Complete set of extrachromosomal DNAs from three pathogenic lines of onion yellows phytoplasma and use of PCR to differentiate each line

 Two lines of onion yellows phytoplasma with reduced pathogenicity have been isolated from the original wild-type line (OY-W). One is a line with mild symptoms (OY-M) and the other is a non-insect-transmissible line, also with mild symptoms (OY-NIM). We previously reported heterogeneity in extrachromosomal DNA (EC-DNA) species in these lines. In this report, another EC-DNA, EcOYNIM, from OY-NIM was cloned and sequenced, providing a complete set of EC-DNAs from the three OY lines. To monitor each phytoplasma in synergism or cross-protection experiments, a pair of polymerase chain reaction (PCR) primers that universally amplify a portion of the EC-DNAs that are characteristic of each line was designed. Using this primer set, a line-specific fragment was amplified from the total DNA of each plant inoculated with one or more phytoplasma lines. The PCR product sizes differ for each phytoplasma line, so the lines can be distinguished even in plants infected with multiple lines. Because EC-DNAs are more abundant than chromosomal genes in phytoplasma cells, this primer set will be valuable for detecting and discriminating these phytoplasma lines and for analyzing their interaction. Received: October 21, 2002 / Accepted: January 8, 2003 RID="*" ID="*" The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession number AB097150 Acknowledgments This work was supported partly by Grants-in-Aid of Scientific Research from the Japan Society for the Promotion of Science (JSPS) (09460155 and 13306004), a Grant-in-Aid of Scientific Research on Priority Areas (C) “Genome Biology” from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and the Program for the Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN) of the Bio-oriented Technology Research Advancement Institution.     

The development of a sensitive enzyme immunoassay for the determination of estrone and estradiol-17beta in bovine blood plasma based on the same homologous combination with antiserum and steroid-enzyme conjugate

The development of a sensitive enzymeimmunoassay (EIA) for the determination of estrone (E1) and estradiol-17beta (E2 beta) in bovine plasma is described. The assay is a homologous double-antibody EIA with E2beta 17hemisuccinate (HS) as hapten for the immunoreactive reagent. The antiserum was raised against E2beta 17HS bovine serum albumin conjugate in the rabbit, and E2beta 17HS-horseradish peroxidase was used as steroid-enzyme conjugate. Each estrogen EIA was distinguished only by using the each working standard and sample for the EIA. Bovine plasma E1 and E2beta were extracted and purified before EIA. The antiserum was used at 1:1,750,000 dilutions for EIA. Estrone and E2beta showed high cross-reactivity with the antiserum (E1: 350.7%, E2beta:100%). The sensitivities were <0.03 pg/well for E1 and <0.12 pg/well for E2beta. Recovery rates of E1 and E2beta added to bovine blood plasma were 94.5% and 93.9%, respectively. The precision for EIA of estrogens was below 9.7%. The profiles of either estrogen as determined by EIA corresponded closely well with follicle dynamics in the cow during the estrous cycles and with placental function in pregnant animals. In conclusion, our new EIA can be applied with sufficient sensitivities, recovery and precision for the routine analysis of E1 and E2beta concentrations in bovine plasma.     

First report of ‘Candidatus Phytoplasma asteris’ infecting hydrangea showing phyllody in Japan

Phytoplasma-induced floral malformations such as virescence, phyllody, and proliferation were observed on hydrangeas in Gunma Prefecture, Japan. Phylogenetic analyses based on 16S rRNA, secY, groEL, and amp gene sequences indicated that the affected hydrangea plants were associated with phytoplasmas belonging to ‘Candidatus Phytoplasma asteris’, but not to ‘Ca. P. japonicum’, which occurs in hydrangeas showing phyllody in Japan. This is the first molecular evidence of an association of ‘Ca. P. asteris’ with hydrangea plants in Japan.     

Seasonal changes in otolith and somatic growth in age-0 Pacific saury <Emphasis Type="Italic">Cololabis saira</Emphasis>

We evaluated the seasonal changes in otolith and somatic growth of age-0 Pacific saury Cololabis saira in 223 fish collected between June and November 2002. We calculated the age in days of each individual by measuring otolith growth increments under a scanning electron microscope. The age was correlated with body length and otolith radius. We also observed seasonal changes in the rate of increase in body length and otolith radius and in the pattern of otolith growth. Until August, both body length and otolith radius increased with age. Thereafter, the otolith radius continued to increase, whereas the rate of somatic growth decreased. As a result, the ratio of otolith radius to body length increased. After August, the percentage of otoliths with unreadable increments on their edge increased due to the formation of hyaline zones. Otoliths grew both radially and in thickness until July, but gradually stopped growing in thickness after August. Beginning in October, more than 80% of otoliths only grew radially. After August, the otolith not only continued growing but the morphological growth pattern also changed.     

A novel waxy allele in sorghum landraces in East Asia

A loss of granule‐bound starch synthase I (GBSS I) activity results in starch granules that contain mostly amylopectin and little or no amylose, a phenotype described as waxy. Previously, two phenotypic classes of waxy alleles, wx^a, associated with no detectable GBSS I, and wx^b, associated with apparently inactive GBSS I in the endosperm, were reported in sorghum (Sorghum bicolor (L.) Moench). In this study, the waxy alleles in a sorghum core collection were investigated using DNA markers. Of the 337 sorghum accessions examined, 17 accessions that were confirmed to be waxy by a negative iodine staining result and 16 were found to be wx^a. A novel waxy allele, wx^c, was found in a Taiwanese landrace. This allele consists of a +1G to C mutation in the 5′ splice site at the intron 10–exon 11 boundary, a mutation that most likely resulted in the suppression of GBSS I gene expression. A DNA marker specific for wx^c was produced to distinguish the wx^c allele from other alleles, allowing the identification of heterozygous non‐waxy plants.