利用流式细胞仪快速鉴定棉花倍性的方法比较  

Comparison of Methods for Rapid Determination of Cotton Ploidy by Flow Cytometry

作　　者：王娅丽 周利利 王娜 程红梅[1] WANG Ya-li;ZHOU Li-li;WANG Na;CHENG Hong-mei(Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081)

机构地区：[1]中国农业科学院生物技术研究所,北京100081

出　　处：《生物技术通报》2022年第12期144-148,共5页Biotechnology Bulletin

基　　金：现代种业科技创新专项(21326314D)。

摘　　要：利用流式细胞仪快速检测棉花DNA倍性,为棉花的倍性鉴定和育种研究提供基础数据。以陆地棉珂字棉312(Gossypium hirsutum Linn.)的老叶和嫩叶为试材,用WPB和LB01解离液提取细胞核,经PI染色后使用流式细胞仪检测细胞倍性。结果发现,用WPB解离液提取棉花嫩叶获得的细胞核用于倍性检测效果更好,表现为主峰离子团清晰而集中、背景碎片少,可以得到明显的主峰、杂峰更少,且收集到的细胞核数目更多。初步建立了利用流式细胞仪快速检测棉花DNA倍性的方法。Rapid detection of cotton DNA ploidy by flow cytometry provides basic data for cotton ploidy identification and breeding research.The old and young leaves of Gossypium hirsutum Linn.Coker 312 were used as test material,and cell nuclei were extracted using WPB and LB01 dissociation solution and stained with PI for the ploidy detection by flow cytometry.The result showed that nuclei extracted from the cotton young leaves using WPB dissociation solution were more effective for ploidy detection,which had clear and focused peak particle,little background debris and obtained clear peak,less miscellaneous peak,less cellular debris and more nuclei.In conclusion,a method for rapid detection of cotton DNA ploidy by flow cytometry is initially established.

关 键 词：棉花 流式细胞术 细胞核DNA含量 染色体倍性 

分 类 号：S562[农业科学—作物学]