Transient expression and analysis of fluorescent reporter proteins in plant pollen tubes.

Abstract:

:The pollen tube is an excellent single-cell model system for studying cellular processes in plant cell biology. This protocol describes a detailed step-by-step procedure with optimized conditions for introducing various fluorescent reporter proteins into lily, tobacco and Arabidopsis pollen grains by means of biolistics for their transient expression and subsequent analysis in germinating pollen tubes. The whole experiment consists of four major stages: coating gold microcarriers with DNA constructs, preparation of pollen grains, transformation of plasmid DNA into pollen grains by particle delivery system and germination of bombarded pollen grains in optimized germination media to obtain pollen tubes for protein trafficking, protein localization, drug treatment and organelle dynamics analysis. This protocol takes about 4-12 h from pollen preparation to protein detection.

journal_name

Nat Protoc

journal_title

Nature protocols

authors

Wang H,Jiang L

doi

10.1038/nprot.2011.309

subject

Has Abstract

pub_date

2011-04-01 00:00:00

pages

419-26

issue

4

eissn

1754-2189

issn

1750-2799

pii

nprot.2011.309

journal_volume

6

pub_type

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