Muscle fiber types,growth and development in the whole myotome of cultured Pacific bluefin tuna Thunnus orientalis

Following the successful development of Pacific bluefin tuna (PBT) aquaculture, it is of considerable importance to determine the muscle fiber types and their growth patterns for future development. Muscle fiber profiles of dorsal ordinary, lateral ordinary (LO) and dark muscles and their growth patterns in PBT from 3.0 to 54.3 kg body weight were studied. Muscle fibers were histochemically stained for NADH-diaphorase and myosin adenosine triphosphatase activity (mATPase), and immunohistochemically stained with S-58 slow-muscle myosin antibody. All muscle fibers in dorsal and LO muscles showed low NADH-diaphorase activity, and acid-labile (pH 4.0 or 4.3) and alkali-stable mATPase activity. In LO muscle adjacent to dark muscle, three intensities of mATPase activity were observed after acid pre-incubation at pH 4.5 or 5.0, and the activity was related to the muscle fiber diameter. In dark muscle, all small and some large fibers stained intensely for NADH-diaphorase activity, related to their high aerobic metabolism. The high-active fibers with NADH-diaphorase in dark muscle were positive for S-58 antibody. Some large fibers in dark muscle showed intermediate NADH-diaphorase activity and high mATPase activity after alkali pre-incubations. These are fast-twitch oxido-glycolytic fibers in dark muscle and transformed to red muscle fibers with increasing body weight.     

Evaluation of rolling shear strength of plywood by flexural vibration method

The rolling shear strength of plywood was evaluated using a flexural vibration test. Test specimens were lauan and Douglas fir three-ply plywoods made from thick veneers. The dynamic shear and Young's moduli were determined using the flexural vibration method, which involved in-plane and out-of-plane flexural vibration. The rolling shear strength was determined using the static destructive method, which is dependent on the direction of the lathe check in the core veneer. Before and after accelerated aging treatments were conducted, there were relations between out-of-plane dynamic properties (out-of-plane shear and Young's moduli) and its rolling shear strength. It was concluded that the rolling shear strength is related not only to the shear property of the core but the flexural stiffness of two faces when the deformation of out-of-plane plywood was not restrained.Part of this work was presented at the 47th Annual Meeting of The Japan Wood Research Society, Kochi, April 1997     

Influence of intergeneric/interspecies mitochondrial injection; parthenogenetic development of bovine oocytes after injection of mitochondria derived from somatic cells

Interspecies/intergeneric mitochondrial heteroplasmy can occur in interspecies/intergeneric hybrid embryos or following nuclear transfer. In the present study, intergeneric buffalo (Bubalus bubalis) mitochondria (WB-mt) or interspecies murine (Mus spretus) mitochondria (M-mt) were injected into bovine (Bos taurus) oocytes, and the subsequent embryonic development was characterized. Fibroblast mitochondria (WB-mt or M-mt) were microinjected into in vitro matured bovine oocytes followed by oocyte activation by a combination of electrical stimulation and 6-dimethylaminopurine treatment. After seven days of culture, embryo development was evaluated. The copy number of specific mtDNA populations (introduced and native mtDNA) from heteroplasmic oocytes was estimated using real-time PCR. The results illustrated that oocytes injected with either WB-mt or M-mt can develop to the blastocyst stage (20.6% and 19.6%). Cleavage division rates and development to the morula stage in oocytes injected with WB-mt were lower (76.2% and 45.9%, respectively) in comparison with uninjected oocytes (89.2% and 59.1%, respectively) (P<0.05). However, no differences were found in comparing M-mt injected oocytes and controls (P>0.05). An increase in bovine mtDNA copy number was observed at the expanded blastocyst stage of injected embryos (P<0.01), while the number of injected mtDNA was stable throughout development. This study demonstrates that interspecies/intergeneric mitochondrial injected bovine oocytes have the ability to develop to the blastocyst stage after parthenogenetic activation and that injected mtDNA was neither selectively destroyed nor enhanced through development. Moreover, injected intergeneric mitochondria had a demonstrated influence on bovine parthenogenetic development and mtDNA replication.     

Primers for the Amplification of the Circular Chloroplast DNA from the A-genome Group of Cultivated Cotton

The availability of the plastid genome sequences is one of the bases for comparative,functional,and structural genomic studies of plastid-containing living organisms,in addition to the application of plastid genetic engineering technology.The past efforts to sequence plastid genomes involve complicated preparation protocols.One procedure starts with the isolation of plastids,which was tiresome and time wasting that followed by a second step to extract plastid DNA from the isolated plastids,then finally the build up of plasmid or bacterial artificial chromosome (BAC) library.     

Evaluation of Loads and Sources of Heavy Metals in Tama River, Tokyo

The loads and sources of heavy metals (Cd, Co, Cu, Mn, Mo, Ni, Pb, V, and Zn) in the midstream of Tama River were evaluated on the basis of their chemical analyses in ordinary and stormwater runoff from July 2007 to November 2008. Tama River is one of the three major rivers flowing into Tokyo Bay. The total annual water discharges differed largely for 2007 and 2008, depending on the scales of typhoon rainfalls and other heavy rainfalls in each year. The concentrations of the metals other than Mo in the river did not change markedly at a flow rate of less than approximately 200 m³ s^?1, but at higher flow rates, the concentrations of all the metals increased linearly with the increase in the log of flow rate (r ²?=?0.94–0.99). The annual loads of heavy metals for 2007 and 2008 were estimated using regression equations between the above parameters and the hourly flow rate data for each year. For the metals other than Mo, the contribution of the loads at higher flow rates (>200 m³ s^?1) was much larger than that at lower flow rates (<200 m³ s^?1), showing the importance of the particulate loads (primarily crustal materials) during storm runoff following typhoon rainfalls and other heavy rainfalls. In contrast, the loads of Mo at lower flow rates accounted for major portions (56–78%) of the total loads, because of a relatively small contribution of particulate load during storm runoff. The contribution of the loads of Mo, Zn, Cd, Cu, and Ni at lower flow rates to the annual loads was larger than that of other metals. It was found that the concentrations of these metals in ordinary runoff are strongly affected by the discharge of treated water from sewage treatment plants which are located along the catchment. Thus, treated water from sewage treatment plants may be the primary source contributing to the present pollution of Mo, Zn, Cd, Cu, and Ni in Tokyo Bay.     

The genes encoding glycoside hydrolase family 6 and 7 cellulases from the brown-rot fungus <Emphasis Type="Italic">Coniophora puteana</Emphasis>

Four genes encoding glycoside hydrolase (GH) family 6 and 7 cellulases (cel6A, cel6B, cel7A, and cel7B) were obtained from the brown-rot fungus Coniophora puteana by genomic polymerase chain reaction (PCR) using consensus degenerate hybrid oligonucleotide primers (CODEHOPs) designated from the amino acid sequence of cellobiohydrolases (CBHs) from white-rot fungi. The nucleotide sequences of four genes showed high homology with basidiomycetes CBHs, suggesting the fi rst cloning of the genes encoding Cel6 and Cel7 from brown-rot fungi. PCR using CODEHOP pairs at the catalytic domain successfully amplifi ed both cel6A and cel6B, whereas only cel6A fragment was obtained using the primers including the carbohydrate-binding modules (CBMs), suggesting lack of CBM in Cel6B. Moreover, both cel7A and cel7B were amplified by the PCR using CODEHOP pairs at the catalytic domain, but not by those including CBM, suggesting the absence of Cel7 with CBM in the fungus. From these results, three of four cellulases from C. puteana may not carry CBM, which has an important role for the degradation of crystalline cellulose.     

A novel Tos17 insertion upstream of Hd1 alters flowering time in rice

Heading date 1 (Hd1) is one of the major determinants of flowering time and regional adaptability in rice (Oryza sativa L.), because there is a significant correlation between Hd1 allelic diversity and phenotypic differences in rice cultivars. In this study, we isolated a novel rice mutant with a Tos17 retrotransposon insertion upstream of Hd1. The Tos17 insertion mutant of Hd1 [designated as Tos17(Hd1)] had decreased photoperiod sensitivity and mRNA expression of Hd1, as compared with wild‐type Nipponbare. Between Tos17(Hd1) and Nipponbare, expression levels of OsPRR1 and OsGI were similar, whereas those of two florigen genes Hd3a and RFT1, were significantly different under both short and long‐day conditions. Tos17(Hd1) showed photoperiod sensitivity intermediate between that of Nipponbare and a near‐isogenic line having a non‐functional allele of Hd1, indicating that Tos17(Hd1) has an attenuated but functional Hd1 allele. Our results revealed that allele mining of Hd1 may provide good opportunities to develop novel rice cultivars showing different levels of photoperiod sensitivity.