采用流式细胞仪鉴定紫花苜蓿花药愈伤组织的变异

草业学报 ›› 2011, Vol. 20 ›› Issue (3): 156-161.

采用流式细胞仪鉴定紫花苜蓿花药愈伤组织的变异

耿小丽¹,魏臻武^2*,姚喜红¹

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

收稿日期:2011-02-23 出版日期:2011-03-25 发布日期:2011-06-20
通讯作者: E-mail:zhenwu_wei@yahoo.com.cn
作者简介:耿小丽(1979-),女,甘肃兰州人,博士。
基金资助:
国家自然科学基金(30671486)和国家863计划项目(2008AA10Z149)资助。

Genetic variation of Medicago sativa callus revealed by the ploidy analyser

GENG Xiao-li¹, WEI Zhen-wu², YAO Xi-hong¹

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

Received:2011-02-23 Online:2011-03-25 Published:2011-06-20

摘要/Abstract

摘要： 本试验采用流式细胞仪,对3个基因型8个继代周期的苜蓿愈伤组织DNA含量进行测定。通过DPAC软件(data pool application of cytometre)分析出细胞DNA含量变异百分率和处于S期细胞的百分率,并采用SPSS 10.0软件分析出愈伤组织继代时间与DNA含量变异率之间的关系。结果表明,本试验所采用的3个基因型8个继代周期24个紫花苜蓿愈伤组织样本DNA含量均发生了变化,且全都出现了DNA加倍的现象。在24个样本中,DNA含量变异率最大的是自选系大叶0510的愈伤组织继代15周时的样本,变异率达到了19.45%。通过SPSS 10.0软件分析,表明紫花苜蓿愈伤组织DNA含量变异细胞百分率与继代时间存在相关性,随着继代时间的延长,染色体变异率有增加的趋势,但这种趋势不是无限增大的。继代0～12周是愈伤组织DNA含量变异的集中发生期,以后随着继代次数的增加,DNA含量变异率趋向于稳定。

Abstract: DNA contents of callus of three genotype and eight culture time, which derived from anther culture in Medicago sativa, were detected by flow-cytometric analysis. The percentage of S stage cells and percentage of DNA content variation cells were analysed by the Data Pool Application of Cytometre software, the relationship between the callus cultivating time and the DNA content variation were analysed by the SPSS 10.0 software. One purpose of the study is to seek the relationship between DNA content variation and the culture time, and another is to provide the basic study of the law of DNA content variation in M. sativa for further research. DNA content of 24 samples were all changed and all DNA doubled. By the SPSS 10.0 software analysed, the correlativity between percentage of DNA content variation cells and cultivating time in M. sativa were found. With the increase of cultivating time, chromosome mutation were added. However, the trend was not always unchanged. The 0-12 week was the concentrated occurring stage of M. sativa callus DNA content variation, whereafter, DNA content variation become smooth with the increase of subculture times.

中图分类号:

耿小丽,魏臻武,姚喜红. 采用流式细胞仪鉴定紫花苜蓿花药愈伤组织的变异[J]. 草业学报, 2011, 20(3): 156-161.

GENG Xiao-li, WEI Zhen-wu, YAO Xi-hong. Genetic variation of Medicago sativa callus revealed by the ploidy analyser[J]. Acta Prataculturae Sinica, 2011, 20(3): 156-161.

参考文献

[1] 杨磊, 王彦荣, 余进德.干旱荒漠区土壤种子库研究进展[J]. 草业学报, 2010, 19(2): 227-234.
[2] Larkin P J, Scowcroft W P. Somalia variation a novel source of variability from cell-cultures for plant improvement[J]. Theoretical and Applied Genetics, 1981, 60: 197-214.
[3] Yoshida S Y, Kazuhiko W, Morihiro F J. Non-random game to clone variation in rice regenerate from callus subcultures for a prolonged period under high somatic stress[J]. Euphytica, 1998, 104: 87-94.
[4] Muller E, Brown P T, Harked S, et al. DNA variation in tissue-culture-derived rice plants[J]. Theoretical and Applied Genetics, 2001, 80: 673-679.
[5] 谷明光. 玉米花粉单倍体植株染色体上异染色质的变异[J]. 遗传学报, 1991, 18(3): 235-238.
[6] 崔广荣, 张子学, 张从宇. 文心兰多倍体诱导及其鉴定[J]. 草业学报, 2010, 19(1): 184-190.
[7] 云岚, 云锦凤, 李俊琴. 新麦草愈伤组织多倍体诱导与倍性鉴定[J]. 草业学报, 2010, 19(6): 126-131.
[8] 陈智勇, 胡忠红, 蒋建雄. 早熟禾族3种基因型草坪草的染色体核型分析[J]. 草业学报, 2010, 19(6): 281-285.
[9] 李士生. 小麦愈伤组织及再生植株的染色体变异[J]. 遗传学报, 1991, 18(4): 332-338.
[10] 梁虹, 陈耀锋, 王睿辉, 等. 小麦幼胚体细胞无性系及其继代过程的微观结构观察[J]. 西北农林科技大学学报(自然科学版), 2005, 6(9): 62-67.
[11] Phillips R L, Kaepler S M, OLhoft P. Genetic Instability of Plant Tissue Culture: Breakdown if Normal Controls[M]. Proceedings of the National Academy of Sciences USA, 1998, 91: 5222-5226.
[12] Barcaccia G, Rosellini D, Falcinelli M, et al. Reproductive behaviour of tetraploid alfalfa plants obtained by unilateral and bilateral sexual polyploidization[J]. Euphyatica, 2002, 99(3): 18-24.
[13] 胡钠梅, 韩素英, 梁国鲁. 中间锦鸡儿染色体变异研究[J]. 生物技术通报, 2009, 10: 75-79.
[14] 张玉静. 分子遗传学[M]. 北京: 科学出版社, 2001: 27.
[15] 商效民. 植物离体培养中染色体的变异[J]. 细胞生物学杂志, 1984, (1): 5-12.
[16] 黄晓梅, 李桂英, 梁艳, 等. 组织培养中大葱染色体倍性变异的研究[J]. 中国农学通报, 2005, 21(1): 50-56.
[17] Myers J M, Simon P W. Regeneration of garlic callus as affected by colonel variation, plant growth regulators and culture conditions over time[J]. Plant Cell Report, 1999, 19(1): 32-36.
[18] Bouman H G, De Klert J. Measurement of the extent of soma clone variation in begonia plants regenerated under various conditions, comparison of three assays[J]. Theoretical and Applied Genetics, 2001, 12(1): 111-117.
[19] Hao Y J, You C X, Deng X X. Analysis of ploidy and the patterns of amplified fragment length polymorphism and methyl nation sensitive amplified polymorphism in strawberry plants recovered from cypresses ration[J]. Cryoletters, 2006, 21: 37-46.
[20] Hao Y J, You C X, Deng X X. Effects of cryopreservation on developmental competence, cytological and molecular stability of citrus callus[J]. Cryoletters, 2006, 23(1): 27-35.
[21] Hao Y J, Deng X X. Occurrence of chromosomal variations and plant regeneration from long-term cultured citrus callus[J]. Biologia Plantarum, 2002, 38(5): 32-38.