利用流式细胞术鉴定桦木染色体倍性和DNA含量  

Identification of Chromosomal Ploidy and DNA Content in Betula by Flow Cytometry

作　　者：黄茂根 沈凝 刘雪羽 吴兴盛 陈爱平 孟岩 胡现铬 童再康[2] 黄华宏[2] 楼雄珍[2] Huang Maogen;Shen Ning;Liu Xueyu;Wu Xingsheng;Chen Aiping;Meng Yan;Hu Xiange;Tong Zaikang;Huang Huahong;Lou Xiongzhen(Nature Reserve Management Station of Luoboyan,Sanming Fujian 365050,China;The State Key Laboratory of Subtropical Silviculture,Zhejiang A&F University,Linan Zhejiang 311300,China;Guanzhuang State-owned Forest Farm,Sanming Fujian 365050,China;Xinyang Forestry Science Institute,Xinyang Henan 464000,China)

机构地区：[1]罗卜岩自然保护区管理站,福建三明365050 [2]浙江农林大学亚热带森林培育国家重点实验室,浙江临安311300 [3]官庄国有林场,福建三明365050 [4]信阳市林业科学研究所,河南信阳464000

出　　处：《西南林业大学学报（自然科学）》2023年第1期49-57,共9页Journal of Southwest Forestry University：Natural Sciences

基　　金：国家自然科学基金青年科学基金(32001327)资助;浙江省自然科学基金项目(LQ21C160002)资助。

摘　　要：基于流式细胞术(FCM)研究体系,构建能快速、准确鉴定桦木属植物倍性和测定DNA含量的研究体系,是测定不同桦木属植物倍性与基因组大小的有效方法。使用流式细胞术检测7种桦树属类物种,以物种不同生长状态(嫩叶、幼叶、成熟叶)为实验材料,不同存储方式(常温、冷藏和聚乙烯吡咯烷酮保存PVP)进行实验处理,对9种常用的裂解液进行筛选与优化。结果表明:桦木属植物DNA含量测定中,新鲜幼叶以1%PVP浸泡常温保存,并使用MgSO_(4)裂解液制备细胞核悬浮液上机效果最佳;桦木属植物中,基因组倍性依次为黑桦((2.09±0.04)Gb,8倍体)>坚桦((1.53±0.08)Gb,6倍体)>光皮桦((0.87±0.05)Gb,4倍体)=岳桦((0.80±0.07)Gb,4倍体)>垂枝桦((0.46±0.03)Gb,2倍体)=河桦((0.51±0.01)Gb,2倍体)。因此,桦木属植物DNA倍性和DNA含量可以通过FCM研究体系进行测定,且3倍体((0.66±0.02)Gb)子代的鉴定结果表明光皮桦与河桦之间不存在生殖隔离。This study aims to conduct flow cytometry(FCM)research that can accurately determine the ploidy level,DNA content,and genome size of different Betula species.Experimental materials with different growth stages(tender leaves,young leaves,mature leaves)were treated with varying methods of storage(room temperature,cold storage and polyvinylpyrrolidone,commonly referred to as PVP).Then,FCM was used to screen and optimize 9 kinds of commonly used dissociation fluids,and the cell ploidy and DNA content of the experimental materials were detected.The results showed that fresh young leaves,soaked in 1%PVP at room temperature and the nuclear suspension prepared by MgSO_(4) dissociation solution,is the best dissociation fluid.Of all 7 tested Betula spp.,the sequence of genomic ploidy was Betula dahurica(2.09±0.04 Gb,octaploid)>Betula chinensis(1.53±0.08 Gb,hexaploid)>Betula luminifera(0.87±0.05 Gb,tetraploid)=Betula ermanii(0.80±0.07 Gb,tetraploid)>Betula pendula(0.46±0.03 Gb,diploid)=Betula nigra(0.51±0.01 Gb,diploid).The ploidy level of Betula luminifera×Betula nigra hybrid progeny was found to be triploid(0.66±0.02 Gb),which indicated that there was no reproductive isolation between the 2 species.

关 键 词：流式细胞术 桦木 染色体倍性 DNA含量 

分 类 号：S792[农业科学—林木遗传育种]