Phylogenetic relationships of species in <Emphasis Type="Italic">Pseudoroegneria</Emphasis> (Poaceae: Triticeae) and related genera inferred from nuclear rDNA ITS (internal transcribed spacer) sequences

To evaluate the phylogenetic relationships of species in Pseudoroegneria and related genera, the nuclear ribosomal internal transcribed spacer (ITS) sequences were analyzed for eighteen Pseudoroegneria (St), two Elytrigia (E ^e St), two Douglasdeweya (StP), three Lophopyrum (E ^e and E ^b ), three Agropyron (P), two Hordeum (H), two Australopyrum (W) and two Psathyrostachys (Ns) accessions. The main results were: (i) Pseudoroegneria gracillima, P. stipifolia, P. cognata and P. strigosa (2x) were in one clade, while P. libanotica, P. tauri and P. spicata (2x) were in the other clade, indicating there are the differentiations of St genome among diploid Pseudoroegneria species; (ii) P. geniculata ssp. scythica, P. geniculata ssp. pruinifera, Elytriga caespitosa and Et. caespitosa ssp. nodosa formed the E ^e St clade with 6-bp indel in ITS1 regions; and (iii) Douglasdeweya wangii, D. deweyi, Agropyron cristatum and A. puberulum comprised the P clade. It is unreasonable to treat P. geniculata ssp. scythica and P. geniculata ssp. pruinifera as the subspecies of P. geniculata, and they should be transferred to a new genus Trichopyrum, which consists of species with E ^e St genomes. It is also suggested that one of the diploid donor of D. wangii and D. deweyi is derived from Agropyron species, and it is reasonable to treat tetraploid species with StP genomes into Douglasdeweya.     

Phylogenetic analysis of questionable tetraploid species in Roegneria and Pseudoroegneria (Poaceae: Triticeae) inferred from a gene encoding plastid acety1-CoA carboxylase

To evaluate the phylogenetic relationships of questionable tetraploid species Roegneria alashanica Keng, Roegneria magnicaespes (D.F. Cui) L.B. Cai, Roegneria elytrigioides C. Yen et J.L. Yang, Roegneria grandis Keng and Pseudoroegneria geniculata (Trin.) Á. Löve, the single copy sequences of the plastid acetyl-CoA carboxylase gene (Acc1) were analyzed among the five species and the related diploid and tetraploid species. The results indicated that: (a) R. alashanica contained one set of modified St genome which was closely related to the E^e genome, and the other set of genome was closely related to the P genome; (b) R. magnicaespes contained one set of St genome, the other set of genome might be closely related to the P genome. There are close affinities between R. magnicaespes and R. alashanica; (c) R. elytrigioides contained two sets of St genomes, and it is reasonable to be treated as Pseudoreogneria elytrigioides (C. Yen et J.L. Yang) B.R. Lu; (d) the genome of R. grandis should be designed as St^gY. The St^g genome was a differentiated form of the St genome in Pseudoroegneria and was homoeologous with the Y genome in Roegneria; (e) the genomic constitution of P. geniculata was similar to that of R. magnicaespes and R. alashanica and distinctly related to P. geniculata ssp. scythica (E^eSt). They should be treated as different species in different genera; and (f) the Y genome was possibly originated from the St genome, and was sister to the St, E^e, E^b and W genomes.     

Phylogeny and maternal donors of the tetraploid species with St genome (Poaceae: Triticeae) inferred from CoxII and ITS sequences

Mitochondrial CoxII and ITS sequences of 29 tetraploid species with St genome were compared with their related genera and species of Pseudoroegneria (Nevski) Á. Löve (St), Hordeum L. (H), Thinopyrum bessarabicum (Savul. & Rayss) Á. Löve (E^b), Lophopyrum elongatum (Host) Á. Löve (E^e), Agropyron Gaertner (P), Australopyrum (Tzvelev) Löve (W) and Psathyrostachys Nevski (Ns). The results indicate that: (1) the maternal donors of North American and Eurasian StStHH tetraploid species may have acquired their St genome from distinct Pseudoroegneria gene pools, with Pse. spicata (Pursh) Á. Löve potentially the maternal donor of North American species; (2) Lophopyrum is the maternal donor of StStE^eE^e tetraploid species, hence, Pse. geniculata ssp. scythica (Nevski) Á. Löve, Elytrigia caespitosa (K. Koch) Nevski and El. caespitosa ssp. nodosa (Nevski) Tzvelev should be identified as species of Trichopyrum Á. Löve; (3) tetraploid species with the same maternal donors were more closely related to each other than those with different maternal donors.     

Molecular cytogenetic study of three common Mediterranean limpets, Patella caerulea, P. rustica and P. ulyssiponensis (Archaeogastropoda, Mollusca)

The present paper shows the results of chromosome banding and rDNA-FISH study performed on several specimens of different populations of Patella caerulea, Patella rustica and Patella ulyssiponensis. The taxonomic attribution of specimens was ascertained by the molecular phylogenetic analysis of the mitochondrial 16S rRNA gene. P. caerulea and P. rustica had 2n = 18 chromosomes with first seven of biarmed pairs and the remaining two uniarmed pairs. P. ulyssiponensis had 2n = 16 with all biarmed chromosomes. Ag-NOR loci were on the short arms of the first metacentric pair in the three studied limpets, whereas they showed a different pattern of heterochromatin distribution and composition. A chromosome mosaicism was observed in several P. caerulea specimens, which exhibited an unpaired metacentric element and loss of a telocentric pair. The obtained results suggest that in the genus Patella specific diversification was accompanied by variations in heterochromatin distribution and composition and reduction of chromosome number by Robertsonian centric fusion.     

Molecular phylogeny of RNA polymerase II gene reveals the relationships of tetraploid species with St genome (Triticeae: Poaceae)

To evaluate phylogeny of tetraploid with St genome, phylogenetic analyses of RNA polymerase II (RPB2), a member of the nuclear gene family encoding the second largest subunit, were performed. Our results showed that: (1) Roegneria magnicaespes and Roegneria alashanica are related to Pseudoroegneria. (2) Roegneria elytrigioides has StStStSt genomes and should therefore be classified as Pseudoroegneria elytrigioides. (3) Pseudoroegneria tauri and Pseudoroegneria deweyi which have StStPP genomes should be transferred to Douglasdeweya and be renamed as Douglasdeweya wangii and Douglasdeweya deweyi, respectively. (4) Pseudoroegneria geniculata ssp. scythica is related to Pseudoroegneria and Lophyrum, and hence should be identified as a species of Trichopyrum. (5) Pseudoroegneria libanotica might be a parental donor for Elytrigia caespitosa rather than Elytrigia caespitosa ssp. nodosa. It is unreasonable to recognize El. caespitosa ssp. nodosa as a subspecies of El. caespitosa. (6) Interspecific and intergeneric variations are detected in St genome of these tetraploid species.     

Conservation genetics of Butte County meadowfoam (Limnanthes floccosa ssp. californica Arroyo), an endangered vernal pool endemic

The endangered annual plant Limnanthes floccosa ssp. californica Arroyo is restricted to vernal pools in Butte County, California. To identify populations with unique genetic resources, guide reintroduction efforts, and design seed collection scenarios for long-term ex situ seed storage we determined extant genetic diversity and structure by surveying 457 individuals from 21 distinct populations using nine polymorphic microsatellite markers. We found low within population genetic diversity: low allelic diversity (1.9 [0.06 SE] alleles/locus); low heterozygosity (H _obs = 0.10 ± 0.018, H _exp = 0.19 ± 0.015), and a high fixation index (0.556 ± 0.044). The number of polymorphic loci ranged between 11 and 89%. Bayesian ordination determined 20 distinct populations and we found high genetic structure among these (F _st = 0.65, P < 0.0001). We identified notable gene flow barriers across populations, confirming regional structuring between three previously defined population density centers and two outlying populations (F _st = 0.21, P < 0.0001). Population size estimates ranged between ~50 and >5,000 extant plants per site. Our study confirms previous isozyme-based results and suggests that the loss of any population would represent a significant loss in the species’ genetic diversity. Recovery requires active restoration of existing populations and permanent habitat protection. We recommend close comparison of microhabitats of declining populations with genetically similar populations, to determine the potential for human assisted gene flow via seed movement to recover declining populations.     

Quantitative analysis of a stand of Pinus densiflora undergoing succession to Quercus mongolica ssp. crispula : II. Growth and population dynamics of Q. mongolica ssp. crispula under the P. densiflora canopy

We report here the results of an 8-year study of the growth and population dynamics of Quercus mongolica ssp. crispula in a Pinus densiflora stand in a state of succession. In 1998, there were 169 Q. mongolica ssp. crispula individuals in a 400-m² plot under the P. densiflora canopy. This number remained nearly constant between 1998 and 2005. Mean recruitment of new individuals was 11 year⁻¹, while mean mortality was 12 year⁻¹. Of the 35 individuals ≥60 cm in height existing in 1998, 30 were still surviving in 2005. We were able to represent the height growth of Q. mongolica ssp. crispula individuals as H=30 [1+21.96 exp(−0.0839t)]⁻¹, with t = years since 1998 and H = height in meters. Using this equation we predict that by 2015 the mean height of Q. mongolica ssp. crispula trees in the stand will exceed those of understory trees, such as Rhus trichocarpa and Prunus maximowiczii. Once above the understory stratum, the Q. mongolica ssp. crispula trees can be expected to grow more rapidly due to the better light conditions, thereby rapidly reaching the canopy stratum of the P. densiflora stand.     

Development and characterization of microsatellite loci in <Emphasis Type="Italic">Ixeridium dentatum</Emphasis> (Asteraceae,Lactuceae)

We isolated and characterized seven microsatellite loci for the perennial herb Ixeridium dentatum ssp. dentatum, an apomictic triploid distributed throughout the lowland areas of East Asia. The number of alleles ranged from two to seven in 32 screened individuals of I. dentatum ssp. dentatum from Japan. The observed and expected heterozygosities were 0.000–0.950 and 0.000–0.891, respectively, calculated using genotypes of 20 individuals of I. dentatum ssp. nipponicum. One locus (msid4) deviated significantly from Hardy–Weinberg equilibrium (P = 0.0001). These microsatellites were also tested for cross-amplification in 11 other taxa of Lactuceae, including five endangered taxa. These primers should be useful genetic tools not only for Ixeridium but also for other Lactuceae taxa.     

The transfer RNA genes in Oryza sativa L. ssp. indica

The availability of the draft genome sequence of Oryza sativa L. ssp. indica has made it possible to study the rice tRNA genes. A total of 596 tRNA genes, including 3 selenocysteine tRNA genes and one suppressor tRNA gene are identified in 127551 rice contigs. There are 45 species of tRNA genes and the revised wobble hypothesis proposed by Guthrie and Abelson is perfectly obeyed. The relationship between codon usage and the number of corresponding tRNA genes is discussed. Redundancy may exist in the present list of tRNA genes and novel ones may be found in the future. A set of 33 tRNA genes is discovered in the complete chloroplast genome of Oryza sativa L. ssp. indica. These tRNA genes are identical to those in ssp. japonica identified by us independently from the origional annotation.     

First report on the molecular prevalence of Mycoplasma capricolum subspecies capripneumoniae (Mccp) in goats the cause of contagious caprine pleuropneumonia (CCPP) in Balochistan province of Pakistan

Contagious caprine pleuropneumonia (CCPP) caused by Mycoplasma capricolum subspecies capripneumoniae (Mccp) is a disease of goats which causes high morbidity and mortality and is reported in many countries of the world. There are probably no reports on the molecular prevalence of Mccp, Mycoplasma capricolum subsp. capricolum (Mcc) and Mycoplasma putrefaciens (Mp) in Balochistan and any other part of Pakistan. Thirty goats (n = 30) with marked respiratory symptoms were selected and procured from forty goat flocks in Pishin district of Balochistan in 2008. The genomic deoxyribonucleic acid (DNA) from the lung samples (n = 30) of the slaughtered goats was purified and subjected to polymerase chain reaction (PCR) assays for the presence of Mycoplasma mycoides cluster members and Mp. The PCR-RFLP (restriction fragment length polymorphism) was also used to further confirm the Mccp. Of the thirty lung samples 17 (56.67%) were positive for the molecular prevalence of Mcc, Mccp and Mp. In total the molecular prevalence was observed as 17.65% for Mccp (n = 3), 70.59% for Mcc (n = 12) and 11.76% for Mp (n = 2). The RFLP profile has also validated the PCR results of Mccp by yielding two bands of 190 and 126 bp. The results of PCR-RFLP coupled with the presence of fibrinous pleuropneumonia and pleurisy during postmortem of goats (n = 3) strongly indicated the prevalence of CCPP in this part of world. Moreover the prevalence of Mcc and Mp is also alarming in the study area. We report for the very first time the molecular prevalence of Mcc, Mccp, and Mp in the lung tissues of goats in the Pishin district of Balochistan, Pakistan.     

Primary production and nutrient budgets of Sarcocornia perennis ssp. alpini (Lag.) Castroviejo in the salt marsh of the Palmones River estuary (Southern Spain)

Biomass, primary production and nutrient budgets associated to Sarcocornia perennis subspecies (ssp.) alpini were studied in the Palmones River estuary salt marsh (Southern Spain) to evaluate the nutrient sequestration capacity of the low marsh. Above- and belowground living and dead biomass, as well as carbon, nitrogen and phosphorus content were monitored during 1 year. Additionally, the fate of aboveground detritus was evaluated in an experiment on litter decomposition. The detritus production of S. perennis ssp. alpini was almost equivalent to its annual primary production indicating a rapid turnover of biomass. We calculated that only 12% of the aboveground detritus was exported out of the low marsh while the rest was decomposed in the sediment with a rate of 0.8 year⁻¹. Changes in concentrations of total carbon, nitrogen and phosphorus in the sediment showed patterns related to S. perennis ssp. alpini belowground biomass. Our results suggested that the sediment functions as a net sink for nutrients accumulating 550 g C m⁻² year⁻¹, 55 g N m⁻² year⁻¹, and 13 g P m⁻² year⁻¹.     

Ricerche Embriologiche Sul Genere Thalictrum. II. Embriologia e Cariologia Di Thalictrum Lucidum L. e di Thalictrum Minus L. Subsp. Minus

Abstract

Embryological researches in the «Thalictrum» genus. II. Embryology and caryology of «Thalictrum lucidum» L. and «Thalictrum minus» L. ssp. «minus». — In their development the female gametophytes of Thalictrum lucidum L. and Thalictrum minus L. ssp. minus follow the Normal type. In the female gametophytes of these species several types of antipodal cells occurs such as: their considerable enlargement, formation of many antipodals polyploid or polinucleated antipodal cells. In these various types, however, a rapid regression of the antipodals occurs. In T. lucidum some tendency to a disposition of the tetrapolarized type has been verified. The chromosome number has been confirmed to ben 2n=28 in besides, in the pollen development, many cases of regression of the microspores has been verified. The chromosome number has been confirmed to ben 2n=28 in Thalictrum lucidum L., and has been found to ben 2n=14 in Thalictrum minus L. ssp. minus, a new number in this species, which was earlier reported to have 2n=42.     

The allotetraploid <Emphasis Type="Italic">Arabidopsis thaliana–Arabidopsis lyrata</Emphasis> subsp. <Emphasis Type="Italic">petraea</Emphasis> as an alternative model system for the study of polyploidy in plants

Polyploidy is known to be common in plants and recent work has focused on the rapid changes in genome structure and expression that occur upon polyploidization. In Arabidopsis, much of this work has been done on a synthetic allotetraploid obtained by crossing a tetraploid Arabidopsis thaliana (2n = 4x = 20) with A. arenosa (2n = 4x = 32). To explore an alternative route to polyploidy in this model species, we have developed a synthetic allopolyploid by crossing two diploid species: A. thaliana (2n = 2x = 10) and Arabidopsis lyrata subsp. petraea (2n = 2x = 16). F₁ hybrids were easy to obtain and phenotypically more similar to A. lyrata. Spontaneous chromosome doubling events occurred in about 25% of the F₁s, thus restoring fertility. The resulting allotetraploids (2n = 26) exhibited many genomic changes typically reported upon polyploidization. Nucleolar dominance was observed as only the A. lyrata rDNA loci were expressed in the F₁ and allotetraploids. Changes in the degree of methylation were observed at almost 25% of the loci examined by MSAP analysis. Finally, structural genomic alterations did occur as a large deletion covering a significant portion of the upper arm of chromosome II was detected but no evidence of increased mobility of transposons was obtained. Such allotetraploids derived from two parents with sequenced (or soon to be sequenced) genomes offer much promise in elucidating the various changes that occur in newly synthesized polyploids.     

Grazer interactions with four species of Lyngbya in southeast Florida

Blooms of the toxic cyanobacteria Lyngbya spp. have been increasing in frequency and severity in southeast Florida in recent years. Lyngbya produces many active secondary metabolites which often act as feeding deterrents to generalist herbivores, possibly increasing the longevity of these nuisance blooms. Whilst diverse arrays of small invertebrate consumers are often found in association with Lyngbya, little is known of their grazing selectivity among species of Lyngbya. This study examines the feeding preference of grazers for four local Lyngbya species (Lyngbya majuscula, Lyngbya confervoides, Lyngbya polychroa and Lyngbya spp.). Stylocheilus striatus and Haminoea antillarum showed no dietary selectivity between L. polychroa, L. majuscula and Lyngbya spp. in multiple choice feeding assays, whereas Bulla striata showed a distinct preference for L. polychroa (P < 0.001). To determine whether preference might be related to species-specific secondary metabolites, L. majuscula, L. confervoides and L. polychroa non-polar and polar extracts were incorporated into artificial diets and offered to a range of mesograzers. No significant difference was noted in feeding stimulation or deterrence amongst extracts and the controls for any of the grazers. When fed a monospecific diet of L. polychroa, S. striatus consumed more (P < 0.001) and attained a higher daily biomass (P = 0.004) than S. striatus fed L. confervoides. As L. polychroa and L. confervoides often co-exist on local coral reefs and yield dense numbers of S. striatus, host switching to a more palatable species of Lyngbya may have important implications regarding top-down control of local blooms leading to proliferation of one species and decimation of another. S. striatus fed a diet of L. polychroa consumed more (P = 0.003), had a greater increase in body mass (P = 0.020) and higher conversion efficiency (P = 0.005) than those fed L. confervoides regardless of host origin. Possible explanations for host switching between species of Lyngbya related to morphology, toxicity and nutrient requirements for growth are discussed.     

Hosts and distribution of Viscum album L. ssp. album in Croatia and Slovenia

Abstract

Viscum album L. ssp. album is semi-parasitic on deciduous trees and shrubs. In order to identify hosts and map the distribution of V. album ssp. album in Croatia and Slovenia, field research was carried out, and herbaria were surveyed. In Croatia and Slovenia, V. album ssp. album occurred on 59 taxa. In Croatia, there were 52 hosts (33 autochthonous and 15 allochthonous species, two cultivars and two hybrids). In Slovenia, there were 25 hosts (21 autochthonous and four allochthonous species). There were 18 hosts common to both countries, 34 hosts were found only in Croatia, and seven hosts only in Slovenia. The hosts belonged to 13 families. The majority of these (19 species) belong to the Rosaceae, followed by Salicaceae, Aceraceae, Betulaceae, Fagaceae, Juglandaceae, Tiliaceae, Hippocastanaceae, Ulmaceae, Oleaceae, Fabaceae, Moraceae and Viscaceae. All hosts have been previously recorded in the literature, except Alnus japonica (Thunb.) Steud., Amelanchier lamarckii F.G. Schroed. and Crataegus nigra Waldst. et Kit. The distribution of this mistletoe was scattered, due to the scattered distribution of hosts, local conditions, movement of bird-vectors, etc. A continuous distribution was found only in part of the distribution area of narrow-leaved ash (Fraxinus angustifolia Vahl).     

Karyotype and genomic in situ hybridization pattern in ×Brassicoraphanus, an intergeneric hybrid between Brassica campestris ssp. pekinensis and Raphanus sativus

The karyotype and genomic in situ hybridization (GISH) of an intergeneric hybrid Baemoochae, ×Brassicoraphanus, which originated from hybridization between Chinese cabbage, Brassica campestris (synonym, rapa) ssp. pekinensis, and radish, Raphanus sativus, were analyzed to determine its chromosome complement. In the karyotype analysis, B. campestris was verified to have 2n = 20 chromosomes, including a particular pair of the subtelomeric chromosomes with the nucleolar organizer; R. sativus to have 2n = 18 chromosomes, including a particular pair of the submetacentric chromosomes with the secondary constriction of nucleolar organizer; and ×Brassicoraphanus to have 2n = 38 chromosomes, including both the subtelomeric chromosomes of Brassica and the secondary constriction chromosome pair of Raphanus. These findings indicate that ×Brassicoraphanus is a polyploid between Brassica and Raphanus. In the GISH analysis using chromosomes of B. campestris and R. sativus as the probe and blocking DNA, respectively, only 20 chromosomes of Brassica had hybridization signals. This result reveals that ×Brassicoraphanus is an intergeneric hybrid consisting of the complete genomes of both Brassica and Raphanus. However, the nucleolar organizers of Brassica and Raphanus were not identified because the hybridization signals appeared to be centering mainly around the centromere, becoming weak at the edges.     

Description of Brenneria roseae sp. nov. and two subspecies, Brenneria roseae subspecies roseae ssp. nov and Brenneria roseae subspecies americana ssp. nov. isolated from symptomatic oak

Gram-negative, facultatively anaerobic bacteria were isolated from symptomatic oak tissue in the UK and USA. Partial gyrB sequencing placed ten strains in the genus Brenneria, with B. goodwinii as the closest phylogenetic relative. The strains were investigated further using a polyphasic approach including MLSA (based on partial gyrB, rpoB, infB and atpD gene sequences), 16S rRNA gene sequencing, DNA–DNA relatedness studies and both phenotypic and chemotaxonomic assays. The MLSA and 16S rRNA gene analyses separated the strains into two groups based on origin, suggesting that they belong to Brenneria as two novel species. However, the DNA–DNA relatedness values revealed a closer relationship between the groups and indicated that they should belong to the same species. As the two groups of strains from the UK and USA can be differentiated from each other phenotypically and by ERIC PCR fingerprints, it is proposed to classify them as novel subspecies of a novel Brenneria species. The name Brenneria roseae sp. nov. (FRB 222^T = LMG 27714^T = NCPPB 4581^T) is proposed, with Brenneria roseae subsp. roseae ssp. nov. (FRB 222^T = LMG 27714^T = NCPPB 4581^T) for the strains from the UK and Brenneria roseae subsp. americana ssp. nov. (FRB 223^T = LMG 27715^T = NCPPB 4582^T) for the strains from the USA.     

Additions to the cytologically investigated species of <Emphasis Type="Italic">Potentilla</Emphasis> L. (Rosaceae) from India

At present 14 species of Potentilla L. have been cytologically worked out from different geographical areas of Kashmir and Himachal Pradesh in the Western Himalayas. New chromosome numbers in nine species—Potentilla argyrophylla (n = 14), P. atrosanguinea (n = 7, 14), P. desertorum (n = 7), P. gerardiana (n = 14), P. indica (n = 14), P. micropetala (n = 14), P. nepalensis (n = 14), P. sibbaldia (n = 14) and P. thomsonii (n = 7)—have been reported on a worldwide basis for the first time. Additional chromosomal races of polyploid cytotypes for P. argyrophylla (n = 28) and P. desertorum (n = 14) along with a diploid cytotype for P. micropetala (n = 7) plus diploid cytotypes for the five species as P. fulgens (n = 7), P. gelida (n = 7), P. kleiniana (n = 7), P. sibbaldia (n = 7) and P. sundaica (n = 7) as well as a tetraploid cytotype for P. fruticosa (n = 14) all have been cytologically worked out from India for the first time. The course of meiosis varies from normal to abnormal in different populations of the majority of the species, such as P. argyrophylla, P. atrosanguinea, P. desertorum, P. fruticosa, P. fulgens, P. gelida, P. indica, P. nepalensis, P. sibbaldia and P. sundaica, except for normal meiosis observed in P. gerardiana, P. kleiniana, P. micropetala and P. thomsonii. The anomalous taxa are marked with meiotic abnormalities in the form of cytomixis, chromosomal stickiness, unoriented bivalents, formation of laggards and bridges resulting in abnormal microsporogenesis, and production of heterogenous-sized fertile pollen grains along with reduced pollen fertility. All the taxa with normal meiotic courses show nearly one hundred percent pollen fertility.     

Serological Evidence of <Emphasis Type="Italic">Paracoccidioides brasiliensis</Emphasis> Infection in Chickens from Paraná and Mato Grosso do Sul States,Brazil

The objective of this study was to detect antibodies against Paracoccidioides brasiliensis in free-range and caged chickens Gallus domesticus. Initially, the humoral immune response of two chickens immunized with P. brasiliensis was evaluated. Both animals showed the production of antibodies to gp43, the major P. brasiliensis antigen. The seroepidemiological survey was conducted in chickens from the Pantanal region in Mato Grosso do Sul State (free-range n = 40) and from northern region of Paraná State (free-range n = 100, caged n = 43). The serum samples were analyzed by indirect ELISA using gp43 as antigen. The positivity observed in free-range chickens from Mato Grosso do Sul (55%) was significantly higher (P = 0.0001) than in free-range chickens from Paraná State (16%). In contrast to the free-range chickens, no positivity was observed in the caged chickens (P = 0.003). This is the first report showing serological evidence of P. brasiliensis infection in chickens. The results suggest that free-range chickens are more frequently infected by P. brasiliensis, probably due to the constant contact with soil than caged chickens and could be useful as epidemiological markers of paracoccidioidomycosis.     

Nucleotide divergence and genetic relationships of Pseudoroegneria species

Nucleotide variation in 8 diploid Pseudoroegneria species was characterized using two single copy nuclear genes, the second largest subunit of RNA polymerase II (RPB2) and the translation elongation factor G (EF-G), and one chloroplast TrnD/T intergenic region. Among the Pseudoroegneria species studied, the estimates of nucleotide diversity (π) ranged from 0.04577 (RPB2) and 0.00183 (TrnD/T) for Pseudoroegneria tauri to 0.10667 (RPB2), 0.06174 (EF-G) and 0.03743 (TrnD/T) for Pseudoroegneria spicata. The highest nucleotide diversity of the RPB2 data set was found for P. spicata among the taxa analyzed. Our phylogenetic analyses separated the accessions of P. spicata into several groups, with 7 accessions of P. spicata forming a highly supported subclade (BS = 100%, PP = 1.00). The phylogenetic analysis also suggests that P. spicata, Pseudoroegneria gracillima and Pseudoroegneria stipifolia have a closer relationship than the other species within Pseudoroegneria. Pseudoroegneria libanotica and P. tauri were also found to exhibit a high level of sequence homology, however, only nuclear gene data (RPB2 and EF-G) clearly indicated the differentiation between the P. libanotica + P. tauri group with other St genome species.