Plant Molecular Biology Reporter

, Volume 20, Issue 1, pp 19–26 | Cite as

Single-embryo RT-PCR assay to study gene expression dynamics during embryogenesis inArabidopsis thaliana

Protocols

Abstract

We have developed a single-embryo RT-PCR protocol for studying gene expression during plant embryogenesis. Four genes,glyceraldhyde-3-phosphate dehydrogenase (GAPC), shoot-meristemless (STM), monopteros (MP), andshaggy-like kinase etha (ASKη), fromArabidopsis thaliana were used to test the sensitivity and reliability of this method by analyzing the differential signal intensities of their RT-PCR products. The method could detect genes expressed during embryogenesis at a single-embryo level and, therefore, can be used to identify phenotypes. When in vitro, embryogenesis also is used to control the time course of zygote development exactly. The single-embryo RT-PCR protocol becomes a powerful method to survey the dynamics of specific gene expression.

Key words

Arabidopsis thaliana embryogenesis gene expression RT-PCR single embryo 

Abbreviations

ASKη

shaggy-like kinase etha

DAP

days after pollination

DEPC

diethylpyrocarbonate

GAPC

glyceraldhyde-3-phosphate dehydrogenase

MP

monopteros

RT-PCR

reverse transcriptase-polymerase chain reaction

RVC

ribonucleoside vanadyl complexes

STM

shoot-meristemless

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Copyright information

© International Society for Plant Molecular Biology 2002

Authors and Affiliations

  1. 1.Key Laboratory of MOE for Plant Developmental BiologyWuhan UniversityWuhanPR. China

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