Protocol for evaluating salt and alkaline tolerance in maize seedlings using a sand-bed germination assay

Here, we present a protocol to independently and concurrently evaluate salt and alkaline tolerance in maize at the seedling stage using a controlled sand-bed germination assay. We describe steps for preparing standardized stress solutions, establishing the sand-bed assay, applying stresses either independently or in combination, and measuring multiple phenotypic traits. Furthermore, we detail procedures for ranking germplasm using a comprehensive scoring system based on a membership function. This protocol enables reproducible, large-scale screening of maize germplasm at the seedling stage.