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.
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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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Zou, J.W., Sun, MX. & Yang, H.Y. Single-embryo RT-PCR assay to study gene expression dynamics during embryogenesis inArabidopsis thaliana . Plant Mol Biol Rep 20, 19–26 (2002). https://doi.org/10.1007/BF02801929
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DOI: https://doi.org/10.1007/BF02801929