Anther Culture of Medicago sativa L. and Ploidy detection.

doi:10.11733/j.issn.1007-0435.2010.05.019

›› 2010, Vol. 18 ›› Issue (5): 714-718.DOI: 10.11733/j.issn.1007-0435.2010.05.019

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Anther Culture of Medicago sativa L. and Ploidy detection.

GENG Xiao-li¹, WEI Zhen-wu², YAO Xi-hong², ZHAO Yan²

1. College of Grassland Science, Gansu Agricultural University, Lanzhou, Gansu Province 730070, China;
2. College of Animal Science & Technique, Yangzhou University, Yangzhou, Jiangsu Province 225009, China

Received:2010-03-11 Revised:2010-09-21 Online:2010-10-15 Published:2010-10-15

苜蓿花药培养再生植株染色体倍性检测研究

耿小丽¹, 魏臻武², 姚喜红², 赵艳²

1. 甘肃农业大学草业学院, 甘肃, 兰州, 730070;
2. 扬州大学动物科学与技术学院, 江苏, 扬州, 225009

通讯作者: 魏臻武,E-mail:zhenwu_wei@yahoo.com.cn
作者简介:耿小丽(1979- ),女,甘肃兰州人,博士,主要从事牧草遗传育种学研究,E-mail:gxli-1979@163.com
基金资助:
国家“863”计划项目(2008AA10Z149);国家自然科学基金项目(30972136)资助

Abstract

Abstract: Ploidy levels of F₁ hybrid plants,derived from anther culture in Medicago sativa L.,were determined by flow-cytometric analysis.Purpose of the study was to seek a fast and reliable method for DNA ploidy detection to establish an effective system of anther culture while improving haploid induction in Medicago sativa L.There were 42 tetraploids(2n=4x=32),5 mixoploids of diploid and tetraploid(2n=4x +2n=2x),2 diploids,1 triploid in 50 regeneration plants of anther culture.The optimization of both cell suspension buffer and anther culture system in Medicago sativa L.was also discussed in the paper.

Key words: Medicago sativa L., Anther culture, Ploidy detection, Flow cytometry

摘要： 本试验采用流式细胞仪对苜蓿(Medicago sativa L.)F₁代杂种花药培养再生植株进行染色体倍性检测。旨在建立高效的苜蓿花药培养体系,提高苜蓿单倍体的诱导效率,寻求一种快速简便的苜蓿倍性检测方法。检测结果为流式细胞仪矩形图上清晰地检测出二倍体、三倍体、四倍体以及二倍体加四倍体的嵌合体。50株苜蓿花药再生植株中有42株四倍体(2n=4x),2n=4x与2n=2x的混合体为5株,二倍体(2n=2x)为2株,三倍体(2n=3x)为1株。本文还讨论了苜蓿花药培养体系的建立,优化,以及筛选出了适宜制作苜蓿细胞悬浮液的缓冲液(15 mmol·L^-1Tris-HCl(pH7.5);80 mmol·L^-1KCl;20 mmol.L^-1NaCl;2 mmol·L^-1EDTA-Na₂;15 mmol·L^-1β-Mercaptoethanol;0.1%(V/V)TritonX-100;0.5 mmol·L^-1Spermine·4HCl)及其流式细胞仪检测苜蓿倍性的适应性程序。

关键词: 苜蓿花药培养, 再生植株, 倍性检测, 流式细胞仪

CLC Number:

GENG Xiao-li, WEI Zhen-wu, YAO Xi-hong, ZHAO Yan. Anther Culture of Medicago sativa L. and Ploidy detection.[J]. , 2010, 18(5): 714-718.

耿小丽, 魏臻武, 姚喜红, 赵艳. 苜蓿花药培养再生植株染色体倍性检测研究[J]. , 2010, 18(5): 714-718.

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Anther Culture of Medicago sativa L. and Ploidy detection.

苜蓿花药培养再生植株染色体倍性检测研究

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