流式细胞仪鉴定猕猴桃倍性技术研究

doi:10.7525/j.issn.1673-5102.2014.06.022

植物研究 ›› 2014, Vol. 34 ›› Issue (6): 845-849.doi: 10.7525/j.issn.1673-5102.2014.06.022

流式细胞仪鉴定猕猴桃倍性技术研究

1.上海市农业科学院林果所,上海　201106；
2.上海市设施园艺重点实验室,上海　201106；
3.淮阴工学院,淮安　223003

收稿日期:2013-11-20 出版日期:2014-11-20 发布日期:2015-01-06
基金资助:

Identification of Kiwifruit Ploidy Using Flow Cytometry

1.Forestry and Pomology Research Institute, Shanghai Academic of Agricultural Science,Shanghai　201106；
2.Shanghai Key Lab of Protected Horticultural Tech,Shanghai　201106；
3.Huaiyin Institute of Technology,Huai’an　223003

Received:2013-11-20 Online:2014-11-20 Published:2015-01-06
Supported by:

摘要/Abstract

摘要： 利用流式细胞仪（Flow cytometer,FCM）对不同倍性猕猴桃DNA的相对含量进行测定，建立一套简易、快速的鉴定猕猴桃倍性方法。以二倍体“红阳”猕猴桃为对照，‘81-5’、‘皖翠’猕猴桃为试材，通过筛选最优鉴定叶片部位；改良解离液配方；提高滤膜的目数；过滤次数；增加离心次数等优化鉴定技术。结果表明，露地栽培条件下，茎尖下初展叶是倍性鉴定的最佳部位；两次离心；解离液中2%PVP-30；500目滤膜抽滤两次可以有效解决仪器堵管现象，有效去除杂质峰并获得清晰样品峰。通过优化后的鉴定技术可实现同时测定2个以上倍性的猕猴桃混合样品。

关键词: 流式细胞仪, 猕猴桃, 倍性鉴定, 峰值

Abstract: The contents of nuclear DNA in different ploidy kiwifruit cultivars were measured by flow cytometry, a simple, rapid method for the identification of kiwifruits ploidy was established. Diploid “Red Sun” Kiwi was used as control, ‘81-5’, ‘Wan Chui’ Kiwi as material, the ploidy was identified by screening the optimal leaf area, improving dissociation liquid formulations, improving membrane mesh and the number of filters, increasing the number of centrifugation. The results showed that open conditions of field cultivation, the early exhibition leaf under the stem tip is the best part of ploidy identification; twice centrifugation; dissociation solution 2% PVP-30; 500 mesh membrane filtration twice can effectively resolve the blocking of tube, effectively remove impurities and obtain a clear peak sample peak. The optimized technology enables the simultaneous determination of two or more kiwifruit mixed samples.

Key words: flow cytometer, kiwifruit, ploidy identification, peak

中图分类号:

施春晖1,2；徐兰兰3；王晓庆1,2；骆军1,2*. 流式细胞仪鉴定猕猴桃倍性技术研究[J]. 植物研究, 2014, 34(6): 845-849.

SHI Chun-Hui1,2；XU Lan-Lan3；WANG Xiao-Qing1,2；LUO Jun1,2*. Identification of Kiwifruit Ploidy Using Flow Cytometry[J]. Bulletin of Botanical Research, 2014, 34(6): 845-849.

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流式细胞仪鉴定猕猴桃倍性技术研究

Identification of Kiwifruit Ploidy Using Flow Cytometry

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