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学科主题: Biochemical Research Methods; Cell Biology
题名: Genetic heterogeneity and ploidy level analysis among different gynogenetic clones of the polyploid gibel carp
作者: Wei, WH ; Zhang, J ; Zhang, YB ; Zhou, L ; Gui, JF
通讯作者: Gui, JF, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan Ctr Dev Biol, Wuhan 430072, Peoples R China
关键词: gibel carp ; blood cell ; sperm ; DNA content ; polyploid ; flow cytometry
刊名: CYTOMETRY PART A
发表日期: 2003
卷: 56A, 期:1, 页:46-52
收录类别: SCI
部门归属: Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Inst Hydrobiol, Wuhan Ctr Dev Biol, Wuhan 430072, Peoples R China
摘要: Background: Some triploid and tetraploid clones have been identified in the gynogenetic gibel carp, Carassius auratus gibelio Bloch, by karyotypic and cytologic analyses over many years. Further, 5-20% males and karyotypic diversity have been found among their natural and artificial populations. However, the DNA contents and the relation to their ploidy level and chromosome numbers have not been ascertained, and whether normal meiosis occurs in spermatogenesis needs to be determined in the different clones. Methods: The sampled blood cells or sperms were mixed with blood cells from chicken or individual gibel carp and fixed in 70% pre-cooled ethanol overnight at 4degreesC. The mixed cell pellets were washed 2-3 times in 1x phosphate buffered saline and then resuspended in the solution containing 0.5% pepsin and 0.1 M HCl. DNA was stained with propidium iodide solution (40 mug/mL) containing 4 kU/ml RNase. The measurements of DNA contents were performed with Phoenix Flow Systems. Results: Triploid clones A, E, F, and P had almost equal DNA content, but triploid clone D had greater DNA content than did the other four triploid clones. DNA content of clone M (7.01 +/- 0.15 pg/nucleus) was almost equal to the DNA content of clone D (5-38 +/- 0.06 pg/nucleus) plus the DNA content of common carp sperm (1.64 +/- 0.02 pg/nucleus). The DNA contents of sperms from clones A, P, and D were half of their blood cells, suggesting that normal meiosis occurs in spermatogenesis. Conclusions: Flow cytometry is a powerful method to analyze genetic heterogeneity and ploidy level among different gynogenetic clones of polyploid gibel carp. Through this study, four questions have been answered. (a) The DNA content correlation among the five triploid clones and one multiple tetraploid clone was revealed in the gibel carp, and the contents increased with not only the ploidy level but also the chromosome number. (b) Mean DNA content was 0.052 pg in six extra chromosomes of clone D, which was higher than that of each chromosome in clones A, E, F, and P (about 0.032 pg/ chromosome). This means that the six extra chromosomes are larger chromosomes. (c) Normal meiosis occurred during spermatogenesis of the gibel carp, because DNA contents of the sperms from clones A, P, and D were almost half of that in their blood cells. (d) Multiple tetraploid clone M (7.01 +/- 0.15 pg/nucleus) contained the complete genome of clone D (5.38 +/- 0.06 pg/nucleus) and the genome of common carp sperm (1.64 +/- 0.02 pg/nucleus). Cytometry Part A 56A:46-52, 2003. (C) 2003 Wiley-Liss, Inc.
WOS记录号: WOS:000186285400006
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内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/9624
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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Wei, WH; Zhang, J; Zhang, YB; Zhou, L; Gui, JF.Genetic heterogeneity and ploidy level analysis among different gynogenetic clones of the polyploid gibel carp,CYTOMETRY PART A,2003,56A(1):46-52
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