Comparison of kinetic properties of a hydrophilic form of acetylcholinesterase purified from strains susceptible and resistant to carbamate and organophosphorus insecticides of green rice leafhopper (Nephotettix cincticeps Uhler)

A hydrophilic form of acetylcholinesterase (AChE) was purified from N-methyl carbamate susceptible (SA) and highly N-methyl carbamate-resistant (N3D) strains of the green rice leafhopper (GRLH), Nephotettix cincticeps Uhler. Both of purified AChE from SA and N3D strains displayed the highest activities toward acetylthiocholine (ATCh) at pH 8.5. In the SA strain, the optimum concentrations for ATCh, propionylthiocholine (PTCh), and butyrylthiocholine (BTCh) were about 1 × 10⁻³, 2.5 × 10⁻³, and 1 × 10⁻³ M, respectively. However, in the N3D strain, substrate inhibition was not identified for ATCh, PTCh, and BTCh to 1 × 10⁻² M. The K_m value in the SA strain was 51.1, 39.1, and 41.6 μM and that in the N3D strain was 91.8, 88.1, and 85.2 μM for ATCh, PTCh, and BTCh, respectively. The K_m value in the N3D strain indicated about 1.80-, 2.25-, and 2.05-fold lower affinity than that of the SA strain for ATCh, PTCh, and BTCh, respectively. The V_max value in the SA strain was 70.2, 30.5, and 4.6 U/mg protein and that in the N3D strain was 123.0, 27.0, and 14.5 U/mg protein for ATCh, PTCh, and BTCh, respectively. The V_max value in the N3D strain was 1.75- and 3.15-fold higher for ATCh and BTCh than that in the N3D strain. However, it was 1.13-fold lower for PTCh. The increased activity of AChE in the N3D strain is due to the qualitatively modified enzyme with a higher catalytic efficiency. The bimolecular rate constant (k_i) for propoxur was 27.1 × 10⁴ and 0.51 × 10⁴ M⁻¹ min⁻¹ in the SA and N3D strain and that for monocrotophos was 0.031 × 10⁴ and 2.0 × 10⁴ M⁻¹ min⁻¹ in the SA and N3D strain. AChE from the N3D strain was 53-fold less sensitive than SA strain to inhibition by propoxur. In contrast, AChE from the N3D strain was 65-fold more sensitive to inhibition by monocrotophos than AChE from the SA strain. This indicated negatively correlated cross-insensitivity of AChE to propoxur and monocrotophos.     

Fibrodysplasia ossificans progressiva-like condition in a cat

Fibrodysplasia ossificans progressiva (FOP)-like condition was diagnosed in a Japanese domestic cat with stiffness, marked atrophy of the muscles, and limited mobility of all joints in both the pelvic limbs. Etretinate, a retinoid, was used for medical management; however, no improvement in the clinical signs was observed. Inheritance of the disorder has not yet been demonstrated. Furthermore, the clinical signs and histopathological findings of feline FOP-like condition in the present case differed from those of the previously reported cases.     

QTL analysis of panicle morphology response to irrigation regime in aerobic rice culture

In water-efficient rice production, grain yield is often constrained by panicle size. The objective of this study was to genetically dissect the response of panicle morphology to irrigation regimes in aerobic rice culture. We grew ‘Akihikari’ (a lowland japonica cultivar) × ‘IRAT109’ (an upland japonica cultivar) backcross inbred lines in aerobic soils with full or limited irrigation for 2 years, and examined 4 panicle traits—number of florets per panicle (FPP), number of primary branches per panicle (BPP), number of florets per primary branch (FPB), and frequency of pre-flowering floret abortion (%FA)—and grain yield. QTLs for BPP were detected in both the irrigation regimes but QTLs for FPB and %FA were detected mostly only in either of the irrigation regimes. The QTL for FPP on chromosome 2 (RM3421–RM213) coincided with that for yield under full irrigation, showing that this QTL is related to sink capacity and yield potential in aerobic rice culture. On the other hand, the QTL for FPB on chromosome 1 (RM3148–RM243) coincided with that for yield under limited irrigation, when water deficit was moderate. The QTL for root axis length at vegetative stage, previously identified in the same mapping population, was located near this region. This study unravelled the complicated genetic control on panicle morphology in aerobic rice culture, and suggested the positive roles of the dehydration avoidance mechanism by vigorous root growth on panicle size and yield under dry soil conditions.     

Effect of the timing of first cleavage on in vitro developmental potential of nuclear-transferred bovine oocytes receiving cumulus and fibroblast cells

The aim of the present study was to examine whether cumulus and fibroblast cell nuclear-transferred oocytes, which have high and low potential to develop into normal calves, respectively, are different in terms of in their patterns of timing of first cleavage and in their relationships between timing of first cleavage and in vitro developmental potential. The timing of first cleavage was similar in both types of nuclear-transferred and in vitro fertilized oocytes. More than 86% of the oocytes cleaved within 24 h after activation or in vitro fertilization; these oocytes contributed to more than 98% of the total number of blastocysts in all three groups. The potential of oocytes that cleaved at different intervals to develop into blastocysts differed among the groups. The developmental potential of the cumulus cell nuclear-transferred oocytes and in vitro fertilized oocytes decreased with the increase in time required for cleavage. Fibroblast cell nuclear-transferred oocytes that cleaved at 20 h, an intermediate cleaving time, had higher potential to develop into blastocysts. The results of the present study suggest that the type of donor nucleus used for nuclear transfer affects the timing of first cleavage.     

Polyacrylamide gel electrophoresis of S-RNase fragments for identification of S-genotypes of Japanese pear (Pyrus pyrifolia)

Japanese pear (Pyrus pyrifolia) exhibits gametophytic self-incompatibility (GSI) controlled by a complex and multiallelic S locus. The pistil-part product of the S locus is the polymorphic ribonuclease, S-RNase. Information on S-genotypes is important for the production and breeding of Japanese pears. Molecular analyses of S-genotypes of Japanese pear have been conducted with the CAPS (cleaved amplified polymorphic sequence) system; PCR amplification of S-RNase fragments by a common primer pair followed by digestion with restriction enzymes each of which cleaves a specific S haplotype. Here, we show that the separation of S-RNase fragments by polyacrylamide gel electrophoresis (PAGE) distinguishes four out of nine S haplotypes of Japanese pear without restriction digestion. S³-, S⁵-, S⁶- and S⁸-RNases were identified as distinct bands by PAGE. S³- and S⁵-RNases were separated by PAGE despite their identical fragment sizes. Using this system, three Japanese pear lines with unknown S-genotypes were analyzed. The newly determined S-genotypes of the lines were confirmed by CAPS analysis.     

Effect of genome composition and cytoplasm on petal colour in resynthesized amphidiploids and sesquidiploids derived from crosses between Brassica rapa and Brassica oleracea

The effect of genome composition and cytoplasm on petal colour was studied in Brassica. Three accessions of yellow‐petalled B. rapa (2n= 20, AA) were crossed with a white‐petalled B. oleracea var. alboglabra (2n= 18, CC) and with three cream‐yellow‐petalled B. oleracea var. gongylodes (2n= 18, CC) to produce resynthesized B. napus (2n= 38, AACC or CCAA) and sesquidiploids (2n= 29, AAC or CAA). Petal colour was measured with a Hunter automatic colour difference meter. The results revealed that petal colour in Brassica is controlled by nuclear genes and by cytoplasmic factors. Additive and epistatic gene effects were involved in the action of nuclear genes. When crosses were made between yellow‐petalled B. rapa and white‐petalled B. oleracea var. alboglabra, significant additive, epistatic and cytoplasmic effects were found. White petal colour was partially epistatic over yellow petal colour. When crosses were made between yellow‐petalled B. rapa and cream‐yellow‐petalled B. oleracea var. gongylodes, only epistatic effects were detected. Yellow petal colour was epistatic over cream‐yellow.     

A Computer Program for Automatic Watering Based on Potential Evapotranspiration by Penman Method and Predicted Leaf Area in Miniature Pot Rose Production

In this study, we developed a computer program for automatic prediction of watering time point by considering the environmental factors such as solar radiation, air temperature and relative humidity based on the multiple linear regression equation of leaf area and Penman Method. The experiments were carried out for a year in two watering experimental plots, one of which was controlled by pF value, and the other by the computer program. After comparing the results of the two plots, the following findings were obtained. In the computer program plot, the observed and predicted values of both leaf area and evapotranspiration indicated significant correlation at the 1% level, which suggested that the computer program had high prediction accuracy. In addition, no significant difference was observed between the two experimental plots with respects to the plant height, plant diameter, leaf area, leaf number, fresh weight, and dry weight, which demonstrated that the plants in the computer program plot had normal growth. On the other hand, although the number of flower buds and flowering shoots showed higher values at the end of certain cultivations in the computer program plot than those in pF value plot, we proposed that it was due to the effect of cumulative daily solar radiation in the greenhouse, rather than the watering. Thus, we have reached the conclusion that the computer program for automatic prediction of watering time point developed by this study has high applicability in miniature pot rose production.     

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.