Abstract

Context Estimating chloroplast pigment content by using chlorophyll meters is faster and cheaper than by using traditional pigment-extraction protocols. However, the reliability of chlorophyll meters requires a preliminary test of the relationship between the estimated and actual concentrations. Aims To establish an optimized protocol for extracting leaf chloroplast pigments from Eucalyptus urophylla and Khaya senegalensis seedlings. Moreover, we evaluated the relationship between chlorophyll-content estimation by using chlorophyll meters with the SPAD index by using SPAD-502 and with Falker chlorophyll index (FCI) by using ClorofiLOG, and that by using the optimal chloroplast pigment-extraction protocol. Methods The effectiveness of dimethylsulfoxide (DMSO) saturated with CaCO3 at 25°C and 65°C and 80% acetone (v/v) was assessed as extractors, by using different incubation times. Chlorophyll-content estimation was assessed using chlorophyll meters with the SPAD index by using SPAD-502 and with Falker chlorophyll index (FCI) by using ClorofiLOG. Key results For both Eucalyptus urophylla and Khaya senegalensis seedlings, the optimal protocol for extracting pigments from chloroplasts was incubation for 4 h by using DMSO saturated with CaCO3 at 65°C. Chlorophyll-content estimation using SPAD-502 and ClorofiLOG showed a strong positive correlation with measurements from the optimized pigment-extraction protocol. Furthermore, the regression equations of the chlorophyll-content estimation with chlorophyll meters allowed predicting the real content, because the values were very close to those measured using traditional extraction protocols. Conclusions The SPAD index and the FCI were reliable methods for estimating chlorophyll content in seedlings of both species. This could make measuring the chloroplast pigments faster and cheaper, especially regarding experimentation under field conditions.

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