Seasonal variation in pigmentation and anthocyanidin phenetics in commercial Eustoma flowers

The seasonal change in petal color and pigmentation of 29 commercial Eustoma cultivars was studied. The flowers are basically divided into four groups according to the major anthocyanidin phenotype in association with petal coloration, i.e., delphinidin (Dp)-based (purple flower), cyanidin (Cy)-based (reddish purple flower), pelargonidin (Pg)-based (pink flower), and none (white flower) groups. The constitution of petal anthocyanidins was not changed by forcing treatment in most of the flowers. Lightness (L^*) and chroma (C^*, color saturation) showed a change along with the increase/decrease of hue angle difference (ΔH^*), thus simultaneously the chromatic tonalities tended to move to redder and bluer, respectively. Floral pigment clustering described two flower groups in a dendrogram, based on anthocyanidin constitutions as phenetic markers, which are apparently the Dp- and Pg-based phenotypes of anthocyanidin syntheses. The Cy-based flowers made a subcluster with the Pg-based flowers, indicating a close relationship in the biosynthesis of the two anthocyanidins, and suggesting the Dp- and Pg-syntheses complement one another.     

Pine Wilt Disease Caused by the Pine Wood Nematode: The Induced Resistance of Pine Trees by the Avirulent Isolates of Nematode

Since the beginning of the 20th century, pine trees in Japan have been seriously damaged by the pine wilt disease. This disease is caused by the pine wood nematode, Bursaphelenchus xylophilus, which is transmitted by the Japanese pine sawyer, Monochamus alternatus. The control of disease depends to a large extent on chemicals, but the public is now demanding environmentally friendly control methods. The virulence of B. xylophilus varies very widely. Pre-inoculation of young pine trees in a nursery with avirulent B. xylophilus has induced systemic resistance of trees against a subsequent inoculation with virulent B. xylophilus. This induced resistance was considered a hopeful means for developing a biological control for the disease. The induced resistance by the avirulent nematodes was also expressed in mature pine trees in a forest where the disease was naturally epidemic. However, the effects of induced resistance were not satisfactory for practical biological control. Since the inoculation with higher concentrations of the avirulent B. xylophilus induced the resistance more effectively, the pre-inoculation method will need to be improved to develop the biological control. The induced resistance of pine trees by avirulent B. xylophilus should be one of the candidate biological control methods against pine wilt disease. This induced resistance also provides an experimental system to clarify physiological interactions between the nematodes and pine trees.     

Caprine somatic cell nuclear transfer using in vivo matured oocytes collected by laparoscopic follicular aspiration

In vivo matured oocytes collected by laparoscopic follicular aspiration (LFA) from hormone treated female goats were used as recipient ooplasts for somatic cell nuclear transfer (SCNT). Japanese native (Shiba) goats were used as donor females and some donor females were used repeatedly (two or three times) at intervals of a few months. To induce synchronization of estrus, a sponge containing 0.5 g of progesterone was inserted into the vagina of each goat for 14 days. These animals were also treated with follicle stimulating hormone (FSH) in a series of 8 injections over 4 days. The first FSH injection was administered on the morning of day 9 of sponge insertion. On the morning of day 13, 50 µg of gonadotropin‐releasing hormone (GnRH) was injected into each animal. Twenty‐nine hours after GnRH injection, LFA was performed. After removal of cumulus cells, collected oocytes with the first polar body were selected and enucleated for nuclear transfer. Anterior pituitary cells isolated from an adult male Shiba goat were transfected with a DNA fragment containing the enhanced green flourescent protein gene and the puromycin resistance gene. A single donor cell was inserted into the perivitelline space of each enucleated oocyte and fusion was induced with one electric pulse of 20 V for 10 µs. The SCNT goat eggs were cultured in chemically defined medium at 38.5°C in 5% CO₂, 5% O₂, 90% N₂ for 9 days. By LFA, 396 oocytes were collected from a total of 30 females. After removal of cumulus cells, 64% of them extruded the first polar body. The percentage of SCNT goat eggs produced using in vivo matured oocytes which developed to the blastocyst stage (20–21%) was significantly higher (P < 0.05) than that produced with in vitro matured oocytes (3–8%).     

Long-term growth analyses of Japanese cedar trees in a plantation: neighborhood competition and persistence of initial growth deviations

The individual growth of tree diameter at breast height (dbh) is analyzed in an even-aged plantation of Cryptomeria japonica from stand age of 45 to 94 years, to examine how the growth of individual trees has been affected by the changes in spacing resulting from thinning operations. At any age, a significant proportion (0.37–0.46) of the variation in dbh growth during a 5–11-year period was explained by dbh at the beginning of the period, probably due to greater leaf mass of larger trees. Next, either one-sided or two-sided competition was added to the model, by calculating the basal area (BA) of neighboring trees around each tree within a given radius or BA for trees having larger dbh than the focal tree within the radius. After preliminary analyses, a radius of 8 m was selected as the critical range for tree competition. Although both types of competition explained a significant proportion (0.09–0.43) of growth variation, one-sided competition was not significant at ages greater than 54 years. Based on the model at 45 years of age, the initial deviation of growth rate for each tree from the predicted rate was calculated and added to the models as a third variable. This raised the coefficient of determination up to 0.50–0.74. These findings have practical significance for forest plantation management, particularly for controlling the growth of standing trees via thinning, to produce high-quality timber in the future.     

Does future climate change facilitate expansion of evergreen broad-leaved tree species in the human-disturbed landscape of the Korean Peninsula? Implication for monitoring design of the impact assessment

To quantitatively assess future change of evergreen broad-leaved tree species’ distributions in human-disturbed landscapes of the Korean Peninsula under climate change, potential habitats (PHs) were projected for four important evergreen broad-leaved tree species (Quercus acuta, Castanopsis sieboldii, Machilus thunbergii, and Neolitsea sericea) by species distribution models (SDMs). The distribution data (presence/absence) of the target species in Korea and Japan were used as response variables for SDMs, and climatic data were used as explanatory variables. Three general circulation models under A2 emission scenarios were used as future climate scenarios for the years 2070–2099. Potential habitats masked by land-use data (PHLUs) were projected to assess the impact of anthropogenic activities. Highly accurate SDMs were obtained for all the target species. The current PHs were decreased to 21–35 % by the anthropogenic activities. Future PHLUs for all the target species were projected to increase by 2.0–18.5 times of current PHLUs. These results suggest that all the target species are applicable as indicator species for monitoring in the Korean Peninsula, even if anthropogenic effects are incorporated. Variation of the increasing rate was caused by the differences in the response to temperature changes. M. thunbergii responded sensitively to the increase of minimum temperature of coldest month and had a largest increase in PHLUs under future climate. Therefore, M. thunbergii is considered to be most appropriate species for monitoring the changes of horizontal distributions above all focal evergreen broad-leaved tree species.     

A size-structured model for a stock-recovery program for an exploited endemic fisheries resource

The size of harvests temporarily decreases following stock-recovery programs for overexploited resources. The process of stock recovery is quite uncertain, because of unknown life-history parameters, changes in environmental conditions, and human impact. To project future stock recovery, we considered a size-structured matrix model of a stock-recovery program, in which the minimum body size in the first harvest is increased. We applied some common concepts, including yield per recruitment, spawning per recruitment, and reproductive value, to a size-structured matrix model that incorporated economic discounting. The size-structured model predicted the following characteristics in stock-recovery processes: several years must pass for the harvest to increase, and the time-lag between the inception of the management program and the increase in the catch is caused by intergenerational momentum in demography. We also investigated the effects on the recovery process of temporal environmental fluctuations. We applied our model to a stock-recovery program of the corbiculid Seta clam (Corbicula sandai) in Lake Biwa, Japan.     

QTL analysis of photoperiod sensitivity in common buckwheat by using markers for expressed sequence tags and photoperiod-sensitivity candidate genes

Photoperiod sensitivity is an important trait related to crop adaptation and ecological breeding in common buckwheat (Fagopyrum esculentum Moench). Although photoperiod sensitivity in this species is thought to be controlled by quantitative trait loci (QTLs), no genes or regions related to photoperiod sensitivity had been identified until now. Here, we identified QTLs controlling photoperiod sensitivity by QTL analysis in a segregating F₄ population (n = 100) derived from a cross of two autogamous lines, 02AL113(Kyukei SC2)LH.self and C0408-0 RP. The F₄ progenies were genotyped with three markers for photoperiod-sensitivity candidate genes, which were identified based on homology to photoperiod-sensitivity genes in Arabidopsis and 76 expressed sequence tag markers. Among the three photoperiod-sensitivity candidate genes (FeCCA1, FeELF3 and FeCOL3) identified in common buckwheat, FeELF3 was associated with photoperiod sensitivity. Two EST regions, Fest_L0606_4 and Fest_L0337_6, were associated with photoperiod sensitivity and explained 20.0% and 14.2% of the phenotypic variation, respectively. For both EST regions, the allele from 02AL113(Kyukei SC2)LH.self led to early flowering. An epistatic interaction was also confirmed between Fest_L0606_4 and Fest_L0337_6. These results demonstrate that photoperiod sensitivity in common buckwheat is controlled by a pathway consisting of photoperiod-sensitivity candidate genes as well as multiple gene action.     

Involvement of mammalian Mus81 in genome integrity and tumor suppression

Mus81-Eme1 endonuclease has been implicated in the rescue of stalled replication forks and the resolution of meiotic recombination intermediates in yeast. We used gene targeting to study the physiological requirements of Mus81 in mammals. Mus81-/- mice are viable and fertile, which indicates that mammalian Mus81 is not essential for recombination processes associated with meiosis. Mus81-deficient mice and cells were hypersensitive to the DNA cross-linking agent mitomycin C but not to gamma-irradiation. Remarkably, both homozygous Mus81-/- and heterozygous Mus81+/- mice exhibited a similar susceptibility to spontaneous chromosomal damage and a profound and equivalent predisposition to lymphomas and other cancers. These studies demonstrate a critical role for the proper biallelic expression of the mammalian Mus81 in the maintenance of genomic integrity and tumor suppression.     

Nuclear transfer using a bovine mammary epithelial cell line (BMEC)

The developmental potential of nuclei from a bovine mammary epithelial cell line (BMEC) in nuclear transfer was investigated. For nuclear transfer donors, BMEC cells (passage 15) were cultured for 4–5 days after seeding at cell densities of 1.0 × 10⁵ cells/cm² (high‐density group) or 0.8 × 10⁴ cells/cm² (low‐density group). First, the effective electric stimulation for fusion of enucleated oocytes with BMEC cells was examined. Fusion rates reached maximum with two DC pulses of 30 V/150 µm for 20 µs in the high‐density group and with two DC pulses of 25 V/150 µm for 10 µs in the low‐density group. The fusion rate (37.5%) in the high‐density group was significantly (P < 0.005) lower than in the low‐density group (71.4%). Second, the in vitro developmental potential of nuclear transfer embryos derived from BMEC cells was examined. In the high‐density and low‐density groups, 18.8% and 24.1% of fused oocytes, respectively, developed to blastocyst stage. The results of this study indicate that nuclei from BMEC cells support the development of nuclear transfer embryos to the blastocyst stage and that the efficiency of oocyte–cell fusion is affected by the culture conditions of the donor BEMC cells before nuclear transfer.