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Maize is an excellent nutritional source and is consumed as a staple food in different parts of the world, including India. Developing a maize genotype with a combination of higher lysine and tryptophan, along with β-carotene, can help alleviate the problem of protein-energy malnutrition (PEM) and vitamin A deficiency (VAD). This study is aimed at improving lysine and tryptophan content by transferring  opaque-2  ( o2 ) gene from donor HKI163 to β- carotene -rich inbred lines  viz ., UMI1200β +  and UMI1230β + . For this purpose, F 1 , BC 1 F 1 , BC 2 F 1 , BC 2 F 2 , and BC 2 F 3  plants were developed using an o2 line HKI163 and two β-carotene-rich inbred lines, UMI1200β +  and UMI1230β + , as the parents. Foreground selection using the associated marker umc1066 for the  o2  gene and the marker  crtRB1  3′TE for the  crtRB1  gene was used to select the target genes. A total of 236 simple sequence repeat (SSR) markers distributed evenly across the maize genome were employed for the background selection. To fix the  crtRB1  allele in the BC 1 F 1  stage, individual plants homozygous at the  crtRB1  locus and heterozygous at the  o2  locus were selected and used for backcrossing to produce BC 2 F 1  plants. Furthermore, the selected heterozygous BC 2 F 1  plants from both crosses were selfed to obtain the BC 2 F 2  plants, which were then selected for the target gene and selfed to generate the BC 2 F 3  lines. From each cross, five improved lines with homozygous marker alleles for the  crtRB1  and  o2  genes with a recurrent parent genome (RPG) recovery ranging from 86.75 to 91.21% in UMI1200β + ×HKI163 and 80.00 to 90.08% in UMI1230β + ×HKI163 were identified. The improved lines had good agronomic performance and possessed high lysine (0.294–0.332%), tryptophan (0.073–0.081%), and β- carotene  (6.12–7.38 µg/g) content. These improved lines can be used as genetic resources for maize improvement.

Marker-Assisted Selection to Pyramid the Opaque-2 (O2) and β-Carotene (crtRB1) Genes in Maize