The flavonoids of ferns in the isolated genera Loxsoma and Loxsomopsis

Kaempferol and quercetin 3-O-glucosides and 3-O-rhamnoglucosides are common to both Loxsoma cunninghamii and Loxsomopsis costaricensis, but in the former the new flavonoid glycosides, kaempferol and quercetin 3-O-glucoside-7-O-arabinoside have also been identified. The data are consistent with the proposed taxonomic relationship between these geographically isolated genera. Comparative flavonoid chemistry indicates that the Loxsomaceae may be a primitive family, not closely related to the Hymenophyllaceae or the Cyatheaceae.     

A comparative study of the flavonoids of Medicago radiata with other Medicago and related Trigonella species

The flavonoid glycosides of Medicago radiata as well as M. arabica, M. polymorpha, M. sativa, Trigonella coerulescens, T. foenum-graecum and T. spicata were studied in detail. Major glycosides identified were the 7-glucuronides of apigenin, luteolin, chrysoeriol and tricin, as well as lesser amounts of di- and triglucuronides of chrysoeriol and tricin. Also identified were the 3-robinobioside and 3,7-diglucoside of kaempferol, along with lesser amounts of quercetin-3,7-diglucoside, 4′,7-dihydroxyflavone, 3′,4′,7-trihydroxyflavone, formononetin and daidzein. Twelve other Medicago and Trigonella species were also studied for their flavonoid aglycones. The systematic position of M. radiata is discussed.     

The occurrence of acylated flavonol glycosides in the cruciferae

Nine acylated glycosides of kaempferol or quercetin were identified in Sisymbrium gilliesii, and in three Crambe spp. They were usually present together with the related unacylated glycosides. Acylation is a very common characteristic of the four crucifer species studied.     

Distribution of flavonoids and their systematic significance in Clematis subsection Viornae

Fifteen flavonoids were identified in the eight taxa of Clematis subsection Viornae. Flavonols, flavones, and C-glycosyl flavones were present. The compounds were based primarily on luteolin, apigenin, quercetin, and kaempferol. The systematic significance of the distribution of these compounds among the taxa is discussed.     

Flavonoid glycosides and the chemosystematics of Eucalyptus camaldulensisis

Leaf flavonoid glycosides of Eucalyptus camaldulensis were identified as kaempferol 3-glucoside and 3-glucuronide; quercetin 3-glucoside, 3-glucuronide, 3-rhamnoside, 3-rutinoside and 7-glucoside, apigenin 7-glucuronide and luteolin 7-glucoside and 7-glucuronide. Two chemical races were observed based on the flavonoid glycosides. These races correspond to the northern and southern populations of species growing in Australia. The Middle Eastern species examined were found to belong to the southern Australian chemical race. The major glycosides of E. occidentalis proved to be quercetin and myricetin 3-glucuronide.     

The systematic significance of flavonoids in Persea of the Southeastern United States

The systematic significance of flavonoid distribution in the southeastern United States taxa of Persea is discussed. The compounds thus far elucidated are glycosides based on quercetin, kaempferol, apigenin or luteolin.     

Antioxidant Properties of the Major Polyphenolic Compounds in Broccoli

We have examined the antioxidant activity of the major phenolic compounds in Broccoli: two flavonol glycosides (quercetin 3-O-sophoroside and kaemp-ferol 3-O-sophoroside) and four hydroxycinnamic acid esters (1,2'-disinapoyl-2-feruloyl gentiobiose, 1-sinapoyl-2-feruloyl gentiobiose, 1,2,2'-trisinapoyl gentiobiose and 1,2-disinapoyl gentiobiose). The Trolox C equivalent antioxidant capacity (TEAC) and inhibition of iron/ascorbate-induced lipid per-oxidation of phosphatidyl choline vesicles were measured. In the aqueous phase TEAC assay, the two flavonol glycosides were less active than their respective aglycones. TEAC values for the hydroxycinnamic acid esters were less than the sum of their constituent hydroxycinnamic acids on a molar basis. Quercetin 3-O-sophoroside was a potent inhibitor of lipid peroxidation, in contrast to kaempferol 3-O-sophoroside. The hydroxycinnamic acid esters were highly effective at preventing lipid damage with the exception of 1,2,2'-trisinapoyl gentiobiose. The six compounds analysed herein demonstrate the antioxidant activity of the major phenolics in broccoli and indicate the effect on antioxidant activity of sugar substitutions in the phenolic B ring.     

Air pollution effects on structure,proteins and flavonoids in pollen grains of Thuja orientalis L. (Cupressaceae)

Increase in the levels of air pollution due to the increase in industrial and agricultural technology has prompted investigation of mechanisms that contribute to air pollution tolerance in plants. Pollen grains of Thuja orientalis were collected from controlled (less polluted) and polluted areas (mainly SO₂, NO₂, CO, HC and APM). Thuja pollen is considered inaperturate and granulate. The exine is shed during rehydration, leaving the male gametophyte naked. The pollen grains collected from polluted areas are smaller and more fragile compared to control ones. The exine splitting is faster and higher in polluted pollen grains. SDS–PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) pattern do not show significant differences in polluted pollen than those in the control group. HPLC (high pressure liquid chromatography) analysis demonstrates that air pollution induces flavonoids accumulation to significantly higher levels in polluted pollen than in controlled ones. These observations suggest that plants try to respond suitably by adjusting their metabolism so that minimum damage is done due to air pollutants. Their protective responses may include an increase in antioxidant enzymes and metabolites and induction of protection-related secondary metabolite genes especially flavonoids.     

New views of tapetum ultrastructure and pollen exine development in Arabidopsis thaliana

Background and Aims

The Arabidopsis thaliana pollen cell wall is a complex structure consisting of an outer sporopollenin framework and lipid-rich coat, as well as an inner cellulosic wall. Although mutant analysis has been a useful tool to study pollen cell walls, the ultrastructure of the arabidopsis anther has proved to be challenging to preserve for electron microscopy.

Methods

In this work, high-pressure freezing/freeze substitution and transmission electron microscopy were used to examine the sequence of developmental events in the anther that lead to sporopollenin deposition to form the exine and the dramatic differentiation and death of the tapetum, which produces the pollen coat.

Key Results

Cryo-fixation revealed a new view of the interplay between sporophytic anther tissues and gametophytic microspores over the course of pollen development, especially with respect to the intact microspore/pollen wall and the continuous tapetum epithelium. These data reveal the ultrastructure of tapetosomes and elaioplasts, highly specialized tapetum organelles that accumulate pollen coat components. The tapetum and middle layer of the anther also remain intact into the tricellular pollen and late uninucleate microspore stages, respectively.

Conclusions

This high-quality structural information, interpreted in the context of recent functional studies, provides the groundwork for future mutant studies where tapetum and microspore ultrastructure is assessed.     

A BAHD acyltransferase is expressed in the tapetum of Arabidopsis anthers and is involved in the synthesis of hydroxycinnamoyl spermidines

BAHD acyltransferases catalyze the acylation of many plant secondary metabolites. We characterized the function of At2g19070 , a member of the BAHD gene family of Arabidopsis thaliana . The acyltransferase gene was shown to be specifically expressed in anther tapetum cells in the early stages of flower development. The impact of gene repression was studied in RNAi plants and in a knockout (KO) mutant line. Immunoblotting with a specific antiserum raised against the recombinant protein was used to evaluate the accumulation of At2g19070 gene product in flowers of various Arabidopsis genotypes including the KO and RNAi lines, the male sterile mutant ms1 and transformants overexpressing the acyltransferase gene. Metabolic profiling of flower bud tissues from these genetic backgrounds demonstrated a positive correlation between the accumulation of acyltransferase protein and the quantities of metabolites that were putatively identified by tandem mass spectrometry as N ¹, N ⁵, N ¹⁰-trihydroxyferuloyl spermidine and N ¹, N ⁵-dihydroxyferuloyl- N ¹⁰-sinapoyl spermidine. These products, deposited in pollen coat, can be readily extracted by pollen wash and were shown to be responsible for pollen autofluorescence. The activity of the recombinant enzyme produced in bacteria was assayed with various hydroxycinnamoyl-CoA esters and polyamines as donor and acceptor substrates, respectively. Feruloyl-CoA and spermidine proved the best substrates, and the enzyme has therefore been named spermidine hydroxycinnamoyl transferase (SHT). A methyltransferase gene ( At1g67990 ) which co-regulated with SHT during flower development, was shown to be involved in the O -methylation of spermidine conjugates by analyzing the consequences of its repression in RNAi plants and by characterizing the methylation activity of the recombinant enzyme.     

Olea Europea 3-year pollen record in the area of Thessaloniki,Greece and its sensitizing significance

Summary The observations of airborne pollen ofOlea europea and the incidence of clinical manifestations in patients allergic to this pollen type have not been registered so far in the city of Thessaloniki. The purpose of this study was: 1. to assess theO. europea pollen circulation in the area of our city, and 2. to detect the percentage of sensitivity toO. europea pollen in patients with pollinosis. We collected daily pollen samples during a 3-year period (February '87-January '90), using a Burkard volumetric trap, located on a high level area in the centre of the city. The pollen counts were then registered. The O.europea pollen grains were not differentiated microscopically from the other Oleaceae, but identified through phenological criteria. The patients included in the assessment of the sensitivity toO. europea came from the out-patient clinic of bronchial asthma of the General Hospital ?G. Papanicolaou?. They had a seasonal pollinosis and they were submitted to prick test using a battery of 22 groups and an O.europea extract. Pollen ofO. europea appears first in the atmosphere of Thessaloniki at the beginning of May, shows a peak in the end of May and continues to be present till the end of June. The quantity ofO. europea pollen ranked 6th in the list of the total pollen count and its flowering period coincided with that of grasses. In a sample of 360 patients with seasonal pollinosis, we detected anO. europea pollen sensitivity combined with other alleargens in 37% of the patients and a monosensivity in 4%. We conclude that pollen ofO. europea results to be present over a relative short period of time (May–June) in the area of Thessaloniki. The percentage of patients' sensitization toO. europea pollen was a little less frequent than sensitization to grasses, even if their flowering time coincides and their presence in the air shows about the same concentration values.     

Phenolamides during the ripening of dormant and non-dormant seeds in different Petunia lines

Phenolamides were extracted from reproductive organs and different capsule parts (fruit wall, placenta and seeds) of dormant and non-dormant lines derived from the species Petunia hybrida hort,. Petunia axillaries Lam. B.S.P. and Petunia parodii Steere. The different compounds were separated by high-pressure liquid chromatography and analyzed before pollination, during ripening, at ripeness and after 2 weeks of after-ripening of the seeds. The extracts were examined particularly for the presence of an alkaline compound (X), which is a phenolic derivative of tryptophanamide and appears to be a biochemical marker for the dormancy of ripe dry seeds of Petunia. During capsule growth and seed ripening, compound X was not a marker for dormancy, although it occurred in the reproductive organs of seed-parent plants. After pollination and fertilization, accumulation of compound X started mainly in the fruit wall, but in ripe dry capsules it was present entirely in the seeds. In contrast to compound X, neutral phenolamides were not detected in extracts from the fruit wall: and from the early stages of the capsule growth substantial accumulation took place only in the seeds.
Absence of compound X from ripe dry seeds appears as a necessary but not a sufficient condition for non-dormancy.     

Distribution of loosely-bound calcium in the vegetative and generative cells of the pollen grains in Chlorophytum elatum

We used chlorotetracycline fluorescence, alizarin staining and potassium pyroantimonate methods, as well as X-ray microanalysis, to demonstrate the differential localization of Ca2+ during pollen maturation. Level of loosely-bound Ca2+ ions was higher in the generative cell than in the vegetative cell of the mature pollen grain which is one of the symptoms of metabolic differentiation of the two sister cells. This revised version was published online in July 2006 with corrections to the Cover Date.     

Transmission of sowbane mosaic virus by Thrips tabaci in the presence and absence of virus-carrying pollen

When pollen of sowbane mosaic sobemovirus (SoMV)-infected Chenopodium amaranticolor was dusted onto C. amaranticolor and C. quinoa test seedlings which were then infested with 5–10 adult Thrips tabaci, SoMV was transmitted to 25%C. amaranticolor and 88% of C. quinoa plants. Five hours access of T. tabaci to pollen-contaminated C. quinoa seedlings was sufficient for SoMV transmission, but 1 h was not. SoMV was also transmitted when T. tabaci was mixed with pollen of C. amaranticolor and then placed on C. quinoa test seedlings. Further, plant-to-plant transmission of SoMV occurred in the absence of virus-carrying pollen when T. tabaci was caged on infected non-flowering C. amaranticolor or C. quinoa for 1–8 days and then adults transferred to C. quinoa test seedlings. This is the first time that thrips have been shown to be a vector of SoMV, and that a virus outside the ilarvirus group is reported to be transmitted using pollen and thrips.     

Wettable and unsinkable: the hydrodynamics of saccate pollen grains in relation to the pollination mechanism in the two New Zealand species of Prumnopitys Phil. (Podocarpaceae)

The pollination mechanism of most genera of the Podocarpaceae involves inverted ovules, a pollination drop and bisaccate pollen grains. Saccate grains have sometimes been referred to as 'non-wettable' due to their buoyant properties, while non-saccate pollen grains have been described as 'wettable'. The hydrodynamic properties of saccate pollen grains of seven podocarp species in five genera, Dacrydium Sol. ex G. Forst., Dacrycarpus (Endl.) de Laub., Manoao Molloy, Podocarpus L'Hér. ex Pers. and Prumnopitys Phil. have been tested in water, together with saccate and non-saccate pollen of four other conifer genera, Cedrus Trew (Pinaceae), Cephalotaxus Siebold & Zucc. ex Endl. (Cephalotaxaceae), Cupressus L. (Cupressaceae) and Phyllocladus Rich. ex Mirb. (Phyllocladaceae), and spores of three fern species and one lycopod species. All four spore types studied were non-wettable, whereas the bisaccate and trisaccate pollen types, like all other conifer pollen types, were wettable, enabling the grains to cross the surface tension barrier of water. Once past this barrier, grain behaviour was governed by presence or absence of sacci. Non-saccate and vestigially saccate grains sank, whereas saccate grains behaved like air bubbles, floating up to the highest point. In addition, the grains were observed to float in water with sacci uppermost, consistent with the suggestion that distally placed sacci serve to orientate the germinal furrow of the pollen grain towards the nucellus of an inverted ovule. Observations of pollen grains in the pollen chambers of naturally pollinated Prumnopitvs ovules confirmed this. The combination of buoyancy and wettability in saccate pollen has implications for the efficiency of the typical podocarp pollination mechanism.     

Numerical analysis of modern and fossil pollen spectra as a tool for elucidating the nature of fine-scale human activities in boreal areas

Modern pollen assemblages from the major vegetation units (both natural and anthropogenic) on the island of Hailuoto, Finland are studied from 29 surface moss samples. A total of 59 pollen and spore taxa are recorded. The pollen data-set is related by redundancy analysis (RDA) to six external synthetic variables that characterize in general terms the different major ecological situations relevant to the land-use on the island (sand, humus, forest, field, deforested, treeless). Weighted average (WA) optima are calculated to identify which pollen taxa are most indicative of four of these external variables. Two fossil pollen diagrams from Hailuoto are re-interpreted by positioning their samples on the RDA plot of the modern data-set and by classifying the modern and fossil spectra together in a minimum-variance cluster analysis. The RDA plot shows that the strongest features in the modern pollen assemblages are the contrast between spectra from dry sandy soils and those from damper soils with more organic content, and between assemblages from forested and unforested areas. For anthropogenic situations the numerical analyses detect a distinction in the pollen assemblages characterizing fields from those representing farms and trackways but is less successful in separating the latter two. The WA optima indicate that most pollen taxa considered as indicator types primarily represent a mixed field/farm category of land-use. The RDA and minimum-variance cluster analysis confirm that the two fossil sites have had different vegetational histories. In neither is there any phase equivalent to the modern farm situation on Hailuoto. The initial phase in each diagram represents shore meadows and fields but because Poaceae pollen is identifiable only to family level it is not possible to separate these two habitat types.