基于流式细胞术的乌饭树核DNA含量(2C-值)测定

Determination of nuclear DNA content(2C-value)of Vaccinium bracteatum by flow cytometry

关键词:  乌饭树, 核DNA含量, 2C-值, 流式细胞术

Key words:  Vaccinium bracteatum, nuclear DNA content, 2C-value, flow cytometry(FCM)

摘要:

核DNA含量(2C-值)是描述植物生物多样性的一个重要特征参数。该研究利用流式细胞仪检测越橘属植物乌饭树核DNA含量,建立了适合乌饭树的流式细胞术测定方法:以野生乌饭树的嫩叶为材料,以已知核DNA含量的水稻品种‘日本晴'为内标,采用GPB解离液,细胞核悬液加入50 μL 瘙 簚 mL^-1碘化丙啶染色5 min即可上机检测。结果表明:(1)9个乌饭树单株的核DNA含量平均值为(1.22±0.03)pg,最小值为1.18 pg,最大值为1.27 pg。(2)检测结果与已知的越橘属二倍体植株的2C-值含量相似,且不同地理来源的单株DNA含量没有显著差异(P>0.05),推测这9个单株为二倍体植株。(3)测定的乌饭树核DNA含量(2C-值)可丰富越橘属植物的C-值库; 基于流式细胞术建立的乌饭树核DNA含量测定方法可为该属其他植物的相关研究提供借鉴。

Abstract:

The plant nuclear DNA content(2C-value)is a principal characteristic parameter to describe biodiversity of plant species. For better understanding the nuclear DNA information of this plant, the nuclear DNA content of Vaccinium bracteatum wild plants were estimated by flow cytometry(FCM). In order to establish the optimal FCM method, young fresh leaves were taken as samples, and Oryza sativa subsp. japonica ‘Nipponbare'with known nuclear DNA content was set as internal standard. The nucleus of the mixed leaf cells was isolated by GPB dissociation solution and stained with 50 μL 瘙 簚 mL^-1 propidium iodide(PI)with 5 min, then the PI emission fluorescence intensity was measured by flow cytometry. The results were as follows:(1)The average nuclear DNA content of nine Vaccinium bracteatum plants was(1.22±0.03)pg, the minimum value was 1.18 pg and the maximum was 1.27 pg.(2)The results were similar to the nuclear DNA content of the known diploid plants of Vaccinium, and there was no significant differences in DNA content of plants from different geographical origins(P>0.05). It is speculated that these nine V. bracteatum plants are diploid plants.(3)The detected nuclear DNA contents(2C-value)of V. bracteatum can enrich the C-value database of Vaccinium. The nuclear DNA content detection method established based on FCM for V. bracteatum can provide reference for related research of other Vaccinium species.