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BRCA2 is a breast tumour susceptibility factor with functions in maintaining genome stability through ensuring efficient double-strand DNA break (DSB) repair via homologous recombination. Although best known in vertebrates, fungi, and higher plants also possess BRCA2-like genes. To investigate the role of Arabidopsis BRCA2 genes in DNA repair in somatic cells, transposon insertion mutants of the AtBRCA2a and AtBRCA2b genes were identified and characterized. atbrca2a-1 and atbrca2b-1 mutant plants showed hypersensitivity to genotoxic stresses compared to wild-type plants. An atbrca2a-1/atbrca2b-1 double mutant showed an additive increase in sensitivity to genotoxic stresses compared to each single mutant. In addition, it was found that atbrca2 mutant plants displayed fasciation and abnormal phyllotaxy phenotypes with low incidence, and that the ratio of plants exhibiting these phenotypes is increased by gamma-irradiation. Interestingly, these phenotypes were also induced by gamma-irradiation in wild-type plants. Moreover, it was found that shoot apical meristems of the atbrca2a-1/atbrca2b-1 double mutant show altered cell cycle progression. These data suggest that inefficient DSB repair in the atbrca2a-1/atbrca2b-1 mutant leads to disorganization of the programmed cell cycle of apical meristems.

Guineagrass (Panicum maximum Jacq.) is one of the most important tropical forage grasses due to its high productivity and good forage qualities. Although P. maximum is important forage, basic genomic information such as genome size and ribosomal DNA (rDNA) ideogram information have not been reported. In this study, the DNA contents of one diploid strain and three tetraploid cultivars of P. maximum were examined by flow cytometry and rDNA loci were physical mapped by fluorescence in situ hybridization (FISH). The DNA content of the diploid strain was determined to be approximately 500 Mbp, which is of similar size to Oryza sativa and represents a relatively small genome size when compared to other members of the plant kingdom. The tetraploid cultivars were determined to exhibit a genome size of approximately 1000 Mbp. FISH with rDNA probes was applied to diploid and tetraploid strains. The numbers of 5S rDNA loci in the diploid and the tetraploids were two and four, respectively. The number of 45S rDNA loci varied from four to eight among the tetraploids. Constructed quantitative ideograms of diploid strain by image analysis indicated that the difference in the number of 45S rDNA is dependent on the presence or absence of the locus on chromosome 2. The determined DNA content and constructed ideograms in this study are useful for future research efforts on P. maximum in areas focused on cultivar development, gene isolation of apomixis loci and mutation research.

A genomic DNA containing a rice (Oryza sativa L., cv. Norin-8) gene coding for a pullulanase-type starch debranching enzyme (EC 3.2.1. 41) was sequenced (EMBL/GenBank/DDBJ accession number AB012915). Along the 15, 248 bp DNA, the pullulanase gene is split into 26 exons. The four pullulanase consensus regions are positioned in the middle portion of the sequence and are separated by long introns and 1-3 exons. Comparison of the rice cv. Norin-8 pullulanase genomic structure with that of barley pullulanase (limit dextrinase) (F. Lok et al., EMBL/GenBank/DDBJ accession number AF022725) indicates that most of the pullulanase exons are highly conserved. Alignment of the nucleotide bases of rice exon 8 with those of barley exon 8-intron 8-exon 9 fragment suggests that the 85 bp internal sequence of rice exon 8 was originally an intron, a possibility further indicated by the absence in barley and spinach (A. Renz et al., EMBL/GenBank/DDBJ accession number X83969) pullulanases of amino acid residues encoded by the 85 bp fragment.

The effects of amino acid medium (AA medium) on the dissociation of rice callus tissues were examined in suspension-cultured cells, because a finely dispersed cell suspension had been obtained previously from rice callus tissues in this medium. The level of extracellular polysaccharides formed in cultured AA medium was much higher than that of those formed in cultured B5 medium. The polysaccharides were mainly composed of higher levels of arabinose, xylose, and galactose, suggesting the solubilization of arabinoxylan and (arabino)galactan. Nevertheless, wall polysaccharides in cells cultured in AA medium contained the same levels of arabinosyl-, xylosel-, and galactosyl-linkages as those in B5 medium. Based on amino acid analysis, rice cells in AA medium incorporated the amino acids in 3 days and formed ornithine and urea during the early stages of cultivation, and secreted urea into the culture medium. Transfer of rice tissue cultured in AA medium to B5 medium composed of inorganic nitrogen source caused...

Two isoforms of starch branching enzyme (Q-enzyme), QEI and QEII, have been purified to honlogeneity from developing rice endosperm. QEI and QEII, with molecular weights of about 80 and 85 kDa, respectively, could be fully separated by anion-exchange or hydrophobic chromatography. The peptide maps obtained after V8 proteinase digestion were quite different between the two enzymes. Antibodies prepared against QEI showed no immunological cross-reaction with the QEII protein in Western blot experiments, and anti-QEII serum did not react with the QEI protein. The data indicate that QEI and QEII are distinct proteins encoded by different genes in rice plants.