We describe a major new release of the EoRNA database, a gene expression database for barley based on public data, first published in 20211. EoRNA v.2 (https://ics.hutton.ac.uk/eorna2/index.html) features an order of magnitude more samples and is based on a new automated workflow of sample discovery and processing which has enabled a dramatic scale-up the original database. EoRNA v.2 also features a major rebuild of the web user interface with rich new functionality. All infrastructure-related code and database schemas and web components are now species agnostic and publicly available for reuse with other taxa. A dedicated new reference transcript dataset has been created for EoRNA v.2 which is largely based on the recently published barley pan-transcriptome and represents the most comprehensive dataset of its kind to date.