鱼类染色体组操作的研究 Ⅱ.静水压处理和静水压与冷休克结合处理诱导水晶彩鲫四倍体

STUDIES ON GENOME MANIPULATION IN FISH 鈪? TETRAPLOIDY INDUCED BY HYDROSTATIC PRESSURE TREATMENT AND A COMBINATION OF HYDROSTATIC PRESSURE AND COLD TREATMENTS IN TRANSPARENT COLORED CRUCIAN CARP

  • 摘要: 采用静水压处理和静水压与冷休克结合处理两种方法进行了抑制第一次卵裂诱导水晶彩鲫四倍体的研究。水晶彩鯽受精卵在受精后(发育水温14-15℃)50、54、55和60min时的静水压(650kg/cm2)处理(3min)组中都出现了四倍化胚胎,而在受精后48和48min以前的相同处理组中都没有观察到四倍化胚胎。在受精后(发育水温16-18℃)48、50、52、55、56和60min时的静水压与冷休克结合处理组中,也都出现了四倍化胚胎,且四倍化率比仅用静水压处理高,但存活率更低。在经过处理而发育形成的胚胎中,既有四倍体、次四倍体和4n/2n嵌合体,也有二倍体和次二倍体,并在一些次二倍体和次四倍体中期相中还观察到休克处理致使染色体断裂的痕迹--染色体片段。在抑制卵裂诱发加倍的效应期内,可能存在瞬间对休克耐受性较强的时期,而在此前后,对休克更为敏感。在少数处理组中,已筛选出了数尾四倍体鱼。文中还对鱼类四倍体的诱导技术、抑制卵裂诱发染色体组加倍的效应期以及四倍化胚胎的存活率和生命力等有关问题进行了分析和讨论。

     

    Abstract: The studies on the induction of tetraploidy, using either hydrostatic pressure treatment alone or a combination of hydrostatic pressure and cold treatment for blocking first cleavage, were attempted in transparent colored crucian carp (Carassius auratus transparent colored variety). The shock treatments involving hydrostatic pressure of 650 kg/cm2 lasting 3 minutes were applied at 35, 38, 40, 42, 45, 48, 50, 54, 55 or 60 min after fertilization. Tetraploidy embryos that include tetraploids, hypotetraploids and tetraploid/diploid mosaics were found in the pressure treatment groups at 50, 54, 55 and 60 min after fertilization, whereas the tetraploidy embryos were not found in the pressure treatment groups at 48, 45, 42, 40, 38 and 35 min after fertilization. The results suggest that hydrostatic pressure treatment at 50-60 min post-fertilization arrest the first cleavage and induce tetraploidy. The shocks involving a combination of hydrostatic pressure and cold treatment were applied at 40, 48, 50, 52, 55, 56 or 60 min post-fertilization. The tetraploidy embryos of tetraploids, hypotetraploids and tetraploid/diploid mosaics were observed in the combined treatment groups at 48, 50, 52, 55, 56 and 60 min post-fertilization. The tetraploidy rates in the combined treatment groups were higher than those in the pressure treatment groups. Both methods of treatments, especially the dombined treatment, strongly affected the development of fertilized eggs and decreased the survival rates of embryos. The fertilized eggs subjected to pressure shocks and the combined shocks not only produced tetraploids, hypotetraploids and tetraploid/diploid mosaics, but also developed diploids and hypodiploids. Some chromosome fragments caused by the suboptimal treatments were obviously observed in the metaphases of some hypotetraploids and hypodiploids. We also found a short stage at which fertilized eggs have stronger tolerance towards the shock treatments during the effective period for the inhibition of first mitosis and induction of tetraploidy. Before and after this short stage, the fertilized eggs were more sensitive to the shock treatments. Artificial tetraploid transparent colored crucian carp were identified and selected from the individuals developed from some shock groups through chromosome observation of tail-fin tissue cells of fingerlings and blood cultured cells of adult fish. The problems about the techniques of inducing tetraploidy, the effective period of blocking first mitosis for the doubling of chromosome set and the survival rates and viability of retraploidy embryos in fish were analysed and discussed based on the results obtained from this study and those reported by other investigators.

     

/

返回文章
返回