An efficient method for flow cytometric analysis of pollen and detection of 2n nuclei in Brassica napus pollen.

Abstract:

:A simple and reliable method was developed for isolating pollen nuclei from Brassica napus and Triticum aestivum for DNA analysis using flow cytometry. The nuclei were released from pollen by ultrasonic treatment. The isolated nuclei following filtration through nylon mesh and a purification procedure were suitable for flow cytometric analysis as well as for isolating genomic DNA. Ultrasonic treatment time was optimized for B. napus pollen at different developmental stages. The method is effective and suitable for the preparation of many samples. We analyzed the nuclear DNA levels in pollen of B. napus at three major developmental stages as well as in mature wheat pollen. Only a single 1C peak representing the haploid DNA level was detected in the nuclei isolated from Brassica uninucleate microspores as well as in mature Triticum pollen. Interestingly, diploid nuclei were detected in both binucleate and mature pollen of B. napus. The possible origins of the diploid nuclei are discussed.

journal_name

Plant Cell Rep

journal_title

Plant cell reports

authors

Pan G,Zhou Y,Fowke LC,Wang H

doi

10.1007/s00299-004-0830-y

keywords:

subject

Has Abstract

pub_date

2004-10-01 00:00:00

pages

196-202

issue

4

eissn

0721-7714

issn

1432-203X

journal_volume

23

pub_type

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