Essential oil chemotypes of Aloysia citrodora (Verbenaceae) in Northwestern Argentina

Chemical biodiversity of essential oils of natural populations of Aloysia citrodora Palau (“lemon verbena”, “cedrón”) in Northwestern Argentina was assessed by collecting in the same sites through different years. A total of 36 samples were collected in the Provinces of Salta (El Maray, La Paya, El Sunchal, El Alisal, Chorrillos), Jujuy (Chilcayo, San Roque), Catamarca (Mutquin, Colana) and Tucumán (Amaicha del Valle) in Argentina.Essential oils were obtained by hydrodistillation (Clevenger) of naturally air-dried plant material. Yields ranged from 0.16% to 1.93% (v/w), being the highest those of the collections of Mutquin. More than 65 compounds were identified by CG-FID-MS. Only 19 of these constituents, accounting from 77.3 to 98.9% of the total oil, present in more than 4.0% in at least one sample, were considered as variables for statistical analysis. Agglomerative Hierarchical Cluster analysis was conducted, showing at 65% of similarity, five groups. This grouping was in direct accordance to the biosynthetic pathways of main compounds (chemotypes).In the two sites of Jujuy, 21 collections evidenced four different chemotypes, named after the dominant component as follows: thujones, citronellal, carvone, and citral (neral + geranial). In the populations of Salta and Catamarca, linalool appeared as a new different chemotype.Though cedron is considered as a citral-bearing plant, curiously, in the 36 samples collected in the NW of Argentina, only two samples contained citral as main constituent. On the other hand, a dominance of citronellal and thujones compositions were found in the bulk of the samples collected, while others had very high content of linalool or carvone and its derivatives. Northwestern Argentina has repeatedly been mentioned as the center of biodiversity of this species.The new evidences found on the chemical biodiversity of essential oils of Aloysia citrodora in natural populations in this region, reinforce firmly this idea.     

Genetic diversity and population structure in the Brazilian Cattleya labiata (Orchidaceae) using RAPD and ISSR markers

Brazilian orchids are currently threatened with extinction due to habitat loss and, because of their high ornamental value, intense collecting pressure. Genetic diversity can play a key role in the survival of endangered orchid species. Here we provide the first data on genetic diversity and structure of wild populations in the genus Cattleya, in particular C.?labiata, using random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) markers. We studied 130 individuals, 117 belonging to Cattleya?labiata and 13 from 10 other species in the same genus. Data generated from 12 ISSR and 12 RAPD primers were used to determine genetic variability via a model-based Bayesian procedure (Structure) and molecular variance analysis. In addition, Shannon index, genetic diversity and Jaccard coefficients were also estimated. The marker data indicated that C. labiata has a high level of polymorphism, and five reconstructed populations were identified by Structure. The unweighted pair group method with arithmetic mean dendrogram did not group the samples by origin, which was also confirmed by Bayesian analysis, demonstrating the complex genetic structure of C.?labiata. Other Cattleya species showed no relationship with any C.?labiata sample. This genetic characterization of Cattleya from northeast Brazil contributes to knowledge of the genetic structure of the species and can be used to define strategies for conservation and breeding programmes.     

Genetic Relationships within Tripsacum as Detected by RAPD Variation

Genetic diversity within species of Tripsacum was surveyed basedon randomly amplified polymorphic DNA (RAPD) variation, as detectedwith the polymerase chain reaction (PCR). Thirteen of the 16Tripsacum species, including both temperate and tropical species,were included in this study using 56 decamer oligonucleotideprimers. All of the 56 primers generated repeatable RAPD profilesand 53 of them detected polymorphic bands among the Tripsacumspecies. These 53 primers generated 350 repeatable bands rangingin size from 150–1600 bp, each primer generating an averageof seven scoreable bands. Cluster analysis of polymorphic RAPDsindicated four major clusters. Cluster 1 consists of North AmericanTripsacum species, cluster 2 consists of South American Tripsacumspecies, cluster 3 includes T. zopilotense and T. latifoliumfrom Mexico, and cluster 4 consists of Mesoamerican Tripsacumspecies. Cluster analysis does not reveal the division of twotaxonomic sections (Fasciculata and Tripsacum). Copyright 1999Annals of Botany Company Tripsacum, RAPD, genetic relationships, genetic diversity.     

Variation of the chemical composition of essential oils in Brazilian populations of Cunila menthoides Benth. (Lamiaceae)

Cunila menthoides Benth. is characterized as a xilopodiferous subshrub with strong and pleasant smell. This species is commonly used in South Brazilian folk medicine. Air-dried samples of four Brazilian populations of C. menthoides were extracted by steam distillation and analyzed using GS and GS–MS. A total of 15 volatile compounds were detected and identified. Two groups, which can be considered as chemotypes, were formed by the average linkage cluster analysis. The first group was characterized by high concentrations of pulegone, with some individuals showing variations to menthone; and the second group was characterized by high concentrations of linalool.     

Molecular signatures of sexual communication in the phlebotomine sand flies

Phlebotomine sand flies employ an elaborate system of pheromone communication wherein males produce pheromones that attract other males to leks (thus acting as an aggregation pheromone) and females to the lekking males (sex pheromone). In addition, the type of pheromone produced varies among populations. Despite the numerous studies on sand fly chemical communication, little is known of their chemosensory genome. Chemoreceptors interact with chemicals in an organism’s environment to elicit essential behaviors such as the identification of suitable mates and food sources. Thus, they play important roles during adaptation and speciation. Major chemoreceptor gene families, odorant receptors (ORs), gustatory receptors (GRs) and ionotropic receptors (IRs) together detect and discriminate the chemical landscape. Here, we annotated the chemoreceptor repertoire in the genomes of Lutzomyia longipalpis and Phlebotomus papatasi, major phlebotomine vectors in the New World and Old World, respectively. Comparison with other sequenced Diptera revealed a large and unique expansion where over 80% of the ~140 ORs belong to a single, taxonomically restricted clade. We next conducted a comprehensive analysis of the chemoreceptors in 63 L. longipalpis individuals from four different locations in Brazil representing allopatric and sympatric populations and three sex-aggregation pheromone types (chemotypes). Population structure based on single nucleotide polymorphisms (SNPs) and gene copy number in the chemoreceptors corresponded with their putative chemotypes, and corroborate previous studies that identified multiple populations. Our work provides genomic insights into the underlying behavioral evolution of sexual communication in the L. longipalpis species complex in Brazil, and highlights the importance of accounting for the ongoing speciation in central and South American Lutzomyia that could have important implications for vectorial capacity.     

Chemical composition of flavonoids and styrylpyrones and the genetic variability of isozymes in natural populations of Cryptocarya mandioccana Meisner (Lauraceae)

The chemical composition of leaves of 57 trees of Cryptocarya mandioccana from three populations of southeastern Brazil was investigated through HPLC, assaying six flavonoids, seven styrylpyrones, and seven unidentified compounds. From 51 of the former trees, genotypes were obtained from 40 polymorphic loci of 19 isozymes. Cluster analyses of the phytochemical and genetical variation revealed that trees exhibited four chemotypes and five clusters from isozymes, respectively. Discriminant analyses from selected variables of the isozymic and chemical data sets were performed, respectively, in relation to the four chemotypes and the five isozyme clusters. The classification of individuals presented respective error estimates of 9.16% and 13.57%, indicating that the genetic data could explain the clusters from chemotypes and vice versa at acceptable error levels. Linear regressions with Dummy variable showed significant association of locus Skdh-2 with quercetin-3-O-β-d-glucopyranoside and cryptofolione, indicating that its alleles would be responsible for the chemotype variation between individuals.     

Genetic diversity of Histoplasma capsulatum strains in Brazil

This study establishes the genetic relatedness among Brazilian Histoplasma capsulatum samples obtained from different sources. A PCR-based random amplified polymorphic DNA (RAPD) assay was used to delineate polymorphisms among isolates in geographically diverse regions in Brazil. RAPD fingerprints revealed distinct DNA profiles and provided a high level of discrimination among H. capsulatum strains from different locations. Cluster I was composed of H. capsulatum isolates from the northeast region. The majority of strains from southeast and south were categorized as major cluster II. The strain 84564 from Rio de Janeiro State showed no genetic correlation to any of the isolates from the same state. The RAPD patterns of H. capsulatum isolates from Goias (Cluster III) were unrelated to DNA fingerprints observed among the other H. capsulatum strains (48% similarity). This study is the first report that stratifies the clusters of H. capsulatum strains from Brazil by molecular typing and associates them with the geographical origin.     

Molecular Characterization of Yarrowia lipolytica and Candida zeylanoides Isolated from Poultry

Yeast isolates from raw and processed poultry products were characterized using PCR amplification of the internally transcribed spacer (ITS) 5.8S ribosomal DNA region (ITS-PCR), restriction analysis of amplified products, randomly amplified polymorphic DNA (RAPD) analysis, and pulsed-field gel electrophoresis (PFGE). ITS-PCR resulted in single fragments of 350 and 650 bp, respectively, from eight strains of Yarrowia lipolytica and seven strains of Candida zeylanoides. Digestion of amplicons with HinfI and HaeIII produced two fragments of 200 and 150 bp from Y. lipolytica and three fragments of 350, 150, and 100 bp from C. zeylanoides, respectively. Although these fragments showed species-specific patterns and confirmed species identification, characterization did not enable intraspecies typing. Contour-clamped heterogeneous electric field PFGE separated chromosomal DNA of Y. lipolytica into three to five bands, most larger than 2 Mbp, whereas six to eight bands in the range of 750 to 2,200 bp were obtained from C. zeylanoides. Karyotypes of both yeasts showed different polymorphic patterns among strains. RAPD analysis, using enterobacterial repetitive intergenic sequences as primers, discriminated between strains within the same species. Cluster analysis of patterns formed groups that correlated with the source of isolation. For ITS-PCR, extraction of DNA by boiling yeast cells was successfully used.     

Variability in essential oil composition of Turkish basils (Ocimum basilicum L.)

Sweet basil (Ocimum basilicum L.), one of the most popular aromatic plants, shows great variation in both morphology and essential oil components. In this study, the composition of 18 Turkish basil essential oils was investigated by GC and GC–MS. Variation of essential oils in the landraces was subjected to cluster analysis, and seven different chemotypes were identified. They were (1) linalool, (2) methyl cinnamate, (3) methyl cinnamate/linalool, (4) methyl eugenol, (5) citral, (6) methyl chavicol (estragol), and (7) methyl chavicol/citral. Methyl chavicol with high citral contents (methyl chavicol/citral) can be considered as a “new chemotype” in the Turkish basils. Because methyl eugenol and methyl chavicol have structural resemblance to carcinogenic phenylpropanoids, chemotypes having high linalool, methyl cinnamate or citral contents and a mixture of these is suitable to cultivate for use in industry.     

Genetic diversity of Persian shallot (Allium hirtifolium Boiss.) populations based on morphological traits and RAPD markers

Mooseer or Persian shallot (Allium hirtifolium Boiss.) is a bulbous perennial herb belonging to Alliaceae family. Genetic diversity of 16 mooseer populations collected from different parts of Iran were evaluated using 14 morphological traits as well as antioxidant activities and RAPD markers. Statistical analysis showed significant differences among populations for all morphological traits. In a principal component analysis, four components showed 77.5 % of the total morphological variation. Based on the morphological traits, populations were divided into five groups by clustering. Antioxidant capacities differed among all populations, with the Sahneh and Nahavand populations having the highest and the lowest activities, respectively. Genetic diversity of populations was also investigated through RAPD technique using 14 random primers. Cluster analysis based on RAPD markers plus 47 % coefficient of similarity divided the genotypes into six distinct groups. The highest and lowest similarities between populations were detected to be 0.63 and 0.21, respectively. These results indicated that the assessment of genetic diversity in Iranian mooseer populations could be appropriately carried out using RAPD technique which correlates with morphological traits.     

An Examination of Morphometric Variations in a Neotropical Toad Population (Proceratophrys cristiceps,Amphibia, Anura,Cycloramphidae)

The species Proceratophrys cristiceps belongs to the genus Proceratophrys within the family Cycloramphidae. These amphibians are found exclusively in South America in the morphoclimatic domain of the semi-arid depression zones in northeastern Brazil known as the Caatinga. We examined intrapopulational variation using univariate and multivariate statistics with traditional and geometric morphometrics, which supported the existence of two morphotypes of this species. Our results indicated significant degrees of variation in skeletal characteristics between some natural populations of this species. Careful analyses of variability levels are fundamental to avoid taxonomic errors, principally in populations that demonstrate characteristics intimately associated with their area of occurrence, as is the case of Proceratophrys cristiceps.     

Genetic diversity in populations of Dalbulus maidis (DeLong and Wolcott) (Hemiptera: Cicadellidae) from distant localities in Brazil assessed by RAPD-PCR markers

Populations of Dalbulus maidis (DeLong and Wolcott) from the northeastern and central-southern regions of Brazil differ morphologically, suggesting that they could be genetically isolated. Here we used the random amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) technique to estimate genetic structuring of this leafhopper species among five geographically distant localities across those regions and to estimate gene flow between populations. Ten specimens were sampled per population and genotyped with RAPD markers generated from amplification with nine oligonucleotides. The percentage of polymorphic loci was 78% in relation to the total number of amplified loci, and genetic similarity either between or within populations was higher than 0.72. Cluster analysis grouped specimens from the northeastern population (Mossoró/RN) into a single group, whereas central-southern specimens were not grouped in relation to their places of origin. Overall, the genetic subdivision index (Fst) was low (or= 0.192 and Nm 相似文献   

A chemical perspective on the evolution of variation in Eucalyptus globulus

It is becoming increasingly easy to generate genotypic data but much harder to gather an equivalent amount of phenotypic information, particularly for chemical traits. In this study of Eucalyptus globulus ssp. globulus, we measured about 60 chemical leaf traits of trees growing in a common garden to address the following questions: (1) how much variation is there between geographic regions, populations within regions and within populations? (2) How do chemical traits vary over the species’ geographic range? (3) If so, does E. globulus ssp. globulus exhibit distinct chemotypes – plants that are morphologically similar but which differ chemically? (4) Are the affinities between E. globulus ssp. globulus and closely related subspecies apparent in the chemical variation? Variation among trees within populations contributed most variation in leaf chemistry followed by variation between geographic regions. For many traits, variation among populations within proposed races and variation among proposed races within geographic regions explained little of the total variation. There was a cline in the concentration of secondary chemicals with the lowest concentrations in Tasmanian populations and the highest in those from eastern Victoria, with intermediate concentrations in populations from Bass Strait Islands. We identified three chemotypes, characterised by specific terpenes and formylated phloroglucinol compounds. The frequency of occurrence of these chemotypes showed a geographic pattern also, with “chemotype 1” predominating in Tasmania, while “chemotypes 2 and 3” occurred at highest frequency in eastern Victoria. We suggest that the chemotypes reflect introgression between E. globulus ssp. globulus and the three closely related subspecies – E. globulus ssp. bicostata, E. globulus ssp. maidenii and E. globulus ssp. pseudoglobulus. Although the formation of land-bridges with fluctuating sea levels has no doubt shaped the evolutionary history of all four subspecies, we propose that the migratory swift parrot (Lathamus discolor), an important pollinator and a species closely associated with E. globulus, has helped shape the evolution of the four tree subspecies.     

Evaluation of genetic diversity of <Emphasis Type="Italic">Clinacanthus nutans</Emphasis> (Acanthaceaea) using RAPD,ISSR and RAMP markers

Three polymerase chain reaction (PCR) techniques were compared to analyse the genetic diversity of Clinacanthus nutans eight populations in the northern region of Peninsular Malaysia. The PCR techniques were random amplified polymorphic deoxyribonucleic acids (RAPD), inter-simple sequence repeats (ISSR) and random amplified microsatellite polymorphisms (RAMP). Leaf genomic DNA was PCR amplified using 17 RAPD, 8 ISSR and 136 RAMP primers . However, only 10 RAPD primers, 5 ISSR primers and 37 RAMP primers produced reproducible bands. The results were evaluated for polymorphic information content (PIC), marker index (MI) and resolving power (RP). The RAMP marker was the most useful marker compared to RAPD and ISSR markers because it showed the highest average value of PIC (0.25), MI (11.36) and RP (2.86). The genetic diversity showed a high percentage of polymorphism at the species level compared to the population level. Furthermore, analysis of molecular variance revealed that the genetic diversity was higher within populations, as compared to among populations of C. nutans. From the results, the RAMP technique was recommended for the analysis of genetic diversity of C. nutans.     

Genetic Diversity Analysis of Starchy Curcuma Species Using RAPD Markers

The genetic variability in starchy Curcuma species was assessed using Random Amplified Polymorphic DNA (RAPD) technique. The RAPD pattern generated by 20 primers revealed a high degree of polymorphism. A total of 274 bands were generated of which 264 were polymorphic. All the species were separated into 3 clusters using UPGMA. C. aromatica, C. leucorrhiza, and C. brog formed a cluster within which C. longa and C. zedoaria formed a subgroup. C. harita was genetically distinct from all the other Curcuma species. Since it is difficult to distinguish different species by leaf morphology, the RAPD pattern has high utility in identification of starchy Curcuma species.     

RAPD Analysis of the Genome in Species of the Genus Hordeum

Random amplification of polymorphic DNA (RAPD) was used to analyze six species, three populations, and seven regional cultivars of barley. A unique pattern of amplified DNA products was obtained for each species of the genus Hordeum.High polymorphism of barley species was revealed. Specific fragments were found in most RAPD patterns; the fragments can be used as molecular markers of corresponding species and subspecies. Several other DNA fragments were shown to serve as molecular markers of the H genome. Specific RAPD patterns were obtained for each population and each cultivar of H. vulgaresensu lato. In total, variation between the populations and between the cultivars was substantially lower than between species. Cluster analysis (UPGMA) was used to estimate genetic distances between theHordeumspecies, between the H. spontaneumpopulations, and between regional H. vulgarecultivars and a dendrogram was constructed.     

Random Amplified Polymorphic DNA Analysis of Heterodera cruciferae and H. schachtii populations

Heterodera schachtii and H. cruciferae are sympatric in California and frequently occur in the same field upon the same host. We have investigated the use of polymerase chain reaction (PCR) amplification of nematode DNA sequences to differentiate H. schachtii and H. cruciferae and to assess genetic variability within each species. Single, random oligodeoxyribonucleotide primers were used to generate PCR-amplified fragments, termed RAPD (random amplified polymorphic DNA) markers, from genomic DNA of each species. Each of 19 different random primers yielded from 2 to 12 fragments whose size ranged from 200 to 1,500 bp. Reproducible differences in fragment patterns allowed differentiation of the two species with each primer. Similarities and differences among six different geographic populations of H. schachtii were detected. The potential application of RAPD analysis to relationships among nematode populations was assessed through cluster analysis of these six different populations, with 78 scorable markers from 10 different random primers. DNA from single cysts was successfully amplified, and genetic variability was revealed within geographic populations. The use of RAPD markers to assess genetic variability is a simple, reproducible technique that does not require radioisotopes. This powerful new technique can be used as a diagnostic tool and should have broad application in nematology.     

Low Genetic Polymorphism in Natural Populations of Crotalaria Longipes

Natural genetic variation present in two populations of the critically endangered legume C. longipes was revealed by random amplified polymorphic DNA (RAPD) analysis. Out of the 30 primers used to test the intra-specific genetic polymorphism (between individuals from isolated populations) only 21 gave amplification. Eight primers produced monomorphic bands and 13 primers produced polymorphism. But the range of percent polymorphism was only 0 to 33 %. There was close similarity between individuals within and between populations. Cluster analysis based on Nei's indices did not reveal any population differentiation and individuals of both populations clustered with each other. These results point to a very low genetic polymorphism in C. longipes populations.     

Continent-wide tracking to determine migratory connectivity and tropical habitat associations of a declining aerial insectivore

North American birds that feed on flying insects are experiencing steep population declines, particularly long-distance migratory populations in the northern breeding range. We determine, for the first time, the level of migratory connectivity across the range of a songbird using direct tracking of individuals, and test whether declining northern populations have higher exposure to agricultural landscapes at their non-breeding grounds in South America. We used light-level geolocators to track purple martins, Progne subis, originating from North American breeding populations, coast-to-coast (n = 95 individuals). We show that breeding populations of the eastern subspecies, P. s. subis, that are separated by ca. 2000 km, nevertheless have almost completely overlapping non-breeding ranges in Brazil. Most (76%) P. s. subis overwintered in northern Brazil near the Amazon River, not in the agricultural landscape of southern Brazil. Individual non-breeding sites had an average of 91 per cent forest and only 4 per cent agricultural ground cover within a 50 km radius, and birds originating from declining northern breeding populations were not more exposed to agricultural landscapes than stable southern breeding populations. Our results show that differences in wintering location and habitat do not explain recent trends in breeding population declines in this species, and instead northern populations may be constrained in their ability to respond to climate change.     

Genetic relationships among NE Turkish Lilium L. (Liliaceae) species based on a random amplified polymorphic DNA analysis

Random amplified polymorphic DNA (RAPD) fingerprinting was used to study species boundaries in six closely related NE Turkish Lilium (Liliaceae) taxa of the section Liriotypus. The investigated taxa were L. ciliatum, L. akkusianum, L. ponticum, L. kesselringianum, L. armenum, and L. szovitsianum. Of the 108 primers screened, 11 provided polymorphic and reproducible bands. A total of 93 polymorphic bands were scored for 122 individuals from 18 populations of the six Lilium taxa and principle coordinate analysis and neighbour-joining cluster analysis based on these RAPD profiles were performed. The results demonstrate a clear distinction between the two species L. ciliatum and L. akkusianum, and the other four species. While populations of the two species groups are found to be allopatrically distributed, the two species groups overlap in their geographical ranges. Analysis of molecular variance (AMOVA) indicated that nearly half of the total molecular variance is found within the individual populations and that the molecular variance among species is as high as the variance within the individual species, indicating that genetic differentiation of the species is rather weak.