Background and aims Plant adaptation to waterlogged conditions requires a set of morphological and physiological/biochemical changes. The formation of aerenchyma is one of the most crucial adaptive traits for waterlogging tolerance. Enzymatic scavenging may also potentially contribute to waterlogging tolerance by providing detoxification of reactive oxygen species (ROS).

Methods Changes of root porosity (as an indicator of aerenchyma formation) and activities in leaves of four major antioxidant enzymes, γ-amino butyric acid (GABA) and lactic acid contents in roots were evaluated in six barley genotypes contrasting in waterlogging tolerance.

Results Soil waterlogging caused significant increases in adventitious root porosity in all genotypes. Waterlogging-tolerant genotypes showed not only significantly higher adventitious root porosity than sensitive genotypes but also much faster development of aerenchyma. The greatest difference in adventitious root porosity among genotypes was observed after 7 days of waterlogging treatment. At the same time, antioxidant enzyme activities in leaves, GABA and lactic acid contents in roots did not correlate with waterlogging tolerance.

Conclusions A faster formation of aerenchyma in adventitious roots is one of the key factors for waterlogging tolerance in barley. This protocol is recommended to be applied in future studies to identify molecular markers linked to this trait using appropriate mapping populations.