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ARS Home » Southeast Area » Dawson, Georgia » National Peanut Research Laboratory » Research » Publications at this Location » Publication #424752
Research Project: Integration of Traditional Methods and Novel Molecular Strategies for Improving Disease Resistance and Input-use Efficiency in Peanut

Location: National Peanut Research Laboratory

Title: Determination of chlorophyll content in in vitro peanut leaves

Author
item Faustinelli, Paola
item Massa, Alicia
item SORIA, NESTOR - Catholic University Of Córdoba
item LOPEZ-COLOMBA, ELIANA - Instituto Nacional De Tecnologia Agropecuaria
item SUAREZ, PAOLA - Instituto Nacional De Tecnologia Agropecuaria
item Lamb, Marshall

Submitted to: Current Protocols in Plant Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/5/2025
Publication Date: 5/22/2025
Citation: Faustinelli, P.C., Massa, A.N., Soria, N.W., Lopez-Colomba, E., Suarez, P.A., Lamb, M.C. 2025. Determination of chlorophyll content in in vitro peanut leaves. Current Protocols in Plant Biology. 5, e70150. https://doi.org/10.1002/cpz1.70150.
DOI: https://doi.org/10.1002/cpz1.70150

Interpretive Summary: This article describes a simple and efficient chlorophyll quantification procedure optimized from an existing method (Monteoliva et al. 2019). The optimization involved using a very small leaf sample (less than 2 mg) from in vitro peanut shoots, and 3 µL of chlorophyll extraction. The method allows for the measurement of different types of chlorophyll (a, b, and total) using a UV-Vis spectrophotometer. This protocol can help evaluate gene editing by linking plant color changes, such as albinism, to changes in the gene causing albino phenotype (phytoene desaturase gene - PDS).

Technical Abstract: Detection of chlorophyll is a convenient method for assessing gene editing efficiency when generating mutations in the phytoene desaturase gene (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 obtain enough leaf tissue for chlorophyll content analysis. In this study, we optimized an existing chlorophyll quantification method (Monteoliva et al. 2019) by using a minimal amount of in vitro leaf tissue (less than 2 mg) and a microvolume of chlorophyll extraction solution (3 µL) to quantify chlorophyll a, b, and total chlorophyll on a UV-Vis spectrophotometer. The 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.