Gil Hyun Woo, Kong Hee Jeong, An Cheul Min, Kim Bong-Seok, Lim Sang-Gu, Park In-Seok
Division of Marine Bioscience, College of Ocean Science and Technology, Korea Maritime and Ocean University, Busan, 606-791 Korea.
Biotechnology Research Division, National Institute of Fisheries Science (NIFS), Busan, 619-705 Korea.
Springerplus. 2016 Feb 27;5:186. doi: 10.1186/s40064-016-1830-4. eCollection 2016.
In this study, we induced tetraploidy in Korean rose bitterling, Rhodeus uyekii, by applying various hydrostatic pressure shock conditions. Tetraploidy was not induced under 4500 psi pressure treatment in any experimental group. Instead, the induction rate of tetraploidy was highest under 7500 psi hydrostatic pressure treatment. As a result, when the processing method was similar and as the process time increased, the induction rate of each experimental group increased; however, there was no significant difference (P > 0.05). The production rate was 3.1 %, which was highest in all experimental groups exposed to 6000 psi for 10 min after being fertilized for 100 min. The production rate was highest in the experimental groups treated with hydrostatic pressure alone, whereas the production rate was lowest in groups treated under hydrostatic pressure with chemical treatment. The abnormal rate of all experimental groups treated with 7500 psi for 20 min was very high, at about 5 %. Based on these studies, only hydrostatic pressure shock was considered effective at inducing tetraploidy based on the calculated hatching, abnormal, and induction rates. The most effective condition for inducing tetraploidy was 6000 psi of hydrostatic pressure shock for 10 min after being fertilized for 100 min. The chromosome number of the induced tetraploid Korean rose bitterling was 4n = 96, while that of the diploid was 2n = 48. In the diploid, there were 1 or 2 nucleoli in the cells, whereas the induced tetraploids contained 1, 2, 3, or 4. The DNA content of tetraploids and diploids were 3.68 ± 0.009 pg/nucleus and 1.84 ± 0.019 pg/nucleus, respectively, according to flow cytometric analysis. The DNA content and chromosome number of the tetraploids were twice that of the diploids.
在本研究中,我们通过施加各种静水压力冲击条件,诱导韩国玫瑰无须魮(Rhodeus uyekii)产生四倍体。在任何实验组中,4500磅力/平方英寸压力处理下均未诱导出四倍体。相反,在7500磅力/平方英寸静水压力处理下四倍体诱导率最高。结果表明,当处理方法相似且处理时间增加时,各实验组的诱导率均有所提高;然而,差异不显著(P>0.05)。产率为3.1%,在受精100分钟后接受6000磅力/平方英寸处理10分钟的所有实验组中最高。单独施加静水压力处理的实验组产率最高,而静水压力与化学处理相结合的组产率最低。所有接受7500磅力/平方英寸处理20分钟的实验组异常率都非常高,约为5%。基于这些研究,根据计算出的孵化率、异常率和诱导率,仅静水压力冲击被认为对诱导四倍体有效。诱导四倍体的最有效条件是受精100分钟后施加6000磅力/平方英寸的静水压力冲击10分钟。诱导出的四倍体韩国玫瑰无须魮的染色体数为4n = 96,而二倍体的染色体数为2n = 48。在二倍体细胞中,有1个或2个核仁,而诱导出的四倍体含有1个、2个、3个或4个核仁。根据流式细胞术分析,四倍体和二倍体的DNA含量分别为3.68±0.009皮克/细胞核和1.84±0.019皮克/细胞核。四倍体的DNA含量和染色体数是二倍体的两倍。