Biochemical analysis of the allergenic pollen ofMadhuca indica Gmel. andSpathodea campanulata Beauv

Summary The present paper deals with the biochemical analysis in the form of quantitative estimation of carbohydrates, proteins, nucleic acids, minerals, ash and moisture of the pollen of two allergenic plants, i.e.Manhuca indica andSpathodea campanulata. Carbohydrates constitute the major dry matter fractions in both the pollen types with highest amount inM. indica (25.37%). Soluble proteins were estimated along with lipid content.S. campanulata showed 12.74% protein andM. indica 11.58%. The estimation of nucleic acids revealed the presence of DNA from 0.36 to 0.72% and RNA from 0.28 to 1.03%. Minerals were found in very low amounts ranging from 0.14 to 0.19% for Calcium and 0.15 to 0.20% for Magnesium. The ash and moisture content were also found variable. The present findings revealed that these chemical constituents may be responsible for the control of the rate and type of allergic reaction caused by the pollen of these two plants.     

Study of pollen brush in selected species of Astragalus L. subgenus Pogonophace Bunge (Leguminosae)

Nine species of Lippia (Verbenaceae) were studied by RAPD markers in order to evaluate the degree of genetic diversity. The following species were collected at the Cadeia do Espinhaço Mountains, Southeast Brazil: L. corymbosa, L. diamantinensis, L. filifolia, L. florida, L. hermannioides, L. lupulina, L. rotundifolia, L. rosella and L. sidoides. The analysis was performed using 18 primers that generated 490 fragments and only one primer was found to be monomorphic in all individuals. The average interspecific genetic distances were similar for all species and higher than the intraspecific genetic distances. Species with narrow occurrence did not show low intraspecific diversity. The molecular data were used to generate an UPGMA dendrogram that showed two major groups with a clear distribution among the species. RAPD analysis was efficient to address the genetic diversity of Lippia species and contributed to understand the adaptation to the environment, conservation and taxonomic implications.     

Biochemical and serological characterization of b-proteins fromNicotiana species

Summary Proteins associated with the hypersensitive response (b-proteins) were purified from variousNicotiana species and compared biochemically and serologically. The method developed to purify proteins b₁, b₂ and b₃ ofN. tabacum cv. Xanthi-nc was used to purify b-proteins present inN. sylvestris (b₀, b₁ and b₃) andN. tomentosiformis (b₂), the parental species ofN. tabacum, and b₁″ from bothN. glutinosa andN. debneyi. Ultracentrifugation and amino acid analysis of some of these proteins has shown that they are very similar and that they are all monomers in their native form (mol wt = 15 700 for b₀, b₁, b₂ and b₃; mol wt = 13 800 for b₁″). Based on their reactions to an antiserum produced against protein b₁ ofN. tabacum cv. Xanthi-nc, 3 serological groups can be recognized which are independent of the source species (I) b₀ and b₁, (II) b₁″ and b₂, (III) b₃. Thus, proteins in the same serological group but from different species are more closely related than the b-proteins in different serological groups but present in the same species. The implication of this site on the possible phylogeny of b-proteins is discussed. Serological tests confirmed the b-protein present as a constitutive component in the virus resistant interspecific hybrids ofN. glutinosa ×N. debneyi as protein b₁″.     

Fatty acid analysis of four Azospirillum species reveals three groups

Cellular fatty acid composition of 14 strains from the four species of Azospirillum was determined by gas chromatographic analysis. All strains of Azospirillum lipoferum and Azospirillum brasilense were similar in fatty acid data, thus not revealing an expected distinction between the two long established species. Strains of both Azospirillum halopraeferens and Azospirillum amazonense, however, differed significantly from this first group of strains.     

Allozymic variation in four Indian species of genusMus: A comparative analysis

The present study focusses on allozyme variation in the commensal house mouseMus musculus, the pygmy field miceM. booduga andM. terricolor, and the spiny mouseM. platythrix. Genetic heterozygosity was estimated using a set of 24 polymorphic biochemical genetic markers. The extent of variability present inM. booduga, M. terricolor andM. platythrix has been compared with that in theM. musculus complex. Levels of allozyme variation at species level indicate thatM. musculus has the maximum heterogeneity, followed byM. booduga andM. terricolor, whileM. platythrix shows comparatively homogeneous genetic make-up. Gene frequency data have been used to trace phylogenetic relationships among these four species.     

A preliminary pachytene analysis of two species of Arachis L.

Summary The morphology of pachytene chromosomes was studied in A. glabrata Benth. and A. pusilla Benth. belonging respectively to the sections Rhizomatosae and Triseminale. These two species can not be crossed with the cultivated groundnut A. hypogaea L. All 20 chromosomes of A. glabrata could be identified individually and further classified into 5 basic types. The features that enabled the identification of chromosomes were: total length, arm ratios, nucleolus attachment and position and extent of heterochromatin. A simple key has been proposed for classifying different chromosomes to facilitate their easy identification. The genomes of A. glabrata did not resemble those of A. hypogaea except for the presence of an A chromosome, 2 euchromosomes and 2 nucleolus organisers. A. glabrata did not appear to be an amphidiploid but rather an allopolyploid hybrid. The genome of A. pusilla contained chromosomes unlike those of any other species of section Arachis. It was concluded that both these species are quite unrelated to other species of the section Arachis.     

Stable transformation of the food yam Dioscorea alata L. by particle bombardment

Summary A biolistic particle gun was used to deliver genetic material into intact yam cells. Cultured suspension cells of D. alata were bombarded with microprojectiles coated with pBI221.2 DNA and histochemical assays were carried out to show transient GUS expression in bombarded cells. Stably transformed D. alata cells were recovered from cultured cells after bombardment with microprojectiles coated with pRT99gus harbouring both the nptII and uidA genes. Bombarded cells were selected on a medium containing geneticin (G418). Two months after bombardment, calli resistant to G418 were assayed for GUS expression. There was a 100% correlation between resistance to G418 and GUS expression. From these calli, four cell lines were established and GUS activity in each line was determined fluorometrically. The use of a specific GUS inhibitor showed that the GUS activity was due to the introduced uidA gene rather than to any intrinsic GUS-like activity originating from the plant. Incorporation of the introduced DNA into the plant genomic DNA was confirmed by Southern analysis.Abbreviations GUS -glucuronidase - MU 4-Methylumbelliferone - MUG 4-Methylumbelliferyl--D-glucuronide - PVP Polyvinylpyrrolidone - SDS Sodium dodecyl sulphate - TAE Tris-acetate-EDTA buffer - X-Gluc 5-Bromo-4-chloro-3-indolyl--D-glucuronide     

Genetic variation of Carthamus tinctorius L. and related species revealed by SRAP analysis

Genetic diversity of 23 populations of Carthamus tinctorius L. and two populations of Carthamus lanatus L. in China was investigated using Sequence-related Amplified Polymorphism (SRAP). All populations could be uniquely distinguished by 30 primer combinations with 483 bands and 274 polymorphic bands which generated 57% of polymorphic ratio. Unweighed pair-group method of with arithmetical averages (UPGMA) cluster analysis enabled construction of a dendrogram for estimating genetic distances among different populations. The extreme variation was observed when No. 4 cultivated and No. 13 wild population of C. lanatus were grouped at GS = 0.58, and separated from 23 populations of C. tinctorius at GS = 0.10. The result suggested that the cultivated and wild populations of C. lanatus had close relationship with each other and far relationship with C. tinctorius. Dendrogram also revealed a large genetic variation in 23 C. tinctorius populations; different primer combinations allowed them distinctly distinguished one from others with relatively low genetic similarity. Furthermore, five typical representative fragments in C. lanatus were obtained by four most informative primer combinations, which provided a possibility to distinguish C. lanatus from the C. tinctorius evidently.     

Antifeedant and larvicidal activities of Rhein isolated from the flowers of Cassia fistula L.

Antifeedant and larvicidal activities of rhein (1,8-dihydroxyanthraquinone-3-carboxylic acid) isolated from the ethyl acetate extract of Cassia fistula flower were studied against lepidopteron pests Spodoptera litura and Helicoverpa armigera. Significant antifeedant activity was observed against H. armigera (76.13%) at 1000 ppm concentration. Rhein exhibited larvicidal activity against H. armigera (67.5), S. litura (36.25%) and the LC₅₀ values was 606.50 ppm for H. armigera and 1192.55 ppm for S.litura. The survived larvae produced malformed adults.     

Immunoelectrophoretic analysis of seed proteins from Pisum sativum L.

Summary Soluble proteins of pea seed were investigated by quantitative immunological methods. Vicilin, legumin, pea seed lectin (PEA), 26 albumins and a globulin (B₁) were detected and observed during seed development, germination and under different extraction and fractionation procedures. Vicilin and legumin were found to be immunologically distinctly different. Legumin was found to be comprised of two similar proteins, Legumin species I and II. Vicilin, but no legumin, was detected in the embryonic axis.Three albumins, B₁ and PEA were found to be synthesized after the onset of legumin synthesis.Among the pea lines investigated, one line exhibited distinct differences with respect to the albumins and PEA.Some observations indicate that PEA might interact with other seed proteins of pea.     

Clinical significance of allergic sensitization toOlea europaea L. pollen in Naples area,Italy

Summary The aim of this study is the evaluation of clinical significance of the allergic sensitization toOlea europaea L. pollen allergens in the Naples area, because in the last 10 years an increasing frequency of IgE mediated sensitization to these allergens has been observed. 4124 patients of both sexes (2989 adults and 1153 children) consecutively examined in our Allergological Center from January 1, 1988 to December 30, 1989 have been considered in this study. The patients were selected on the basis of a history of rhinitis (R) and/or asthma (A) and were evaluated by clinical examination, SPT, RAST and specific or aspecific BPT. Among 2267 adults and 879 children sensitized to almost one aeroallergen we found respectively 306 (13.49%) and 75 (8.53%) subjects with an allergic sensitization toOlea europaea L. pollen. The frequency of clinical manifestation was respectively R+A (58.16%), R (39.54%), A (2.28%) in adults; R+A (61.33%), R (26.66%), A (12%) in children. A monosensitization toOlea europaea L. pollen was found in 1.33% of children and in 2.28% of adults examined. The association with other allergens especially with mites and/or other pollen was very common.     

Thin layer chromatographic analysis of glucose and maltose in estivated Biomphalaria glabrata snails and those infected with Schistosoma mansoni

Thin layer chromatography was used to analyze the glucose and maltose concentrations of the digestive gland–gonad complex (DGG) of uninfected-estivated Biomphalaria glabrata snails and estivated B. glabrata patently infected with Schistosoma mansoni. All snails were estivated in a most chamber at a relative humidity of 98 ± 1% and a temperature of 23 ± 1 °C for 14 days. Carbohydrates were extracted from the DGG with 70% aqueous ethanol, and extracts were analyzed on silica gel preadsorbent plates using ethyl acetate–glacial acetic acid–methanol–water (60:15:15:10) mobile phase, α-naphthol–sulfuric acid detection reagent, and quantification by densitometry. The concentrations of glucose and maltose were significantly reduced in both uninfected-estivated snails and infected-estivated snails.     

四种栎属青冈亚属植物叶绿体基因组特征及系统发育研究

栎属青冈亚属植物的系统发育地位长期存在着争议,部分种的种间关系不明确。为揭示宁冈青冈(Quercus ningangensis)、曼青冈(Q.oxyodon)、毛曼青冈(Q.gambleana)、竹叶青冈(Q.neglecta)的叶绿体基因组特征及系统发育关系,该研究选择以上4种栎属青冈亚属植物的成熟叶片进行二代测序,对其叶绿体基因组结构和特征进行分析,并结合相关类群进行系统发育研究。结果表明：(1)宁冈青冈、曼青冈、毛曼青冈、竹叶青冈的叶绿体基因组序列长度分别为160 906、160 883、160 832、160 784 bp,均编码133个基因,包括88个蛋白质编码基因、37个tRNA基因、8个rRNA基因。(2)4种栎属青冈亚属植物偏好以A/T结尾的密码子,质体基因组变异区域主要存在于非编码序列。(3)通过IR边界分析得出,4种栎属青冈亚属植物存在ycf1假基因且在IRb/SSC区域发生扩张。(4)系统发育分析显示,在壳斗科中,水青冈属(Fagus)和轮叶三棱栎属(Trigonobalanus)较早分化出来,栎亚属(subg.Quercus)未形成一个单系群,叶绿体基因组建树结...     

Pollination of four sympatric species ofAngelonia (Scrophulariaceae) by oil-collecting bees in NE. Brazil

The manner whereby the oil-producing bisaccate flowers ofAngelonia (Scrophulariaceae) are pollinated by female oil-collecting bees is reported for the first time. Observations were made in the Caatinga formation of Pernambuco, NE. Brazil, on four synchronopatric species. These differ in sizes and structural details of the corolla, level of flower exposition, and habitat preferences. All legitimate visitors wereCentris spp. (Anthophoridae):Angelonia hirta was mainly pollinated byC. fuscata andA. pubescens byC. hyptidis; A. bisaccata andA. hookeriana shared an unidentified species. Several exomalopsine, tetrapediine and meliponid bees exploit the flowers less descriminately for oil or pollen, respectively, without regularly contacting anthers and stigma. The flowers are protandrous, and are self-incompatible except those of the annualA. pubescens. After alighting, theCentris bees introduce their front legs simultaneously into each of the pouches and start alternate collecting movements to gather the oil from the trichome elaiophores. While doing so, they are forced by projections of the corolla floor to press their head under the anthers and stigma, whereby pollen is transferred with their frons or clypeus. On account of their collector type and behaviour,C. fuscata andC. spec. are not specialized toAngelonia but may equally exploit other nonrelated taxa for oil, whereasC. hyptidis exhibits oligolecty onA. pubescens. It possesses relatively elongate forelegs with padlike collectors suitable for sweeping the lipids from the scattered glandular hairs inside the divergent spurs of its host. It is the only species that also collects pollen (by buzzing) from the oil host.A. hirta and relatives, provided with dense elaiophore carpets, are, for their part, adapted to scrapingCentris species with typical oil collectors. Flower and bee phenologies, although largely dependent on the irregular rainfalls, are not always coincident.     

Development of structure-less pollen wall inCeratophyllum demersum L. (Ceratophyllaceae)

Developmental process of structure-less exine is studied in a hydrophilous plant,Ceratophyllum demersum L., with electron microscopy. The plant shows a characteristic feature in tetrad formation. A callose wall is not synthesized and exine initiation does not occur during the tetrad stage. After release of microspores, a trilaminar layer with two electron-dense lines is formed in the surface of each microspore. The trilaminar layer develops to a thin structure-less exine that is considered to consist of only an endexine. The unusual exine would be an adaptive feature for submersed pollination in fresh water.     

Phylogenetic analysis of Fosterella L.B. Sm. (Pitcairnioideae, Bromeliaceae) based on four chloroplast DNA regions

The about 31 species of Fosterella L.B. Sm. (Bromeliaceae) are terrestrial herbs with a centre of diversity in the central South American Andes. To resolve infra- and intergeneric relationships among Fosterella and their putative allies, we conducted a phylogenetic analysis based on sequence data from four chloroplast DNA regions (matK gene, rps16 intron, atpB-rbcL and psbB-psbH intergenic spacers). Sequences were generated for 96 accessions corresponding to 60 species from 18 genera. Among these, 57 accessions represented 22 of the 31 recognized Fosterella species and one undescribed morphospecies. Maximum parsimony and Bayesian inference methods yielded well-resolved phylogenies. The monophyly of Fosterella was strongly supported, as was its sister relationship with a clade comprising Deuterocohnia, Dyckia and Encholirium. Six distinct evolutionary lineages were distinguished within Fosterella. Character mapping indicated that parallel evolution of identical character states is common in the genus. Relationships between species and lineages are discussed in the context of morphological, ecological and biogeographical data as well as the results of a previous amplified fragment length polymorphism (AFLP) study.     

Germination of Pistacia vera L. pollen in liquid medium

Summary The osmotic effect of Polyethylene glycol (PEG) has been shown to be sufficient to induce the germination of Pistacia vera L. pollen in liquid medium. The prehydration of the pollen in a saturated atmosphere for approximately 10 h was necessary to obtain maximum in vitro germination. Imbibition of the pollen in water resulted in the rapid leakage of solutes into the medium. These solutes consisted of approximately 50% carbohydrates, of which sucrose (0.65 mol/mg), glucose (0.77 mol/mg) and fructose (0.78 mol/mg) were the major sugars; the remaining 50% comprised proteins with the following major molecular weights 63 kDa, 60 kDa, 59 kDa, 40 kDa, 36 kDa, 35.5 kDa, 31 kDa, other organic matter and minerals.     

Inter-relationships between Najas marina L. and three other species of aquatic macrophytes

The ability of Najas marina L. to thrive in the presence of the submerged hydrophytes (Myriophyllum spicatum L. and Potamogeton lucens L.) and of an emerged hygrophyte (Scirpus litoralis Schard.) was investigated in Tel Aviv, Israel. Najas plants were unaffected by the presence of Potamogeton lucens and Scirpus litoralis, but were significantly suppressed by Myriophyllum. Bilateral negative relationships exist between Najas and Myriophyllum and these seem to be of an allelophatic nature, depending more on the nature of the accompanying species rather than on their mass.     

Pollinator foraging behavior and pollen collection on the floral morphs of tristylous Pontederia cordata L.

Summary The foraging behavior of the pollinators of tristylous Pontederia cordata was studied to determine if differences in floral morphology would lead to preferential visitation of the floral morphs. Although nectar production is not different in the three floral morphs, differences in the production and size of pollen grains produced by the three anther levels results in the morphs offering variable amounts of resources to pollen-collecting insects. Bumblebees (Bombus spp.) and the solitary bee Melissodes apicata used P. cordata primarily as a nectar source and therefore did not seem to exhibit any morph preference. In contrast, honeybees visited flowers mainly for pollen and preferred to forage on long-level anthers of the short-and mid-styled morphs. An analysis of the composition of corbicular pollen loads indicated that, relative to the frequency of production in the population: 1) honeybees collected an excess of pollen from long-level anthers; 2) bumblebees collected the three types of pollen without any apparent preference; and 3) M. apicata preferentially collected pollen from the short-level anthers — presumably because their proboscides are modified by the presence of tiny hairs. The results suggest that P. cordata in Ontario is serviced by a diverse, unspecialized pollinator fauna which is not co-adapted to the tristylous floral polymorphism.