Conservation and multivariate analysis utility in characterization of ecogegraphical relationships of Trifolium and Lotus species

The maintenance of species in an environment and the durability of rangeland and fallow improvement depend on the choice of populations used. The study of abundance and the relationship between the natural distribution of spontaneous legumes species of forage and/or pastoral interest and environmental factors is an important step for an adequate utilization of local genetic resources. Collection of annual Trifolium species and the genus of Lotus were conducted at 45 sites in Morocco in May/June 2004. Ecogeographical information was recorded at each site. Twelve species of annual Trifolium and two species of Lotus were identified. Most prolific were T. tomentosum and L. corniculatus. A principal components analysis and canonical variate analysis were conducted to group the sites, using the ecogeographical variables collected at each site. The variables that were found to be most important in grouping the sites were mean coldest month temperature, mean hottest month temperature, mean annual rainfall, latitude, longitude and soil pH. None of the species identified were limited to only one ecogeographical group.     

Improving representativeness of genebank collections through species distribution models, gap analysis and ecogeographical maps

An efficient germplasm collecting method was evaluated using six Lupinus species and the Spanish Lupinus collection as a study case. This method includes the application of geographic information systems, ecogeographical land characterization maps, species distribution models and gap analysis to identify prioritized collecting sites. To evaluate the efficiency of this collecting method, field collecting expeditions were carried out focusing on prioritized sites and the results of these collections were analyzed. Prioritized sites were identified using spatial and ecogeographical gaps, and potential species richness maps. The spatial gaps corresponded to populations non-included in the collection but recorded by other information sources while ecogeographical gaps corresponded to spatial gaps that were located in ecogeographical categories (obtained from the ecogeographical map) that were scarcely represented in the collection. A potential Lupinus species richness map was obtained by adding the information of single maps of Lupinus species distribution models. Subsequently, prioritized sites were obtained in ecogeographical gaps with high potential species richness values. Collecting expeditions were made in Spain in 2006, 2007 and 2008. Results showed that using the efficient germplasm collecting methodology was highly positive not only from a quantitative viewpoint (between 7.8 and 11% increase) but also in qualitative terms, focusing collection efforts in ecogeographical categories with low or null representation in the Spanish Lupinus collection (41% of the new accessions). Phenotypic differences related to adaptation to environment were observed in the field between the populations that grow in low or null represented categories and those that grow in highly represented categories.     

Ecotypes and genetic divergence among sympatrically distributed populations of Hordeum vulgare and Hordeum spontaneum from the xeric region of Jordan

Summary The progeny of paired samples of Hordeum vulgare L. and Hordeum spontaneum C. Koch, collected from Jordan's xeric region was used in this study. Statistical analyses of seven easily measured morphometric traits were used to elucidate the relationships and distances between populations of both species, to detect any ecogeographical races, and to study the interrelationships and adjustments in the morphometric traits under study. Flag leaf area and plant height were the two most important discriminating variables which totally separated Hordeum vulgare from Hordeum spontaneum and accounted for 85.3% of total phenotypic variance in the collection. Cluster analysis indicated that the level of divergence among populations of both species was considerably different. Populations of Hordeum vulgare clustered at a maximum Euclidean distance of 2.08, while the maximum distance at which populations of Hordeum spontaneum clustered was 1.49. Three ecotypes each of Hordeum vulgare and Hordeum spontaneum were identified. These ecotypes corresponded to the environmental range of the collection sites. The interrelationships between the seven morphometric traits were adjusted in different ways as revealed by the principal components analysis. Sampling from the different clusters identified in this analysis is expected to increase the allelic diversity for selection and breeding purposes.     

The mycorrhizal status of Welwitschia mirabilis

This is the first report of the mycorrhizal status of Welwitschia mirabilis, a gymnosperm endemic to the Namib Desert. Like all other gymnosperms except the Pinaceae and Gnetaceae, W. mirabilis is associated with vesicular-arbuscular mycorrhizal (VAM) fungi. Mycorrhizal colonization of roots and the diversity and abundance of VAM species were determined at seven sites. Six sites received annual rainfall of 0–100 mm, varying widely from year to year. The seventh site experienced more predictable annual rainfall of 150–200 mm. Perennial vegetation was sparse at the six low-rainfall sites. Dry annual grasses from previous rain events were present at only three of these six sites and mean mycorrhizal colonization levels of W. mirabilis at these three sites were as high as 18%. W. mirabilis was not mycorrhizal at sites where grasses were absent. The seventh site, receiving higher rainfall, supported small trees and annual grasses in addition to W. mirabilis. Mycorrhizal colonization levels of W. mirabilis at this site were significantly higher than at the other six sites, closely paralleling those of the surrounding annual grasses. The mycorrhizal flora of W. mirabilis consisted of four Glomus species. These taxa were not unique to W. mirabilis, having been found with Stipagrostis and Cladoraphis grasses throughout the Namib and Kalahari deserts.     

RAPD polymorphism of wild emmer wheat populations, Triticum dicoccoides, in Israel

 Genetic diversity in random amplified polymorphic DNAs (RAPDs) was studied in 110 genotypes of the tetraploid wild progenitor of wheat, Triticum dicoccoides, from 11 populations sampled in Israel and Turkey. Our results show high level of diversity of RAPD markers in wild wheat populations in Israel. The ten primers used in this study amplified 59 scorable RAPD loci of which 48 (81.4%) were polymorphic and 11 monomorphic. RAPD analysis was found to be highly effective in distinguishing genotypes of T. dicoccoides originating from diverse ecogeographical sites in Israel and Turkey, with 95.5% of the 100 genotypes correctly classified into sites of origin by discriminant analysis based on RAPD genotyping. However, interpopulation genetic distances showed no association with geographic distance between the population sites of origin, negating a simple isolation by distance model. Spatial autocorrelation of RAPD frequencies suggests that migration is not influential. Our present RAPD results are non-random and in agreement with the previously obtained allozyme patterns, although the genetic diversity values obtained with RAPDs are much higher than the allozyme values. Significant correlates of RAPD markers with various climatic and soil factors suggest that, as in the case of allozymes, natural selection causes adaptive RAPD ecogeographical differentiation. The results obtained suggest that RAPD markers are useful for the estimation of genetic diversity in wild material of T. dicoccoides and the identification of suitable parents for the development of mapping populations for the tagging of agronomically important traits derived from T. dicoccoides. Received: 13 July 1998 / Accepted: 13 August 1998     

Recruitment of Eucheuma and Kappaphycus on a farm in Tawi-Tawi,Philippines

Vegetative regeneration is the only farming method for Eucheuma and Kappaphycus, the world's most important carrageenophytes. Lack of seedstock has been a problem following natural calamities, overharvesting, grazing and outbreak of "ice-ice" disease. Hence, studies to help develop an alternative method of generating Eucheuma and Kappaphycus sporelings on a seaweed farm in Tawi-Tawi, Philippines were carried out from May 1993 to July 1994. Mactan stone blocks were deployed in two positions at two sites in Tawi-Tawi, one of the biggest farming areas in the Philippines. Recruits were first observed five months after deployment. Nested ANOVA was used to investigate the effect of current speed (C), water movement (M), number of days with minus tide (D), salinity (S), temperature (T), area of deployment (A) and position of blocks (P) on the recruitment of sporelings (R). Since C, M, D, S, and T were always on the same mean level within an area, the effects of these variables were assumed by the effect of area and thus, the A and P variables were the ones fitted in the ANOVA model. Repeated measures ANOVA showed no significant difference in the effects of A and P on visible R from May to November 1993 and March to July 1994. The Helmert transformation showed that mean recruitment of Eucheuma and Kappaphycus between May 1993 and subsequent time points was significantly different. This was also true for recruitment in September 1993, December 1993, January 1994 and February 1994, which were highly significantly different compared to the means of subsequent time points. Multiple regression analysis showed that C, D, and S had significant effects on recruitment of Eucheuma and Kappaphycus in both areas while effect of T was significant only in area I.     

Conspecific Sharing of Breeding Sites by Anopheline Female Mosquitoes (Diptera: Culicidae) Inferred from Microsatellite Markers

The number of Anopheles gambiae and Anopheles arabiensis females that used each of the 33 sampled breeding sites in west Kenya was estimated by microsatellite markers and related statistics to test the hypothesis that conspecific females share aquatic sites. Totally, 166 An. gambiae and 168 An. arabiensis larvae were identified and were genotyped. The mean number of larvae per breeding site was 8.3 for An. gambiae and 8.4 for An. arabiensis. The likelihood method estimated that, for An. gambiae, the mean number of females that would have laid eggs per breeding site was 5.2 and ranged from 2 to 9, and for An. arabiensis, the mean was 5.0 with a range of 2–10. The clustering method estimated that the mean number of females laying eggs per breeding site was 6.8 for An. gambiae. The results provide molecular evidence that females of one or both species share breeding sites.     

Phylogenetic relationships of JapaneseAster (Asteraceae, Astereae) sensu lato based on chloroplast-DNA restriction site mutations

RFLPs of cpDNA were examined for 18 species ofAster, six species ofKalimeris, two species ofMiyamayomena and one species and one variety ofHeteropappus from Japan, using 16 restriction endonucleases. Approximately 275 restriction sites were surveyed, and a total of 74 restriction site mutations was detected, and 31 of these were phylogenetically informative. Sixteen most parsimonious trees constructed from Wagner parsimony analysis indicated the polyphyly ofKalimeris andMiyamayomena sensu Kitamura;K. miqueliana belongs to a different clade from the remaining species ofKalimeris, and two species ofMiyamayomena did not make a single clade. This result suggests that the shortening or loss of pappus have happened parallelly in different evolutionary lineages. We must be careful to assess the pappus character in taxonomy and phylogeny, and it is desirable to examine their phylogenetic relationships using a molecular data.     

Development of a RIVPACS-type predictive model for bioassessment of wadeable streams in Wyoming

RIVPACS models produce a community-level measure of biological condition known as O/E, which is derived from a comparison of the observed (O) biota with those expected (E) to occur in the absence of anthropogenic stress. We used benthic macroinvertebrate and environmental data collected at 925 stream monitoring stations, from 1993 to 2001, to develop, validate, and apply a RIVPACS model to assess the biological condition of wadeable streams in Wyoming. From this dataset, 296 samples were identified as reference, 157 of which were used to calibrate the model, 46 to validate it, and 93 to examine temporal variability in reference site O/E-values. We used cluster analyses to group the model development reference sites into biologically similar classes of streams and multiple discriminant function analysis to determine which environmental variables best discriminated among reference groups. A suite of 14 categorical and continuous environmental variables best discriminated among 15 reference groups and explained a large proportion of the natural variability in biota within the reference dataset. Eleven of the predictor variables were derived from GIS. As expected, mean O/E-values for reference sites used in model development and validation were near unity and statistically similar. Temporal variability in O/E-values for reference sites was low. Test site values ranged from 0 to 1.45 (mean = 0.73). The model was accurate in both space and time and precise enough (S.D. of O/E-values for calibration data = 0.17) to detect modest alteration in biota associated with anthropogenic stressors. Our model was comparable in performance to other RIVPACS models developed in the United States and can produce effective assessments of biological condition over a broad, ecologically diverse region. We also provide convincing evidence that RIVPACS models can be developed primarily with GIS-based predictor variables. This framework not only simplifies the extraction of predictor variable information while potentially reducing expenditures of time and money in the collection of predictor variable information, but opens the door for development and/or application of RIVPACS models in regions where there is a paucity of local-scale, abiotic information.     

Abundance,body size,and morphology of bumblebees in an area where an exotic species, <Emphasis Type="Italic">Bombus terrestris</Emphasis>, has colonized in Japan

An exotic bumblebee species, Bombus terrestris, has colonized in Japan and becomes dominant in some local communities. We examined the effects of land use and bumblebee abundance on the number and body size of bumblebees collected using window traps in a lowland area in the southern Ishikari district, Hokkaido. In 2004, we collected 922 bumblebees of six species using 70 traps at 17 sites. A statistical model fitted to the data demonstrated that dispersion from commercial B. terrestris colonies used in greenhouses positively affected the number of B. terrestris caught by each trap. This exotic species was abundant in sites where paddy fields were prevalent, but three native species, B. hypocrita, B. ardens, and B. diversus, were abundant in sites where farms and woodlands were widespread. The local abundance of B. terrestris was not associated negatively with the number and body size of native bumblebees. Thus, we did not find any competitive interactions between exotic and native bumblebees although habitat conditions seem to be common determinants of the bumblebee populations. A morphological analysis showed that B. terrestris had intermediate tongue length between B. hypocrita and B. ardens.     

Variation in the summer diet of age-0 brown trout in a regulated mountain stream

We describe the age-0 brown trout (Salmo trutta) collected from a mountain stream downstream from a reservoir in June, July, and August 1990. Sample sites were 1, 12, and 20 km downstream from the reservoir at elevations of 2840, 2600, and 2230 m above mean sea level. The mean length of fish in the samples increased with downstream progression and from June through August. Zooplankton predominated the diet of fish collected in June from the most upstream sampling site, while Diptera larvae predominated the diet of fish from more downstream sampling sites in June. In July and August, the diet of fish from the upstream site was predominated by Diptera larvae, while the diet of fish from the other two sites was predominated by Ephemeroptera nymphs. Variation in diet among sampling sites and sampling months was related not only to increasing fish sizes over the summer but also to the availability of zooplankton flushed from the reservoir.     

Natural levels of fungal infections in grasshoppers in Northern Benin

The infection of grasshoppers by naturally occurring, entomopathogenic fungi was monitored at two sites in Malanville, northern Benin, Africa. Grasshoppers were collected and recorded from the sites between June and December 1992 and all of them, barring the first instars, were incubated in ventilated cages. At the first site, 1343 individuals of 35 grasshopper species were incubated, and at the second site, 857 individuals of 36 grasshopper species were incubated. Three hyphomycete fungi (Deuteromycotina: Hyphomycetes), Metarhizium flavoviride Gams and Rozsypal, Beauveria bassiana (Bals.) Vuillemin and Sorosporella sp. were found infecting grasshoppers. The average incidence of M. flavoviride infection was 2.9% and 1.8% at the two sites for all host species. M. flavoviride sporulated on most grasshopper cadavers within 10 days of collection. B. bassiana and Sorosporella sp. were only collected from one and five grasshopper individuals respectively. A significant difference was noted in the time to death of small grasshopper species infected with M. flavoviride compared to larger species. At one site, M. flavoviride infection was positively correlated with rainfall during the 10‐day period in which samples were taken.     

Length–weight relationships of four fish species from the upper Paraná River basin,Southeastern Brazil

Here the length–weight relationships (LWR, hereafter) of Pareiorhina pelicicei, Trichomycterus candidus, T. pirabitira, and T. piratymbara are provided. Samples of each species were collected between April and July 2017 with a hand net (1.5 mm mesh) in three sites from the upper Paraná River basin, Southeastern Brazil. These results increase knowledge about the LWR of Neotropical freshwater fish.     

Movements and Site Fidelity of two Juvenile Fish Species Using Surf Zone Nursery Habitats Along The Southeastern North Carolina Coast

We examined the extent of movements of juvenile Florida pompano, Trachinotus carolinus, and gulf kingfish, Menticirrhus littoralis, along an open ocean beach. Fishes were collected by seine at three sites along Masonboro Island and Carolina Beach, NC between 7 June and 7 July 1995. All specimens 40 mm standard length (SL) were tagged with coded wire tags and released at the capture sites. Between 7 July and 9 August and on 15–16 September Masonboro Island and northern Carolina Beach were surveyed for tagged fish. A controlled tag mortality/retention study was conducted for both species. Overall, 1569 Florida pompano (40–135 mm SL) were tagged. Sixty-one (3.9%) of these were recaptured, and only eight moved away from the original tagging sites. The largest movements by two Florida pompano were 2.1 and 10.5 km. Many fish remained at their original tagging sites for 21–27 days. Of 488 gulf kingfish (36–158 mm SL) tagged, 16 (3.3%) were recaptured. Gulf kingfish also exhibited little movement away from tagging sites during the study, with individuals remaining at original tagging sites up to 21 days. Stock size estimates for Florida pompano ranged from 3354 to 4670 among the tagging sites, with densities ranging from 1.9 to 2.6 fish m^–2. The remarkable site fidelity exhibited by these two species suggests that resources were not limiting or that predation pressure was not high enough to cause large scale movements during the study. This implies that local disturbances could impact behavior or survival of juvenile fishes in the surf zone.     

Climatic signals in tree rings of <Emphasis Type="Italic">Burkea africana</Emphasis> and <Emphasis Type="Italic">Pterocarpus angolensis</Emphasis> from semiarid forests in Namibia

Tree-ring studies contribute worldwide to the understanding of climate and its relation to tree growth. Long tree-ring chronologies serve as climate proxies for the reconstruction of past, pre-instrument climate and its recent change. In tropical regions, the availability of exactly dated tree-ring chronologies is limited. The dendroclimatic potential of two dominant species from dry forests in northern Namibia was examined in the study presented in this paper. Both species (Burkea africana Hook and Pterocarpus angolensis DC) were sampled at two sites (ca. 900 km apart), and the response to several climatic variables, including ENSO indices, is studied. All specimens showed distinct growth rings and cross-dating between radii was successful for all trees. Species-specific mean curves were built for both sites. The mean curves of different species of the same site synchronised significantly, allowing the construction of a site-specific chronology. Synchronisation between sites was not possible, but spectral analysis of the chronologies implied that both show similar long-term (6.7 year) oscillation patterns. B. africana is more sensitive to rainfall variation than P. angolensis at both sites. Growth response to rainfall was positive, but a time-lag in the reaction occurred between the sites, corresponding to the time-lag of the beginning of the rainy season. Air temperature showed a negative correlation with stem increment at both sites. The response at the westernmost site to two ENSO indices indicates a tree growth decrease during El Niño years, which are generally dry in southern Africa.     

Ecological aspects of syllids (Annelida: Polychaeta: Syllidae) on Thalassia testudinum beds in Venezuela

The spatial and temporal variations of syllids associated with the tropical Thalassia testudinum were studied at four seagrass beds in the Morrocoy National Park. The epifaunal syllids were collected at quarterly intervals throughout one year using a modified suction sampler. Possible relationships between sampling sites, months and species were evaluated by Principal Components Analysis. The spatial-temporal variability of the predominant species was tested by means of a 2-factor ANOVA with the sampling sites and months as factors. A total of 1138 individual syllids were collected, belonging to 41 species and 12 genera, of which Branchiosyllis, Exogone, Odontosyllis, Sphaerosyllis and Syllis showed the highest diversity. Spatial variations were defined by the species of Branchiosyllis (B. exilis, B. lorenae, B. oculata) whereas temporal variability was defined by certain species of Syllis(S. beneliahui, S. broomensis and S. prolifera), the latter also being the most species rich genus present. Of the 41 species found, only these six showed consistent presence–absence and abundance patterns. Species richness and abundance were significantly higher in March at all sampling sites. Collectively, the syllid fauna of the Morrocoy seagrass beds was richer than recorded from other similar habitats. These findings are discussed in relation to the physical and biotic factors that may affect the variability of syllid populations in these seagrass habitats.     

Character displacement in Hydrobia

Summary Fenchel's study of size variation in Hydrobiid snails in the Limfjord, Denmark, provides one of the most convincing cases of ecological character displacement available. In order to assess the generality of the phenomena within the Hydrobia genus, allopatric and sympatric Hydrobia ventrosa, H. neglecta and H. ulvae were collected from 24 coastal sites around Eastern England in July and October, 1982. Shell heights of 5,850 snails from 55 samples were recorded. These data were analysed for intraspecific allopatric-sympatric, and interspecific height differences. Such differences were not significant for H. ventrosa/H. neglecta, but the species exhibited parallel variation at sympatric sites. H. ventrosa/H. ulvae showed only significant interspecific differences, but in both allopatry and sympatry. The pattern of mean size variation for these species resembles that in the Limfjord. However the statistical analyses fail to provide support for character displacement. Differences in character states attributed to the process of character displacement may result from a number of other causes. Environmental conditions at sympatric and allopatric sites may act differentially on the heights of H. ulvae and H. ventrosa. The conditions that prevail at sympatric H. ulvae sites appear to lead to increased size in this species irrespective of the presence of H. ventrosa.     

Phylogenetic relationships among cultivated Allium species from restriction enzyme analysis of the chloroplast genome

Summary The genus Allium contains many economically important species, including the bulb onion, chive, garlic, Japanese bunching onion, and leek. Phylogenetic relationships among the cultivated alliums are not well understood, and taxonomic classifications are based on relatively few morphological characters. Chloroplast DNA is highly conserved and useful in determining phylogenetic relationships. The size of the chloroplast genome of Allium cepa was estimated at 140 kb and restriction enzyme sites were mapped for KpnI, PstI, PvuII, SalI, XbaI, and XhoI. Variability at restriction enzyme sites in the chloroplast DNA was studied for at least three accessions of each of six cultivated, old-world Allium species. Of 189 restriction enzyme sites detected with 12 enzymes, 15 mutations were identified and used to estimate phylogenetic relationships. Cladistic analysis based on Wagner and Dollo parsimony resulted in a single, most-parsimonious tree of 16 steps and supported division of the species into sections. Allium species in section Porrum were distinguished from species in sections Cepa and Phyllodolon. Two species in section Rhiziridium, A. schoenoprasum and A. tuberosum, differed by five mutations and were placed in separate lineages. Allium cepa and A. fistulosum shared the loss of a restriction enzyme site and were phylogenetically closer to each other than to A. schoenoprasum. This study demonstrates the usefulness of restriction enzyme site analysis of the chloroplast genome in the elucidation of phylogenetic relationships in Allium.     

Genetic identification of larvae and juveniles reveals the difference in the spawning site among Cyprininae fish species/subspecies in Lake Biwa

To understand the differences in the spawning sites among Cyprininae fishes in Lake Biwa, we conducted periodic sampling of larvae and juveniles at three sites (irrigation ditch, St. 1; river, St. 2; and satellite lake, St. 3). On the basis of species/subspecies identification by using RAPD analysis, we examined the species composition of the larvae and juveniles at these three sampling sites. The number of specimens was 616, 68, and 117 at St. 1, St. 2, and St. 3, respectively. Based on morphological and genetic identification, the specimens were found to include nine fish species/subspecies, namely, Carassius auratus grandoculis, Carassius cuvieri, Carassius auratus langsdorfii, Cyprinus carpio, Sarcocheilichthys sp., Silurus asotus, Oryzias latipes, Odontobutis obscura obscura, and Rhinogobius sp. The species composition at the three sites also differed. Among the Cyprininae fishes, C. auratus grandoculis, C. auratus langsdorfii, and Cyprinus carpio were found in abundance at St. 1; C. cuvieri was not collected from St. 1 but was found at the other two sites, particularly St. 3. Among the other fishes, Rhinogobius sp. was collected at St. 1 and St. 3, whereas the other four occurred only at St. 1. These results suggest that the selection of spawning sites by C. cuvieri differs to a certain extent from that of the other Cyprininae fishes, and the irrigation ditch in the lake is an important habitat for the larvae and juveniles of native fish species.