Cellular and humoral immune defenses of Oxya japonica (Orthoptera: Acrididae) to entomopathogenic fungi Metarhizium anisopliae

One Indonesian isolate of the fungus Metarhizium anisopliae, named Majalengka strain, was evaluated not only for its virulence but also for the immune response of rice grasshopper Oxya japonica (Orthoptera: Acrididae) as a target organism. Five aqueous suspensions with different conidia concentrations in logarithmic series were prepared. The fungus showed high virulence as it caused 100% mortality at low conidia concentration (1.5 × 10² conidia/mL). Remarkable changes in the cellular and humoral responses were also observed when adult grasshoppers were infected with the fungus. The number of hemocytes decreased significantly within 12 h after infection. In addition, the total number of granulocytes increased rapidly in the first 12 h then gradually decreased 24 and 48 h after infections, while the number of coagulocytes fluctuated over time. The infection influenced the humoral response by increasing the phenoloxidase activity.     

Taxonomy and geographical distribution of Dugesia japonica and D. ryukyuensis in the Far East

Dugesia japonica Ichikawa et Kawakatsu, 1964, is a common and polymorphic species of freshwater planarian distributed widely in the Far East. In 1976 the geographic populations were separated into 2 subspecies (D.j.japonica and D.j. ryukyuensis). The taxonomy of this species is reconsidered once again from the morphological, anatomical, histological, and karyological viewpoints. From the result of these studies, D.j. ryukyuensis is elevated to the rank of species: D. ryukyuensis Kawakatsu, 1976. D. japonica (n = 8, 2x = 16, 3x = 24) differs from D. ryukyuensis (n = 7, 2x = 14, 3x = 21) in having an asymmetrical penis papilla without a well-developed valve surrounding its basal part, and a well-developed vagina (distribution: the Japanese Islands, Taiwan, the Korean Peninsula, China, and Primorskiy, Northeast Siberia, in Russia). D. ryukyuensis is characterized by an asymmetrical penis papilla with a well-developed valve surrounding its basal part, and a less-developed vagina (distribution: the Southwest Islands of Japan).     

Spatial genetic structure within two populations of a self-incompatible perennial,Chionographis japonica var.japonica (Liliaceae)

Intrapopulational spatial genetic structure was examined in two populations ofChionographis japonica var.japonica, a self-incompatible perennial, by spatial autocorrelation analysis of enzyme polymorphism. Although most spatial autocorrelation indices (Moran'sI) in the shortes distance class were significantly positive, most in the other distance classes did not significantly deviate from the values expected from random distributions of genotypes in both populations. This contrasts with a spatial genetic pattern previously reported for a population of the predominantly selfing congener,C. japonica var.kurohimensis, indicating that pollen-mediated gene flow highly impedes genetic substructuring within populations of outcrossingC. japonica var.japonica. Genetic similarity in very proximate distance found in outcrossingC. japonica var.japonica is probably due to restricted dispersal of seeds.     

中华稻蝗四个种群的遗传分化

采用水平淀粉凝胶电泳技术研究中华稻蝗(Oxya chinensis)四个种群的12个基因座位,探讨其遗传分化.这四个种群分别采自内蒙古呼和浩特、山西代县、山西太原和陕西西安.Ck和Mdh-2在四个种群中均为单态,其余的基因座位至少在一个种群内有两个以上的等位基因;在Ldh和Mdh-1的等位基因频率呈现梯度分布趋势.多态基因座位百分率(P)和平均每个基因座位的等位基因数目(A)分别为58.3%-66.7%和2.2-2.8,平均杂合度为Ho=0.173-0.240.除Gpi,Hk-2,Idh,Ldh和Mdh-1在部分蝗虫种群符合Hardy-Weinberg平衡外,其余绝大多数基因座位的基因型频率显著偏离Hardy-Weinberg平衡.四个种群间FST平均值不存在显著差异(FST=0.0510,P＞0.05),结合高的Nei's遗传一致度(I＞0.97)可知,种群之间遗传分化不明显.我们认为人类的农业活动可能促进了种群间的基因交流,从而降低了分化程度[动物学报50(2)187-192,2004].     

Genetic diversity of six populations of Atractomorpha sinensis and Atractomorpha peregrina in Shanxi Province, China

Ten enzymes (AAT, CK, G3PDH, HEX, IDH, LDH, MDH, ME, PGI, PGM) were examined using horizontal starch gel electrophoresis to estimate the levels of genetic variation within and among six natural populations of two grasshopper species Atractomorpha sinensis and A. peregrina from Shanxi, China. The collecting sites were geographically distant from each other from south to north: Quwo district, Linfen city; Xiangyuan county, Changzhi; Jinyuan district, Taiyuan city; Yuanping county, Xinzhou city and Fanshi county of Xinzhou.A. sinensis showed 43 alleles at 16 loci but A. peregrine showed 39 alleles at 15 loci (Idh-1 was deficient). The zymograms showed that some common alleles were shared at several loci in these two species (Aat-1-b, Aat-2-b, G3pdh-a, Ck-1-b and Ldh-b). However, Hex-1-a, Hex-2-a, Hex-3-a, Idh-2-b, Mdh-2-b, Mdh-1-f, Pgi-b, Pgm-b had common alles in A. sinensis and Hex-1-b, Hex-2-b, Hex-3-b, Idh-2-a, Mdh-2-a, Mdh-1-d, Pgi-a, Pgm-c were of high frequency in A. peregrine instead. Most of the observed genotype frequencies were found to significantly deviate from the Hardy-Weinberg expectations in both species. A tendency of clinal distribution of allele frequency was observed at three loci. The frequency of the moderately migrating allele Me-c (0.318–0.740) in A. peregrina, Hex-1-a (0.800–1.000) and Ldh-b (0.487–0.750) in A. sinensis demonstrated increased frequency from north to south. Such tendency suggests that the allele frequency in these three loci may be correlated with the species’ geographic distributions. A. sinensis showed higher genetic diversity than A. peregrina as indicated by higher mean number of alleles per locus (A = 1.9–2.3 in A. sinensis and 1.7–2.2 in A. peregrina), percentage of polymorphic loci (56.3%–68.8% in A. sinensis and 43.8%–56.3% in A. peregrina), and the observed heterozygosities (H _o = 0.072–0.096 in A. sinensis and 0.70–0.107 in A. peregrina). The observed heterozygosities of the six populations were all noticeably lower than the Hardy-Weinberg expectations, mostly due to heterozygote deficiency in the populations of both species. The overall mean F _ST were small (F _ST = 0.045, P > 0.05 in A. sinensis populations and 0.087, P > 0.05 in A. peregrina populations). Nei’s genetic identity (I) estimates indicate low intraspecific (>0.95) but higher interspecific (0.377–0.447) genetic diversity. The cluster analysis based on modified Roger’s genetic distance (D) showed that the two species were divided into two branches. Both species are of limited dispersal capacity and a moderate geographical barrier might significantly restrict the gene exchange among populations, resulting in accumulation of local genetic differentiations. The A. sinensis populations used in this study were separated from each other by 155.2 to 271.4 km and the A. peregrina populations were separated from each other by 78.8 to 174.9 km with observable physical barriers. The allozyme data showed only minimal genetic differentiation at population level, most likely as a result of gene exchange. It is reasoned that natural factors and human agricultural activities might have facilitated migration and dispersal for the two species. __________ Translated from Acta Ecologica Sinica, 2005, 25(10): 2574–2481 [译自: 生态学报]     

Morphometrics reveal correlation between morphology and bioclimatic factors and population mixture in Tetrix japonica (Orthoptera: Tetrigidae)

The morphology of an organism is limited by genetic and environmental factors, and the precise mechanism is inconsistent between species. Tetrix japonica Bolivar, 1987, is a widely distributed pygmy grasshopper in East Asia. However, the population clustering and relationships between the morphology and bioclimatic factors have not been previously investigated. Here, 32 geographic populations were sampled from China, and morphometrics and multiple statistical analyses were applied to detect the population clustering and relationship between the morphology and bioclimatic factors. The results suggested that T. japonica with females are significantly bigger than males in the eight morphological traits. The 32 populations do not obviously cluster according to the natural geographic area. The body sizes of females are mainly related to the lowest temperature and precipitation; by contrast, males only have a significant relationship with the lowest temperature. The forewing size is significantly related to the maximum precipitation. Furthermore, the Mantel test showed that the morphological size variation of females has a weak positive correlation to geographic distance, but is insignificant in males. It was concluded that Chinese geographic populations of T. japonica mixed and that the size of the morphological structure is limited by bioclimatic factors.     

蝗总科部分种类等位基因酶的比较研究

用水平淀粉凝胶电泳技术对采自山西太原黄陵、山西临猗伍姓湖及山西雁门关两科4种蝗虫的4个等位基因酶位点的基因频率进行了比较研究,并用BIOSYS-Ⅱ软件进行结果分析。结果表明：所研究种群的苹果酸脱氢酶（MDH）都存在两个位点,中华稻蝗的MDH-1还存在两条亚带。在所研究种群中的MDH-1图谱中,一个中等迁移率的谱带存在于所研究的4个种群中。东亚飞蝗在乳酸脱氢酶（LDH）和苹果酸酶（ME）中存在两个固定的等位基因,同时黄胫小车蝗在MDH-1中存在一个固定的等位基因,在MDH-2中存在一个独特的等位基因。在所有4个种群中,中华稻蝗遗传多样性水平最高（每个位点的等位基因数为3个,He=0.220）,黄胫小车蝗遗传多样性水平最低（每个位点的等位基因数为1.5个,He=0.013）。除中华稻蝗的MDH-1和LDH处于哈代-温伯格平衡,其余种群的4个位点的等位基因频率均不同程度的偏离哈代-温伯格平衡。等位酶数据表明这4个种群在系统发育关系方面是相近的,但在遗传多样性水平上却不同。     

Karyological and taxonomic studies of Dugesia japonica Ichikawa et Kawakatsu in the Far East

A review of previous studies on the taxonomy, karyology and chorology of a polymorphic species Dugesia japonica from the Far East is presented. Two subspecies are now known: D. j. japonica (n = 8, 2x = 16, 3x = 24) and D. j. ryukyuensis (n = 7, 2x = 14, 3x = 21). An attempt has also been made to determine the definition of the B-chromosome as LB and SB and the variation of the karyotypes of both subspecies is described. Every known karyotype of D. japonica is classified into six groups (see Table 2). D. japonica from many localities has a diploid karyotype (2x), a triploid karyotype (3x) and an orthoploidic mixoploid karyotype of 2x & 3x. The origin and the karyological significance of these karyotypes are discussed.     

Genetic diversity and multilocus associations in Cunninghamia lanceolata (Lamb.) Hook from The People's Republic of China

Open-pollinated seeds were assayed for allozyme polymorphisms at 24 loci to assess genetic diversity and multilocous associations in 16 populations of Cunninghamia lanceolata (Lamb.) Hook in the People's Republic of China. On average, the percentage of polymorphic loci was 88.0, the number of alleles per locus was 3.0, and the expected heterozygosity was 0.394. The distribution of genetic diversity was not correlated with the geographic and climatic variables of the populations. However, allele frequencies correlated linearly with the mean annual temperature of the populations at Mdh-1, Mdh-2, Mnr-2, Pgi-1, and Skdh-1 and with the altitude of the populations at Aph-4 and 6Pg-2. Of the total gene diversity 6% was attributed to among-population differentiation; 94% resided within populations. Two-locus gametic disequilibria were found in 15 of the 16 populations, and higher-order gametic disquilibria were significant in most populations. The gametic disequilibria did not correlate with geographic and climatic variables. The results suggest that population subdivision, founder effect, occurrence across diverse environments, a mating system dominated by inbreeding, and historical events from 2000 years of cultivation are contributing factors in the generation and maintenance of the multilocus genetic structure in this conifer.     

Genetic variation and population structure in Korean populations of eurya japonica (Theaceae)

We investigated levels of genetic diversity, population genetic structure, and gene flow in Eurya japonica, a widespread and broad-leaved evergreen dioecious tree native to Japan, China, Taiwan, and the southern and southwestern coast of the Korean Peninsula. Starch-gel electrophoresis was conducted on leaves collected from 1,000 plants in 20 Korean populations. All 12 loci examined were polymorphic in at least one population, and the mean number of alleles per locus was 3.79. In addition, mean observed population heterozygosity (Ho_p = 0.425), expected heterozygosity (He_p = 0.462), and total genetic diversity (H_T = 0.496) were substantially higher than average values for species with similar life history traits. Although significant differences in allele frequency were detected between populations at all loci (P < 0.001), <7% of the genetic variation was found among populations (F_ST = 0.069). There was a significant negative correlation between genetic identity and distance between populations (r = -0.341; P < 0.05), but this explained only a small amount of the diversity among populations. Indirect estimates of the number of migrants per generation (Nm) (3.37, calculated from F_ST; 3.74, calculated from the mean frequency of eight private alleles) indicate that gene flow is extensive among Korean populations of E. japonica. Factors contributing to the high levels of genetic diversity found within populations of E. japonica include large and contiguous populations, obligating outcrossing (dioecious plant), high fecundity, and long generation time. Occasional seed dispersal by humans and pollen movement by domesticated honey bees may further enhance gene flow within the species.     

Aspartate aminotransferase isozymes in the genusCapsella (Brassicaceae): Subcellular location,gene duplication,and polymorphism

The subcellular location of aspartate aminotransferase isozymes (EC 2.6.1.1) in the genusCapsella(Brassicaceae) was studied. The diploid speciesC. grandiflora andC. rubella have three AAT isozymes, including one located in the plastids. Each locus is duplicated in the tetraploidCapsella bursa-pastoris. Variation at the plastid-coding locus exceeded that at the other loci.C. bursa-pastoris had some unique alleles not detected in the diploid species. Segregation in open-pollinated families revealed thatCapsella grandiflora was outcrossing, whereasC. rubella was highly inbred, with most populations homozygous or uniform at all three loci. Inheritance in the tetraploid colonizerC. bursa-pastoris is disomic. This species was also predominantly selfing with outcrossing rates between 2% and 10%.Financial support by the German Research Foundation DFG is gratefully acknowledged.     

Karyomorphological study of the Spiraea japonica complex (Rosaceae)

Reported in this paper is a karyomorphological study on one natural population of each of eight varieties in theSpiraea japonica complex. The interphase and mitotic prophase can be classified into the simple chromocenter type and the proximal type, respectively. The metaphase karyotypes of the eight varieties were formulated as follows:S. japonica var.japonica: 2n=18=14m+2sm+2st;S. japonica var.acuta: 2n=18=11m+4sm+3st;S. japonica var.incisa: 2n=18=12m+4sm+2st;S. japonica var.stellaris: 2n=18=15m+1sm+2st;S. japonica var.acuminata: 2n=18=14m+2sm+2st;S. japonica var.ovalifolia: 2n=18=10m+2sm+2st+4t;S. japonica var.glabra: 2n=18=10m+4sm+4st;S. japonica var.fortunei: 2n=36=17m+16sm+3st. Karyomorphological study reveals that the chromosome numbers within the complex are stable, and that karyomorphological divergence between the varieties lies mainly in chromosome size and organization. Based on the karyomorphological data and geographical distribution of the complex, the differentiation pattern as well as evolutionary mechanism of the complex is evaluated.     

Estimation of prey consumption of a mantid,Paratenodera angustipennis (S.) in a natural habitat

Using a relationship between prey consumption and growth rate, field prey consumption of adults of Paratenodera angustipennis (S.) in a paddy field was estimated. Since a great number of grasshoppers (Oxya japonica) lived in the research area and the mantids had frequently eaten O. japonica, we presumed that they consumed only O. japonica. As a result, it was estimated that average prey consumptions for 6 days were 406 mg for the females and 23 mg for the males. From the result of this estimation and the feeding rhythm of the mantids, it is concluded that each female captured one grasshopper on average every a few days. By comparing this actual feeding level of P. angustipennis with those of other predacious arthropods, characteristics of P. angustipennis as a predator were discussed.     

Response of Japanese beech (Fagus japonica Maxim.) sprouts to canopy gaps

The response of Japanese beech (Fagus japonica Maxim.) sprouts to canopy gaps in natural beech forest in central Japan was studied using two contrasted gaps in which tree-ring chronologies of regenerating stems were analyzed. The gaps were created by uprooting of a single Quercus mongolica var. grosseserrata stem (diameter: 50 cm; gap size: 40 m²; 23 years old) and by concurrent uprootings of four F. japonica stools (gap size: 180 m²; 30 years old). Japanese beech sprouts emerged before and after the gap formation and dominated stem populations in both gaps. In gaps, growth of F. japonica sprouts was equal or lower than growth of stems of seed origin, but most sprouts (F. japonica, Acer mono var. marmoratum) appeared a few years before emergence of seedlings. The small gap created by single stem fall was dominated by some beech sprouts from stools adjacent to the gap. The multiple gap was not closed by beech sprouts from stools surrounding the gap, but some dominant beech stems were resprouts from the uprooted beech stools. The existence of a sprout bank under the canopy may play an important role in the closing process of gaps in natural Japanese beech forest.     

Comparative leaf development ofOsmunda lancea andO. japonica (osmundaceae): Heterochronic origin of rheophytic stenophylly

Comparative development of the narrow pinnules of rheophyticOsmunda lancea and of the broad pinnules of a related dryland species,O. japonica, was examined and the origin of rheophytic stenophylly was discussed. The mature leaves and their various parts ofO. lancea are smaller and narrower than those ofO. japonica. The young pinnules ofO. lancea at the initiation of cell expansion are smaller than those ofO. japonica. The growth pattern of the pinnules is fundamentally the same in the two species, but pinnule growth period is shorter inO. lancea than inO. japonica. While the largest growth rate in pinnule length is quite similar, inO. lancea the pinnules are less elongated and much less broadened during ontogeny. Cell expansion in the mesophyll and epidermis proceeds acropetally and toward the margin along the axes of costules and veins. Although the numbers of mesophyll and epidermal cells between two adjacent veinlets are almost the same inO. lancea andO. japonica, during the subsequent growth period inO. lancea, the cells expand to a smaller extent and the veinlets become more narrowly oblique to the costule. This oblique distortion of laminar segments framed by veins causes stenophylly, an allometric modification. The stenophylly ofO. lancea is believed to have arisen by heterochronic evolution, in particular, progenesis.     

Development of microsatellite markers in white birch (Betula platyphylla var. japonica)

Betula platyphylla var. japonica is a typical pioneer tree species in the secondary succession in northern Japan. We describe the cloning and characterization of 13 polymorphic, codominant microsatellite loci isolated from this species. These polymorphic loci had 2–8 alleles per locus and a range of expected heterozygosities from 0.050 to 0.808.     

Systematics of Mielichhoferia (Bryaceae: Musci). III. Hybridization between M. elongata and M. mielichhoferiana

Allozyme variation in mixed populations of Mielichhoferia elongata and M. mielichhoferiana was investigated to determine if interspecific hybridization occurs when these two closely related species grow together. Previous research has shown that M. elongata and M. mielichhoferiana can be distinguished by three diagnostic isozyme loci (Gpi-1, Mdh-2, and Mdh-3) at which the two species do not share alleles in 32 allopatric populations from North America and Europe. The present study shows that in five populations from Colorado, Norway, and Sweden, gametophytes resulting from interspecific hybridization can be recognized by recombinant genotypes combining alleles of the otherwise diagnostic loci. A total of 32 multilocus genotypes was found among the 111 individuals sampled, of which 13 were recombinants. The frequency of recombinants ranged from 12% to 35% within populations, and all but one population contained both parental species. Moreover, recombinant genotypes could be accounted for by the allelic constitution of sympatric parents. In two of the populations, more than one hybridization event was necessary to account for the diversity of recombinant genotypes. Twenty-nine of the 32 genotypes detected in this study were restricted to one population each, two occurred in two Swedish populations separated by approximately 14 km, and one occurred in both Sweden and Norway.     

Genetic structure of Halotydeus destructor and Penthaleus major populations in Victoria (Acari: Penthaleidae)

The redlegged earth mite (Halotydeus destructor) and the blue oat mite (Penthaleus major) are major pests of pastures and crops in southern Australia. Reproductive modes, migration rates and levels of differentiation between populations were investigated using allozyme electrophoresis. Collections were made throughout Victoria and a sample was also obtained from Western Australia. Three enzyme loci were polymorphic in H. destructor (Mdh-1, Mdh-2 and Idh). Genotype frequencies of these loci did not differ between phenotypic males and females, providing no evidence for haplodiploidy. Allele frequencies were in Hardy-Weinberg equilibrium, indicating that H. destructor is diploid and sexual. This was confirmed via crosses between males and females. Allele frequencies differed between Victorian sites, although F statistics indicated little differentiation over all loci. A sample from Western Australia did not differ in allele frequencies from the Victorian sites. Four polymorphic loci were found in P. major (Mdh-1, Mdh-2, Idh and Gpi). Only a few multilocus genotypes occurred in a sample, indicating that P. major is parthenogenic. No male P. major were found in this study. A number of colour morphs were also identified and a genetic association between genital plate colour and clonal type was found in one population of P. major. Two different body colour morphs were associated with different clonal types.     

Isoenzyme Variation in Melipona rufiventris (Hymenoptera: Apidae, Meliponina) in Minas Gerais State, Brazil

The stingless bee Melipona rufiventris is an important pollinator in several Brazilian ecosystems. Originally widely distributed in Minas Gerais (MG) state, this species is becoming very rare. Therefore this species was included in the endangered species list of MG. We used isoenzyme data for a better understanding of the genetic structure of several M. rufiventris colonies. Samples of 35 colonies were collected from 12 localities and evaluated by nine enzymatic systems, which yielded 17 loci. M. rufiventris genetic variation was found to be low, typical of an endangered species. The proportion of polymorphic loci was 5.88% in both ecosystems. Only Est-4 was polymorphic in colonies from the Forest and Mdh-1 in colonies from the Cerrado. The expected heterozygosity ranged from 0.0068 in the Cerrado to 0.0078 in the Forest. Despite this, enzyme electrophoretic analyses provided a good idea of the diversity between samples from Cerrado and Forest which reinforce the existence of two different forms of M. rufiventris in MG, one present in the Cerrado and the other in Forest. This information is of great importance for the conservation of, M. rufiventris in MG.     

Flagelloscypha japonica: a new species of minute basidiomycete (Niaceae) from Japan

A minute basidiomycete belonging to Flagelloscypha (Niaceae, Agaricales) was found on blighted leaves of Rodgersia podophylla in Aomori Prefecture, Japan. The fungus proved to be a new species and was named Flagelloscypha japonica based on its morphological characteristics.