热休克法诱导虹鳟三倍体技术研究
Technical Research on Induced Triploid of Rainbow Trout by Heat Shock

DOI: 10.12677/OJFR.2019.62008, PP. 63-72

Keywords: 虹鳟，热休克法，三倍体，诱导率，综合评分
Rainbow Trout, Heat Shock, Triploid, Induction Rate, Comprehensive Score

Full-Text   Cite this paper   Add to My Lib

Abstract:

本文以虹鳟(Oncorhynchus mykiss)为实验对象，采用热休克法诱导虹鳟三倍体，利用流式细胞仪(CytoFLEX S)以及比较红细胞长径对虹鳟倍性进行鉴定；在热休克法诱导虹鳟三倍体的两个正交重复实验中，主要研究了处理的起始时间、温度、持续时间三个因素对于虹鳟三倍体诱导效果的影响，结果显示，在实验设定的起始时间、温度、以及持续时间内，温度大小对发眼率、诱导率以及综合评分影响程度都是最大(R值最大)，在发眼率结果中，25℃时有最高的发眼率；在诱导率结果中，25℃、27℃明显高于29℃组的诱导率；在综合评分结果中25℃、27℃明显高于29℃组的综合评分。在起始时间为10~20 min内，起始时间对诱导率没有显著影响，但起始时间为20 min时诱导效果最优。在起始时间与处理强度双因素实验结果中，综合评分高于70.00%的共两组，分别为第一组、第二组，热休克的起始时间都是25 min，作用参数分别为25℃持续20 min、27℃持续15 min，是较为理想的诱导三倍体处理强度参数。
In this study, the rainbow trout (Oncorhynchus mykiss) was used as the experimental object. The heat shock method was used to induce the triploid of rainbow trout. The ploidy of rainbow trout was identified by using flow cytometry (CytoFLEX S) and comparison of the long axis of erythrocyte. In the two orthogonal replicate experiments induced by heat shock induced rainbow trout triploid, the effects of three factors, including start time, temperature and duration, on the induction of rainbow trout triploid were studied. The results showed that the temperature had the greatest impact on the eye rate, induction rate and overall score (maximum R value) at the start time, temperature, and duration of the experiment. In the eye rate results, the highest eye rate was obtained at 25？C; in the induction rate results, induction rate of the 25？C, 27？C group was significantly higher than that of the 29？C group; in the comprehensive scoring results; the comprehensive scores of the group at 25？C and 27？C were significantly higher than those at 29？C. In the initial time of 10~20 min, the initial time had no significant effect on the induction rate, but the induction effect was optimal when the starting time was 20 min. In the two-factor experiment results of starting time and treatment intensity, there are two groups whose comprehensive score is higher than 70.00%: the first group and the second group, respectively. The starting time of heat shock is 25 min, and the action parameter is 25？C for 20 min and 27？C for 15 min respectively, which is an ideal parameter for inducing triploid treatment intensity.

References

[1]	王炳谦. 中国鲑鳟鱼养殖[M]. 北京: 中国农业出版社, 2015: 1-15.
[2]	王炳谦, 徐连伟, 贾钟贺, 等. 热休克诱导全雌虹鳟三倍体[J]. 水产学杂志, 2005, 18(2): 22-27.
[3]	Galbreath, P. and Samples, B. (2000) Optimization of Thermal Shock Protocols for Induction of Triploid in Brook Trout. North American Journal of Aquaculture, 62, 249-259. https://doi.org/10.1577/1548-8454(2000)062<0249:OOTSPF>2.0.CO;2
[4]	Chourrout, D. (1980) Thermal Induction of Diploid Gynogenesis and Triploid in the Eggs of the Rainbow Trout (Salmo gairdneri Richardson). Reproduction Nutrition Development, 20, 727-733. https://doi.org/10.1051/rnd:19800415
[5]	张艳萍, 王太, 俞小牧, 等. 热休克诱导虹鳟三倍体的最佳条件[J]. 西北师范大学学报(自然科学版), 2011, 47(6): 69-74.
[6]	贾钟贺, 徐革锋, 牟振波, 等. 金鳟伪雄鱼的制备及全雌三倍体的诱导[J]. 大连海洋大学学报, 2009, 24(1): 8-11.
[7]	陈春山, 魏凯, 韩姝伊, 等. 热休克诱导细鳞鲑三倍体的初步研究[J]. 淡水渔业, 2018, 48(6): 63-68.
[8]	龙强, 宗虎民, 陈冰君. 虹鳟鱼三倍体的育种技术[J]. 北京水产, 2004(5): 36-37.
[9]	Gervai, J., Páter, S., Nagy, A., et al. (1980) Induced Triploid in Carp, Cyprinus carpio L. Journal of Fish Biology, 17, 667-671. https://doi.org/10.1111/j.1095-8649.1980.tb02800.x
[10]	尤锋. 黑鲷三倍体的人工诱导研究[J]. 海洋与湖沼, 1993(3): 248-255+334.
[11]	吴玉萍, 叶玉珍, 吴清江. 热休克诱导斑马鱼异源三倍体的研究[J]. 海洋与湖沼, 2000, 31(5): 465-470.
[12]	林琪, 吴建绍, 曾志南. 静水压休克诱导大黄鱼三倍体[J]. 海洋科学, 2001, 25(9): 6-9.
[13]	王茂林, 姜志强, 李荣. 红鳍东方鲀三倍体诱导的初步研究[J]. 水产科学, 2006, 25(7): 349-352.
[14]	宋立民. 温度休克诱导黄颡鱼三倍体及其倍性鉴定研究[D]: [硕士学位论文]. 武汉: 华中农业大学, 2009.
[15]	王昭明, 王新军, 陈惠, 等. 食盐水洗卵法在鲑科鱼人工繁殖中的应用试验[J]. 水产学杂志, 2000, 13(2): 6-8.
[16]	魏华, 吴垠, 等. 鱼类生理学[M]. 北京: 中国农业出版社, 2011: 99-112.
[17]	楼允东. 国外对鱼类多倍体育种的研究[J]. 水产学报, 1984, 8(4): 343-356.
[18]	宋保坤, 王辉, 叶富良, 等. 水产动物多倍体育种技术研究[J]. 北京水产, 2004(6): 38-41.
[19]	Diter, A., Quillet, E. and Chourrout, D. (1993) Suppression of First Egg Mitosis Induced by Heat Shocks in the Rainbow Trout. Journal of Fish Biology, 42, 777-786. https://doi.org/10.1111/j.1095-8649.1993.tb00383.x
[20]	Solar, I.I., Donaldson, E.M. and Hunter, G.A. (1984) Induction of Triploid in Rainbow Trout (Salmo gairdneri Richardson) by Heat Shock, and Investigation of Early Growth. Aquaculture, 42, 57-67. https://doi.org/10.1016/0044-8486(84)90313-2
[21]	桂建芳, 肖武汉, 梁绍昌, 等. 静水压休克诱导水晶彩鲫三倍体和四倍体的细胞学机理初探[J]. 水生生物学报, 1995, 19(1): 49-55.

Full-Text