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  • 1
    ISSN: 1432-2242
    Keywords: Key words Plasma membrane H+-ATPase gene ; Salt stress ; Salt-tolerant mutant ; Rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Plasma membrane (PM) H+-ATPase plays an important role in the establishment and maintenance of ion homeostasis. To investigate its expression in the rice salt-tolerant mutant M-20 and the original variety 77–170 during salt stress, a cDNA fragment corresponding to the PM H+-ATPase gene was obtained by PCR from rice japonica variety 77–170 and designated as OSA3. Sequence analysis of OSA3 revealed its high homology with two other published PM H+-ATPase genes, OSA1 and OSA2, in rice. Southern-blot analysis detected a RFLP between M-20 and 77–170, and one copy of the OSA3 gene was mapped to a position on rice chromosome 12 where a salt tolerance QTL was closely located. The expression of the PM H+-ATPase gene, as revealed by the OSA3 fragment, was compared between M-20 and 77–170. The results demonstrated that M-20 shoots accumulated less transcripts than 77–170 shoots at a later stage of salt treatment, and M-20 showed high expression at 300 mM NaCl while 77–170 reached its maximum at 200 mM NaCl. In roots, the difference in the level of the PM H+-ATPase gene expression between stressed and non-stressed plants was substantially greater in M-20 than that in 77–170. The relative abundance of PM H+-ATPase gene transcripts in M-20 roots may indicate the active role of this gene in the strict control of Na+ and Cl+ uptake into root symplast and apoplast, and further translocation into the shoot, hence leading to the reduced gene expression of M-20 shoots under salt-stress conditions.
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  • 2
    ISSN: 1432-2242
    Keywords: Key words Genetic map ; Physical map ; Map-based gene cloning ; Disease resistance ; Rice ; DNA markers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The recessive gene, xa13, confers resistance to Philippine race 6 (PXO99) of the bacterial blight pathogen Xanthomonas oryzae pv oryzae. Fine genetic mapping and physical mapping were conducted as initial steps in an effort to isolate the gene. Using nine selected DNA markers and two F2 populations of 132 and 230 plants, xa13 was fine-mapped to a genomic region 〈4 cM on the long arm of rice chromosome 8, flanked by two RFLP markers, RG136 and R2027. Four DNA markers, RG136, R2027, S14003, and G1149, in the target region were used to identify bacterial artificial chromosome (BAC) clones potentially harboring the xa13 locus from a rice BAC library. A total of 11 BACs were identified, forming four separate contigs including a single-clone contig, 29I3, associated with the RG136 STS marker, the S14003 contig consisting of four clones (44F8, 41O2, 12A16, and 12F20), the G1149 contig with two clones, 23D11 and 21H18, and the R2027 contig consisting of four overlapping clones, 42C23, 30B5, 6B7 and 21H14. Genetic mapping indicated that the xa13 locus was contained in the R2027 contig. Chromosomal walking on the R2027 contig resulted in two more clones, 33C7 and 14L3. DNA fingerprinting showed that the six clones of the R2027 contig were overlapping. Clone 44F8 hybridized with a single fragment from the clone 14L3, integrating the R2027 and S14003 contigs into a single contig consisting of ten BAC clones with a total size of approximately 330 kb. The physical presence of the xa13 locus in the contig was determined by mapping the ends of the BAC inserts generated by TAIL-PCR. In an F2 population of 230 plants, the BAC-end markers 42C23R and 6B7F flanked the xa13 locus. The probes 21H14F and 21H14R derived from BAC clone 21H14 were found to flank xa13 at a distance of 0.5 cM on either side, using a second F2 population of 132 plants. Thus, genetic mapping indicated that the contig and the 96-kb clone, 21H14, contained the xa13 locus.
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