容作適當修改。推廣魚苗的部份則完成約44萬尾。另外,本研究 再次分析多雌遺傳之遺傳特性,結果顯示多雌遺傳的特性尚未能穩定的操控,但再 次發現多雌遺傳能促進魚體成長,未來值得更進一步研究。 (四)長硨磲蛤人工繁養殖及中間育成技術建立(澎湖海洋生物研究中心) 本年度共採集長硨磲蛤種貝20顆,進行三批次人工繁殖操作,迄本年度為止,成功 生產超過10000顆長硨磲蛤苗,培育長硨磲蛤1公分稚貝5000顆以上。以溫度刺激法 可成功誘導生殖腺成熟種貝排精排卵,在28℃、33psu的培育條件下,受精卵約需 14-15天可變態成為底棲稚貝。成功設置中間育成場,供種貝及稚貝渡冬及中間育成 使用。

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 for 180 days and one day under the same thawing conditions fertilized test at concentration of 10 2 / mL to 10 3 / mL at 23-25 ° C, comparing the occurrence of embryos in two cell stages, as a reference for evaluating the quality of sperm liquid nitrogen storage. The results showed that the fertilized rate was 4.07±4.09% and 20.74±5.48%. In addition, the use of published Abalone shriveling syndrome associated virus (AbSV) virus-related genes as genetic markers of sperm screening to exclude suspected pathogen-infected samples.

2. In study the cryopreservation of giant grouper ( Epinephelus lanceolatus ) sperm , mature E. lanceolatussemen was used as experimental material, and concentration of various semen diluents, and concentration of cryoprotectant were screened. We used liquid nitrogen and 2.0 ml cryovials to preserve sperms by step cooling procedure. The result showed the cryopreservation of E. lanceolatus semen that Hank’s solution and Marine Fish Ringeras semen diluents was better than MPRS solution as semen duluents, which preserved 90.33% ±0.58% and 91.00% ±1.00% 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 cryopreservation (P>0.05) . The best of all is 12% DMSO, which preserved 74.61% ± 4.57 % sperm motility respectively after unfrozen. Using cryopreserved E. lanceolatus semen to fertilize E. moara  eggs, the rates of fertilization were 90.37 % ± 8.71% , without difference with fresh sperms. This study showed that 12% DMSO using Hank’s solution as the liquid foundation could cryopreserve E. lanceolatus semen .We established a frozen sperm library based 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.(1)The technology transfer of genetically male tilapia.(2)The supply of 500,000 fries of Nile tilapia with the feature of high growth rate.(3)The application of molecular protocols to analysis the reason of low male-ratio 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 female and try to realize the characters. The results show that breeding on the genetic female still cannot breed high female-ratio stably. Otherwise, the genetic female marker could be applied on body development in the future. 4.20 mature giant clamsTridacna maxima were collected for artificial propagations. More than 10 thousands of juveniles were produced in three batches and 5000 of them reached >1 cm in shell length until the end of 2017.Spawning can be successfully induced by dry-out treatment combined with temperature fluctuation application.Fertilized eggs metamorphosed into settled juveniles with water conditions under 28℃ and 33psu within 14-15 days.The grow-out cage was established for broodstock and juveniles to escape from mass mortalities caused by chill event during winter time.