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  • 1
    ISSN: 1617-4623
    Keywords: KeywordsOryza sativa L.  ;  AFLP markers  ;  Selective genotyping  ;  Submergence tolerance  ;  QTL analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract By combining the amplified fragment length polymorphism (AFLP) technique with selective genotyping, we constructed a linkage map for rice and assigned each linkage group to a corresponding chromosome. The AFLP map, consisting of 202 AFLP markers, was generated from 74 recombinant inbred lines (RIL) which were selected from both extremes of the population (250 lines) with respect to the response to complete submergence. Map length was 1756 cM, with an average interval size of 8.5 cM. To assign linkage groups to chromosomes, we used 50 previously mapped AFLP markers as anchor markers distributed over the 12 chromosomes. Other AFLP markers were then assigned to specific chromosomes based on their linkage to anchor markers. This AFLP map is equivalent to the RFLP/AFLP map constructed previously as the anchors were in the same order in both maps. Furthermore, tests with two restriction fragment length polymorphism (RFLP) markers and two sequence-tagged site (STS) markers showed that they mapped in the expected positions. Using this AFLP map, a major gene for submergence tolerance was localized on chromosome 9. Quantitative trait loci (QTL) associated with submergence tolerance were detected on chromosomes 6, 7, 11, and 12. We conclude that the combination of AFLP mapping and selective genotyping provides a much faster and easier approach to QTL identification than the use of RFLP markers.
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  • 2
    ISSN: 1432-2242
    Keywords: Key words Genetic map  ;  Marker-aided selection  ; Oryza sativa  ;  RFLP markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We exploited the newly developed amplified fragment length polymorphism (AFLP) technique to study the polymorphism, distribution and inheritance of AFLP markers with a doubled haploid rice population derived from ‘IR64’/‘Azucena’. Using only 20 pairs of primer combinations, we detected 945 AFLP bands of which 208 were polymorphic. All 208 AFLP markers were mapped and distributed over all 12 chromosomes. When these were compared with RFLP markers already mapped in the population, we found the AFLP markers to be highly polymorphic in rice and to follow Mendelian segregation. As linkage map of rice can be generated rapidly with AFLP markers they will be very useful for marker-assisted backcrossing.
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  • 3
    ISSN: 1432-2242
    Keywords: Key words AFLP ; Fingerprinting ; Genetic diversity ; Hybrid rice ; Oryza sativa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The cytoplasmic genetic male-sterile (CMS) lines developed at the International Rice Research Institute are valuable in producing tropical rice hybrids. Efficient use of CMS lines in hybrid rice production will depend on their level of genetic diversity. Aside from morphological characterization, molecular analysis based on DNA markers can provide information on the genetic diversity of the germplasm. The Amplified Fragment Length Polymorphism (AFLP) technique was used to fingerprint 71 CMS lines and four rice cultivars, ‘IR64’, ‘Azucena’, ‘IR74’, and ‘FR13A’. Eleven primer pair combinations specific to the enzymes PstI and MseI were used to generate 530 AFLP markers, 176 of which were polymorphic. Each CMS line revealed a distinct fingerprint. The AFLP marker-based dendrogram depicted genetic variation among the CMS lines. The CMS lines developed in japonica background grouped with ‘Azucena’, a japonica cultivar. None of the CMS lines clustered with ‘FR13A’, a flood-tolerant traditional indica variety. ‘IR64’ was found to be distinct from the other indica CMS lines and clustered with lines developed in its background. The grouping of CMS lines into a few groups is useful for breeders in selecting genetically diverse CMS lines for hybrid rice production and in avoiding test crossing every CMS line empirically. This study demonstrated that AFLP is a powerful and reliable tool in determining the genetic relationships and in producing distinct fingerprints of rice cultivars.
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  • 4
    ISSN: 1432-2242
    Keywords: Key words AFLP ; Bulked segregant analysis ; Gene tagging ; Marker-assisted selection ; Rice TGMS gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The discovery and application of the thermosensitive genic male sterility (TGMS) system has great potential for revolutionizing hybrid seed production technology in rice. Use of the TGMS system in two-line breeding is simple, inexpensive, efficient, and eliminates the limitations associated with the cytoplasmic-genetic male sterility (CMS) system. An F2 population developed from a cross between a TGMS indica mutant, TGMS–VN1, and a fertile indica line, CH1, was used to identify molecular markers linked to the TGMS gene and to subsequently determine its chromosomal location on the linkage map of rice. Bulk segregant analysis was performed using the AFLP technique. From the survey of 200 AFLP primer combinations, four AFLP markers (E2/M5–600, E3/M16–400, E5/M12–600, and E5/M12–200) linked to the TGMS gene were identified. All the markers were linked to the gene in the coupling phase. All except E2/M5–200 were found to be low-copy sequences. However, the marker E5/M12–600 showed polymorphism in RFLP analysis and was closely linked to the TGMS gene at a distance of 3.3 cM. This marker was subsequently mapped on chromosome 2 using doubled-haploid mapping populations derived from the crosses IR64×Azucena and CT9993×IR62666, available at IRRI, Philippines, and Texas Tech University, respectively. Linkage of microsatellite marker RM27 with the TGMS gene further confirmed its location on chromosome 2. The closest marker, E5/M12–600, was sequenced so that a PCR marker can be developed for the marker-assisted transfer of this gene to different genetic backgrounds. The new TGMS gene is tentatively designated as tms4(t).
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  • 5
    ISSN: 1432-2242
    Keywords: Key words Drought tolerance ; Genetic mapping ; Gene interaction ; Marker-assisted selection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Stay green in sorghum (Sorghum bicolor L. Moench) is characterized by the plant’s ability to tolerate post-flowering drought stress, thereby delaying the premature leaf and plant death. It contributes to normal grain filling and reduces the incidence of stalk lodging and charcoal rot disease during the late stages of grain development. Breeding for improving post-flowering drought tolerance in sorghum hybrids remains an important objective of sorghum breeders. Since evaluation of the stay green response is difficult and unreliable under field conditions, due to the timing and intensity of moisture stress and large environmental interaction, progress in improving drought tolerance by conventional breeding methods has been slow. The objective of the present study was to determine the consistency of quantitative trait loci (QTLs) controlling stay green in sorghum. We re-evaluated the Recombinant Inbred Line (RIL)-mapping population from the cross B35 x Tx7000 in two locations over 2 years and compared it with earlier reports. Analysis using the combined stay green-rating means of seven environments and the expanded molecular map reconfirmed all four stay green QTLs (Stg1, Stg2, Stg3 and Stg4) that were identified earlier by Xu et al. (2000). Similarly, comparison of the stay green QTL locations with earlier reported results indicated that all four stay green QTLs showed consistency across different genetic backgrounds. Examination of the stay green QTL profiles of the best and poorest stay-green lines indicated that three stay green QTLs, Stg1, Stg2 and Stg3, appear to be important for the expression of this trait when the percent phenotypic variation, and the consistency in different backgrounds and different environments, are considered. A significant epistatic interaction involving Stg2 and a region on linkage group C was also identified for the stay green and chlorophyll content. We concluded that Stg2 is the most important QTL controlling stay green, explaining the maximum amount of phenotypic variation. This report further strengthens our view to target the Stg2 QTL region for gene discovery in order to improve the basic understanding of the stay green phenomenon, which might be helpful in manipulating this trait not only in sorghum but also in other cereal crop species.
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