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Determination of Chlorophyll Concentration in In Vitro Peanut Leaves

The detection of chlorophyll is a convenient method for assessing gene editing efficiency when generating mutations in the phytoene desaturase (PDS) gene, which is related to chlorophyll biosynthesis. Various instruments and protocols are available for measuring chlorophyll; however, a minimum leaf...

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Main Authors: Faustinelli, Paola C., Massa, Alicia N., Soria, Néstor W., Lopez Colomba, Eliana, Suarez, Paola Alejandra, Lamb, Marshall C.
Format: info:ar-repo/semantics/artículo
Language:Inglés
Published: Wiley 2025
Subjects:
Arachis hypogaea
Laboratory Experimentation
Chlorophylls
Leaves
In Vitro Experimentation
Experimentación en Laboratorio
Clorofila
Hojas
Experimentación In Vitro
Maní
Peanuts
Online Access:http://hdl.handle.net/20.500.12123/24824
https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpz1.70150
https://doi.org/10.1002/cpz1.70150
Summary:The detection of chlorophyll is a convenient method for assessing gene editing efficiency when generating mutations in the phytoene desaturase (PDS) gene, which is related to chlorophyll biosynthesis. Various instruments and protocols are available for measuring chlorophyll; however, a minimum leaf area is required for precise quantifications. In vitro plant regeneration often requires several months to produce enough leaf tissue for chlorophyll content analysis. In this study, we optimized an existing chlorophyll quantification method by using a minimal amount of in vitro leaf tissue (<2.0 mg) and a microvolume of chlorophyll extraction solution (3 µl) to quantify chlorophyll a, chlorophyll b, and total chlorophyll concentration using a UV-Vis spectrophotometer. This method will serve as a basic protocol for evaluating gene editing efficiency by correlating degrees of albinism (pigment content) with changes in the PDS gene sequence.

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