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Molecular Breeding for the Development of Blast and Bacterial Blight Resistance in Rice cv. IR50

^a Entomology and Plant Pathology Div., IRRI, DAPO Box 7777, Metro Manila, The Philippines
^b Center for Advanced Studies in Botany, Univ. of Madras, Guindy Campus, Chennai-600025, India

View larger version (63K): [in a new window]   Fig. 1. Phenotypic reaction of Piz-5 introgressed IR50 line against rice blast isolates C9240-5, IK81-3, and IK81-25.

View larger version (30K): [in a new window]   Fig. 2. Phenotypic evaluation of blast resistant IR50 containing the Piz-5 gene using three diagnostic strains following the procedure of Bonman et al., 1986.

View larger version (89K): [in a new window]   Fig. 3. The generation of sequence-specific amplicon polymorphism between resistant (C101A51) and susceptible (IR50) rice varieties for marker-aided selection. (A) Gel analysis of polymerase chain reaction (PCR) products before restriction digestion. (B) Gel analysis of PCR products after digestion with restriction enzyme HaeIII. Lanes 1 and 2 represent the susceptible parent IR50 and donor resistant line C101A51, respectively. The molecular weight of PCR products as banding patterns is indicated in kb (kilobases). M = Molecular weight marker (1-kb ladder).

View larger version (30K): [in a new window]   Fig. 4. Evaluation of isogenic lines and gene stacked lines after 15 to 30 d with 5-d interval in the rice blast nursery at IRRI, The Philippines, based on the standard evaluation system of rice 15, 20, 25, and 30 d after inoculation.

View larger version (194K): [in a new window]   Fig. 5. Blast nursery showing higher infection in susceptible check (IR50) and less infection on isolines of IR50 with Piz-5 gene.

View larger version (111K): [in a new window]   Fig. 6. Bacterial blight reactions to (A) Race PXO61 and (B) PXO86 in IR24 (1), IRBB4 (2), and IR50 (3) to examine endogenous Xa-4 gene in rice cv. IR50.

View larger version (76K): [in a new window]   Fig. 7. Polymerase chain reaction analysis of transgenic IR50 T0 Plants 13, 14, and 15 showing a 1.4-kilobase (marked with arrow) Xa-21-specific DNA fragment amplified by primers U1 and I1. M = molecular weight marker; Lanes 1, 2, and 3 represent nontransformed IR50, IRBB21, pC822, and transgenic IR50 Plants 13, 14, and 15, respectively.

View larger version (82K): [in a new window]   Fig. 8. Southern analysis of transgenic T0 plants. The arrow represents the expected 3.8-kilobase (kb) band corresponding to the Xa21 transgene. Lanes 1 to 5 represent nontransformed IR50, transgenic T0 IR50-13, 14, and 15, and plasmid pC822, respectively. Five- to 7-µg plant genomic DNA and 30 pg of plasmid DNA were digested with EcoRV. The 1.4-kb polymerase chain reaction amplified fragment of pC822 was labeled with -[32P]-dCTP using the Rediprime Labeling Kit (Amersham Heights, IL) and was used as the hybridization probe.

View larger version (48K): [in a new window]   Fig. 9. Southern analysis of transgenic T1 plants. A total of 5 to 7 µg of plant genomic DNA and 30 pg of plasmid DNA were digested with EcoRV and hybridized with the same enzyme-digested plasmid DNA fragment. The arrow marks the expected 3.8-kilobase hybridizing band corresponding to Xa21 transgene in T0 and T1 plants. T0 = primary transgenic plant, T1 = selfed progenies of T13 (T0).

View larger version (68K): [in a new window]   Fig. 10. Reactions of transgenic T1 IR50 plants carrying Xa21 and Piz-5 introgressed for blast resistance when inoculated with IK81-3. Leaves 1 to 4 represent CO39, IR50, C101A51 (Piz-5), and transgenic IR50 (Piz-5 + Xa-21), respectively.

View larger version (75K): [in a new window]   Fig. 11. Resistant reactions of transgenic T1 IR50 plants introgressed with Xa-21 for bacterial blight resistance inoculated with Races (A) PXO99, (B) PXO86, and (C) PXO341. Leaves 1 to 5 represent IR24, IR50, IRBB4, IRBB21, and transgenic IR50 (T13) (Piz-5 + Xa-21), respectively, scored 14 d after inoculation.