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Conversion of Fertility Restoration of the Sorghum IS1112C (A3) Male-Sterile Cytoplasm from Two Genes to One Gene

USDA-ARS, Crop Genetics and Environment Unit, Department of Plant Pathology, 1453 Fifield Hall, University of Florida, Gainesville, FL 32611 USA

View larger version (28K): [in a new window]   Fig. 1. Transcript analysis of segregants for Mmt1 among B3Tx398/IS1112C F5 progeny. Action of Mmt1 results in enhanced transcript processing activity 5' to urf209, generating a 832 nt transcript from the progenitor 1044 nt transcript. Segregants B, C, E, F, and I represent genotypes Mmt1Mmt1 or Mmt1mmt1, while A, D, G, H, J, K, and L are mmt1mmt1.

View larger version (21K): [in a new window]   Fig. 2. Strategy for selecting lines (B3)FL3 (Rf3Rf3rf4rf4) and (B3)FL4 (rf3rf3Rf4Rf4). B3Tx398/IS1112C F5:6 individuals were scored for Mmt1, tightly linked to Rf3. Rf3rf3 or Rf3Rf3 individuals were used to pollinate A3Tx398 and resulting sterile plants identified the Rf3Rf3rf4rf4 lines, designated (B3)FL3. Segregants that were mmt1mmt1, and thus rf3rf3, were used to pollinate A3Tx398 to verify the genotype. The line A3Tx398/(B3)FL3 (Rf3rf3rf4rf4) was pollinated with the candidate lines; progeny that yielded only sterile F1s (Rf3rf3rf4rf4) identified rf4rf4 lines, while progeny that segregated 1 fertile:1 sterile identified the rf3rf3Rf4Rf4 lines, designated (B3)FL4.

View larger version (16K): [in a new window]   Fig. 3. Identification of Rf4Rf4 lines among rf3rf3 F5:6 lines crossed as male with A3Tx398/(B3)FL3. The codominant STS marker LW9 is 366 bp in IS1112C (A) and five segregants (C-G), while B3Tx398 (B) exhibits a 358-bp fragment.

View larger version (16K): [in a new window]   Fig. 4. Strategy for selecting (A3)FL3 (Rf3Rf3rf4rf4). A3Tx398 was pollinated with Mmt1 lines identified from the B3Tx398/IS1112C F5:6 population. Fertile plants were disregarded. Sterile lines were pollinated with (B3)FL3, backcrossed with (B3)FL3, then pollinated with (B3)FL4. Progeny which were all fertile identified the female (A3)FL3.

View larger version (86K): [in a new window]   Fig. 5. Iodine staining of pollen from line (A3)Rf3Rf3rf4r4 pollinated with A) (B3)FL4 (rf3rf3Rf4Rf4) and B) IS1112C (Rf3Rf3Rf4Rf4), resulting in approximately 25 and 50% staining, respectively. Note partially filled grains in each line, as previously described (Tang et al., 1998).

View larger version (16K): [in a new window]   Fig. 6. Strategy for selecting (A3)FL4 (rf3rf3Rf4Rf4). A3Tx398 was pollinated with (B3)FL4, backcrossed with (B3)FL4, and progeny were examined for presence of the rf4-linked codominant markers LW8 and LW9. Selected females were pollinated with (B3)FL3; the F1s were fertile with 25% iodine-stained pollen, identifying the line as (A3)FL4.