流式细胞仪对甘蓝花粉倍性的测定
Determination of pollen ploidy of cabbage by flow cytometry
杨鼎 1李崇娟 1吕凤仙 1和江明 2兰梅 2胡靖峰 2徐学忠2
作者信息
- 1. 云南省农业科学院园艺作物研究所,昆明 650205;云南大学农学院,昆明 650504
- 2. 云南省农业科学院园艺作物研究所,昆明 650205;国家蔬菜改良中心云南分中心,昆明 650205
- 折叠
摘要
利用流式细胞仪测定甘蓝(Brassica oleracea var.capitata)花粉的倍性,以甘蓝GL21028的花粉为材料,以叶片为对照,比较花粉2种处理方式、3种不同细胞裂解液、2种核酸染料的效果差异.通过B缓冲液处理后获取的花粉细胞可以直接染色,为最简便处理方式;采用"Aru"buffer和Y2种裂解液,染色剂DAPI,处理甘蓝叶片和花粉,通过流式细胞仪检测,样本细胞DNA成峰集中,细胞碎片很少;配制的Y裂解液能够适用甘蓝材料流式细胞仪的观察.甘蓝花粉通过B缓冲液处理后,可以直接作为流式细胞仪倍性鉴定的材料,不再需要其他处理,配制的Y裂解液可以得到较好的试验结果.
Abstract
Pollen ploidy of Brassica oleracea var.capitata was determined by flow cytometry.Pollen of cabbage GL21028 was used as the material,and leaves were used as the control to compare the effects of two pollen treatments,three different cell lysates,and two nucleic acid dyes.Pollen cells obtained through B buffer could be directly stained,which was the easiest treatment method.Cabbage leaves and pollen were treated with"Aru"buffer and Y lysates and stains DAPI,and flow cytometry was used for detection.The DNA of the sample cells was concentrated,and there were few cell fragments.The prepared Y lysate could be used for the observation of cab-bage material by flow cytometry.Cabbage pollen treated with B buffer could be directly used as the material for flow cytometry ploidy identification,no other treatment was required,and the prepared Y lysate could obtain better test results.
关键词
甘蓝(Brassica/oleracea/var.capitata)/流式细胞仪/花粉/测定/倍性/裂解液Key words
Brassica oleracea var.capitata/flow cytometry/pollen/determination/ploidy/lysate引用本文复制引用
基金项目
国家大宗蔬菜产业技术体系建设项目昆明综合试验站项目(CARS-23-G33)
云南省"万人计划"产业领军人才项目(YNWR-CYJS-2018-040)
云南省通海县蔬菜科技特派团项目(2022)(202204BI090023)
绿色食品品牌打造科技支撑行动(果蔬)专项()
出版年
2024
NETL
NSTL
万方数据