EMA-based chromosome preparation and fluorescent banding as an aid to detect adulteration in cultivated Lens culinaris.

India, the world's largest consumer and importer of lentils, is concerned about intentional or unintentional adulteration of this high-protein staple, which compromises food security, local price stability, and efforts to boost per capita lentil production to reduce costly imports. In this study, cost-effective EMA-based chromosomal analysis using Giemsa and fluorescence staining confirms the presence of a mixed population of lentils growing in farmers' fields in West Bengal, raising quality concerns for lentil farmers in these areas. Moreover, the adulterant is identified as a very closely related germplasm of wild-type lentils, L. ervoides, by cluster analysis based on comparative chromosome analysis against the previously reported Lentil database. The karyomorphological features, such as the terminal secondary constrictions, 18-20 DAPI-positive signals in all chromosomes, and CMA-positive signals at nucleolar regions, along with seed and flower morphology and the presence of tendrils, effectively differentiate cultivated lentils from wild-type (adulterant) lentil germplasm. Cluster analysis also indicates that the adulterant is more closely related to wild-type lentil L. ervoides. This method provides an important tool for quality control and seed authentication in lentils and helps to enhance farmers' productivity, boost the economy, improve consumers' nutrition, and contribute to national productivity and GDP. Moreover, it emphasizes the need to create a national database of Indian lentil germplasm and develop region-specific certified germplasm programs to optimize per capita lentil production, considering the diverse agronomic conditions across India.