<Emphasis Type="Italic">In vitro</Emphasis> screening of sugarcane to evaluate smut susceptibility

Tissue-cultured plantlets of three sugarcane (Saccharum spp.) cultivars having a known field smut reaction were screened for susceptibility to Ustilago scitaminea H&P Sydow. Plantlets were inoculated with 0.5 l of a suspension of equally mixed quantities of plus and minus mating type sporidia of U. scitaminea at concentrations ranging from 1×10¹ to 1×10⁶ cells. Fungal sori (whips) were produced in cultivar N12 (intermediate) 6weeks following inoculation with 1×10⁵ mixed sporidia and thereafter in cultivar NCo310 (susceptible) but not in cultivar N19 (resistant). Sori bearing teliospores were produced up to 3months following inoculation and incubation at 26°C. No sori were produced at mixed sporidial concentrations lower than 1×10⁵cells. The in vitro soral production in cultivars N19, N12 and NCo310 was 0, 27.5 and 47.5% respectively. Plantlets inoculated with 1×105sporidia of only one mating-type did not produce sori in any of the three cultivars tested. Blind scoring of an unknown sugarcane cultivar by this method corresponded exactly with its field smut rating.     

Competition between <Emphasis Type="Italic">Eucalyptus victrix</Emphasis> seedlings and grass species

Competition in a natural system may be interspecific or intraspecific. In semiarid ecosystems, competition for resources between established neighboring grass species and newly recruited seedlings is very high. To examine the effects of grass species density, growing space and time of establishment on Eucalyptus victrix seedlings (interspecific competition), and the effect of density and growing space within E.victrix (intraspecific competition) we conducted an experiment under controlled conditions. We tested four hypotheses (i) E.victrix seedling growth is not affected by grass density; (ii) there is no difference in E.victrix survival and growth between early and later grass establishment; (iii) interspecific competition is not more intense than intraspecific competition in E.victrix; and (iv) growth of E.victrix seedlings is not dependent on available growing space. In a monoculture of E.victrix, seedling mortality was higher (10%) in large pots. In mixed culture pots, where E.victrix seedlings and grass seedlings were planted on the same day, E.victrix seedlings survived for up to 4weeks, but started to die after week five in the smallest pots. However, mortalities occurred in pots of all sizes when grass was established before E.victrix seedlings. Results also indicated that the resources necessary for the growth of individual E.victrix seedlings were more limiting under conditions of increased density of neighboring grass species rather than intraspecific competition. In particular, photosynthetic area of E.victrix seedlings was drastically reduced in mixed cultures. Although density, pot size and time of planting had impacts on E.victrix seedlings, the patterns of these impacts were variable.     

Assessment of within-population genetic structure in <Emphasis Type="Italic">Quercus crispula</Emphasis> and <Emphasis Type="Italic">Q. dentata</Emphasis> by amplified fragment length polymorphism analysis

The spatial genetic structure was assessed by amplified fragment length polymorphism (AFLP) technique for Quercus crispula Blume (37 individuals) and Q.dentata Thunberg (54 individuals) growing along a 550-m transect in Ishikari, Hokkaido, northern Japan. The simple Mantel test revealed significant negative correlation between genetic similarity and geographic distance in Q.dentata and negative but insignificant correlation in Q.crispula. Spatial autocorrelation analysis revealed that Mantels r generally decreased from positive to negative values with the increase of spatial distance in both oak species with significant deviation from zero for the 50–100-m (positive) and 250–300-m classes (negative) in Q.dentata. Thus, significant spatial genetic structure was revealed at least in Q.dentata     

Development of high frequency multiple shoot formation in Persian clover (<Emphasis Type="Italic">Trifolium resupinatum</Emphasis> L.)

An efficient and reliable micropropagation system for Persian clover (Trifolium resupinatum L.) was developed using different explants and media. Node, hypocotyl and cotyledonary node explants were cultured on Murashige and Skoog (MS) medium supplemented with combinations of either 6-benzyladenine (BA) and indole-3-butyric acid (IBA) or BA, Kinetin (KIN) and IBA. Direct multiple shoots developed within 6weeks in all explants in most media tested. The best shoot multiplication capacity was obtained from cotyledonary node explants on MS medium containing 7.1M BA and 1M IBA or 14.1M BA and 1M IBA. Elongated shoots were rooted on either MS medium alone or combination with different concentrations of indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) and -naphthaleneacetic acid (NAA). High rooting was achieved in half strength MS medium containing 8M IBA.     

Early development of the self-fertilizing mangrove killifish <Emphasis Type="Italic">Rivulus marmoratus</Emphasis> reared in the laboratory

The mangrove killifish Rivulus marmoratus was reared at 25°±1°C and 17ppt salinity from 0 to 100 days after hatching (DAH), and its early development was described by examining growth and morphometric parameters, meristic characters (vertebral and fin-ray counts), bone-cartilage development, and pigmentation. Growth was isometric for preanal length, head length, snout length, body depth, pectoral-fin length, dorsal-fin length, anal-fin length, and caudal-peduncle depth. Negative allometric growth was observed in eye diameter and gape size. Meristic counts (mean±SD) for vertebrae (34.2±0.4) and dorsal- (8.6±0.5), anal- (11.4±0.5), and caudal-fin rays (30.2±0.8) were complete at 0 DAH (n=5), whereas pectoral-fin rays and pelvic-fin rays were complete by 30 DAH (14.5±0.4, n=5) and 60 DAH (4.2±0.8, n=5). Full ossification of meristic elements proceeded in the following sequence: vertebrae (by 30 DAH), caudal-, dorsal-, and anal-fin rays (by 60 DAH), pectoral-fin rays (between 60 DAH and 100 DAH), and pelvic-fin rays (by 100 DAH). Both morphological characters and meristic counts indicate that this species can be considered to be a juvenile after 9.8mm in standard length (20 DAH).     

Efficiency of herbivore exclusion by ants attracted to aphids on the vetch <Emphasis Type="Italic">Vicia angustifolia</Emphasis> L. (Leguminosae)

The efficiency of herbivore exclusion by ants on the vetch Vicia angustifolia L. (Leguminosae) with extrafloral nectary, mediated by ant attraction to aphids was investigated in a field census and laboratory experiments. In the field, workers of Lasius japonicus Santschi and Tetramorium tsushimae Emery frequently visited plants of the vetch parasitized by aphids of Aphis craccivora Koch, but only a few workers visited plants without aphids. An increase in the number of ants visiting a plant with increasing numbers of aphids caused a decrease in the number of larvae of the weevil, Hypera postica Gyllenhal. Therefore, the efficiency of herbivore exclusion by ants was higher on plants parasitized by Ap.craccivora aphids than that on plants unparasitized by aphids. In the laboratory experiments, L.japonicus workers frequently patrolled not only shoots with Ap.craccivora aphids but also shoots without them. However, T.tsushimae workers visited mainly shoots with Ap.craccivora aphids but less frequently on shoots without aphids. Therefore, L.japonicus workers excluded herbivores more efficiently on plants of the vetch than T.tsushimae workers. Consequently, the efficiency of herbivore exclusion by ants on the vetch can be influenced directly by differences in ant species and indirectly by the presence of aphids on plants. The present study highlights the significance of indirect interactions between ants and plants with extrafloral nectary, mediated by ant attraction to aphids for herbivore exclusion of plants.     

Fine root biomass,production and turnover in a fertilized <Emphasis Type="Italic">Larix leptolepis</Emphasis> plantation in central Korea

The effects of fertilization [control (C), 200kgNha^–1+25kgP ha^–1 (LNP) and 400kgNha^–1+ 50kgP ha^–1 (HNP)] on fine root dynamics were examined in a 40-year-old Larix leptolepis plantation in central Korea. The average fine root biomass during the growing season for C, LNP and HNP was 957, 934 and 814kgha^–1, respectively, whereas the fine root production for C, LNP and HNP was 2103, 2131 and 2066kgha^–1, respectively. Nitrogen and P inputs into the soil via fine root turnover for C, LNP and HNP were 23.0 and 1.2, 23.3 and 1.2 and 22.6 and 1.2kgha^–1, respectively. There were no significant differences in fine root biomass, production and N and P inputs through fine root turnover between the fertilization treatments during the first growing season after fertilization.     

Carbon dynamics and budget in a <Emphasis Type="Italic">Miscanthus sinensis</Emphasis> grassland in Japan

We investigated the carbon dynamics and budget in a grassland of Miscanthus sinensis, which is widely distributed in Japan, over a 2-year period (2000–2001). Plant biomass began to increase from May and peaked in September, then decreased towards the end of the growing season (October). Soil respiration rates also exhibited seasonal fluctuations that reflected seasonal changes in soil temperature and root respiration. The contribution of root respiration to total soil respiration was 22–41% in spring and summer, but increased to 52–53% in September. To determine the net ecosystem production (carbon budget), we estimated annual net primary production, soil respiration, and root respiration. Net primary production was 1207 and 1140gCm^–2 in 2000 and 2001, respectively. Annual soil respiration was 1387gCm^–2 in 2000 and 1408gCm^–2 in 2001; root respiration was 649 and 695gCm^–2 in 2000 and 2001, respectively. Moreover, some of the carbon fixed as net production (457–459gCm^–2) is removed by mowing in autumn in this grassland. Therefore, the annual carbon budget was estimated to be –56gCm^–2 in 2000 and – 100gCm^–2 in 2001. These results suggest that the Miscanthus sinensis grassland in Japan can act as a source of CO₂.     

Consequences of harvesting for genetic diversity in American ginseng (<Emphasis Type="Italic">Panax quinquefolius</Emphasis> L.): a simulation study

American ginseng, Panax quinquefolius L., is one of the most heavily traded medicinal plants in North America. The effect of harvest on genetic diversity in ginseng was measured with a single generation culling simulation program. Culling scenarios included random harvest at varying levels, legal limit random harvest and legal limit mature plant harvest. The legal limit was determined by the proportion of legally harvestable plants per population (% mature plants per population). Random harvest at varying levels resulted in significant loss of genetic diversity, especially allelic richness. Relative to initial levels, average within-population genetic diversity (H _e) was significantly lower when plants were culled randomly at the legal limit (Mann–Whitney U=430, p<0.001) or when only mature plants were culled (Mann–Whitney U=394, p<0.01). Within-population genetic diversity was significantly higher with legal limit mature plant harvest (H _e=0.068) than when plants were culled randomly at the legal limit (H _e=0.064; U=202, p<0.01). Based on these simulations of harvest over one generation, we recommend that harvesting fewer than the proportion of mature plants could reduce the negative genetic effects of harvest on ginseng populations.     

Effects of sex and age of a dioecious tree, <Emphasis Type="Italic">Forchhammeria pallida</Emphasis> (Capparaceae) on the performance of its primary herbivore, <Emphasis Type="Italic">Murgantia varicolor</Emphasis> (Hemiptera: Pentatomidae)

The effects of sex and age of a dioecious tree, Forchhammeria pallida, on the survival and development of a Mexican harlequin bug, Murgantia varicolor, were examined. In the laboratory, bugs reared on fruits or fruits and leaves developed rapidly and had a high rate of survivorship, whereas bugs reared on leaves alone developed slowly after the second instar and died by the fifth instar. Murgantia varicolor showed a preference for leaves from juvenile plants over leaves from mature plants, and bugs reared on leaves from juvenile plants survived significantly longer than those reared on leaves from mature plants. Murgantia varicolor showed no preference for feeding on leaves of mature male versus mature female plants. In the field, M.varicolor occurred on male, female and juvenile plants. Although nymphs were less common on male than female F.pallida, their presence in substantial numbers shows that M.varicolor oviposits on both male and female plants. However, adult M.varicolor were rare on male plants. This result is supported by the inability of nymphs to complete their development on vegetative parts of F.pallida in the laboratory. These observations suggest that dioecy in F.pallida may be disruptive to populations of M.varicolor.     

<Emphasis Type="Italic">Uncisudis posteropelvis</Emphasis>, a new species of barracudina (Aulopiformes: Paralepididae) from the western North Pacific Ocean

Six specimens (2 flexion larvae: 9.5–10.4mm in notochord length; 4 postflexion larvae: 12.3–18.2mm in standard length) collected from the western North Pacific are tentatively ascribed to the genus Uncisudis of the tribe Lestidiini of the subfamily Paralepidinae (Paralepididae) in sharing remarkably elongate and filamentous pelvic fin rays, their tips reaching the origin of the anal fin. They are described as Uncisudis posteropelvis sp. nov. in uniquely having the insertion of pelvic fins closer to the origin of anal fin than to the posterior end of dorsal fin base among lestidiine species. Addition to this character, the new species has remarkably elongate and filamentous dorsal fin rays, the short distance between anus and origin of anal fin (4.2–6.1% of standard length, SL), the posteriorly located pelvic fins (prepelvic length 69.4–71.5% SL), dorsal fin rays 10, anal fin rays 28–29, myomeres 41–42+38–40=80–81 (vertebrae 38+41=79), and peritoneal pigment spots 11–12. The occurrence of larvae differing in pigment pattern from the present new species suggests another undescribed species of Uncisudis in the western South Pacific.     

A novel psychrotrophic fungus, <Emphasis Type="Italic">Mortierella minutissima</Emphasis>, for <Emphasis Type="BoldSmallCaps">D</Emphasis>-limonene biotransformation

Of 98 strains of moulds, isolated from arctic soils, Mortierella minutissima 01, grew the best on agar plates with limonene vapor. Perillyl alcohol and perillic acid were the main products of limonene biotransformation. Maximal yield of perillyl alcohol (125mgl^–1) occurred in medium containing 0.8% substrate, at 15°C, pH 6 and after 4–5 d. Revisions requested 27 October 2004; Revisions received 27 November 2004     

Larval development and food habits of the marbled parrotfish, <Emphasis Type="Italic">Leptoscarus vaigiensis</Emphasis>, associated with drifting algae

The larval development and food habits of the marbled parrotfish, Leptoscarus vaigiensis (Scaridae) associated with drifting algae were studied. In this study, 628 L. vaigiensis of various developmental stages ranging from postflexion larvae (9.4mm in standard length, SL) to adults (192.0mmSL) were sampled from drifting algae at two fishing ports in Nakagusuku Bay of Okinawa Island. In 3969 fish comprising 65 taxa in 34 families of Teleostei collected with drifting algae, L. vaigiensis occupied 15.8% of samples and occurred generally throughout the whole year. A large number of L. vaigiensis were collected from July to October accompanied by an occurrence of drifting algae composed of Sargassum spp. Larvae and early juveniles ranging from 11.1 to 14.9mmSL appeared sporadically throughout the year, and postflexion larvae 11mmSL occurred from July to November. Their food shifted from planktonic copepods in postflexion larvae and juveniles ranging from 10.0 to 14.9mmSL to seaweed in the juveniles ranging from 15.0 to 24.9mmSL. Furthermore, adults and young over 25mmSL fed almost exclusively on seaweed, with Sargassum spp. constituting the drifting algae. These facts indicate that drifting algae may have a role concerning food and habitat, and may act as a nursery for L. vaigiensis.     

Effects of domestication and agronomic selection on phytoalexin antifungal defense in <Emphasis Type="Italic">Phaseolus</Emphasis> beans

Systems of wild and cultivated relatives are good experimental systems to test chemical defense theory because they provide closely related varieties that differ in discrete traits. To determine the relationship between resistance and chemical defense diversity among wild, landrace and cultivar accessions of Phaseolus vulgaris, we measured resistance to fungal infection in laboratory and field experiments, quantified phytoalexin diversity, and assessed the effectiveness of phytoalexin mixtures extracted from live tissue. Results show a gradient of resistance to fungal infections between wild, landrace and cultivar varieties. In the laboratory, wild seedlings were more resistant (93% non-infected) than landrace seedlings (80% non-infected) and modern cultivar seedlings (68% non-infected). Under field conditions wild seedlings were more resistant (97% non-infected) than cultivar seedlings (71% non-infected). Wild seedlings presented the highest phytoalexin diversity (H=1.11), while those of the landrace presented an intermediate level (H=0.97) and cultivar seedlings presented the lowest diversity (H=0.93). No differences were found in total concentrations. The in vitro inhibitory properties on hyphal growth of the isoflavonoid mixtures produced by individual seedlings showed a similar trend. Our results are consistent with similar gradients in other species of Phaseolus beans and resistance to Colletotrichum sublineolum in sorghum.     

Effects of population size on genetic diversity and seed production in the rare <Emphasis Type="Italic">Dictamnus albus</Emphasis>(Rutaceae) in central Germany

It is widely assumed that population size significantly affects the dynamics of plant populations. Smaller populations are threatened by genetic drift and inbreeding depression, both of which may result in a decrease of genetic variation and a resulting negative impact on plant fitness. In our study we analysed the patterns of random amplified polymorphic DNA (RAPD) variation among 10 Dictamnus albuspopulations of varying size. The aim was to examine local differentiation in relation to spatial isolation resulting from limited population size and geographical distancing between populations. Significant correlations were noted between population size and both percentage of polymorphic loci (P <0.01) and genetic diversity (P<0.01). The matrix correlation between genetic and geographical distances revealed that geographical differentiation was reflected in the RAPD profile (Mantel test: r²=0.34, P<0.001). We found the highest level of molecular variance of RAPD patterns among individuals within the populations (72.6%), whereas among-population variation accounted for only 21.6% of variation. These results were highly significant in that they indicated a restricted population differentiation, as would be expected from outcrossing species. An additional analysis of seed production showed that there was significant variation among populations in terms of mean seed number per flower and mean seed mass per population which could be attributed to differences in population size as well as levels of genetic variation.     

Population density and group composition of Japanese sika deer (<Emphasis Type="Italic">Cervus nippon yakushimae</Emphasis>) in an evergreen broad-leaved forest in Yakushima,southern Japan

The population density of Japanese sika deer (Cervus nippon yakushimae Kuroda and Okada) in an evergreen broad-leaved forest in Yakushima, southern Japan, was surveyed over 4years from 1998 to 2001. Two approximately 50ha study sites, Hanyama and Kawahara, were established with a total of 4km of census trails at each site. The estimated densities of sika deer at the two sites were 43–70 deerkm^–2 at Hanyama and 63–78 deerkm^–2 at Kawahara, although these values might be underestimates. The adult sex ratio (number of adult males:number of adult females) ranged from 0.6 to 1.0 at Hanyama, and from 0.4 to 0.9 at Kawahara. Mean group size was 1.9 deer (male group, 1.5 deer; female group, 1.6 deer; mixed group, 3.6 deer). The population density of sika deer was relatively high compared to other sites in Japan, with the exception of very small (<10km²) islands. Possible explanations for this naturally high density of sika deer in an evergreen broad-leaved forest in Yakushima are discussed.     

Descriptive morphology of the reared eggs,larvae, and juveniles of the marine atherinid fish <Emphasis Type="Italic">Atherinomorus duodecimalis</Emphasis>

Embryonic, larval, and juvenile development of the marine atherinid Atherinomorus duodecimalis are described from laboratory-reared specimens. The eggs, measuring 1.15–1.24mm in diameter, were demersal and almost spherical in shape with numerous chorionic filaments. Hatching occurred 7 or 8 days after spawning, the newly hatched larvae measuring 4.5–5.1mm in body length (BL) and having 5+34=39 myomeres. Notochord flexion started at 6.9mm BL and finished at 8.3mm BL. Aggregate numbers of all fin rays were completed by 14mm BL. Eggs of this species could be distinguished from other known atherinid eggs on the basis of egg diameter and the arrangement, length, and number of chorionic filaments. Larvae were also distinctive in myomere counts, pigmentation, and locations of the anus and second dorsal fin origin.     

Mycelial growth of strains of the genera <Emphasis Type="Italic">Suillus</Emphasis> and <Emphasis Type="Italic">Boletinus</Emphasis> in media with a wide range of concentrations of carbon and nitrogen sources

Suillus and Boletinus were studied using Ohta medium. In media with glucose or trehalose, all tested strains grew well. With mannose and cellobiose, strains generally grew well, except for one strain of Suillus. Utilization of dextrin and soluble starch differed with each strain, and that of sucrose and glycerol was low for all strains. Utilization of four amino acids, arginine, glutamic acid, aspartic acid, and alanine, was similar to that of ammonium tartrate for Suillus strains, but mycelial growth with amino acids was clearly lower than with ammonium tartrate for the Boletinus strain. The effect of glucose and ammonium tartrate concentrations for nine strains of the genera Suillus and Boletinus was studied with ranges for glucose of 1–100 and 200g/l, respectively, and for ammonium tartrate of 0.2–5 and 20g/l, respectively. Six strains showed maximal growth at a glucose concentration greater than 25g/l, and one strain showed maximal growth at 70g/l. The results indicate that these fungi are adapted to relatively high concentrations of carbon sources. In general, glucose concentration at mycelial growth maximum decreased as ammonium tartrate concentration increased, and at higher concentrations of glucose, mycelial growth decreased more rapidly in higher concentrations of ammonium tartrate.     

Milk-clotting protease production by <Emphasis Type="Italic">Nocardiopsis</Emphasis> sp. in an inexpensive medium

The milk-clotting and proteolytic activities of extracellular enzyme preparations from Nocardiopsis sp. were investigated under different culture conditions. A soybean flour medium was used, with concentrations of soybean flour and of glucose varying from 0.25 to 1% w/v and from 0 to 1% w/v, respectively. Growth was monitored with 2ml samples withdrawn from the culture medium at 8-h intervals, for determination of total protein, proteolytic activity, milk-clotting activity and sugar reduction. The best milk-clotting protease production, with a specific activity of 24.49U/mg at 40h, was obtained in the glucose-free medium containing soybean flour 1% w/v.     

Significance of flower exploding pollination on the reproduction of the Scotch broom, <Emphasis Type="Italic">Cytisus scoparius</Emphasis> (Leguminosae)

The significance of single flower visits by pollinators on the reproductive success of the Scotch broom, Cytisus scoparius, with explosive flowers was investigated. Unexploded flowers (intact flowers) produced no fruit, implying that autonomous selfing and apomixis do not occur and that the explosion of flowers by insects is necessary for the fertilization of flowers. The fruit set in the natural condition was intermediate between that in artificial selfing or exploding and that in artificial outcrossing. Seed set and the ratio of seeds to ovules did not differ significantly between the natural condition and artificial outcrossing. The fruit set by a single flower visit from Apis mellifera was similar to natural conditions. Apis mellifera was the most frequent visitor and the number of flowers tripped open per hour was the highest among all the flower visitors. Approximately 75% of pollen on a flower disappeared in a single flower visit by A.mellifera. The small amount of pollen remaining on already-exploded flowers appears to be the reason why A.mellifera do not frequently visit already-exploded flowers. The higher seed set by outcrossing was believed to contribute to the seed set in both natural conditions and single flower visits by A.mellifera, and did not differ between them. Therefore, at least in Japan, A.mellifera appears to be the most important pollinator for C.scoparius, resulting in a high outcrossing rate with each single flower visit.