草莓组培苗遗传稳定性的ISSR检测及DNA甲基化变异  

ISSR Detection of Genetic Stability and DNA Methylation Variation of Tissue Culture Seedling in Strawberry

作　　者：苗徐静 文壮 文晓鹏[2] Miao Xujing;Wen Zhuang;Wen Xiaopeng(College of Life Science,Guizhou University,Gniyang,550025;Institute of Agro-bioengineering,The Key Laboratory of Plant Resources Conservation and Germplasm Innovation in Mountainous Region (Ministry of Education),Guizhou University,Guiyang,550025)

机构地区：[1]贵州大学生命科学学院,贵阳550025 [2]贵州大学农业生物工程研究院山地植物资源保护与种质创新教育部实验重点实验室,贵阳550025

出　　处：《分子植物育种》2019年第2期531-538,共8页Molecular Plant Breeding

基　　金：贵州省高层次创新型人才培养项目(黔科合人才((2016)4010))资助

摘　　要：以贵州主栽草莓品种-‘红颜’和‘章姬’,继代次数为第0、5、7、9、10、12、14、18次以及移栽时长为1月、2月、3月的试管苗为材料,利用ISSR标记和MSAP检测,对其遗传变异情况作了探讨。红颜采用49个ISSR引物扩增出207个标记,章姬采用49个ISSR引物扩增出214个标记。用MS+0.5 mg/L BA+0.1 mg/L IBA+30 g/L蔗糖+7 g/L琼脂(pH 6.0)培养基继代红颜至12代及章姬至18代DNA水平突变率为0,仍然保持其遗传稳定性,可采用组培快繁;利用MSAP技术得出随继代次数的增加,组培苗的甲基化敏感多态性呈现出下降趋势。但在田间无性繁殖过程中,组培苗DNA甲基化水平又逐渐升高,且两个品种的DNA甲基化变异程度大致相同。推测在组培环境下诱导草莓基因组DNA发生的去甲基化并不稳定,可能与适应组培环境有关。Based on ISSR markers and MSAP detection, we explored effects of the times of subculture(0, 5, 7,9, 10, 12, 14, 18) and transplantation(1, 2, 3 months) on the genetic diversity of strawberry(Fragariag ananassa‘Benihoppe’ and ‘Akihime’) which is widely cultivated in Guizhou Province. The results showed that 207 bands were amplified from 49 ISSR primers of ‘Benihoppe’, 214 bands were amplified from 49 ISSR primers of ‘Akihime’.After subculture with MS+0.5 mg/L BA+0.1 mg/L IBA+30 g/L sucrose+7 g/L agar(pH 6.0) culture medium, the mutation rate of DNA level of ‘Benihoppe’ from 0 to 12 generation and ‘Akihime’ from 0 to 18 generation were 0,and their genetic stability still could be maintained, so tissue culture could be used for rapid propagation.Methylation-sensitive amplified polymorphism of tissue culture seedlings showed a downward trend with the increase of subculture times by MSAP. However DNA methylation levels of tissue culture seedlings during the asexual propagation in the field increased gradually,and the DNA methylation variation was similar between the two strawberry cultivars. We speculated that the demethylation of strawberry genomic DNA induced by tissue culture might be unstable, which might be related to the adaptation to tissue culture environment.

关 键 词：草莓 继代次数 遗传变异 ISSR MSAP 

分 类 号：S668.4[农业科学—果树学]