diff --git a/tox.ini b/tox.ini
index f68e1eeb6f4..7da146a097d 100644
--- a/tox.ini
+++ b/tox.ini
@@ -49,6 +49,7 @@ passenv =
 setenv =
     coverage: GALAXY_TEST_COVERAGE=1
     first_startup: GALAXY_CONFIG_DATABASE_AUTO_MIGRATE=true
+    first_startup: GALAXY_PYTHON={env_python}
     mulled: marker=external_dependency_management
     mulled,unit: GALAXY_VIRTUAL_ENV={envdir}
     mypy,test_galaxy_packages,test_galaxy_packages_for_pulsar: PIP_INDEX_URL=https://wheels.galaxyproject.org/simple