Warm Temperature Promotes Shoot Regeneration in Arabidopsis thaliana

Many plants are able to regenerate upon cutting, and this process can be enhanced in vitro by incubating explants on hormone-supplemented media. While such protocols have been used for decades, little is known about the molecular details of how incubation conditions influence their efficiency. In this study, we find that warm temperature promotes both callus formation and shoot regeneration in Arabidopsis thaliana. We show that such an increase in shoot regenerative capacity at higher temperatures correlates with the enhanced expression of several regeneration-associated genes, such as CUP-SHAPED COTYLEDON 1 (CUC1) encoding a transcription factor involved in shoot meristem formation and YUCCAs (YUCs) encoding auxin biosynthesis enzymes. ChIP-sequencing analyses further reveal that histone variant H2A.Z is enriched on these loci at 17 degrees C, while its occupancy is reduced by an increase in ambient temperature to 27 degrees C. Moreover, we provide genetic evidence to demonstrate that H2A.Z acts as a repressor of de novo shoot organogenesis since H2A.Z-depleted mutants display enhanced shoot regeneration. This study thus uncovers a new chromatin-based mechanism that influences hormone-induced regeneration and additionally highlights incubation temperature as a key parameter for optimizing in vitro tissue culture.

Automated summary

This study reveals that higher temperature promotes callus formation and shoot regeneration in Arabidopsis thaliana. The increased shoot regenerative capacity at higher temperatures is correlated with enhanced expression of several regeneration-associated genes and reduced occupancy of a histone variant at specific loci. The study also provides genetic evidence that the histone variant acts as a repressor of shoot organogenesis. These findings uncover a chromatin-based mechanism and highlight the importance of incubation temperature for optimizing tissue culture.