Comprehensive Physio-Biochemical Evaluation Reveals Promising Genotypes and Mechanisms for Cadmium Tolerance in Tibetan Hull-Less Barley

Cadmium (Cd) toxicity in agricultural soil is increasing globally and significantly impacts crop production and food safety. Tibetan hull-less barley (<i>Hordeum vulgare</i> L. var. <i>nudum</i>), an important staple food and economic crop, exhibits high genetic diversity and is uniquely adapted to the harsh conditions of the Qinghai–Tibet Plateau. This study utilized hydroponic experiments to evaluate the genotypic differences in Cd tolerance among 71 Tibetan hull-less barley genotypes. Physiological assessments revealed significant reductions in various growth parameters under Cd stress compared to normal conditions: soil–plant analysis development (SPAD) value, shoot height, root length, shoot and root fresh weight, shoot and root dry weight, of 11.74%, 39.69%, 48.09%, 52.88%, 58.39%, 40.59%, and 40.52%, respectively. Principal component analysis (PCA) revealed key traits contributing to Cd stress responses, explaining 76.81% and 46.56% of the variance in the preliminary and secondary selection. The genotypes exhibited varying degrees of Cd tolerance, with X178, X192, X215, X140, and X162 showing high tolerance, while X38 was the most sensitive based on the integrated score and PCA results. Validation experiments confirmed X178 as the most tolerant genotype and X38 as the most sensitive, with observed variations in morphological, physiological, and biochemical parameters, as well as mineral nutrient responses to Cd stress. Cd-tolerant genotypes exhibited higher chlorophyll content, net photosynthesis rates, and effective photochemical capacity of photosystem II, along with an increased Cd translocation rate and reduced oxidative stress. This was accompanied by elevated activities of antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), indicating a robust stress response mechanism. These findings could facilitate the development of high-tolerance cultivars, with X178 as a promising candidate for further research and cultivation in Cd-contaminated soils.