流式细胞术鉴定苹果花药培养再生植株倍性的方法  

Ploidy Identification of Regenerated Plants from Anther Culture of Apple by Flow Cytometry

作　　者：邓舒 张春芬 肖蓉 曹秋芬 DENG Shu;ZHANG Chunfen;XIAO Rong;CAO Qiufen(Institute of Pomology,Shanxi Agricultural University,Taiyuan 030031,China;College of Life Sciences,Shanxi Agricultural University,Taiyuan 030031,China)

机构地区：[1]山西农业大学果树研究所,山西太原030031 [2]山西农业大学生命科学学院,山西太原030031

出　　处：《山西农业科学》2020年第9期1379-1382,共4页Journal of Shanxi Agricultural Sciences

基　　金：国家自然科学基金面上项目(31372033);山西省回国留学人员科研资助项目(2016-131,2016-132);山西省重点研发计划国际合作项目(201803D421023);山西省农业科学院优势课题组项目(YCX2018DZYS15);山西省农业科学院农业科技创新-农谷研发专项(YCX2020201)。

摘　　要：苹果花药培养再生获得的纯合基因型植株倍性各异。为了进一步在育种和遗传分析中应用这些株系,需要准确鉴定其倍性,而流式细胞术测定植物细胞DNA含量是一种简单、快速、精确的倍性鉴定方法。试验用4种不同的解离液对苹果花药培养再生植株的叶片样品进行解离,并进行了DNA含量检测。结果表明,WPB解离液解离样品后上机检测,主峰高且明显。利用WPB解离液对苹果花药培养再生植株叶片进行解离,解离后不经过离心,直接染色,是用流式细胞术鉴定苹果花药培养再生植株倍性的最佳方法。Apple homozygous plants obtained from anther culture have different ploidy levels.Plant ploidy analysis is essential for breeding and genetic analysis used in further research.Flow cytometric method is a rapid,convenient and accurate way to measure ploidy levels of plant cell.In this study,four different nuclear isolation buffers were used to prepare samples for analysing nuclear DNA content of apple plantlets from anther culture.The results showed that the main peaks of samples isolated by WPB buffer were high and obvious.Therefore,isolation by WPB buffer without centrifugation was the best method for ploidy flow cytometric analysis of apple homozygous plants from anther culture.

关 键 词：苹果 花药培养 倍性鉴定 流式细胞术 

分 类 号：S661.1[农业科学—果树学]