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草业科学  2014 

流式细胞术分析苜蓿相对DNA含量的样品处理方法

DOI: 10.11829\j.issn.1001-0629.2014-0096, PP. 1718-1723

Keywords: 苜蓿,流式细胞术,酶解法,细胞核提取缓冲液

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Abstract:

?为制备用于流式细胞仪分析的高纯度苜蓿(Medicagosativa)细胞核悬液,筛选最适合流式细胞仪分析的苜蓿细胞核提取方法,选用江苏地方品种淮阴苜蓿为供试材料,分别采用酶解法和机械解离法提取苜蓿细胞核悬液;用5种细胞核分离缓冲液及两种混合酶解液制备样品,根据各自优缺点筛选最适宜的苜蓿细胞核提取法。在机械法所用的5种细胞核提取缓冲液中,LB01缓冲液提取效果最好,提取的细胞核形态及内部结构完整,获得的细胞核量多。利用两种混合酶液制备苜蓿细胞核悬液,均出现倍性明显的峰,混合酶液Ⅰ制备的单细胞核悬液优于混合酶液Ⅱ,且酶解法制备细胞核悬液,可获得数量更多更完整的原生质体。但利用酶解法制备细胞核原生质体,其悬浮液中杂质多,需时较长。结果表明,LB01缓冲液为最适合制备苜蓿细胞核悬液的提取缓冲液。

References

[1]  洪绂曾.苜蓿科学.北京:中国农业出版社,2009.
[2]  Doleel J,Binarova P,Lucretti S.Analysis of nuclear DNA content in plant cells by flow cytometry.Biologia Plantarum,1989,31:113-120.
[3]  Galbraith D W,Harkins K R,Maddox J M,Ayres N M,Sharma D P,Firoozabady E.Rapid flow cytometric analysis of the cell cycle in intact plant tissues.Science,1983,220:1049-1051.
[4]  Doleel J.Application of flow cytometry for the study of plant genomes.Journal of Applied Genetics,1997,38:285-302.
[5]  Galbraith D W.Flow cytometric analysis of plat genomes.Methods Cell Biology,1990,33(48):549-562.
[6]  Arumuganathan K,Earle E D.Estimation of nuclear DNA contents of plants by flow cytometry.Plant Molecular Biology Reporter,1991,9(3):229-233.
[7]  Marie D,Brown S C.A cytometric exercise in plant DNA histograms,with 2C values for 70 species.Biology of the Cell,1993,78:41-51.
[8]  Biradar D P,Rayburn A Lane.Heterosis and nuclear DNA content in maize.Heredity,1993,71:300-304.
[9]  Awoleye F,Duren van M,Doleel J.Nuclear DNA content and in vitro induced somatic polyploidzation cassava(Manihot esculenta Crantz) breeding.Euphytica,1994(76):195-202.
[10]  Fréderique Ollitrault-Sammarcelli,Legavea J M,Nicole Michaux-Ferriereb.Use of flow cytometry for rapid determination of ploid level in the genus Actinidia.Scientia Horticulturae,1994,57(4):303-313.
[11]  Baird W,Vance,Estager Agnes S.Estimating nuclear DNA content in Peach and related diploid species using laser flow cytomety and DNA hybridization. Journal of the American Society for Horticultural Science,1994,119(6):1312-1316.
[12]  耿小丽.利用花药培养诱导苜蓿单倍体的研究.兰州:甘肃农业大学,2007.
[13]  耿小丽,魏臻武,程鹏舞,刘欢.苜蓿花药培养及倍性鉴定.草原与草坪,2008(5):1-5.
[14]  耿小丽.苜蓿单倍体、纯合四倍体的诱导及愈伤组织DNA含量变异的研究.兰州:甘肃农业大学,2010.
[15]  柳青慕,王赟文,王小山.流式细胞仪快速检测鸭茅与多花黑麦草染色体倍性的研究.草业科学,2012,29(3):403-410.
[16]  Doleel J.Flow cytometric analysis of nuclear DNA content in higher plants.Phytochem Analysis,1991(2):143-154.
[17]  Doleel J,Greilhuber J,Suda J.Estimation of nuclear DNA content in plants using flow cytometry.Nature Protocols,2007,2(9):2233-2244.
[18]  王娟.苜蓿原生质体分离与培养的研究.兰州:甘肃农业大学,2010.
[19]  Doleel J,Bartos J.Plant DNA flow cytometry and estimation of nuclear genomesize.Annals of Botany,2005,95:99-110.
[20]  汪静儿,孙玉强,祝水金.棉花原生质体培养与体细胞杂交研究进展.棉花学报,2007(2):139-144.
[21]  孙永星,李素花,魏小丽,韩榕.适合流式细胞仪分析的小麦细胞核提取方法比较.生物学杂志,2012,29(3):88-91.
[22]  李铁军,石俊英,王丽珍,史世娟,杜雪.长春花同源四倍体的诱导与鉴定.中国现代中药,2010,12(2):25-28.

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