Genetic improvements of iron (Fe) and zinc (Zn) content in pearl millet [Pennisetum glaucum (L.) R. Br.] may reduce the problems of anemia and stunted growth among millet dependent staple food consumers. The availability of variation in diverse-breeding lines is essential to improve grain micronutrients in high-yielding cultivars. This study aimed to determine the extent of variability, heritability and diversity for grain Fe, Zn and protein, along with key agronomic traits, in 281 advanced breeding lines bred at ICRISAT and evaluated across two seasons (environments). A pooled analysis of variance displayed significant variation for all these traits. Highest variability was recorded for Fe (35–116 mg kg⁻¹), Zn (21–80 mg kg⁻¹), and protein (6–18%), and a three-fold variation was observed for panicle length, panicle girth and 1000-grain-weight (TGW). Diversity analysis showed 10 clusters. Cluster-III had maximum lines (25%) and Cluster-V showed the highest mean values for Fe, Zn, protein and TGW. These results highlight the success of breeding program that aimed both the maintenance and creation of genetic variability and diversity. A significant positive correlation among Fe, Zn, protein and TGW indicated the potential for simultaneous improvement. Grain yield had a non-significant association with Fe and Zn, while protein showed a negative correlation. These results suggest that significant variability exists in elite-breeding lines, thus highlighting an opportunity to breed for biofortified varieties without compromising on the grain yield. The lines with high Fe, Zn and protein content can be used as hybrid parents and may also help in further genetic investigations.

珍珠小米[ Pensetsetum glaucum（L.）R. Br。]中铁（Fe）和锌（Zn）含量的遗传改良可减少依赖小米的主食消费者的贫血和生长迟缓问题。多样化育种系中变异的可用性对于改善高产品种的谷物微量营养素至关重要。这项研究旨在确定在ICRISAT育成并在两个季节（环境）中进行评估的281个高级育种品系中谷物铁，锌和蛋白质的变异性，遗传性和多样性程度以及关键的农艺性状。汇总的方差分析显示了所有这些特征的显着差异。Fe（35–116 mg kg ^-1），Zn（21–80 mg kg ^-1）记录的最大变异性）和蛋白质（6-18％），并且穗长，穗周长和1000粒重（TGW）观察到三倍变化。多样性分析显示10个群集。聚类III的谱线最大（25％），聚类V的Fe，Zn，蛋白质和TGW的平均值最高。这些结果突出了旨在维持和创造遗传变异性和多样性的育种计划的成功。Fe，Zn，蛋白质和TGW之间的显着正相关表明了同时改善的潜力。谷物产量与铁和锌无显着关联，而蛋白质则呈负相关。这些结果表明，在优良育种系中存在显着的变异性，从而突出了在不损害谷物产量的情况下进行生物强化品种育种的机会。铁含量高的线