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Induction of haploids in eastern little tuna, Euthynnus affinis, and application to fish breeding materials

东部小金枪鱼 Euthynnus affinis 单倍体的诱导及其在鱼类育种材料中的应用

基本信息

批准号：
21K14916
负责人：
遠藤充
金额：
$ 2.91万
依托单位：
Fisheries Research and Education Agency (2022)Ehime University (2021)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Early-Career Scientists
财政年份：
2021
资助国家：
日本
起止时间：
2021-04-01 至 2024-03-31
项目状态：
已结题

项目摘要

今年度は、雄性発生半数体誘起のための卵核の不活性化と、雌性発生半数体誘起のための精子核の不活性化に適した紫外線照射量の検討を目的とした。照射量の異なる実験区を設定し、各実験区について受精率、受精18時間後の体節形成期および受精36時間後の孵化期における生残率、孵化率、奇形仔魚率を調査し、紫外線照射による胚発生能力への影響を調べた。孵化期まで生残した胚は、セルアナライザーを用いた倍数性解析に供し、各実験区の半数体誘起効率を調べた。各実験区の胚発生成績については、雌性発生では照射量の増加に伴い受精率が低下し、雄性発生では生残率が低下する傾向が見られた。半数体の作出条件については、雌性発生における精子への紫外線照射では、試みた全ての照射量で半数体が高効率に得られた。しかし精子に一定量以上の照射を行った実験群では受精率が低下したため、照射が過剰と考えられた。また雄性発生における卵への紫外線照射では、孵化期まで胚が生残し、半数体が高効率に得られる照射量を明らかにした。しかし卵に一定量以上の照射を行った実験群では、受精胚が得られたもののその後の生残率が極めて低かった。なお具体的な紫外線照射量は、論文執筆中のため記述を控えた。雄性発生処理における卵への紫外線照射は、卵を水平方向に振盪しながら行うため、卵同士の接触による物理的ダメージも考えられたが、振盪が受精率などの胚発生成績に悪影響を及ぼさないことを合わせて確認した。また、雄性発生については、紫外線照射を用いない方法として知られる低温処理を試みたが、受精胚が得られなかった。今年度は低温雄性発生の条件決定には至らなかったが、低温雄性発生法は受精後の卵に対する処理法であるため、人工授精が容易ではないスマに対しては、今後も検討していく必要があると考えられる。

This year, half of the males and half of the males have inactivated egg nuclei, and half of the females have inactivated sperm nuclei. The radiation dose, fertilization rate, incubation rate, singularity rate, fertilization rate, fertilization rate, incubation rate, During the incubation period, the embryo was born and disabled, the embryo was analyzed by ploidy analysis, and the rate of half of the body weight in each area was measured. The embryos in each area were divided into four groups: female, male, female, male, male, female, female Half of the body samples were conditioned, the spermatozoa of the female pregnant women were exposed to ultraviolet rays, and the whole exposure dose was tested. The sperm were irradiated with more than a certain amount of radiation, the fertilization rate was low, the fertilization rate was low, and the exposure rate was low. The male eggs were exposed to UV radiation, the embryos were disabled during the incubation period, and the exposure dose was obtained for half of the body weight. After the eggs were irradiated with more than a certain amount of radiation, the fertilized embryos were divided into two groups, and the rate of life and disability was significantly lower than that of the fertilized embryos. Check the specific UV exposure, and record how to control the UV exposure in the text. In the male, the eggs were irradiated with ultraviolet rays, the eggs were exposed to ultraviolet rays in the horizontal direction, the eggs were exposed to each other in contact with the eggs, and the fertilization rate of the eggs was measured. The embryo produces the shadow of the egg, and the egg cooperates to confirm it. The fish, the male and the ultraviolet radiation were used to know the low temperature, the fertilized embryo and the fertilized embryo. The conditions of low-temperature male breeding this year determine the conditions of low-temperature male fertility, low-temperature male fertilization, artificial insemination, artificial insemination and artificial insemination.