Diurnal and seasonal variations in stomatal conductance of rice at elevated atmospheric CO2 under fully open‐air conditions

Understanding of leaf stomatal responses to the atmospheric CO₂ concentration, [CO₂], is essential for accurate prediction of plant water use under future climates. However, limited information is available for the diurnal and seasonal changes in stomatal conductance (g_s) under elevated [CO₂]. We examined the factors responsible for variations in g_s under elevated [CO₂] with three rice cultivars grown in an open‐field environment under flooded conditions during two growing seasons (a total of 2140 individual measurements). Conductance of all cultivars was generally higher in the morning and around noon than in the afternoon, and elevated [CO₂] decreased g_s by up to 64% over the 2 years (significantly on 26 out of 38 measurement days), with a mean g_s decrease of 23%. We plotted the g_s variations against three parameters from the Ball‐Berry model and two revised versions of the model, and all parameters explained the g_s variations well at each [CO₂] in the morning and around noon (R² > 0.68), but could not explain these variations in the afternoon (R² < 0.33). The present results provide an important basis for modelling future water use in rice production.     

Sympatric speciation in a genus of marine reef fishes

Sympatric speciation has been contentious since its inception, yet is increasingly recognized as important based on accumulating theoretical and empirical support. Here, we present a compelling case of sympatric speciation in a taxon of marine reef fishes using a comparative and mechanistic approach. Hexagrammos otakii and H. agrammus occur in sympatry throughout their ranges. Molecular sequence data from six loci, with complete sampling of the genus, support monophyly of these sister species. Although hybridization occurrs frequently with an allopatric congener in an area of slight distributional overlap, we found no F₁ hybrids between the focal sympatric taxa throughout their coextensive ranges. We present genetic evidence for complete reproductive isolation based on SNP analysis of 382 individuals indicating fixed polymorphisms, with no shared haplotypes or genotypes, between sympatric species. To address questions of speciation, we take a mechanistic approach and directly compare aspects of reproductive isolation between allopatric and sympatric taxa both in nature and in the laboratory. We conclude that the buildup of reproductive isolation is strikingly different in sympatric vs. allopatric taxa, consistent with theoretical predictions. Lab reared hybrids from allopatric species crosses exhibit severe fitness effects in the F₁ or backcross generation. No intrinsic fitness effects are observed in F₁ hybrids from sympatric species pairs, however these treatments exhibited reduced fertilization success and complete pre‐mating isolation is implied in nature because F₁ hybrid adults do not occur. Our study addresses limitations of previous studies and supports new criteria for inferring sympatric speciation.     

Ultrastructure of the Harmful Unarmored Dinoflagellate Cochlodinium polykrikoides (Dinophyceae) with Reference to the Apical Groove and Flagellar Apparatus

ABSTRACT. The external and internal ultrastructure of the harmful unarmored dinoflagellate Cochlodinium polykrikoides Margalef has been examined with special reference to the apical groove and three‐dimensional structure of the flagellar apparatus. The apical groove is U‐shaped and connected to the anterior sulcal extension on the dorsal side of the epicone. The eyespot is located dorsally and composed of two layers of globules situated within the chloroplast. A narrow invagination of the plasma membrane is associated with the eyespot. The nuclear envelope has normal nuclear pores similar to other eukaryotes but different from the Gymnodinium group with diagnostic nuclear chambers. The longitudinal and transverse basal bodies are separated by approximately 0.5–1.0 μm and interconnected directly by a striated basal body connective and indirectly by microtubular and fibrous structures. Characteristic features of the flagellar apparatus are as follows: (1) a nuclear extension projects to the R1 (longitudinal microtubular root) and is connected to the root by thin fibrous material; (2) fibrillar structures are associated with the longitudinal and transverse flagellar canal; and (3) a striated ventral connective extends toward the posterior end of the cell along the longitudinal flagellar canal. We conclude, based on both morphological and molecular evidence, that Cochlodinium is only distantly related to Gymnodinium.     

Isolation and characterization of highly polymorphic microsatellite loci in a pelagic‐spawning reef fish Hypodytes rubripinnis

Group spawning (1 female and more than 2 males) and streaking behaviour of subordinate males are common in pelagic‐spawning reef fishes such as the temperate tetrarogidid fish Hypodytes rubripinnis. Five microsatellite loci were isolated and characterized for paternity analysis of this species. Four of the five loci were highly polymorphic, having 11–34 alleles with observed heterozygosities between 0.89 and 1.00 in 37 unrelated individuals. These four loci conformed to Hardy–Weinberg equilibrium. These loci should be useful in future studies that assess how fertilization is partitioned among male H. rubripinnis in multimale spawning events.     

Temperature-Sensitive Responses in Blepharisma

Temperature sensitivity of Blepharisma cultured at 23°C was investigated in a temperature range between 18.5°C and 33.5°C. The cells accumulated in an optimal temperature (ca. 27°C) region when they were placed in a chamber with a temperature gradient, although a certain population of the cells accumulated at much higher temperatures. The quantitative analysis of behavioral responses exhibited by the cells revealed that three types of thermal response were responsible for thermoaccumulation of the cells in an optimal temperature: (1) an increase in the frequency of thermophobic response in the cells swimming away from the optimal temperature region; (2) acceleration of forward swimming velocity of the cells swimming toward the optimal temperature region; and (3) higher frequency of spontaneous ciliary reversal of the cells in higher temperature regions.     

Production of a mouse strain with impaired glucose tolerance by systemic heterozygous knockout of the glucokinase gene and its feasibility as a prediabetes model

Exon II of glucokinase (Gk) was deleted to produce a systemic heterozygous Gk knockout (Gk^+/−) mouse. The relative expression levels of Gk in the heart, lung, liver, stomach, and pancreas in Gk^+/− mice ranged from 0.41–0.68 versus that in wild (Gk^+/+) mice. On the other hand, its expression levels in the brain, adipose tissue, and muscle ranged from 0.95–1.03, and its expression levels in the spleen and kidney were nearly zero. Gk knockout caused no remarkable off-target effect on the expression of 7 diabetes causing genes (Shp, Hnf1a, Hnf1b, Irs1, Irs2, Kir6.2, and Pdx1) in 10 organs. The glucose tolerance test was conducted to determine the blood glucose concentrations just after fasting for 24 h (FBG) and at 2 h after high-glucose application (GTT2h). The FBG-GTT2h plots obtained with the wild strain fed the control diet (CD), Gk^+/− strain fed the CD, and Gk^+/− strain fed the HFD were distributed in separate areas in the FBG-GTT2h diagram. The respective areas could be defined as the normal state, prediabetes state, and diabetes state, respectively. Based on the results, the criteria for prediabetes could be defined for the Gk^+/− strain developed in this study.     

Determination of effective light pulse and classical Bünsow experiment in the larval diapause of the Indian meal moth Plodia interpunctella

Plodia interpunctella Hübner (Lepidoptera: Pyralidae) comprises a model insect to analyse the photoperiodic time‐measuring system controlling its larval diapause. In the present study, the effective length of light pulse in night interruption experiments is determined at 25 °C. Various lengths of light pulse are tested by inserting them at the midnight of an LD 12 : 12 h photoperiod. When the light pulse is 15 or 30 min, the incidence of diapause is 86%. To inhibit the induction of diapause effectively, a light pulse of 1.75–2 h is needed. The incidence of diapause is 12% under an LD 12 : 5 : 2 : 5 h photoperiod. To determine the precise role of the light pulse, 2‐h light pulses placed at the midnight of an LD 12 : 12 h photoperiod are disrupted systematically by darkness. When a 2‐h light pulse is disrupted by 15 min of darkness, diapause is generally prevented (< 29%) regardless of the temporal position of darkness. Longer disruption by darkness induces diapause moderately (37–67%). A Bünsow experiment is also conducted at 25 and 20 °C, in which the main photophase of 12 h of light is combined with 24–72‐h scotophases scanned by a 2‐h light pulse. The photoperiodic cycle length tested, therefore, varies in the range 36–84 h. In each cycle length, the incidence of diapause fluctuates as the light pulse moves toward dawn. However, no regular and circadian changes in the percentage diapause are observed in relation to diapause determination.     

Potential agronomic options for energy-efficient sugar beet-based bioethanol production in northern Japan

Sugar beet (Beta vulgaris L. subsp. vulgaris) is deemed to be one of the most promising bioethanol feedstock crops in northern Japan. To establish viable sugar beet‐based bioethanol production systems, energy‐efficient protocols in sugar beet cultivation are being intensively sought. On this basis, the effects of alternative agronomic practices for sugar beet production on total energy inputs (from fuels and agricultural materials during cultivation and transportation) and ethanol yields (estimated from sugar yields) were assessed in terms of (i) direct drilling, (ii) reduced tillage (no moldboard plowing), (iii) no‐fungicide application, (iv) using a high‐yielding beet genotype, (v) delayed harvesting and (vi) root+crown harvesting. Compared with the conventional sugar beet production system used in the Tokachi region of Hokkaido, northern Japan, which makes use of transplants, direct drilling and no‐fungicide application contributed to reduced energy inputs from raising seedlings and fungicides, respectively, but sugar (or ethanol) yields were also reduced by these practices, to a greater equivalent extent than the reductions in energy inputs. Consequently, direct drilling (6.84 MJ L^?1) and no‐fungicide application (7.78 MJ L^?1) worsened the energy efficiency (total energy inputs to produce 1 L of ethanol), compared with conventional sugar beet production practices (5.82 MJ L^?1). Sugar yields under conventional plow‐based tillage and reduced tillage practices were similar, but total energy inputs were reduced as a result of reduced fuel consumption from not plowing. Hence, reduced tillage showed improved energy efficiency (5.36 MJ L^?1). The energy efficiency was also improved by using a high‐yielding genotype (5.23 MJ L^?1) and root+crown harvesting (5.21 MJ L^?1). For these practices, no major changes in total energy inputs were noted, but sugar yields were consistently increased. Neither total energy inputs nor ethanol yields were affected by extending the vegetative growing period by delaying harvesting.