摘要
基于流式细胞技术对兰属中莲瓣兰和墨兰的基因组大小进行测定,采用Galbraith′s缓冲液解离并制备莲瓣兰和墨兰嫩叶材料的细胞核悬浮液,利用鸡红细胞核(chicken erythocyte nuclei,CEN)作为内标,建立和优化了基于流式细胞技术的两种兰属植物基因组大小的测定方法。研究发现Galbraith’s解离液对莲瓣兰和墨兰样品解离效果最好,测得莲瓣兰的2 C值为(8.15±0.34)pg,墨兰基因组DNA的2 C值为(8.34±0.36)pg。研究结果表明,基于流式细胞技术,优化解离和测定方法可精确测定莲瓣兰和墨兰的基因组大小。本研究丰富了兰科植物的基因库,为揭示该属植物C值大小和物种进化关系提供重要依据,也为其基因组学、分子细胞遗传学等研究奠定基础。
This study aims to determine the genome size of two Cymbidium orchids species C.tortisepalum and C.sinense by the flow cytometry method.Fresh leaves of two species were collected and dissociated to extract their nuclei using the Galbraith′s buffer.Chicken erythocyte nuclei(CEN)was used as the internal standard for cytometrical measurements.The study shows the 2 C value is(8.15±0.34)pg for C.tortisepalum,and(8.34±0.36)pg for C.sinense.By optimizing the dissociation and measurements,our study accurately estimated the genome sizes of both species.The results will enrich the gene pool of orchid family and provide the footing-stone for investigating the genome and species evolution,and further genomics and molecular cytogenetics studies.
作者
林瀚
韩晓文
兰思仁
马晓开
LIN Han;HAN Xiaowen;LAN Siren;MA Xiaokai(UIUC-SIB Joint Center for Genomics and Biotechnology,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;College of Forestry,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;The Key Laboratory of National Forestry and Grassland Administration for Orchid Conservation and Utilization,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;College of Landscape,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)
出处
《森林与环境学报》
CSCD
北大核心
2019年第6期616-620,共5页
Journal of Forest and Environment
基金
福州市市级科技计划项目(2017-G-88)
中国科学院科技服务网络计划项目(KFJ-3W-No1)
关键词
莲瓣兰
墨兰
流式细胞
基因组大小
Cymbidium tortisepalum Fukuy.
Cymbidium sinense Willd.
flow cytometry
genome size
