Comparative analyses of extreme dry seed thermotolerance in five Cactaceae species

Abstract

Extreme dry seed thermotolerance is an important trait in the context of plant adaptation to desert environments, yet the mechanisms underlying interspecific differences in seed extremophily remain little understood. Seeds of a selection of desert Cactaceae species (Leucostele litoralis, Leucostele skottsbergii, Eulychnia breviflora, Eriosyce paucicostata and Ferocactus wislizeni) were subjected to comparative biophysical and molecular analyses prior to (mature dry seeds) and post (imbibing seeds) heat treatment (103 °C).Biophysical properties of mature dry seeds, such as seed coat hardness and thermal transitions, were characterized through puncture force resistance and differential scanning calorimetry, respectively. In addition, the protein composition of dry seeds was compared by 2-D electrophoresis and de novo sequencing. Finally, oxidative DNA damage repair and gene expression during seed imbibition were analysed via ELISA (8-oxo-dG levels) and comparative transcriptomics.Results revealed that enhanced dry seed thermotolerance could not be explained by seed coat hardness, seed thermal transitions or (ROS-induced oxidative) DNA damage repair. Comparative proteomic analysis showed that the majority of overlapping proteins were more abundant in relatively heat sensitive species, suggesting that higher basal levels of (stress) proteins do not underly improved heat resilience in Cactaceae. However, comparative transcriptomics during imbibition following heat stress detected consistent expression patterns of heat- and drying-responsive genes that correlated with dry seed thermotolerance. These patterns included upregulation of positive (HSP101/CLPB1, MGE2, MBF1c, HSFB2A, RPN1a, RPT2a, DEGP14/PARK13, XPO1A, MAIGO2/MAG2, CAMTA1, ADH1) and downregulation of negative (RCF3, VOZ1, FAD7, ZAT12, RNP1) heat and drought tolerance regulators.Thus, our comparative analyses of extremophile Cactaceae species suggest that higher dry seed thermotolerance is realised through enhanced recovery from heat stress during imbibition, which is a critical stage in the plant life cycle.