Viral RNA extraction using an automatic nucleic acid extractor with magnetic particles and genetic characterization of bovine viral diarrhea virus in Tokachi Province,Japan, in 2016–2017

In this study, the viral genome extraction performance of automatic nucleic acid extractors and manual nucleic acid extraction kits was compared. We showed that compared with manual kits, the automatic extractors showed superior genome extraction performance using bovine viral diarrhea virus (BVDV) genome-positive cattle sera and bovine coronavirus/infectious bovine rhinotracheitis virus-spiked cattle nasal swabs. In addition, the subgenotyping of BVDV strains detected in Tokachi Province in Japan during 2016–2017 was performed. Results showed that most of these BVDV strains belonged to subgenotype 1b, while few strains belonged to subgenotypes 1a and 2a. This study showed the high applicability of automatic nucleic acid extractors in extracting multiple viral genomes and the dominant subgenotype of BVDV in Tokachi.     

Influence of volcanic ash sediments on spore adhesion of the red alga Gelidium elegans from the coastal seabed around Miyakejima Island, Japan

The populations of red alga Gelidium elegans along the coast of Miyakejima Island were severely damaged by a volcanic eruption in 2000. The effect of this volcanic eruption has been long lasting, and populations of this red alga still have not recovered. We investigated the effect of seabed sediment particles derived from volcanic ash on the substrate adhesion of G.?elegans spores. The analysis provides evidence that increasing amounts of sediment particles result in lower adhesion rates of G.?elegans spores, and that smaller sediment particles have a greater influence on adhesion. The amount of seabed sediment particles around Miyakejima Island was 9.3?C1815.4?mg/cm². This amount has changed greatly from year to year. The adhesion rate of G.?elegans spores in water around Miyakejima Island was 0?% at all points in 2008 and 2010, but it was estimated as 6.3?C38.6?% in 2009. These results suggest that there is significant inhibition of algal spore adhesion by seabed sediment particles derived from volcanic ash around Miyakejima Island.     

Effects of maturity genes E2 and E3 on yield formation in soybean cultivar Enrei in warm region,Fukuyama in Japan

ABSTRACT

Understanding how maturity genes affect soybean yield formation will provide important information for crop management decisions. This study aimed to reveal how maturity genes E2 and E3 in the soybean cultivar ‘Enrei’ affect yields and yield formation in warm regions of Japan. ‘Enrei’ (e2e3) and three near-isogenic lines of ‘Enrei’ (e2E3, E2e3, and E2E3) were cultivated in 2016 and 2017 in Fukuyama, Japan (34°30′N, 133°23′E). Two sowing dates were set in each year (June sowing and July sowing). E2 extended the period from emergence to R1 and also the period from R1 to R7, whereas E3 extended only the period from emergence to R1. Interaction between E2 and E3 did not affect duration of the period from emergence to R1, but did affect the period from R1 to R7. Although seed yield did not differ between genotypes in the June sowings, the effects of E2 and E3 on seed yield in July sowing were both significant and interaction between E2 and E3 also observed. The total number of nodes increased in E3 genotypes in both sowing dates, especially in E2E3. Pod-set ratio was lower in E2 and E3 genotypes than in e2 and e3 genotypes in the June sowings, but did not differ between genotypes in the July sowings. The high yield of E2E3 genotypes in the July sowings was attributed to increased number of nodes and flower production while maintaining pod-set ratio. Appropriate choice of sowing date is suggested to be essential when using E3 genotypes.Abbreviations: HI: harvest index; NIL: near-isogenic line; RUE: radiation use efficiency; TDM: total above-ground dry matter; TRI: total solar radiation intercepted     

The effects of distance from coral reefs on seagrass nursery use by 5 emperor fishes at the southern Ryukyu Islands,Japan

Whilst the importance of seagrass beds as nurseries for coral reef fishes has been clearly recognized, the vast majority of early studies on fish nursery habitats emphasized the close proximity of the latter to coral reefs. To determine the potential nursery role of isolated seagrass beds, we investigated the degree to which juvenile emperor fishes (Lethrinidae) utilized seagrass beds in the presence/absence of adjacent coral habitats at Ishigaki Island (southern Japan), such fishes being known to use seagrass beds as nurseries. Seagrass beds in close proximity to coral habitats (distance between the two habitats of 50–200 m) had greater densities of lethrinid juveniles than those without adjacent coral habitats (2.5–4 km) for 3 different sites investigated, although a significant difference was obtained only for 1 site. Juveniles of Lethrinus atkinsoni, L. obsoletus, L. harak, and L. nebulosus were observed in seagrass beds with and without adjacent coral habitats, whereas L. ornatus occurred only in the former. Overall, most lethrinid juveniles utilize seagrass beds irrespective of the presence of adjacent adult coral habitat, suggesting that both types of seagrass beds would contribute to lethrinid adult populations. Therefore, management efforts for lethrinid populations should be applied not only to contiguous coral-seagrass habitat systems but also to isolated habitats.     

Thermal behaviors and transitions of wood detected by temperature-modulated differential scanning calorimetry

The thermal properties and transitions of solid and ground wood samples conditioned at different humidity conditions were investigated by temperature-modulated differential scanning calorimetry. A time-dependent transition was detected as an endothermic peak in the total and non-reversing heat flows and as a step change in the reversing heat flow during the first heating run of samples with moisture contents above 5?%, but it disappeared in the second heating run. These different thermal behaviors indicate that the effect of heat and moisture on the thermal properties of wood is history dependent. This step change in the reversing heat flow is considered to be a glass transition of moist wood. Other relaxation processes (e.g., enthalpy relaxation) occur simultaneously with this glass transition. The temperature ranges of the transition and the relaxation decreased drastically as the moisture content increased up to 11?%, while they remained almost constant at higher moisture contents. In addition, the transitions of the ground wood occurred at lower temperatures than those of the solid wood at similar moisture contents. Kissinger plots revealed that the apparent activation energy for the glass transition of the solid wood with a moisture content of 11?% was about 600?kJ/mol, whereas that of the ground wood was 220?kJ/mol.     

Comparison of stand dynamics after dieback caused by pine wilt disease among pine forests with different management regimes in western Japan

In pine forests damaged by pine wilt disease, in western Japan, the effect of protection regimes of pine trees on the stand dynamics were examined in the following four stands: (1) lightly damaged stand (age 30–40 years) with no procedure in operation for protecting pine trees; (2) severely damaged stand (age 30–40 years) with no procedure in place for protecting pine trees; (3) severely damaged stand (age 50 years) with a selective cutting of infected trees; (4) severely damaged stand (age 30–40 years) with a selective cutting of infected trees. All the stands had been abandoned before the pine wilt disease damage. The understory structure of the severely damaged stand with no protection procedure was similar to that of the lightly damaged stand. Frequent invasion by tree species and acceleration in the growth of understory trees occurred after the dieback in the selective cutting stand. These results suggest that a deficiency in the canopy layer caused by the dieback resulted in low disturbance intensity in the early stages after the dieback, but the selective cutting increased the intensity by the reduction in the understory as well as the canopy layer. The intensity of the disturbance in the selective cutting stands was larger in the younger stand because it had a higher density of selectively cut pine trees. The different stand structure of pine forests occurred after the dieback because the intensity of the disturbance varied as a result of the selective cutting operation and the stand age.     

Differential overstory leaf flushing contributes to the formation of a patchy understory

Understory individuals were found to form patches in a 100-year-old deciduous broad-leaved forest. The closed forest canopy was uniform, and so the light conditions at various locations across the forest floor differed little after the leaf flush of the overstory. To explain the distribution pattern in the understory, a hypothesis was proposed: in spring, the forest floor is divided into patches according to the timing of leaf flush of the overstory individuals, and the light conditions are more favorable for understory plants under the crowns of trees with later-flushing leaves. In the plot, three groups of early, intermediate, and late, were recognized in the overstory concerning the timing of leaf flush. As for the start of leaf flush, a difference of 31.6 days was recognized among tree species, and for the end of leaf flush, a difference of 40.3 days. In the spring of 1998, the relative photosynthetic-photon-flux density under an intensively studiedCastanea crenata tree (late-flushing species) usually showed higher values than that under a similarly studiedAcer mono tree (early-flushing species). Analysis of the spatial-distribution pattern using Morisita’s1δ index revealed that the understory community had an aggregated distribution. In the overstory, the late- and the intermediate-flushing-species groups showed aggregated distributions, while the early-flushing-species group showed random distribution. Spatial correlation between the understory and the overstory was analyzed by using Morisita’sRδ index. The distribution of whole understory community spatially co-occurred with that of the late-flushing-species group of the overstory. In contrast, the understory community was less developed below the members of the early-flushing-species group of the overstory. We consider that the data presented here support our hypothesis, and we suggest that the growth and survival of understory individuals were promoted in the places receiving light for long periods in spring.     

Impaired female fertility in tubulointerstitial antigen-like 1-deficient mice

Tubulointerstitial nephritis antigen-like 1 (Tinagl1, also known as adrenocortical zonation factor 1 [AZ-1] or lipocalin 7) is a matricellular protein. Previously, we demonstrated that Tinagl1 expression was restricted to extraembryonic regions during the postimplantation period and detected marked expression in mouse Reichert’s membranes. In uteri, Tinagl1 is markedly expressed in the decidual endometrium during the postimplantation period, suggesting that it plays a physical and physiological role in embryo development and/or decidualization of the uterine endometrium during pregnancy. In the present study, in order to determine the role of Tinagl1 during embryonic development and pregnancy, we generated Tinagl1-deficient mice. Although Tinagl1^–/– embryos were not lethal during development to term, homologous matings of Tinagl1^–/– females and Tinagl1^–/– males showed impaired fertility during pregnancy, including failure to carry pregnancy to term and perinatal lethality. To examine ovarian function, ovulation was induced with equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG); the number of ovulated oocytes did not differ between Tinagl1^–/– and Tinagl1^flox/flox. In vitro fertilization followed by embryo culture also demonstrated the normal developmental potential of Tinagl1-null embryos during the preimplantation period. Our results demonstrate that Tinagl1 deficiency affects female mice and results in subfertility phenotypes, and they suggest that although the potential of Tinagl1^–/– oocytes is normal, Tinagl1 is related to fertility in adult females but is not essential for either fertilization or preimplantation development in vitro.     

Gene silencing of cyclooxygenase-2 mRNA by RNA interference in bovine cumulus-granulosa cells

Inhibition of specific gene expression using RNA interference (RNAi) is a valuable tool for functional analysis of a target gene. However, there is little information available concerning RNAi for analysis of gene function in relation to the reproductive physiology of follicular cells in ruminants. Thus, the aim of this study was to evaluate the interfering effect of small interference RNA (siRNA) on expression of cyclooxygenase-2 (Cox-2) mRNA and prostagrandin F(2alpha) (PGF(2alpha)) production in bovine cumulus-granulosa (CG) cells. Bovine CG cells were collected from aspirated follicles and cultured. After reaching confluency, two experiments were conducted. In experiment 1, to investigate the effective concentration of siRNA, 0, 100, 250 and 500 pM of Cox-2 siRNA was introduced into the CG cells, respectively. After 24 h, the amount of Cox-2 mRNA expression was measured by RT-PCR and real-time PCR. In experiment 2, to investigate the time required for effective interference of siRNA and Cox-2 activity, 250 pM siRNA was introduced for 0, 3, 6, 12 and 24 h. After culture, the amount of Cox-2 mRNA expression was measured and the culture medium was collected to determine the PGF(2alpha) concentration by enzyme immunoassay. The Cox-2 mRNA expression was not affected by introduction of 100 pM siRNA into CG cells for 24 h, but 250 and 500 pM Cox-2 siRNA significantly reduced the Cox-2 mRNA expression. Moreover, the significant suppressive effect of 250 pM siRNA was observed 6 h after introduction, and the reduction of mRNA expression by RNAi became more obvious over 12 h. On the other hand, the PGF(2alpha) concentration in the culture medium was not significantly different 12 h after siRNA introduction; however, the PGF(2alpha) concentration 24 h after siRNA introduction was significantly decreased compared with the control at the same time point. These results suggest that gene silencing of Cox-2 with siRNA is capable of analyzing the function and expression of specific genes in bovine CG cells.     

Maize defence mechanisms against the European corn borer,Ostrinia nubilalis Hübner (Lepidoptera: Crambidae)

Maize is arguably the most widely grown crop in the world, but it is often associated with one of its major insect pests, the European corn borer (ECB). The damage caused by this species to maize production is generally variable, but in many cases it is economically significant. This review paper provides an overview of the research findings on the natural plant defence mechanisms against ECB larvae published till now. What is resistance and how it is achieved, what is the chemical response of maize plants to insect feeding and how tolerance can be increased. A short introduction in breeding for resistance and a discussion if the mentioned traits can be used in conventional breeding in order to create maize hybrids less affected by ECB are given.