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Phylogeny and evolution of the Betulaceae as inferred from DNA sequences, morphology, and paleobotany.

Phylogeny of the Betulaceae is assessed on the basis of rbcL, ITS, and morphological data. Based upon 26 rbcL sequences representing most "higher" hamamelid families, the Betulaceae are monophyletic, with Casuarinaceae as its sister group, regardless of whether the outgroup is Cunoniaceae, Cercidiphyllaceae, Hamamelidaceae, or Nothofagus. Within the Betulaceae, two sister clades are evident, corresponding to the subfamilies Betuloideae and Coryloideae. However, with only 13 phylogenetically informative sites, the rbcL sequences provide limited intra-subfamilial resolution. Internal transcribed spacer (ITS) sequences provided 96 phylogenetically informative sites from 491 aligned sites resulting in a single most parsimonious tree of 374 steps (consistency index = 0.791) with two major lineages corresponding to the two traditional subfamilies: Betuloideae (Alnus, Betula) and Coryloideae (Corylus, Ostryopsis, Carpinus, Ostrya). This arrangement is mostly consistent with those from rbcL and morphology and is greatly reinforced by analyses with the three data sets combined. In the Coryloideae, the Ostryopsis-Carpinus-Ostrya clade is well supported, with Corylus as its sister group. The sister-group relationship between Ostryopsis and the Carpinus-Ostrya clade is well supported by ITS, rbcL, and morphological data. Phylogenetic relationships among the extant genera deduced by these analyses are compatible with inferences from ecological evolution and the extensive fossil record.

Journal Article↗

Allergy to pollens from Betulaceae and Corylaceae in a Mediterranean area (Genoa, Italy)--a ten-year retrospective study.

Allergy to pollens from Betulaceae and Corylaceae is becoming a leading problem in Genoa, a northern Mediterranean area in Italy. The results of a 10-year retrospective study on combined observations both on the aerobiological presence of Betulaceae and Corylaceae pollens (Alnus, Corylus, Ostrya) and on the incidence of positive SPT in the allergic population living in the same area are reported. Among 3473 patients suffering from seasonal respiratory allergy with positive SPT to one or more pollens, 558 (16.06%) showed SPT positivity to Betulaceae and/or Corylaceae pollens, both isolated and associated with other allergens. These patients suffered from winter or early spring hay fever, with a high incidence of bronchial asthma. A statistically significant increase (0.02 < p < 0.05) in the number of these sensitizations from 1981 to 1990 has been observed. Some possible explanations for this phenomenon, including an increase in the total amount of local airborne pollens, are suggested. About 24% of the patients with positive SPT to these pollens referred oral allergic syndrome associated with the ingestion of some foods, especially apples and nuts, with or without other additional clinical symptoms.

Adolescent↗

Feeding and attraction of Agelastica coerulea (Coleoptera: Chrysomelidae) to Betulaceae plants.

The feeding and attraction of Agelastica coerulea (Baly) to nine species of Betulaceae in four genera (Alnus, Betula, Carpinus, and Corylus) were examined in the laboratory by using choice and no-choice as well as olfactometer bioassays. In no-choice feeding bioassays with A. coerulea larvae, Alnus hirsuta (Spach) Ruprecht, Alnus japonica Steudel, Corylus sieboldiana variety mandshurica (Max.) c. k. Schneider, and Corylus heterophylla variety thunbergii Blume were the most preferred, whereas Betula davurica Pallas, Betula schmidtii Regel, and Carpinus cordata Blume were the least preferred. The larvae showed moderate preference for Alnus maximowiczii Callier and Carpinus tschonoskii variety brevicalycina Nakai. In choice feeding bioassays, no differences in preference between A. hirsuta and A. japonica were observed. However, there were significant differences in preference between A. hirsuta and each of the other seven plant species. In olfactometer bioassays with adult female A. coerulea, fresh leaf odor from A. hirsuta attracted significantly more adults than that from A. japonica and C. sieboldiana variety mandshurica. These results indicate that A. coerulea differed in its ability to discriminate among the Betulaceae plants at the feeding preference and attraction of host selection.

Animals↗

Complete congruence between morphological and rbcL-based molecular phylogenies in birches and related species (Betulaceae).

Estimations of phylogenies from morphological and molecular data often show contrasting results. We compared morphological and molecular phylogenies in an ancient family of woody dicots, the Betulaceae (birch family). The phylogeny of the family was estimated from parsimony analysis of morphological characters in the genera Alnus, Betula, Carpinus, Corylus, Ostrya, and Ostryopsis and from parsimony and distance-matrix analyses of DNA sequences of the chloroplast gene encoding the large subunit of ribulose-1,5-biphosphate carboxylase (rbcL) in the genera Alnus, Betula, Carpinus, Corylus, and Ostrya and in two outgroups, Quercus and Liquidambar. The topologies obtained by the different methods were completely congruent, and bootstrapping strongly supported the division of the family Betulaceae into two major clades, Betuleae (Alnus and Betula) and Coryleae (other members). Only slightly more homoplasy was present in the rbcL sequence data set than in the morphological set. Relative-rate tests indicated that the Coryleae clade had a faster rate of rbcL evolution than did the Betuleae clade. Heterogeneity of rates of morphological evolution also paralleled those for rbcL.

Base Sequence↗

Differences in spatial autocorrelation between four sub-populations of Alnus trabeculosa Hand.-Mazz. (Betulaceae).

To investigate the spatial structure of Alnus trabeculosa Hand.-Mazz, we compared three sub-populations at Imaichi in Tochigi Prefecture and one sub-population at Juo in Ibaraki Prefecture, Japan. A total of 269 trees, covering 0.71 ha in total, were mapped and genetically analyzed using nine enzyme systems encoding 13 isozyme loci. There were no significant differences between the four sub-populations in terms of Na,Ne, H(o), H(e )and F(IS). However, according to spatial autocorrelation analysis, the Juo sub-population, which is younger than the others, showed an aggregation of multilocus genotypes, especially within 25 m radii. In contrast with the Juo sub-population, those in Imaichi showed no clear spatial structuring. In the three Imaichi sub-populations, gametic phase disequilibrium is attributable to heterogeneity of genotype frequencies, but in the Juo sub-population it seems to be due to other factors, connected with the composition of the trees in the sub-population and/or its founder population, gene flow and asexual propagation. To conserve as much as possible of the genes or genotypes in restricted areas, conservation of populations that do not show clear family structures, such as those in Imaichi, would be most effective.

Betulaceae↗

Ecological and genetic effects of cutting in an Alnus trabeculosa Hand.-Mazz. (Betulaceae) population.

In order to assess the ecological and genetic effects of cutting, we compared two portions of Alnus trabeculosa population at Yuda (Iwate Prefecture, Japan): one that has been cut about 30 years ago and one that has remained uncut. These portions were compared in terms of the degree of sprouting, genetic variation and gene distribution using isozyme markers. First, we determined the multilocus genotype (MLG) of all ramets, then sorted them into individuals according to the distribution of the MLGs. The average (+/- SE) of largest distance between ramets in one individual was 2.1 (+/- 0.18) m, which is consistent with the distance (2.0 (+/- 0.20) m) obtained by tracing physical connections between ramets. We found no significant differences in genetic variation between the two portions, but there were significant differences in their degree of sprouting. Furthermore, there were striking differences in gene distribution: the cut portion showed greater clustering of individuals with identical genetic components, which may be due to regeneration in the gaps made by cutting, reflecting the location of the mother trees, and seed and pollen dispersal from them.

Betulaceae↗

Sequence of 18S rDNA of actinorhizal Alnus glutinosa (Betulaceae).

The small subunit ribosomal DNA for a woody actinorhizal, Alnus glutinosa, was isolated by the PCR method. Amplification products were cloned into the Bluescript SK- vector. Full sequence, 1698 bp, was obtained with NS1 to NS8 primers. Sequence alignments were made by UWGCG sequence data analysis computer programs. 18S rDNA sequence of A. glutinosa was compared to analogous segments of four other angiosperms, tomato, rice, maize and soybean. Sequence homologies are discussed and application for the technique is suggested.

Base Sequence↗

Do pollen donors with fastest-growing pollen tubes sire the best offspring in an anemophilous tree, Betula pendula (Betulaceae)?

The relationship between pollen and progeny performance has been a subject of many studies but the evidence for pollen-tube growth rate as an indicator of progeny fitness is equivocal. We used an anemophilous tree, Betula pendula, to examine the relationship between pollen-tube growth rate and seed and seedling performance. We crossed nine maternal plants with pollen from six pollen donors in a clonal B. pendula seed orchard, measured the pollen-tube growth rates for every cross, and analyzed the performance of the resulting seeds and seedlings. The only significant positive correlation was found between pollen-tube growth rate and seed mass when we controlled for seed number per inflorescence. Using seed mass as a covariate, we found that only maternal parent had a significant effect on the number of seeds per inflorescence, the percentage of germinable and embryonic seeds, and early seedling growth. Both maternal and paternal parents had significant effects on seedling height after 85 d of growth. These results are in concordance with the general view that maternal effects are usually most apparent in seed characters and during early plant growth. This study does not provide strong evidence for the theory of pollen-tube growth rate as an indicator of progeny quality.

Journal Article↗

The potential of Betv1 homologues, a nuclear multigene family, as phylogenetic markers in flowering plants.

Betv1 homologues are a ubiquitous group of genes in flowering plants encoding a class of highly conserved defense-related proteins and containing open reading frames from 465 to 480 bp. Betv1-like genes consist of two exons interrupted by an intron of 76-359 bp, with the intron position highly conserved. The pairwise p distance ranged from 0 to 0.583 among flowering plants. Within plant families, the ranges of the p distance were 0-0.403, 0-0.253, and 0.011-0.369, for Apiaceae, Betulaceae, and Fabaceae, respectively. The most striking feature of the betv1 gene phylogeny was that the multiple sequences from each plant family formed a monophyletic group and sequences from each species were generally more similar to each other than those from other species. The almost exclusive paralogous relationships of genes from the same species suggested that the genes of the multigene family underwent strong concerted evolution. Phylogenies of Betulaceae and Fabaceae inferred from betv1 gene trees were generally congruent with those based on morphology and other molecules. Betv1 homologues constitute potential phylogenetic markers at the intrafamilial level or among closely related families in flowering plants.

Base Sequence↗

Evolution of the mitochondrial rps3 intron in perennial and annual angiosperms and homology to nad5 intron 1.

The plant mitochondrial rps3 intron was analyzed for substitution and indel rate variation among 15 monocot and dicot angiosperms from 10 genera, including perennial and annual taxa. Overall, the intron sequence was very conserved among angiosperms. Based on length polymorphism, 10 different alleles were identified among the 10 genera. These allelic differences were mainly attributable to large indels. An insertion of 133 nucleotides, observed in the Alnus intron was partially or completely absent in the other lineages of the family Betulaceae. This insertion was located within domain IV of the secondary-structure model of this group IIA intron. A mobile element of 47 nucleotides that showed homology to sequences located in rice rps3 intron and in intergenic plant mitochondrial genomes was found within this insertion. Both substitution and indel rates were low among the Betulaceae sequences, but substitution rates were increasingly larger than indel rates in comparisons involving more distantly related taxa. From a secondary-structure model, regions involved in helical structures were shown to be well preserved from indels as compared to substitutions, but compensatory changes were not observed among the angiosperm sequences analyzed. Using approximate divergence times based on the fossil record, substitution and indel rate heterogeneity was observed between different pairs of annual and perennial taxa. In particular, the annual petunia and primrose evolved more than 15 and 10 times faster, for substitution and indel rates respectively, than the perennial birch and alder. This is the first demonstration of an evolutionary rate difference between perennial and annual forms in noncoding DNA, lending support to neutral causes such as the generation time, population size, and speciation rate effects to explain such rate heterogeneity. Surprisingly, the sequence from the rps3 intron had a high identity with the sequence of intron 1 from the angiosperm mitochondrial nad5 gene, suggesting a common origin of these two group IIA introns.

Base Sequence↗

Allergenic pollens and pollinosis in Italy: recent advances.

The authors have mapped the occurrence of allergenic pollens throughout Italy and defined their most common clinical symptoms. To obtain an accurate aerobiological and clinical picture of such a geographically complex country as Italy, a detailed investigation was carried out involving 80 data-gathering stations and 40 clinical centers nationwide. Three main pollination periods can be distinguished: winter-pre-spring (January to March) for Betulaceae, Corylaceae, Cupressaceae, Salicaceae and Ulmaceae: spring-summer (April to June) for Gramineae, Urticaceae, Oleaceae, Plantago, Fagaceae, Pinaceae and Polygonaceae, and summer-autumn (August to September) for Urticaceae, Compositae and Chenopodiaceae. Examination of 49,660 patients affected by pollinosis (conjunctivitis, rhinitis, asthma, with positive skin tests or IgE-specific serum determination: RAST, ELISA) throughout Italy revealed sensitivity to Gramineae in 64.6%, to Parietaria in 36.7% to Olea in 15.8%, to Compositae in 13.2%, to Betulaceae-Corylaceae in 7% and to Fagaceae-Cupressaceae-Plantago in 4%-10%; marked regional variations were observed. The patients suffered from rhino-conjunctivitis (55.7%), rhino-conjunctivitis plus asthma (31.6%) and asthma (12.7%). In monosensitised individuals, Parietaria was seen to be the main cause of the asthmatic syndrome (though our preliminary data also implicate Olea) followed by Gramineae.

Air Pollution↗

Biogeography, host specificity, and molecular phylogeny of the basidiomycetous yeast Phaffia rhodozyma and its sexual form, Xanthophyllomyces dendrorhous.

Phaffia rhodozyma (sexual form, Xanthophyllomyces dendrorhous) is a basidiomycetous yeast that has been found in tree exudates in the Northern Hemisphere at high altitudes and latitudes. This yeast produces astaxanthin, a carotenoid pigment with biotechnological importance because it is used in aquaculture for fish pigmentation. We isolated X. dendrorhous from the Southern Hemisphere (Patagonia, Argentina), where it was associated with fruiting bodies of Cyttaria hariotii, an ascomycetous parasite of Nothofagus trees. We compared internal transcribed spacer (ITS)-based phylogenies of P. rhodozyma and its tree host (Betulaceae, Corneaceae, Fagaceae, and Nothofagaceae) and found them to be generally concordant, suggesting that different yeast lineages colonize different trees and providing an explanation for the phylogenetic distance observed between the type strains of P. rhodozyma and X. dendrorhous. We hypothesize that the association of Xanthophyllomyces with Cyttaria derives from a previous association of the yeast with Nothofagus, and the sister relationship between Nothofagaceae and Betulaceae plus Fagaceae correlates with the phylogeny of X. dendrorhous strains originating from these three plant families. The two most basal strains of X. dendrorhous are those isolated from Cornus, an ancestral genus in the phylogenetic analysis of the host trees. Thus, we question previous conclusions that P. rhodozyma and X. dendrorhous represent different species since the polymorphisms detected in the ITS and intergenic spacer sequences can be attributed to intraspecific variation associated with host specificity. Our study provides a deeper understanding of Phaffia biogeography, ecology, and molecular phylogeny. Such knowledge is essential for the comprehension of many aspects of the biology of this organism and will facilitate the study of astaxanthin production within an evolutionary and ecological framework.

Base Sequence↗

Standardization of an ash (Fraxinus excelsior) pollen allergen extract.

BACKGROUND: Ash tree (Fraxinus excelsior) is the main representative of the Oleaceae family in temperate zones. Diagnosis of ash pollen allergy is made difficult due to (1) an overlapping pollinization period with Betulaceae, (2) non-inclusion in current diagnostic assays, and (3) some cross- reactivity with minor allergens from Betulaceae. The aim of this study was to calibrate an ash pollen in-house reference preparation (IHRP) in allergic patients in order to produce standardized products for diagnosis and immunotherapy purposes. METHODS: Ash pollen IHRP was extracted, ultrafiltered and freeze dried. Allergens in the extract were detected after 2-dimensional PAGE using specific sera and a monoclonal antibody. The Fra e 1 content of IHRP was evaluated by quantitative immunoprint. Forty-eight subjects from the North-East of France exhibiting clinical symptoms, a positive skin test and specific IgE levels > or =class 2 to ash pollen were recruited. IgE immunoprints were performed to select patients sensitized to the ash Fra e 1 allergen as opposed to cross-reacting allergens. Serial 10-fold dilutions of the IHRP were tested by skin prick tests in order to determine the concentration inducing a geometrical mean wheal diameter of 7 mm, said to correspond to an index of reactivity (IR) of 100 per millilitre. RESULTS: IgE-reactive molecules in IHRP comprise Fra e 1, Fra e 2, a 9-kDa molecule (presumably Fra e 3), as well as a doublet at 15 kDa and high molecular weight allergens. The 100 IR concentration of IHRP inducing a geometrical mean wheal diameter of 7 mm in 22 patients sensitized to Fra e 1 corresponds to the 1/126 (w/v) extraction ratio (i.e. 259 microg/ml of protein by Bradford) and contains 17 microg/ml of Fra e 1. The variability in total activity of 5 batches of standardized extracts was found to be significantly reduced when compared with 7 non-standardized extracts. CONCLUSION: An ash pollen IHRP was defined and molecularly characterized. Its successful standardization at 100 IR/ml in patients specifically sensitized to Fra e 1 allowed a skin reactivity-based calibration in properly diagnosed patients. Such a standardized ash pollen extract is a reliable tool to support immunotherapy of ash pollen allergy.

Adolescent↗

Pollen-tube growth and fertilization mode in Gymnostoma (Casuarinaceae): their characteristics and evolution.

A unique mode of fertilization called "chalazogamy", whereby the pollen tube passes through the chalaza instead of the micropyle, is known in several species of derived genera in Casuarinaceae. In this paper we report the occurrence of chalazogamy in Gymnostoma ( G. poissonianum), the most primitive genus in the family. We also show that the pollen tube grows discontinuously from the stigma to ovules in about 3 months. At the time of pollination, the ovules have not yet formed in the ovary, and require a long time to develop. The pollen tube(s) lie in a zigzag line and are branched in the upper region of the style, and their growth is arrested there until the ovary develops further. Studies of the relevant literature further revealed discontinuous pollen-tube growth in relation to a prolonged period between pollination and fertilization, as well as chalazogamy, in Betulaceae, Juglandaceae and/or Fagaceae that are closely related to Casuarinaceae. This feature may have derived early in the evolution of Fagales.

Betulaceae↗

The potential role of orbicules as a vector of allergens.

BACKGROUND: In the locules of anthers of flowering plants, tiny (1.5-2 microm) granules of sporopollenin may occur next to the pollen grains. Those granules, called orbicules, mostly occur on the radial and innermost tangential wall of secretory tapetum cells. METHODS: We have investigated the presence of orbicules in 15 important European allergenic species with scanning electron microscopy (SEM). RESULTS: Orbicules were present in all species investigated of the families Betulaceae, Chenopodiaceae, Fagaceae, Poaceae, Polygonaceae, and Urticaceae. However, in the Asteraceae and Oleaceae species studied, orbicules were lacking. In all Chenopodiaceae, Poaceae, and Urticaceae species, orbicules were attached to the pollen exine. These observations indicate the possibility of the dispersal of orbicules into the atmosphere during anthesis. CONCLUSIONS: The hypothesis of the potential role of orbicules as possible important vectors of allergens is put forward, based on the comparison of our results with recent literature about the evidence of allergenic activity in the smaller micronic atmospheric aerosol fraction. Our results provide evidence that an in-depth investigation of the sites of allergens across the whole anther is required. We suggest that allergen researchers apply immunoelectron microscopy on whole anthers to determine whether orbicules possess allergens.

Allergens↗