《植物生理学报》 2015, 51(12): 2270-2274

大麦小孢子来源单倍体试管苗扩繁中的倍性检测

何婷^1,2,*, 郭桂梅^1,2,*, 陆瑞菊^1,2, 高润红^1,2, 陈志伟^1,2, 徐红卫^1,2, 李颖波^1,2, 黄剑华^1,2, 刘成洪^1,2,**, 王亦菲^1,2,**

¹上海市农业科学院生物技术研究所, 上海201106; ²上海市农业遗传育种重点实验室, 上海201106

通信作者：何；E-mail: chliu001@163.com, cs1@saas.sh.cn；Tel: 021-62202965

摘　要：

以大麦小孢子来源的单倍体试管苗扩繁再生植株为材料, 采用流式细胞术、气孔保卫细胞长度测定和根尖染色体计数三种方法对其倍性进行鉴定, 对大麦单倍体植株增殖继代中的倍性稳定性进行了研究。结果显示, 流式细胞术可对组织培养获得的试管苗做大批量倍性鉴定, 同时气孔保卫细胞长度测定可作为一个辅助检测手段, 两者对幼苗的损伤小、检测方便; 单倍体植株在离体扩繁继代5次后仍能保持单倍体倍性, 表明利用离体扩繁技术可以获得倍性稳定的单倍体再生植株群体。

关键词：大麦; 单倍体; 组织培养; 倍性鉴定

收稿：2015-09-21   修定：2015-11-16

资助：上海市种业发展项目[沪农科种字(2015)第3号]、上海市基础研究项目(14JC1405300)和大麦青稞产业技术体系(CARS-05)。

Ploidy Determination of Plantlets Propagated from Barley Microspore Derived Haploids

HE Ting^1,2,*, GUO Gui-Mei^1,2,*, LU Rui-Ju^1,2, GAO Run-Hong^1,2, CHEN Zhi-Wei^1,2, XU Hong-Wei^1,2, LI Ying-Bo^1,2, HUANG Jian-Hua^1,2, LIU Cheng-Hong^1,2,**, WANG Yi-Fei^1,2,**

¹Biotech Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China; ²Shanghai Key Laboratory of Agricultural Genetics and Breeding, Shanghai 201106, China

Corresponding author: HE Ting; E-mail: chliu001@163.com, cs1@saas.sh.cn; Tel: 021-62202965

Abstract:

In order to determine the ploidy of plants propagated from barley microspore derived, three methods of ploidy identification were used such as the DNA flow cytometry, the length of stomatal guard cell and chromosome counting. The results showed that flow cytometry can be used as a quick identification method for the ploidy identification of a large number of seedlings in tissue culture, and the length of stomatal guard cell can be used as an auxiliary detection means, and both of two methods were easily used to detect the ploidy of plants and did not appear to influence the seedlings. The ploidy of haploids was stable after 5 generations in vitro propagation so that the method can be used in creating the stable haploid population.

Key words: barley; haploid; tissue culture; ploidy identification