Non-invasive Assay for Chlorophyll Biosynthesis Kinetics Determination during Early Stages of Arabidopsis De-etiolation

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Authors

BALAKHONOVA Veronika PUSHKAROVA Nadiia SKALÁK Jan DOBISOVA Tereza BENEDIKTY Zuzana TRTILEK Martin HEJÁTKO Jan

Year of publication 2024
Type Article in Periodical
Magazine / Source Jove-Journal of Visualized Experiments
MU Faculty or unit

Central European Institute of Technology

Citation
Web https://app.jove.com/t/66087
Doi http://dx.doi.org/10.3791/66087
Keywords PLANT GROWTH; LIGHT
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Description Chlorophyll biosynthesis is a hallmark of de-etiolation, one of the most dramatic stages in the plant life cycle. The tightly controlled and highly dynamic process of chlorophyll biosynthesis is triggered during the shift from the dark to the light in flowering plants. At the moment when etiolated seedlings are exposed to the first traces of sunlight, rapid (in order of seconds) conversion of protochlorophyllide into chlorophyllide is mediated by unique light-accepting protein complexes, leading via subsequent metabolic steps to the production of fully functional chlorophyll. Standard techniques for chlorophyll content analysis include pigment extraction from detached plant tissues, which does not apply to studying such fast processes. To investigate chlorophyll kinetics in vivo with high accuracy and spatiotemporal resolution in the first hours after light-induced de-etiolation, an instrument and protocol were developed. Here, we present a detailed procedure designed for statistically robust quantification of chlorophyll in the early stages of Arabidopsis de-etiolation.
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