Morphological identity of a taxonomically unassigned cytochrome c oxidase subunit I sequence from stomach contents of juvenile chum salmon determined using polymerase chain reaction

In a previous report, we analyzed the stomach contents of juvenile chum salmon Oncorhynchus keta by morphological observation and also by molecular identification using the mitochondrial cytochrome c oxidase subunit I (COI) region. However, one of the most frequently detected COI sequences could not be assigned to any specific taxon, even at the phylum level. In the present study, we conducted in situ hybridization (ISH) on the stomach contents of juvenile chum salmon using the COI sequence and polymerase chain reaction amplification of a 18S ribosomal RNA gene from the tissue sections where ISH signals were detected. As a result, the organism that was enigmatic at the phylum level was found to be an appendicularian. Moreover, Oikopleura longicauda collected from the bay where the juvenile chum salmon samples were obtained was shown to have the same COI sequences as this taxonomic “orphan” COI sequence from the stomach contents. The present results suggest that the COI sequences previously deposited in public databases for “Oikopleura” are actually derived from taxonomic groups other than appendicularians, and that this may have hampered our understanding of prey richness in the stomach or gut of certain marine animals based on DNA barcoding.     

Genetic and morphological evidence of hybridization between <Emphasis Type="Italic">Nematalosa japonica</Emphasis> and <Emphasis Type="Italic">N. come</Emphasis> (Clupeiformes: Clupeidae) off Okinawa Island,Ryukyu Archipelago,Japan

Two morphologically similar species of gizzard shad, Nematalosa japonica Regan and N. come (Richardson), sympatrically distributed off Okinawa Island, Japan, were examined using an allozyme locus (SOD*) and two nuclear polymerase chain reaction (PCR)-based DNA markers (ITS-1 and CaM), which provided diagnostic identification of each species. In addition, a multiplex PCR-based mitochondrial DNA (mtDNA) marker (16S) was used to characterize the distribution of mtDNA haplotypes among specimens. The species composition of sympatric and allopatric population samples from Tungkang, southern Taiwan, to Okinawa and the Shikoku Islands, Japan, were also examined. Gizzard shad with hybrid genotypes were detected in three populations from Okinawa Island, with hybrid frequency ranging from 1 to 67%. A backcross level of 2% was detected in the dominant hybrid frequencies of one population sample only. Morphological examination of hybrids showed intermediate forms, with hybrid indices of three meristic characters falling between those of the parental species (range 39–53; mean 45). Although principal component analysis showed differences between N. japonica and N. come based on the first principal component scores, hybrids were difficult to identify. Accordingly, satisfactory identification of species and hybrids could be achieved only using genetic tools. We also discuss the cause of hybridization and its relationship with recently conducted reclamation on Okinawa Island.     

Age,growth and maturation of the redbelly tilapia <Emphasis Type="Italic">Tilapia zillii</Emphasis> introduced into the Haebaru Reservoir on Okinawa-jima Island

ABSTRACT:   The age, growth and maturation of the redbelly tilapia Tilapia zillii introduced into the Haebaru Reservoir on Okinawa-jima Island were studied using 2197 specimens ranging from 7.3 to 168.0 mm standard length (SL). The spawning season was estimated to be from April to August, with a peak in April and May. The first maturation sizes of females and males were estimated to be 38.1 and 33.0 mm SL, respectively. The opaque zones in otoliths that form annually were found to correlate with the spawning season. Maximum ages of females and males were 7 and 6 years, respectively. The von Bertalanffy growth formulae were expressed as: L _t  = 99{1 − exp[−0.67( t  + 0.09)]} for females and L _t  = 155{1 − exp [−0.36( t  + 0.12)]} for males. Males grew to be larger than females from the first year onward. Populations of this species are characterized by early maturation life history parameters and are thought to adapt and become established quickly after being released into new water systems. Furthermore, extermination activity in winter is thought to be an effective strategy before the newly recruited fish begin breeding in the warmer months.     

Molecular analysis of genetic diversity in melon landraces (<Emphasis Type="Italic">Cucumis</Emphasis><Emphasis Type="Italic">melo</Emphasis> L.) from Myanmar and their relationship with melon germplasm from East and South Asia

Genetic diversity of Myanmar melon was evaluated by analysis of 27 RAPD markers and morphological characters using 41 accessions of melon landraces of which 36 accessions were small-seed type. The gene diversity was 0.239, higher than for group Conomon from East Asia and equivalent to Indian melon populations. Melon accessions were classified into six major clusters. The largest cluster IV comprised mainly group Conomon which was closely related to cluster V consisting of mainly group Agrestis. Most of the accessions of group Cantalupensis were grouped into clusters II or VII and were distantly related to groups Conomon and Agrestis. The genetic relationship to melon accessions from neighboring countries was analyzed. The 24 accessions of clusters IV and V were mostly clustered together with small-seed type melon of India, but the 14 accessions of clusters VI and VII were mostly clustered together with large-seed type melon of India. These results indicated that the genetic diversity of Indian melon is conserved in Myanmar. Genetic introgression among melon groups through spontaneous hybridization was also indicated and was considered important to maintain or increase the genetic diversity in Myanmar.     

Microstructure of a Brassica rapa genome segment homoeologous to the resistance gene cluster on Arabidopsis chromosome 4

Genome evolution is a continuous process and genomic rearrangement occurs both within and between species. With the sequencing of the Arabidopsis thaliana genome, comparative genetics and genomics offer new insights into plant biology. The genus Brassica offers excellent opportunities with which to compare genomic synteny so as to reveal genome evolution. During a previous genetic analysis of clubroot resistance in Brassica rapa, we identified a genetic region that is highly collinear with Arabidopsis chromosome 4. This region corresponds to a disease resistance gene cluster in the A. thaliana genome. Relying on synteny with Arabidopsis, we fine-mapped the region and found that the location and order of the markers showed good correspondence with those in Arabidopsis. Microsynteny on a physical map indicated an almost parallel correspondence, with a few rearrangements such as inversions and insertions. The results show that this genomic region of Brassica is conserved extensively with that of Arabidopsis and has potential as a disease resistance gene cluster, although the genera diverged 20 million years ago.     

Quality Control and Its Constraints during the Preparatory-Phase Activities of the Acid Deposition Monitoring Network in East Asia (EANET)

Participating countries of the Acid Deposition Monitoring Network in East Asia (EANET) launched the preparatory-phase activities from April 1998. For the recognition and improvement of the analytical precision and accuracy, the Interim Network Center (INC) carried out the inter-laboratory comparison on the analysis of artificial rainwater samples and soil samples. Relevant laboratories submitted their analytical data to the INC for the evaluation. Submitted data were summarized and evaluated in terms of precision and accuracy, and were compared with the Data Quality Objectives (DQOs) of EANET. These inter-laboratory comparisons made clear the present conditions of laboratories as well as the major constraints that should be solved in the future.     

Effect of intracerebroventricular infusion of urocortin on feed and salt intake in parotid fistulated sheep

The effect of a new peptide, brain urocortin (UCN), in the central regulation of feed and salt intake in parotid fistulated sheep was investigated through a continuous intracerebroventricular (ICV) infusion of the peptide at a small dose for 48 h. Feed and salt intake on day 1 of UCN infusion were similar to those before infusion. However, feed and salt intake on day 2 during UCN infusion decreased significantly. Water intake on days 1 and 2 during UCN infusion was the same as that before infusion. Saliva secretion from the unilateral parotid gland during UCN infusion was unchanged from that before infusion. On the other hand, urine excretion during UCN infusion increased significantly. The present results suggested that UCN acts directly on the brain mechanisms of sheep to decrease feed and salt intake, but does not alter water intake.     

Molecular characterization of South and East Asian melon, Cucumis melo L., and the origin of Group Conomon var. makuwa and var. conomon revealed by RAPD analysis

The genetic diversity and relationship among South and East Asian melon Cucumis melo L. were studied by using RAPD analysis of 69 accessions of melon from India, Myanmar, China, Korea, and Japan. The genetic diversity was large in India, and quite small in Group Conomon var. makuwa and var. conomon from East Asia, clearly indicating a decrease in genetic variation from India toward the east. Cluster analysis based on genetic distance classified 17 groups of accessions into two major clusters: cluster I comprising 12 groups of accessions from India and Myanmar and cluster II that included five groups of accessions of Group Conomon var. makuwa and var. conomon from East Asia. Cluster I was further divided into three subclusters, of which subclusters Ib and Ic included small- and large-seed type populations, respectively. Therefore, this division was based on their seed size, not cultivation area. The large-seed type from east India was differently included in the subcluster of small-seed type (Ib). A total of 122 plants of 69 accessions were classified into three major clusters and subclusters: clusters I and II comprised melon accessions mostly from India and Myanmar, and cluster III comprised Group Conomon var. makuwa and var. conomon from East Asia. The frequency of large- and small-seed types was different between clusters I and II, also indicating genetic differentiation between large- and small-seed types. One plant of the small-seed type from east India was differently included in cluster III, and two plants from east India were classified into subcluster IV. These results clearly showed that South Asian melon is genetically differentiated by their seed size, and that small-seed type melon in east India is closely related to Group Conomon var. makuwa and var. conomon.     

Establishment and characterization of canine rhabdomyosarcoma cell line CMS-C

A novel canine tumor cell line designated as the CMS-C cell line was established from pleomorphic rhabdomyosarcoma (RMS) raised in the prostate gland of a 14-year-old intact male mixed-breed dog. CMS-C cells displayed the same immunohistochemical characteristics (positive for vimentin and desmin and negative for cytokeratin and smooth muscle actin) as the original tumor cells and express myoD1 and UCP3, known as striated muscle-specific molecules, as shown by RT-PCR assay. Therefore, the established CMS-C cell line appears to be of rhabdomyoblast cell origin. The CMS-C cell line established from pleomorphic RMS will be a useful tool for further studies about canine RMS.