Nobumitsu Kawakubo

Relative Role of Flower Color and Scent on Pollinator Attraction: Experimental Tests using F1 and F2 Hybrids of Daylily and Nightlily

The daylily (Hemerocallis fulva) and nightlily (H. citrina) are typical examples of a butterfly-pollination system and a hawkmoth-pollination system, respectively. H. fulva has diurnal, reddish or orange-colored flowers and is mainly pollinated by diurnal swallowtail butterflies. H. citrina has nocturnal, yellowish flowers with a sweet fragrance and is pollinated by nocturnal hawkmoths. We evaluated the relative roles of flower color and scent on the evolutionary shift from a diurnally flowering ancestor to H. citrina. We conducted a series of experiments that mimic situations in which mutants differing in either flower color, floral scent or both appeared in a diurnally flowering population. An experimental array of 6×6 potted plants, mixed with 24 plants of H. fulva and 12 plants of either F1 or F2 hybrids, were placed in the field, and visitations of swallowtail butterflies and nocturnal hawkmoths were recorded with camcorders. Swallowtail butterflies preferentially visited reddish or orange-colored flowers and hawkmoths preferentially visited yellowish flowers. Neither swallowtail butterflies nor nocturnal hawkmoths showed significant preferences for overall scent emission. Our results suggest that mutations in flower color would be more relevant to the adaptive shift from a diurnally flowering ancestor to H. citrina than that in floral scent.

Dioecism of the Genus Callicarpa (Verbenaceae) in the bonin (Ogasawara) Islands

The morphology and function of the flowers of three species of the genusCallicarpa, C. glabra, C. nisnimurae andC. subpubescens, endemic to the Bonin Islands, revealed that all three species were dioecious. Male plants had short-styled flowers with sterile ovaries, while female plants had long-styled flowers with non-germinating inaperturate pollen grains. Such dioecism has never been reported from the family Verbenaceae. This curious sexual system with non-germinating pollen grains may have been maintained as a reward to insect pollinators.

Indigenous utilization of termite mounds and their sustainability in a rice growing village of the central plain of Laos

BackgroundThe objective of this study was to investigate the indigenous utilization of termite mounds and termites in a rain-fed rice growing village in the central plain of Laos, where rice production is low and varies year-to-year, and to assess the possibility of sustainable termite mound utilization in the future. This research was carried out from 2007 to 2009.MethodsThe termites were collected from their mounds and surrounding areas and identified. Twenty villagers were interviewed on their use of termites and their mounds in the village. Sixty-three mounds were measured to determine their dimensions in early March, early July and middle to late November, 2009.ResultsEleven species of Termitidae were recorded during the survey period. It was found that the villagers use termite mounds as fertilizer for growing rice, vegetable beds and charcoal kilns. The villagers collected termites for food and as feed for breeding fish. Over the survey period, 81% of the mounds surveyed increased in volume; however, the volume was estimated to decrease by 0.114 m3 mound-1 year-1 on average due to several mounds being completely cut out.ConclusionIt was concluded that current mound utilization by villagers is not sustainable. To ensure sustainable termite utilization in the future, studies should be conducted to enhance factors that promote mound restoration by termites. Furthermore, it will be necessary to improve mound conservation methods used by the villagers after changes in the soil mass of mounds in paddy fields and forests has been measured accurately. The socio-economic factors that affect mound utilization should also be studied.

Pollinator-Mediated Selection on Flower Color, Flower Scent and Flower Morphology of Hemerocallis: Evidence from Genotyping Individual Pollen Grains On the Stigma

To trace the fate of individual pollen grains through pollination processes, we determined genotypes of single pollen grains deposited on Hemerocallis stigmas in an experimental mixed-species array. Hemerocallis fulva, pollinated by butterflies, has diurnal, reddish and unscented flowers, and H. citrina, pollinated by hawkmoths, has nocturnal, yellowish and sweet scent flowers. We observed pollinator visits to an experimental array of 24 H. fulva and 12 F2 hybrids between the two species (H. fulva and H. citrina) and collected stigmas after every trip bout of swallowtail butterflies or hawkmoths. We then measured selection by swallowtail butterflies or hawkmoths through male and female components of pollination success as determined by single pollen genotyping. As expected, swallowtail butterflies imposed selection on reddish color and weak scent: the number of outcross pollen grains acquired is a quadratic function of flower color with the maximum at reddish color, and the combined pollination success was maximal at weak scent (almost unrecognizable for human). This explains why H. fulva, with reddish flowers and no recognizable scent, is mainly pollinated by swallowtail butterflies. However, we found no evidence of hawkmoths-mediated selection on flower color or scent. Our findings do not support a hypothesis that yellow flower color and strong scent intensity, the distinctive floral characteristics of H. citrina, having evolved in adaptations to hawkmoths. We suggest that the key trait that triggers the evolution of nocturnal flowers is flowering time rather than flower color and scent.

Contribution of Pollinators to Seed Production as Revealed by Differential Pollinator Exclusion in Clerodendrum trichotomum (Lamiaceae)

A diverse assemblage of pollinators, such as bees, beetles, flies, and butterflies, will often visit a single plant species. However, evaluating the effect of several insects on fruit and seed production is difficult in plants visited by a variety of insects. Here, we analyzed the effect of three types of pollinators, Papilio spp., Macroglossum pyrrhosticta, and Xylocopa appendiculata on fruit and seed production in Clerodendrum trichotomum by using a flower visitor barrier experiment with nets of specific mesh sizes. As a result, fruit/flower and seed/ovule ratios were significantly lower under Papilio exclusion than under natural conditions. On the other hand, ratios were not significantly different between Papilio excluded and both Papilio and M. pyrrhosticta excluded treatments. Therefore, Papilio and X. appendiculata are effective pollinators, whereas M. pyrrhosticta, which was the most frequent visitor, of C. trichotomum, is not. From our observations of visiting behaviors, we believe that because M. pyrrhosticta probably promotes self- pollination, this species is a non-effective pollinator. This is the first study to separate and compare the contribution of various visitors to the reproductive success of a plant.

Crisis of Japanese vascular flora shown by quantifying extinction risks for 1618 taxa

Although many people have expressed alarm that we are witnessing a mass extinction, few projections have been quantified, owing to limited availability of time-series data on threatened organisms, especially plants. To quantify the risk of extinction, we need to monitor changes in population size over time for as many species as possible. Here, we present the worlds first quantitative projection of plant species loss at a national level, with stochastic simulations based on the results of population censuses of 1618 threatened plant taxa in 3574 map cells of ca. 100 km2. More than 500 lay botanists helped monitor those taxa in 1994–1995 and in 2003–2004. We projected that between 370 and 561 vascular plant taxa will go extinct in Japan during the next century if past trends of population decline continue. This extinction rate is approximately two to three times the global rate. Using time-series data, we show that existing national protected areas (PAs) covering ca. 7% of Japan will not adequately prevent population declines: even core PAs can protect at best <60% of local populations from decline. Thus, the Aichi Biodiversity Target to expand PAs to 17% of land (and inland water) areas, as committed to by many national governments, is not enough: only 29.2% of currently threatened species will become non-threatened under the assumption that probability of protection success by PAs is 0.5, which our assessment shows is realistic. In countries where volunteers can be organized to monitor threatened taxa, censuses using our method should be able to quantify how fast we are losing species and to assess how effective current conservation measures such as PAs are in preventing species extinction.

Variations in leaf morpho‐anatomy and photosynthetic traits between sun and shade populations of Eurya japonica (Pentaphylacaceae) whose seeds are dispersed by birds across habitats

Eurya japonica occurs in diverse light environments through seed dispersal by birds. As the seed size is extremely small, we hypothesized that newly germinated seedlings with restricted depth of roots and length of the hypocotyl would suffer high mortality due to increased transpiration in sunny habitats and low light in shady habitats. We also expected that surviving seedlings would differ in leaf traits between habitats as a result of selection. We aimed to determine how photosynthetic traits differ between habitats and how leaf structure is related to this difference. We examined photosynthesis and leaf morpho-anatomy for plants cloned from cuttings collected from the forest understory (shade population) and neighboring roadsides and cut-over areas (sun population) and then grown under two irradiances (18.5% and 100% sunlight) in an experimental garden. Under growth in 100% sunlight, cloned plants from the sun population exhibited significantly greater area-based photosynthetic capacity compared to cloned plants from the shade population at a comparable stomatal conductance, which was attributable to a higher area-based leaf nitrogen concentration. On the other hand, mean values of photosynthetic capacity did not significantly differ between the two populations. Cloned plants from the sun population had significantly thicker leaf laminas and spongy tissue and lower stomatal density compared to cloned plants from the shade population. Thickened leaf lamina might have increased leaf tolerance to physical stresses in open habitats. The variation in leaf morpho-anatomy between the two populations can be explained in terms of the economy of leaf photosynthetic tissue.

Fine-scale flower-visiting behavior revealed by using a high-speed camera

Most estimations of the pollination efficiency of insects have been based on observation by the naked human eye. However, insect behaviors are often too rapid to analyze sufficiently this way. Here we demonstrate the use of high-speed cameras to analyze the fine-scale behaviors of Macroglossum pyrrhosticta, Xylocopa appendiculata, and Papilio dehaanii when visiting Clerodendrum trichotomum. The fine-scale nectar drinking time, number of contacts with anthers and/or stigmas, and frequencies of body part contact with anthers and/or stigmas differed significantly among pollinator species. Pollination efficiency was not equal among pollinators. In addition, M. pyrrhosticta made the least number of contacts with anthers and/or stigmas even though it showed the highest visitation frequency. These results demonstrate that when examined from the viewpoint of rapid visitation behaviors, pollination dynamics differ among pollinator species, and flower visits and pollination rates are not equal.

Effects of soil moisture and temperature on decomposition rates of some waste materials from agriculture and agro-industry.

Abstract The effect of soil moisture and temperature on decomposition of waste materials, bagasse, coir dust, rice chaff and rice straw, in soil were examined by measuring the decrease in weight of and CO2 generation from each waste material. The rate of the decrease in weight increased as temperature rose, and was highest in rice straw followed by bagasse, rice chaff and coir dust in this order, irrespective of soil moisture and temperature level. In all waste materials, the rate of decrease in weight was highest in the soil holding the water equivalent to field capacity (saturated soil) followed by submerged soil and dry soil in this order. CO2 generation rate was also highest in rice straw followed by bagasse, rice chaff and coir dust. It was highest in saturated soil followed by half-saturated or submerged soil and dry soil in this order. The rate of CO2 generation from rice straw in saturated soil was highest at the initial period of incubation and it decreased thereafter, but the rate in submerged soil was highest at 40 and 20 days after the start of incubation at 20 and 35ºC, respectively. The rate of CO2 generation from coir dust and rice chaff was very low at all soil moisture levels at either 20 or 35ºC. The content of total N in the waste materials was positively and significantly correlated with the rate of decrease in weight in saturated and submerged soils at a moderate temperature (Oct. – Dec.), and in submerged soil at a high temperature (Aug. – Oct.). It was also significantly correlated with CO2 generation rate in submerged soil at 20ºC. Holocellulose and hemicellulose contents were negatively and significantly correlated with CO2 generation rate in dry soil at 20ºC. Lignin content was also significantly and negatively correlated with CO2 generation in dry soil at 35ºC.

Ramet production byMallotus japonicus, a common pioneer tree in temperate Japan

To clarify the characteristics of clonal growth ofMallotus japonicus, we excavated all roots of this species in a plot and ascertained the connections between shoots. All shoots examined were ramets that originated from roots. Young ramets grew in shady sites as well as sunnier sites, indicating that ramet production was not dependent on local light conditions.