Gene expression analysis related to regulation of life cycle using Porphyra genomic information

点击翻译按钮获取中文摘要

New findings obtained by this study are as follows.1. Screening of highly expressed genes during asexual sporulation through monosporesWe found eight genes encoding enzymes related to metabolism and unknown genes with gametophytic generation-phase-specific expression pattern by cDNA macroarray and RT-PCR or Northern analysis. The eight nucleotide sequences will appear in the GenBank/EMBL/DDBJ databases with accession numbers AB127044 through AB127051.2. Development of a new technique for gene expression analysis and internal standard in Porphyra yezoensis(1) We developed a method for whole-mount in situ hybridization (WISH) by using elongation factor-1α (EF-1α) riboprobes and gametophytic germlings.(2) We developed an internal standard (actin-related protein 4 homologue) in gene expression analyses of differentially expressed genes. A putative bipartite nuclear localization signal (NLS) and an actin motif were found within the PyARP4 amino acid sequences. The expression level of PyARP4 did not change significantly among four developmental stages of the life cycle and was lower than that of a conventional actin.3. Induction of monospore production by chemical treatmentWe found that monospore-like cells with germinability can be obtained when vegetative gametophytic thalli of P.yezoensis were grown in enriched seawater medium that contained 10 mM allantoin. We also found that this chemical treatment is effective to even gametophytic thalli of P.pseudolinearis, which does not have asexual sporulation through monospores in the life cycle.