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. Chlorophyll concentrations of biological soil crust (biocrust) samples arecommonly determined to quantify the relevance of photosynthetically activeorganisms within these surface soil communities. Whereas chlorophyllextraction methods for freshwater algae and leaf tissues of vascular plantsare well established, there is still some uncertainty regarding the optimalextraction method for biocrusts, where organism composition is highlyvariable and samples comprise major amounts of soil. In this study weanalyzed the efficiency of two different chlorophyll extraction solvents, theeffect of grinding the soil samples prior to the extraction procedure, andthe impact of shaking as an intermediate step during extraction. The analyseswere conducted on four different types of biocrusts. Our results show thatfor all biocrust types chlorophyll contents obtained with ethanol weresignificantly lower than those obtained using dimethyl sulfoxide (DMSO) as asolvent. Grinding of biocrust samples prior to analysis caused a highlysignificant decrease in chlorophyll content for green algal lichen- andcyanolichen-dominated biocrusts, and a tendency towards lower values formoss- and algae-dominated biocrusts. Shaking of the samples after eachextraction step had a significant positive effect on the chlorophyll contentof green algal lichen- and cyanolichen-dominated biocrusts. Based on ourresults we confirm a DMSO-based chlorophyll extraction method withoutgrinding pretreatment and suggest the addition of an intermediate shakingstep for complete chlorophyll extraction (see Supplement S6 for detailedmanual). Determination of a universal chlorophyll extraction method forbiocrusts is essential for the inter-comparability of publications conductedacross all continents.

biogeosciences

Revisiting chlorophyll extraction methods in biological soil crusts – methodology for determination of chlorophyll &amp;lt;i&amp;gt;a&amp;lt;/i&amp;gt; and chlorophyll &amp;lt;i&amp;gt;a&amp;lt;/i&amp;gt; + &amp;lt;i&amp;gt;b&amp;lt;/i&amp;gt; as compared to previous methods