Guidelines for using antibodies against dehydrins in agrobiotechnology

Detection of the expression levels of the dehydrin gene in plants is increasingly becoming a widespread technique for testing plant resistance to cold, drought, salinity or high heavy metal concentrations in soils. The simplest way to determine the content of dehydrins consists in using specific antibodies (antibody specificity is critical). The experience of dehydrin detection in various plants provided a basis for this work. In this study, we set out to compare the reactions of antibodies with reactive proteins derived from three most accessible sources, as well as to assess their specificity. The comparability of detected proteins in terms of their quantity and molecular weight was studied for several plants: 1) total and thermostable protein in scots pine needles; 2) mitochondrial protein in peas; 3) total and thermostable protein in winter wheat; 4) total and thermostable protein in arabidopsis. In all the cases, either slight or significant differences in the quantity and molecular weight of the detected protein groups were observed. These differences can be attributed to the three possible reasons: K-segment variability; different carrier proteins used by manufacturers; as well as the presence of shared reactive epitopes in some groups of proteins that are not dehydrins. The analysis of differences in the number of the groups detected using antibodies from different manufacturers reveals their amount to be minimal for cereal plants and becoming larger for dicotyledons and conifers. The largest number of the groups was detected in pea mitochondria. Some of the antibody-binding protein groups are definitely not dehydrins. The abovementioned differences are smaller for thermostable proteins. Thus, obtaining a thermostable protein fraction can be recommended as a simple procedure increasing the dehydrins detection level. The comparison of the dehydrin amounts should not be used as a measure of the relative plant resistance when comparing evolutionarily distant species, since the antibody reactivity to the dehydrins of distant species may differ significantly. At the same time, this method can be used in agrobiology for the molecular-level assessment of the initial or induced resistance of plant varieties (same or related species) to cold, drought, as well as other adverse conditions associated with water deficiency.

dehydrins, plant resistance, western blot, antibodies

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