Contribution of chlorogenic acids to the iron-reducing activity of coffee beverages

The iron-reducing activity of coffee beverages was determined by the ferric reducing antioxidant power (FRAP) assay. The influence on FRAP due to the degree of roasting (light, medium, and dark), species (Coffea arabica and Coffea robusta), and caffeine content (regular and decaffeinated) was investigated using ground and soluble coffee samples. The concentration of specific chlorogenic acids and caffeine in the beverages was determined by high-performance liquid chromatography and related to FRAP using Pearson correlation coefficients. All measurements were expressed per unit of soluble solids. Beverages prepared with ground coffee had, on average, 27% higher FRAP values than those prepared with soluble coffee (p < 0.05). In the former beverages, FRAP of C. robusta samples was significantly higher (on average, 50.3%) when compared to that of C. arabica samples, and FRAP values decreased with increasing degree of roasting (p < 0.05). A strong correlation (r > 0.91) was found between FRAP and the total content of chlorogenic acids, particularly that of the caffeoylquinic acid isomers. The iron-reducing activity of coffee beverages was not influenced by caffeine.     

Genetic diversity of a <Emphasis Type="Italic">Coffea</Emphasis> Germplasm Collection assessed by RAPD markers

Genetic diversity and relationships within and among nine species of Coffea, one species of Psilanthus and the Piatã hybrid from the Coffee Germplasm Collection of Instituto Agronômico de Campinas (IAC), Brazil were assessed using RAPD markers. Genetic diversity and relationships were evaluated by proportion of polymorphic loci (P), Shannon’s genetic index (H′ and G′_ST) and clustering analysis. The overall RAPD variation among all accessions was mostly partitioned between rather than within species. However, C. canephora and C. liberica showed a high genetic diversity within the species (\({\underline{\hbox{H}'}} \) _sp = 0.414 and \({\underline{\hbox{H}'}} \) _sp = 0.380, respectively) and this was highly structured (high \({\underline{\hbox{G}'}} \) _ST). Genetic diversity from C. congensis and C. arabica was also structured, but with lower levels of genetic diversity (\({\underline{\hbox{H}'}} \) _sp = 0.218 and \({\underline{\hbox{H}'}} \) _sp = 0.126, respectively). The results were consistent with agronomic and molecular studies and demonstrated that the IAC Coffea Collection is representative of the phylogenetic structure observed in the genera. This study devises sampling strategies for coffee germplasm collections and provides genetic diversity parameters for future comparisons among them.     

Use of random amplified DNA markers to analyse genetic variability and relationships of Coffea species

Summary The use of random amplified DNA fragments as genetic markers in Coffea was investigated. Arbitrary oligonucleotides were used as primers to amplify genomic DNA of different coffee accessions representing major Coffea species by polymerase chain reaction. Intraspecific variation was easily detected in C. canephora and C. liberica whereas the primers assayed failed to reveal polymorphism between C. arabica accessions. Extensive interspecific variation was observed. Genetic relationships between Coffea species are deduced from the degrees of similarity in amplified product profiles. Random amplified DNA markers appeared to be of high value for characterization, analysis and utilization of coffee genetic resources.     

Antibacterial activity of coffee extracts and selected coffee chemical compounds against enterobacteria

The in vitro antimicrobial activity of commercial coffee extracts and chemical compounds was investigated on nine strains of enterobacteria. The antimicrobial activity investigated by the disc diffusion method was observed in both the extracts and tested chemical compounds. Even though pH, color, and the contents of trigonelline, caffeine, and chlorogenic acids differed significantly among the coffee extracts, no significant differences were observed in their antimicrobial activity. Caffeic acid and trigonelline showed similar inhibitory effect against the growth of the microorganisms. Caffeine, chlorogenic acid, and protocatechuic acid showed particularly strong effect against Serratia marcescens and Enterobacter cloacae. The IC(50) and IC(90) for the compounds determined by the microtiter plate method indicated that trigonelline, caffeine, and protocatechuic acids are potential natural antimicrobial agents against Salmonella enterica. The concentrations of caffeine found in coffee extracts are enough to warrant 50% of the antimicrobial effect against S. enterica, which is relevant to human safety.     

Genetic basis of species differentiation between <Emphasis Type="Italic">Coffea liberica</Emphasis> Hiern and <Emphasis Type="Italic">C. canephora</Emphasis> Pierre: Analysis of an interspecific cross

The phenotypic and genetic differentiation between the two related Coffea species (C. liberica Hiern and C. canephora Pierre) was examined. These species differed markedly in terms of leaf, inflorescence, fruit and seed characters. A genetic map of the interspecific cross Coffea liberica × C. canephora was constructed on the basis of 72 BC1 hybrids. Eighty-three AFLP markers, four inter simple sequence repeats (ISSR) and five microsatellites corresponding to Coffea liberica species-specific markers were mapped into 16 linkage groups. The total length of the map was 1502.5 cM, with an average of 16.3 cM between markers and an estimated genome coverage of 81%. The two species were evaluated relative to 16 quantitative traits and found to be significantly different for 15 of them. Eight QTLs were detected, associated with variations in petiole length, leaf area, number of flowers per inflorescence, fruit shape, fruit disc diameter, seed shape and seed length. Results on segregation distortion and the under-representation of particular markers were interpreted in terms of genome differentiation. The implications for the introgression of QTLs involved in advantageous morphological traits (number of flowers per inflorescence, fruit and seed shape) are discussed.     

Genetic Characterization of Brazilian Annual <Emphasis Type="Italic">Arachis</Emphasis> Species from Sections <Emphasis Type="Italic">Arachis</Emphasis> and <Emphasis Type="Italic">Heteranthae</Emphasis> using RAPD Markers

The genus Arachis is divided into nine taxonomic sections. Section Arachis is composed of annual and perennial species, while section Heteranthae has only annual species. The objective of this study was to investigate the genetic relationships among 15 Brazilian annual accessions from Arachis and Heteranthae using RAPD markers. Twenty-seven primers were tested, of which nine produced unique fingerprintings for all the accessions studied. A total of 88 polymorphic fragments were scored and the number of fragments per primer varied from 6 to 17 with a mean of 9.8. Two specific markers were identified for species with 2n = 18 chromosomes. The phenogram derived from the RAPD data corroborated the morphological classification. The bootstrap analysis divided the genotypes into two significant clusters. The first cluster contained all the section Arachis species, and the accessions within it were grouped based upon the presence or absence of the ‘A’ pair and the number of chromosomes. The second cluster grouped all accessions belonging to section Heteranthae.     

Analysis of Wheat Disease Resistance Data Originating from Screenings of Gatersleben Genebank Accessions during 1933 and 1992

Series of 10,348 accessions belonging to 21 species (hexaploid, tetraploid, diploid) of the genus Triticum and 489 accessions belonging to 20 species of the genus Aegilops were scored for disease resistance during a period of 60 years. Tests were performed at the seedling stage for powdery mildew (Erysiphe graminis DC. f. sp. tritici March.), leaf rust (Puccinia recondita Rob. ex Desm. f. sp. tritici Erikss.), stripe rust (Puccinia striiformis West. f. sp. tritici Erikss.) and eyespot (Pseudocercosporella herpotrichoides (Fron.) Deight.) but also at the adult plant stage considering powdery mildew, leaf rust, stripe rust, eyespot and glume blotch (Septoria nodorum Berk.). About 150,000 disease scores recorded on index cards using different scoring scales were transferred to the computer, converted into a 1–9 scale and used to summarise the results. Within the genus Triticum 20% of the material analysed was classified as heterogeneous. For the accessions without detectable segregation a large variability for resistance/susceptibility was detected. At the adult plant stage resistant accessions without visible infections were identified for all diseases. The percentages of resistant accessions at that growth stage were always higher than the ones found in the material tested at the seedling stage. The probability for finding resistant material was shown to be highest in the diploid species ( > 50%) but decreased with increasing ploidy level to about 10% in the hexaploids. For Aegilops it was shown that most of the accessions were homogeneous and highly resistant against powdery mildew (seedling and adult plant stage), leaf rust (adult plant stage) and eyespot (seedling and adult plant stage/natural infection). The data obtained for the individual accessions are available via Internet (http://www.ipk-gatersleben.de). An erratum to this article is available at .     

Relationships among species of Hystrix Moench and Elymus L. assessed by RAPDs

To assess the generic delimitation and the interspecific relationships between Hystrix and Elymus, three Hystrix and 10 Elymus species were used for random amplified polymorphic DNA(RAPD) assay. Of the 54 primers tested, 26 (48%) produced polymorphic products. A total of 167 products amplified from 16 primers were selected for RAPD analysis, among which 156 (93.4%) amplified products were found to be polymorphic among the 13 species. The polymorphism produced by each primer ranged from 4 to 13, with an average of 9.8. Data were used to generate Jaccard's similarity coefficients and to construct a dendrogram using UPGMA in the NTSYS computer programs. It is concluded from this study that: (1) there were clear differences between Hystrix and Elymus, which possibly suggest that Hystrix is a valid genus; (2) great diversity existed among the species of Hystrix and Elymus; (3) the species similar to each other in morphological characters were grouped together; (4) the species from neighboring geographical regions were clustered; (5) the species with the same genomes and polyploidy level were clustered together; (6) RAPD results are comparable with those obtained from studies on morphology and cytology. It is a useful additional method for assessing the relationships among genera and species in Triticeae.     

Mendeschilis a new genus of Microcoryphia (Insecta, Apterygota, Machilidae) from Spain

A new species and genus of Microcoryphia is described as a result of the detailed comparison of specimens from Italy and Spain which had been initially identified as Praemachilis excelsior. The new genus, named Mendeschilis has as its most important distinctive taxonomic characteristic the presence, in the male, of parameres in the VIII^th and IX^th urosternites, while Praemachilis (from Italy) has only one pair of parameres in the IX^th urosternite. Other anatomical characteristics which allow us to differentiate the new genus and the new species are described.     

Genetic diversity of the complex Paspalum notatum Flügge (Paniceae: Panicoideae)

The genus Paspalum L. comprises approximately 400 species worldwide and about 220 in Brazil. Paspalum is ecologically and economically important, and has been very useful as pasture and P. notatum Flügge (bahiagrass) is a valuable forage grass in the subtropics. This species consists of several sexual (diploid) and apomictic (tetraploid, ocasionally tri and pentaploids) biotypes. In this work, inter simple sequence repeats (ISSR) markers were used to assess the genetic variability of a bahiagrass (P. notatum) collection. Vegetative tissues of 95 bahiagrass accessions were obtained from various locations in South America (Brazil, Argentina and Uruguay). A total of 91 reproducible ISSR fragments were observed and 89 fragments (97.5% of the total observed) were polymorphic. Cluster analyses (UPGMA) were performed from the ISSR data set and the results illustrate the genetic relationships among the 95 accessions of P. notatum. A comparison among molecular, morphological and ploidy levels data were done. ISSR markers were effective in distinguishing the genotypes analyzed, and a wide variability was observed for this species. These results add new information regarding the genetic diversity in P. notatum, thus contributing toward the biological knowledge of this species, and providing with subsides for future plant breeding and conservation programs.     

Genetic diversity and differentiation of <Emphasis Type="Italic">Camellia sinensis</Emphasis> L. (cultivated tea) and its wild relatives in Yunnan province of China,revealed by morphology,biochemistry and allozyme studies

We evaluated morphological, isozyme and biochemical diversity of a total of 87 accessions in the genus Camellia [Camellia sinensis var. sinensis (10), C. talinensis (7), C. sinensis var. dehungensis (3), C. crassicolumna (3) and C. sinensis var. assamica (64)]. Great variation of morphological characters was apparent within each taxa. Across the five taxa, all leaf and most flower quantitative characters showed significant differences while all fruit quantitative characters measured did not differ significantly, and, seven (i.e., life form, bud color, petal texture, pubescence on ovary, style number, stamen location and locule per fruit) of the 33 qualitative characters yield significant differences. As a whole, caffeine content had the highest variation with CV of 22.7%, water extract solid showed the least variation (13.4%) and content of polyphenols (20.0%) and free amino acids (18.8%) showed intermediate variations. Camellia taliensis and C. sinensis var. assamica had significantly higher content of polyphenols and water extract solid than in the other three taxa, while no significant differences were detected for the content of caffeine and free amino acids. For allozyme study, 14 loci presented good resolution, among which, nine loci (64%) were polymorphic in each taxon (AAT-3, FUM-1, 6PDG-1, G6PDH-1, G3PDH-1, ME-1, PGM-1, PGM-2 and SKD-1). The percentage of polymorphic loci (P) for each taxon was 21.4–50.0%. Mean heterozygosity per locus (H _e ) varied 0.114–0.218. F _ST value indicated that only 4.6% of the variations could be ascribable to genetic differences among taxa. Genetic relationships among the five taxa revealed by allozymes, were also exposed by the result of clustering of the morphological and biochemical characters.     

Observations on interspecific compatibility and meiotic chromosome behavior of Capsicum buforum and C. lanceolatum

The wild species, Capsicum buforum Hunz. and C. lanceolatum(Greenm. ex J.D. Sm.) Morton and Standl. were hybridized to nine different Capsicum species to understand their taxonomic and genetic relationships. With C. buforum as the male parent, the compatibility to the nine species varied from species to species and ranged from producing under-developed embryo, seed coat, seedless fruit, to no fruit set. When C. buforum was the female parent, it was incompatible (no fruit set) to the nine Capsicum species tested. When C. lanceolatum was the female parent, the hybridizations to the other species ranged from aborted embryo, seed coat, seedless fruit or no fruit. As a pollen parent, C. lanceolatum was incompatible (no fruit set) to the species investigated. In pollen mother cells (PMCs) of C. buforum, 24 chromosomes (n = 12) paired as 12 bivalents with chromosome lagging at meiotic anaphase-I. Twenty-six chromosomes (n = 13) were detected in PMCs of C. lanceolatum. In C. lanceolatum most chromosomes paired as bivalents, but one quadrivalent was observed in some cells. C. buforum was found to be self-incompatible, while C. lanceolatum was self-compatible.     

Introduction to the wild resources of the genus Boehmeria Jacq. in China

China, the native home of ramie (Boehmeria nivea (L.) Gaud. var. nivea), possesses many wild species in the genus Boehmeria Jacq., including many those rare valuables in some characteristics such as stress resistance and fiber qualities that can be used for biological engineering, genetics and breeding research. From 1995 to 1999, 130 samples of the genus Boehmeria that belong to 22 species and 6 varieties, were collected in 34 counties of 14 provinces in China in the region of N18.8–34.2° and E101–121°. The gene pool of wild species of the genus Boehmeria was constructed at Yichun, Jiangxi Province and 77 samples belonging to 8 species and 4 varieties were conserved alive. The collected species are classified into 5 sections i.e. Boehmeria, Tilocnide Bl., Zollingerianae Satake, Phyllostachys W. T. Wang and Duretia Bl., according to the morphology. They may also be divided into hygric type, moderately hygric type, semi-shade-hygrophyte and xeromorphy, based on their ecological adaptability and primitive growing circumstances. The distribution of the wild species and populations in the genus Boehmeria is related to the altitude.     

Genetic diversity of rhizobia associated with indigenous legumes in different regions of Flanders (Belgium)

We investigated the diversity of rhizobia isolated from different indigenous legumes in Flanders (Belgium). A total of 3810 bacterial strains were analysed originating from 43 plant species. Based on rep-PCR clustering, 16S rRNA gene and recA gene sequence analysis, these isolates belonged to Bradyrhizobium, Ensifer (Sinorhizobium), Mesorhizobium and Rhizobium. Of the genera encountered, Rhizobium was the most abundant (62%) and especially the species Rhizobiumleguminosarum, followed by Ensifer (19%), Bradyrhizobium (14%) and finally Mesorhizobium (5%). For two rep-clusters only low similarity values with other genera were found for both the 16S rRNA and recA genes, suggesting that these may represent a new genus with close relationship to Rhodopseudomonas and Bradyrhizobium. Primers for the symbiotic genes nodC and nifH were optimized and a phylogenetic sequence analysis revealed the presence of different symbiovars including genistearum, glycinearum, loti, meliloti, officinalis, trifolii and viciae. Moreover, three new nodC types were assigned to strains originating from Ononis, Robinia and Wisteria, respectively. Discriminant and MANOVA analysis confirmed the correlation of symbiosis genes with certain bacterial genera and less with the host plant. Multiple symbiovars can be present within the same host plant, suggesting the promiscuity of these plants. Moreover, the ecoregion did not contribute to the separation of the bacterial endosymbionts. Our results reveal a large diversity of rhizobia associated with indigenous legumes in Flanders. Most of the legumes harboured more than one rhizobial endosymbiont in their root nodules indicating the importance of including sufficient isolates per plant in diversity studies.     

The RAPD evidence for the phylogenetic relationship of the closely related species of cultivated apple

The phylogenetic relationship of cultivated apple and its closely related species is still not clear in the taxonomy of genus Malus. To try to find new evidence for the origin and evolution of the cultivated apple, random amplified polymorphic DNA (RAPD) markers of 14 taxa of Malus, among which a reference species (M. toringoides) and six presumably ancestral species of cultivated apple in the genus were investigated. The RAPD data obtained were used to construct both unrooted and rooted trees using TREECON software package. The result showed in our rooted tree that M. sieversii from the Xinjiang Autonomous Region of China is the species which is most closely related to the cultivated apple, M. domesticacv. `Golden Delicious'. The phylogenetic relationships among the species studied are discussed.     

AFLP Analysis of the Phenetic Organization and Genetic Diversity in the Sugarcane Complex, Saccharum and Erianthus

Amplified fragment length polymorphism (AFLP) markers were evaluated for determining the phylogenetic relationships, and the diversity in the Saccharum complex using 30 clones belonging to S. officinarum, S. robustum, S. spontaneum, S. barberi, S. sinense and the related genus Erianthus. The phenetic tree of the species clones based on AFLP data was consistent with the known taxonomical relationships. AFLP gave higher resolution of closely related species into discrete groups than that by RAPD and RFLP markers, reported earlier. The levels of diversity within the various Saccharum species were also found to be higher than those obtained previously with the same set of clones using RAPD markers. The intraspecies similarity in S. barberi and S. sinense was much higher than interspecies similarity suggesting a clear separation of the two, which are considered ‘horticultural species’. The genetic similarity matrix derived from a single primer combination highly correlated (r = 0.980) with that obtained from all the 12 primer combination used in the study, thus highlighting the efficiency of a single primer combination in delineating species relationships. All the primer combinations could identify markers that are specific to each of the species and the genus Erianthus. Among the species, specific markers were highest in S. spontaneum followed by S. robustum, S. barberi, S. officinarum and S. sinense. Erianthus had a distinct profile with 30% of the total amplified fragments being specific to it. This offers great scope for identifying intergeneric hybrids, which has been very difficult using morphological traits and RAPD markers. High degree of correspondence between the results from the cluster analysis based on Jaccard's similarity index, Neighbour Joining tree based on Sokal and Michener distance matrix and AFTD (Analyses Factorielle on Table of Distances) analysis clearly demonstrated that AFLP markers would be an appropriate tool in providing better information about the relationships among the species, estimation of diversity, and in revealing species and genus specific markers that could be directly applied in sugarcane breeding programmes.     

An assessment of the genetic integrity of ex situ germplasm collections of three endangered species of Coffea from Madagascar: implications for the management of field germplasm collections

Madagascar has 59 species of Coffea, of which 42 are listed as Critically Endangered, Endangered, or Vulnerable by criteria of the Red List Category system of the World Conservation Union. In an attempt to assess the conservation value of ex situ collections of Malagasy coffee species, a study was undertaken using the field genebank collections maintained at the Kianjavato Coffee Research Station. Three species were selected for this purpose, C. kianjavatensis, C. montis-sacri, and C. vatovavyensis, and for comparative purposes extant, in situ populations of the same species were studied. Parentage analyses of ex situ propagated offspring of C. kianjavatensis and C. montis-sacri were performed to assess if crossing with other Coffea species maintained in the field genebank is compromising the genetic integrity of the collection. For these three species, higher genetic diversity was observed in the ex situ populations compared to the in situ populations, highlighting the importance of preserving the plants currently in ex situ collections. Parentage analyses of seed-propagated offspring of C. kianjavatensis and C. montis-sacri revealed that cross contamination with pollen from other Coffea species in the ex situ field genebank is occurring. These results have significant implications for the conservation management of wild Coffea species and for the management of ex situ genebanks.     

Electrophoretic variation as a measure of speciesdifferentiation between four species of the genus Lolium

Isozyme electrophoresis was carried out on 423 accessionsbelonging to four species of the genus Lolium toassess the genetic variation within and between the species. Fourenzyme systems (acid phosphatase, glutamate oxaloacetatetransaminase, phospho-gluco isomerase and superoxidedismutase) were used to array allelic diversity at fivepolymorphic loci. Nei's genetic distance analysis andmultivariate analyses were computed and the taxonomic position ofeach species is discussed as a result of these computations.Electrophoresis is shown to clearly differentiate the inbreedingspecies, L. temulentum fromthe outcrossing species, L.perenne, L.multiflorum and L.rigidum, and to separate the species within theout-breeding group. The taxonomic results are discussed inrelation to an earlier paper on morphological variation, concludingthat electrophoresis is a valid taxonomic tool showing distinctseparations between the species, but that current plant breeding andagricultural practises may be increasing the amount of hybridisationbetween the species than occurred in the past.     

Genomic affinities of <Emphasis Type="Italic">Arachis</Emphasis> genus and interspecific hybrids were revealed by SRAP markers

The Arachis genus is native to South America, and contains 70–80 described species assembled into nine sections. A better understanding of the level of speciation and taxonomic relationships is a prerequisite to the effective use of Arachis species in peanut breeding programs. Forty-eight genotypes representing 19 species in 6 sections were evaluated to assay the genetic variability within and among species, and 10 recombinant lines and those parents were identified with introgression of Arachis species chromosome segments into A. hypogaea genome using SRAP markers. Sixty of sixty-four SRAP primers tested were selected for DNA amplification reactions. A dendrogram and principal component analysis were constructed based on 353 SRAP polymorphic bands of the accessions. The number of scored polymorphic bands per each primer combination varied from 1 to 25 with an average of 5.9 per reaction. Estimates of genetic distance among the 48 accessions Arachis species ranged from 0.11 to 0.76. A-genome accessions 475845 (A. duranensis), and 331197 (A. villosa) were most closely associated to A. hypogaea. The first two PCAs accounted for 77.74% (62.02 and 15.72%) of the total variation observed and separated the different genomic groups. SRAPs also identified introgression of Arachis species chromosome segments into A. hypogaea. genome with 10 recombinant lines and those parents. The present results indicated that SRAPs can be used to determine the genetic relationships among species of the different sections of the genus Arachis and to identify introgression of Arachis genus chromosome segments into A. hypogaea genome.     

Molecular phylogeny of genus <Emphasis Type="Italic">Guizotia</Emphasis> (Asteraceae) using DNA sequences derived from ITS

Complete sequences for the internal transcribed spacers of the 18s–26s nuclear ribosomal DNA were generated to establish phylogenetic relationships among five species of the genus Guizotia. Parsimony analysis and pairwise distance data produced a single tree with four clearly distinguished clades that accord with previously reported chromosomal data. The clades produced here have been discussed with reference to existing taxonomic treatments. It appears that Guizotia scabra ssp. scabra, G. scabra ssp. schimperi and Guizotia villosa have contributed to the origin of Guizotia abyssinica, the cultivated species of the genus. The present composition of the species of genus Guizotia and the subtribe the genus presently placed in are suggested to be redefined.