Characterization and molecular marker screening of a rice bacteria-resistant gene Xa-min(t)

来源 :Progress in Natural Science | 被引量 : 0次 | 上传用户:snake_icy1
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To test the resistant spectrum of the Xa min(t) gene introgressed from Oryza minuta , thirty four isolates of different bacterial blight pathogen, Xanthomonas oryzae pv. oryzae (Xoo), from 11 countries were used to inoculate the Xa min(t) introgression line 78 15. Four rice cultivars, IR24, C64 (IRBB21), Nipponbare and Zhonghua 11 were used as controls. The results showed that the Xa min(t) gene was broad spectrum and highly resistant to diverse Xoo isolates. The methods of bulk segregant analysis (BSA), randomly amplified polymorphic DNA (RAPD) and sequence characterized amplified regions (SCAR) were used to analyze F 2 individuals of the hybrid IR24×78 15 and molecular genetic markers linked to Xa min(t) gene were identified. A total of 800 arbitrary decamer oligonucleotide primers were used for RAPD analysis. Two RAPD markers, BE05 300 and BE06 1400 , produced by primers BE05 and BE06 respectively, were closely linked to the Xa min(t) gene. Based on the sequences of these two markers, sequence specific primers were designed and used to screen all F 2 plants. One RAPD marker, BE05 300 , was converted into a stable SCAR marker (ScBE05 300 ). Linkage analysis was carried out using markers ScBE05 300 and BE06 1400 on 948 and 719 F 2 individuals of the hybrid IR24×78 15. Our results indicate that the genetic distances from Xa min(t) to ScBE05 300 and BE06 1400 are 2.2 cM and 3.7 cM respectively on the same side. This study may facilitate the construction of the fine physical map of the Xa min(t) gene. To test the resistant spectrum of the Xa min (t) gene introgressed from Oryza minuta, thirty four isolates of different bacterial blight pathogen, Xanthomonas oryzae pv. Oryzae (Xoo), from 11 countries were used to inoculate the Xa min (t) introgression line 78 15. Four rice cultivars, IR24, C64 (IRBB21), Nipponbare and Zhonghua 11 were used as controls. The results showed that the Xa min (t) gene was broad spectrum and highly resistant to diverse Xoo isolates. The methods of bulk segregant analysis (BSA), randomly amplified polymorphic DNA (RAPD) and characterized amplified regions (SCAR) were used to analyze F2 individuals of the hybrid IR24 × 78 15 and molecular genetic markers linked to Xa min (t) genes were identified. A total of 800 arbitrary decamer oligonucleotide primers were used for RAPD analysis. Two RAPD markers, BE05 300 and BE06 1400, produced by primers BE05 and BE06 respectively, were closely linked Based on the sequences of these two markers, sequence specific primers were designed and used to screen all F2 plants. One RAPD marker, BE05 300, was converted into a stable SCAR marker (ScBE05 300) . Linkage analysis was carried out using markers ScBE05 300 and BE06 1400 on 948 and 719 F2 individuals of the hybrid IR24 × 78 15. Our results indicate that the genetic distances from Xa min (t) to ScBE05 300 and BE06 1400 are 2.2 cM This study may facilitate the construction of the fine physical map of the Xa min (t) gene.
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