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李志, 鲁蒙, 周莉, 王忠卫, 李熙银, 汪洋, 张晓娟, 桂建芳. 外源精子基因组整入雌核生殖银鲫卵子创制异源八倍体的有效方法[J]. 水生生物学报, 2020, 44(1): 26-35. DOI: 10.7541/2020.004
引用本文: 李志, 鲁蒙, 周莉, 王忠卫, 李熙银, 汪洋, 张晓娟, 桂建芳. 外源精子基因组整入雌核生殖银鲫卵子创制异源八倍体的有效方法[J]. 水生生物学报, 2020, 44(1): 26-35. DOI: 10.7541/2020.004
LI Zhi, LU Meng, ZHOU Li, WANG Zhong-Wei, LI Xi-Yin, WANG Yang, ZHANG Xiao-Juan, GUI Jian-Fang. AN EFFECTIVE METHOD FOR CREATING ALLO-OCTOPLOIDS BY INTEGRATING EXOGENOUS SPERM GENOME INTO GIBEL CARP EGGS[J]. ACTA HYDROBIOLOGICA SINICA, 2020, 44(1): 26-35. DOI: 10.7541/2020.004
Citation: LI Zhi, LU Meng, ZHOU Li, WANG Zhong-Wei, LI Xi-Yin, WANG Yang, ZHANG Xiao-Juan, GUI Jian-Fang. AN EFFECTIVE METHOD FOR CREATING ALLO-OCTOPLOIDS BY INTEGRATING EXOGENOUS SPERM GENOME INTO GIBEL CARP EGGS[J]. ACTA HYDROBIOLOGICA SINICA, 2020, 44(1): 26-35. DOI: 10.7541/2020.004

外源精子基因组整入雌核生殖银鲫卵子创制异源八倍体的有效方法

AN EFFECTIVE METHOD FOR CREATING ALLO-OCTOPLOIDS BY INTEGRATING EXOGENOUS SPERM GENOME INTO GIBEL CARP EGGS

  • 摘要: 银鲫(Carassius auratus gibelio Bloth)具有将其他鱼类精子的染色体组、染色体片段并入到卵核中协同发育的能力, 但通过异精雌核生殖自发形成异源八倍体子代的概率极低。研究采用0.25%、0.5%、1%、2%和4%胰蛋白酶溶液处理白鲫精子10min后以及1%胰蛋白溶液处理白鲫精子5min、10min、15min、20min和25min后, 分别与异育银鲫A+系成熟卵子受精; 接着通过比较对照组和不同处理组合中精子结构和活力的变化, 兼顾受精率、孵化率、成活率等繁育指标与较高的子代八倍体率, 建立了一种将外源精子基因组整入雌核生殖银鲫卵子创制异源八倍体的有效方法。即采用1%胰蛋白溶液处理白鲫精子15min后与异育银鲫A+系成熟卵子受精, 子代的平均存活率为(2.4±0.7)%, 平均八倍体率可达(16.3±0.5)%。批量处理并结合流式细胞术筛选6月龄子代, 获得57尾融入有白鲫精子染色体组的异源八倍体成鱼。研究为创制异育银鲫优异异源八倍体种质提供了一个简单易行的方法, 创制的异源八倍体可作为后续培育生长快、抗病能力强的银鲫新品种的核心育种材料。

     

    Abstract: Gibel carp (Carassius auratus gibelio Bloth) possesses a special ability to integrate exogenous sperm genome or chromosome fragments into its eggs for co-development, but the spontaneous formation probability is very low in allogynogenetic offspring. In this study, white crucian carp (C. cuvieri) sperm were treated by 0.25%, 0.5%, 1%, 2% and 4% trypsin solution for 10min, or by 1% trypsin solution for 5, 10, 15, 20 and 25min respectively, and then fertilized with mature eggs of allogynogenetic gibel carp clone A+. By comparing the changes of sperm structure and motility between control group and different treatment groups, also considering reproductive indexes, such as fertilization rate, hatching rate and survival rate, as well as occurrence rate of allo-octoploids, an effective method was established to integrate exogenous sperm genome into allogynogenetic gibel carp eggs to create allo-octoploids. The average survival rate and octoploid rate were (2.4±0.7)% and (16.3±0.5)% respectively when the eggs of gibel carp clone A+ were fertilized with white crucian carp sperm treated by 1% trypsin solution for 15min. The effective method developed in this study will be a valuable way for creating novel genetic resources with excellent economic traits in gibel carp genetic breeding. Subsequently, 57 allo-octoploid adults were obtained by batch processing and flow cytometry screening from 6-month-old offspring, which can be used as core population for breeding of novel gible carp varieties with faster growth and or higher disease-resistance.

     

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