Characterization of a cDNA encoding a homolog of actin-related protein 4 from a marine red alga Porphyra yezoensis

ABSTRACT:   A cDNA ( PyARP4 ) containing an open reading frame for a protein of 573 amino acids was identified in the marine red alga Porphyra yezoensis . The conceptual PyARP4 protein exhibits significant similarity to actin-related protein (ARP) 4 in the terrestrial plant Arabidopsis . Comparison of the deduced amino acid sequence showed moderate sequence identity (30%) to a conventional actin in P. yezoensis , as seen in comparisons between ARP and conventional actins of other organisms. A putative bipartite nuclear localization signal and an actin motif were found within the PyARP4 amino acid sequence. In a phylogenetic analysis, the PyARP4 was found to cluster with the ARP4 of other organisms. The expression level of PyARP4 did not change significantly among four developmental stages of life cycle and was lower than that of a conventional actin. This cDNA therefore may serve as a useful internal standard in gene expression analyses of differentially expressed genes in P. yezoensis .     

Application of a PCR-Based Cytoplasm Genotyping Method for Phylogenetic Analysis in Potato

We previously developed five polymerase chain reaction-based markers (T, S, SAC, D, and A) to distinguish potato cytoplasms into six types (M, P, A, W, T, and D). As the applicability of this genotyping method for phylogenetic studies had been questioned, we applied this method to species distantly related to cultivars (four accessions of two tomato species, and 176 accessions of 29 Solanum species). The T marker uncovered two insertions, which, along with a unique S marker band, supported independency of series Piurana. The A and D markers generated unique bands to A- and D-type cytoplasms, respectively, but the SAC marker generated a similar banding pattern to the cytoplasms of both cultivated and their distantly related species. Consequently, while the cytoplasm type definition is validated only among cultivated potatoes and their closely related wild species, the developed markers, with the exception of the SAC marker, could provide useful phylogenetic information.     

Detection of an isoform of alpha(1)-antitrypsin in serum samples from foals with gastric ulcers

The objective of this study was to find serum indicators of gastric ulcers in foals. By using two-dimensional electrophoresis of serum proteins, three distinct spots were detected in samples from foals with gastric ulcers detected endoscopically. One of them appeared with high frequency and was identified by partial digestion with trypsin and subsequent nano-electrospray ionisation-tandem mass spectrometry (nanoesi-ms/ms) analysis as an alpha(1)-antitrypsin. Western blot analysis, using an antibody against human alpha(1)-antitrypsin, revealed at least two bands, of molecular weight 58 kDa and 55 kDa, in the sera. The 55 kDa band was detected in 44 of 47 serum samples from foals with gastric ulcers, but in only three of 22 serum samples from healthy foals.     

Species relationships in the subgenus Ceratotropis (genus Vigna) as revealed by RAPD analysis

Summary The genetic variation among 23 accessions of 5 species in the subgenus Ceratotropis, genus Vigna, were investigated by random amplified polymorphic DNA (RAPD) analysis. A total of 404 fragments amplified with 24 primers were scored and analyzed by cluster analysis. The accessions used were separated into two main groups with an average of 70% differences. Within the main groups, five subgroups were recognized, which are in complete agreement with taxonomic species. Wild forms were always grouped with their most closely related cultivated forms and they showed variation in each species. The largest intraspecific variation was found in V. radiata (mungbean), in which wild forms (V. radiata var. sublobata) were highly different from each other and from cultivated forms. V. angularis (adzuki bean) showed the least variation and thus, was probably differentiated in relatively recent times.     

Cryptic improvement for fertility by continuous selfing of diploid potatoes using Sli gene

While a diploid potato was continuously selfed by a function of Sli gene, the level of heterozygosity reduced, and the fertility also reduced (inbreeding depression), which might be improved because continuous selfing could eliminate undesirable recessive alleles. To explore what was occurring as advancing self-generations in terms of fertility, 51 plants of an S₃ diploid potato family were evaluated for fertility-related traits and analyzed by RFLP markers. The level of heterozygosity was positively correlated with any of fertility-related traits (r = 0.108−0.333). At least six loci on six chromosome sections were associated with fertility-related quantitative trait loci (QTLs), of which three performed better in the heterozygotes and three in the homozygotes. By further continued selfing, QTLs performing better in heterozygotes were likely fixed to homozygotes with secondly better performance, and those performing better in homozygotes were fixed to the best genotypes. Thus, the selfed progenies were cryptically being improved for fertility by genetic fixation to the best or better genotypes in fertility-related QTLs.     

Mapping genes for deep-seeding tolerance in barley

Deep-seeding tolerance, the emergence of seedlings from deep seeded conditions, is involved in stand establishment in semi-arid regions, where the soil surface is too dry for seed germination. Genes determining deep-seeding tolerance in barley were mapped using two doubled haploid populations derived from the following crosses: Harrington × TR306 (H/T)and Step toe × Morex (S/M). Significant quantitative trait loci (QTLs) for deep-seeding tolerance were found in each population. Two QTL sex plained 40% of the phenotypic variation in the H/T population and one QTL (S/M) 8% of the total phenotypic variance. Multiple QTLs accounting for coleoptile length and first internode length were detected in both populations. In the H/T population, there were coincident QTLs for deep-seeding tolerance, coleoptile length and first internode length on the long arm of chromosome 5H. These QTLs correspond with previously reported QTLs for abscisic acid and gibberellic acid response. QTL coincidence may be due to the pleiotropic effects of alleles at a single locus. This information may be useful for breeding programs manipulating morphological and physiological traits in order to develop varieties for semi-arid regions. This revised version was published online in August 2006 with corrections to the Cover Date.     

麦类引种优选与抗逆性状初步分析

1991年从日本引进国外麦类品种10252个，在我国黄淮海平原盐渍地上种植。经1992--1994年系统观察、分析和鉴定，初步选择出适合该类型区种植的高产、优质品种及大、小麦亲本材料。     

Development of a multiplex PCR method for simultaneous detection of diagnostic DNA markers of five disease and pest resistance genes in potato

Multiplex PCR is practically a reasonable choice for molecular marker-assisted selection in potato breeding. We had developed and were using a multiplex PCR method for selection of resistance genes to cyst nematode (H1), Potato virus X (Rx1) and late blight (R1 and R2). Since then, more reliable and tightly linked markers for H1 and R2, and a new marker for resistance to Potato virus Y (Ry _chc ) were developed. In this article, all these superior markers, including a positive marker to eliminate PCR-failed samples, were incorporated into one multiplex PCR assay. Using the newly developed multiplex PCR technique, five plants potentially harboring all five resistance genes were selected from 96 hybrid plants approximately 5 h after DNA extraction, which is a third of the operation time compared with separate PCR reactions for each marker.     

Reciprocal differences in DNA sequence and methylation status of the pollen DNA between F<Subscript>1</Subscript> hybrids of <Emphasis Type="Italic">Solanum tuberosum</Emphasis> × <Emphasis Type="Italic">S. demissum</Emphasis>

Reciprocal differences, mostly caused by cytoplasmic effects, are frequently observed in interspecific hybrids. Previously, we found that crosses onto Solanum demissum were much successful with the pollen of interspecific hybrids between S. tuberosum as female and S. demissum as male (TD hybrids) than the reciprocal ones (DT hybrids). To elucidate this reciprocally different crossability, we analyzed the pollen DNA of TD and DT using methylation-sensitive amplified polymorphism (MSAP) analysis. Using 126 primer combinations, MSAP analysis revealed 57 different bands between bulked pollen DNA samples of TD and DT. Individual examination of 16 TD and 9 DT plants disclosed eight bands uniformly different between TD and DT. Their sequencing results revealed two pairs of bands to be identical to each other, resulting in six distinct sequences. As expected, one band shared high homology with chloroplast DNA, and another one with mitochondrial DNA. However, one band that was apparently different at DNA sequence level and maternally transmitted from S. demissum, showed no homology with any known sequence. The remaining three bands were of DNA methylation level differences with no or uncertain homology to known sequences. To our knowledge, this is the first report detecting reciprocal differences in DNA sequence or DNA methylation other than those in cytoplasmic DNA.