流式细胞术估测胡椒属植物基因组大小方法的建立及应用  

Establishment and Application of a Method for Genome Size Estimation of Piper L.Using Flow Cytometry

作　　者：伍宝朵[1,3,4] 胡丽松 冯楗雄[2] 范睿 郭芬 吉训志[1,3,4] 郝朝运 闫林 WU Baoduo;HU Lisong;FENG Jianxiong;FAN Rui;GUO Fen;JI Xunzhi;HAO Chaoyun;YAN Lin(Spice and Beverage Research Institute,Chinese Academy of Tropical Agricultural Sciences,Wanning,Hainan 571533,China;College of Tropical Crop Science,Yunnan Agricultural University,Pu’er,Yunnan 665000,China;Hainan Sim Soonliang Aca-demician Workstation,Wanning,Hainan 571533,China;Key Laboratory of Genetic Resources Utilization of Spiced Beverage Crops,Ministry of Agriculture and Rural Affairs,Wanning,Hainan 571533,China)

机构地区：[1]中国热带农业科学院香料饮料研究所,海南万宁571533 [2]云南农业大学热带作物学院,云南普洱665000 [3]海南省SimSoonliang院士工作站,海南万宁571533 [4]农业农村部香辛饮料作物遗传资源利用重点实验室,海南万宁571533

出　　处：《热带作物学报》2023年第11期2265-2272,共8页Chinese Journal of Tropical Crops

基　　金：国家现代农业产业技术体系项目(No.CARS-11);海南省院士创新平台科研专项(No.SQ2021PT0035)。

摘　　要：为建立胡椒属资源基因组大小流式细胞术测定方法,本研究筛选了6种细胞核提取液和3种不同部位叶片,以已知基因组大小的黄瓜为标样,运用流式细胞术对胡椒属资源基因组大小进行了测定。结果表明:改良细胞核提取液和倒一叶是适合胡椒流式细胞术的提取液和取样部位;栽培胡椒品种古晋、Aman和EMAS胡椒的基因组大小为667.94~719.32 Mb,而热引1号、柬埔寨小叶种、班尼约尔1号和73F5的基因组大小为759.69~785.38 Mb,因此不同栽培品种之间基因组大小存在差异;推算基因组大于900 Mb的石楠藤和山蒟的染色体条数应该是大于52且为13的倍数,基因组大小在600~800 Mb范围内种质的染色体数可能是52条。杂交种不但具有与栽培胡椒和黄花胡椒相近的表型,其基因组大小也几乎等于栽培胡椒和黄花胡椒的平均值,推测杂交种可能是栽培胡椒和黄花胡椒的杂交后代。本研究可为胡椒属资源基因组大小测定提供方法,也将为胡椒属种质资源评价及远源杂交等研究提供技术基础。Six nuclear extracts and three leaves of different development stage were screened to establish the flow cytometry(FCM) method for determining the genomic size of Piper L.and Cucumis sativus L.was used as the DNA reference standard.Result showed that the modified nuclear extract and inverted leaf were suitable for the flow cytometry of Piper L.The genome size of cultivated varieties,including Kuching,Aman and EMAS,ranged from 667.94 to 719.32Mb,and Reyin1,microphylla variety from Cambodia,Panniyur-1 and 73F5 ranged from 759.69 to 785.38 Mb.Therefore,the genome size was significantly different among cultivated varieties.Furthermore,it is speculated that the chromosome numbers of P.wallichii and P.hancei,with genome size larger than 900 Mb,should be larger than 52 with a multiple of 13,and the number of chromosomes of germplasm,with genome size ranged from 600 to 800 Mb,may be 52.The hybrid not only had similar phenotypes to cultivated varieties and P.flaviflorum,but also had an average genome size of cultivated varieties and P.flaviflorum.Therefore,it is speculated that the hybrid may be the hybrid offspring of cultivated varieties and P.flaviflorum.This study would provide a method for the determination of the genome size of Piper L.germplasm,and provide a technical basis for the study of Piper L.germplasm evaluation and distant hybridization.

关 键 词：胡椒属 流式细胞术 细胞核提取液 基因组大小 

分 类 号：S573.9[农业科学—作物学]