Androgenetic doubled haploids induced without irradiation of eggs in loach (Misgurnus anguillicaudatus)
Androgenetic doubled haploids induced without irradiation of eggs in loach (Misgurnus anguillicaudatus)
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DOI：
10.1016/j.aquaculture.2013.05.021
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发表时间：
2014-01-20
期刊：
影响因子：
4.5
通讯作者：
Arai, Katsutoshi
中科院分区：
文献类型：
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作者：
Hou, Jilun;Saito, Taiju;Arai, Katsutoshi
在不照射泥鳅卵的情况下，诱导泥鳅雄核发育加倍单倍体。用橙色表型雄性的完整精子激活野生型雌性的卵，并在3 +/-0.5 ℃下处理（激活10 s内）30 min，以消除雌性细胞核。然后将卵在20 +/- 0.5 ℃的水浴中孵育35分钟。最后，恢复二倍体（活化后65分钟）通过在42 +/- 0.5摄氏度下热激处理2分钟。相对于使用的卵子总数，推定的DH（平均值+/- SD）为10.43 +/-1.69%，（P < 0.05）的产率高于在剩余热休克起始条件下（活化后55分钟、60分钟和70分钟）获得的产率。我们用流式细胞仪分析了假定DH的倍性状态。隐性橙子体色性状和微卫星基因型的表达证实了全雄性遗传。在泥鳅的28个基因座（包括27个连锁群）上均未检测到母系遗传的等位基因或杂合基因型，表明所获得的雄激素DH纯合性和父系遗传。(C)2013爱思唯尔有限公司版权所有。
Androgenetic doubled haploids (DHs) were induced in the loach Misgurnus anguillicaudatus (Cobitidae) without irradiation of the eggs. The eggs of wild-type females were activated with the intact sperm of an orange-phenotype male, and treated (within 10 s of activation) at 3 +/- 0.5 degrees C for 30 min, to eliminate the female nucleus. The eggs were then incubated in a water bath at 20 +/- 0.5 degrees C for 35 min. Finally, diploidy was restored (65 min after activation) by heat-shock treatment at 42 +/- 0.5 degrees C for 2 min. Under these conditions, the yield rate (mean +/- SD) of putative DHs relative to the total number of eggs used was 10.43 +/- 1.69%, which was significantly higher (P < 0.05) than the yield rates obtained under the remaining heat-shock initiation conditions (55 min, 60 min, and 70 min after activation). We analyzed the ploidy status of the putative DH by using flow cytometry. All-male inheritance was confirmed by the expression of the recessive orange body color trait and microsatellite genotypes. We detected no maternally derived alleles or heterozygous genotypes at any of the 28 loci (covering 27 linkage groups) of loach, indicating the exclusively paternal inheritance and homozygosity of the obtained androgenetic DHs. (C) 2013 Elsevier B.V. All rights reserved.