Fruit evolution in Hydrophyllaceae.

PREMISE: Fruit type and morphology are tightly connected with angiosperm diversification. In Boraginales, the first-branching families, including Hydrophyllaceae, have one- to many-seeded capsules, whereas most of the remaining families have four-seeded indehiscent fruits. This fact argues for many-seeded capsules as the ancestral condition. However, little is known about the evolution of fruit dehiscence and seed number. The present study investigated the gynoecium and fruit development and morphology and the evolution of seed-numbers in Hydrophyllaceae. METHODS: Gynoecium and fruit development and morphology were studied using scanning electron microscopy and x-ray microcomputed tomography. Ancestral character state reconstruction of seed number was performed using a broadly sampled phylogeny of Boraginales (ndhF and ITS) with an emphasis on Hydrophyllaceae. RESULTS: Our ontogenetic studies not only demonstrate parallel developmental trajectories across Hydrophyllaceae, but also a striking diversity regarding the internal organization of the gynoecium. Ovule number appears to determine ovary structure. Many-seeded capsules are retrieved as the ancestral state of Hydrophyllaceae. At least seven transitions to fruits with (one to) four seeds and four reversals (i.e., from four- to many-seeded fruits) were reconstructed in Hydrophyllaceae. CONCLUSIONS: Several shifts in seed number from "many" to "four" and back to "many" have taken place in capsular-fruited Hydrophyllaceae, a strikingly high number considering that seed number is virtually conserved across the rest of the order. The groups with a conserved seed number of four are characterized by indehiscent schizocarps or drupes and by seeds that are integrated into mericarps. This functional integration probably acts as an evolutionary constraint to shifts in seed number.

Antioxidant properties, phenolic and mineral composition of germinated chia, golden flax, evening primrose, phacelia and fenugreek.

Seeds and sprouts are of considerable interest due to their numerous pro-health benefits. The aim of this study was to investigate the effect of germination on the mineral composition (performed by flame absorption atomic spectroscopy), total phenolic content, antioxidant activity, as well as phenolic profiles (before and after alkaline hydrolysis by high-performance liquid chromatography) of chia, golden flax, evening primrose, phacelia and fenugreek seeds. Generally, significant (p < 0.05) changes in the individual minerals composition of the seeds, improvement of their antioxidant properties, as well as increase in levels of individual phenolic compounds was found after seeds germination. Alkaline hydrolysis allowed to release free forms of phenolics and to confirm (chromatographically) their significantly higher amounts when compared to the nonhydrolyzed fraction. Gallic, protocatechuic, caffeic, p-coumaric, ferulic and sinapic acids, as well as quercetin and kaempferol were identified in analyzed seeds and sprouts. Sprouts exhibited better nutritional values than their un-germinated forms.

Identification and Characterization of Novel Nemophila menziesii Flavone Glucosyltransferases that Catalyze Biosynthesis of Flavone 7,4'-O-Diglucoside, a Key Component of Blue Metalloanthocyanins.

The brilliant blue color of the Nemophila menziesii flower is derived from metalloanthocyanin, which consists of anthocyanin {petunidin 3-O-[6-O-(trans-p-coumaroyl)-ß-glucoside]-5-O-[6-O-(malonyl)-ß-glucoside]}, flavone [apigenin 7-O-ß-glucoside-4'-O-(6-O-malonyl)-O-ß-glucoside] and metal ions (Mg2+, Fe3+). Although the two glucosyl moieties at the apigenin 7-O and 4'-O positions are essential for metalloanthocyanin formation, the mechanism of glucosylation has not yet been clarified. In this study, we used crude protein extract prepared from N. menziesii petals to determine that apigenin is sequentially glucosylated by the catalysis of UDP-glucose:flavone 4'-O-glucosyltrasferase (F4'GT) and UDP-glucose:flavone 4'-O-glucoside 7-O-glucosyltransferase (F4'G7GT). We identified 150 contigs exhibiting homology with a UDP-glucose-dependent GT in the N. menziesii petal transcriptome and isolated 24 putative full-length GT cDNAs which were then subjected to functional analysis. Two GT cDNAs, NmF4'GT and NmF4'G7GT, which are highly expressed during the early stages of petal development and rarely in leaves, were shown to encode F4'GT and F4'G7GT activities, respectively. Biochemical characterization of the recombinant enzymes revealed that NmF4'GT specifically catalyzed 4'-glucosylation of flavonoids and that NmF4'G7GT specifically catalyzed 7-glucosylation of flavone 4'-O-glucosides and flavones. Apigenin 7,4'-O-diglucoside was efficiently synthesized from apigenin in the presence of recombinant NmF4'GT and NmF4'G7GT. Transgenic tobacco BY-2 cells expressing NmF4'GT and NmF4'G7GT converted apigenin into apigenin 7,4'-O-diglucoside, confirming their activities in vivo. Based on these results, we conclude that these two GTs act co-ordinately to catalyze apigenin 7,4'-O-diglucoside biosynthesis in N. menziesii.

Floral neighborhood influences pollinator assemblages and effective pollination in a native plant.

Pollinators represent an important intermediary by which different plant species can influence each other's reproductive fitness. Floral neighbors can modify the quantity of pollinator visits to a focal species but may also influence the composition of visitor assemblages that plants receive leading to potential changes in the average effectiveness of floral visits. We explored how the heterospecific floral neighborhood (abundance of native and non-native heterospecific plants within 2 m × 2 m) affects pollinator visitation and composition of pollinator assemblages for a native plant, Phacelia parryi. The relative effectiveness of different insect visitors was also assessed to interpret the potential effects on plant fitness of shifts in pollinator assemblage composition. Although the common non-native Brassica nigra did not have a significant effect on overall pollinator visitation rate to P. parryi, the proportion of flower visits that were made by native pollinators increased with increasing abundance of heterospecific plant species in the floral neighborhood other than B. nigra. Furthermore, native pollinators deposited twice as many P. parryi pollen grains per visit as did the nonnative Apis mellifera, and visits by native bees also resulted in more seeds than visits by A. mellifera. These results indicate that the floral neighborhood can influence the composition of pollinator assemblages that visit a native plant and that changes in local flower communities have the potential to affect plant reproductive success through shifts in these assemblages towards less effective pollinators.

Evaluating exposure and potential effects on honeybee brood (Apis mellifera) development using glyphosate as an example.

This study aimed to develop an approach to evaluate potential effects of plant protection products on honeybee brood with colonies at realistic worst-case exposure rates. The approach comprised 2 stages. In the first stage, honeybee colonies were exposed to a commercial formulation of glyphosate applied to flowering Phacelia tanacetifolia with glyphosate residues quantified in relevant matrices (pollen and nectar) collected by foraging bees on days 1, 2, 3, 4, and 7 postapplication and glyphosate levels in larvae were measured on days 4 and 7. Glyphosate levels in pollen were approximately 10 times higher than in nectar and glyphosate demonstrated rapid decline in both matrices. Residue data along with foraging rates and food requirements of the colony were then used to set dose rates in the effects study. In the second stage, the toxicity of technical glyphosate to developing honeybee larvae and pupae, and residues in larvae, were then determined by feeding treated sucrose directly to honeybee colonies at dose rates that reflect worst-case exposure scenarios. There were no significant effects from glyphosate observed in brood survival, development, and mean pupal weight. Additionally, there were no biologically significant levels of adult mortality observed in any glyphosate treatment group. Significant effects were observed only in the fenoxycarb toxic reference group and included increased brood mortality and a decline in the numbers of bees and brood. Mean glyphosate residues in larvae were comparable at 4 days after spray application in the exposure study and also following dosing at a level calculated from the mean measured levels in pollen and nectar, showing the applicability and robustness of the approach for dose setting with honeybee brood studies. This study has developed a versatile and predictive approach for use in higher tier honeybee toxicity studies. It can be used to realistically quantify exposure of colonies to pesticides to allow the appropriate dose rates to be determined, based on realistic worst-case residues in pollen and nectar and estimated intake by the colony, as shown by the residue analysis. Previous studies have used the standard methodology developed primarily to identify pesticides with insect-growth disrupting properties of pesticide formulations, which are less reliant on identifying realistic exposure scenarios. However, this adaptation of the method can be used to determine dose-response effects of colony level exposure to pesticides with a wide range of properties. This approach would limit the number of replicated tunnel or field-scale studies that need to be undertaken to assess effects on honeybee brood and may be of particular benefit where residues in pollen and nectar are crop- and/or formulation-specific, such as systemic seed treatments and granular applications.

The effect of floral resources on parasitoid and host longevity: prospects for conservation biological control in strawberries.

The strawberry tortricid, Acleris comariana Lienig and Zeller (Lepidoptera: Tortricidae) is an important pest in Danish strawberry production. Its most common parasitoid is Copidosoma aretas (Walker) (Hymenoptera: Chalcidoidea: Encyrtidae). To identify selective flowering plants that could be used to increase functional biodiversity, the longevity of C. aretas and its host A. comariana was assessed on 5 flowering species: buckwheat, Fagopyrum esculentum Moench (Caryophyllales: Polygonaceae); borage, Borago officinalis L. (Boraginaceae); strawberry, Fragaria x ananassa Duchesne (Rosales: Rosaceae); phacelia, Phacelia tanacetifolia Bentham (Boraginaceae); and dill, Anethum graveolens L. (Apiales: Apiaceae). Dill was only tested with C. aretas. Sucrose and pollen served as positive controls, and pure water as a negative control. In a subsequent field experiment, A. comariana larval density was assessed at 1, 6, and 11 m distances from buckwheat flower strips in 3 fields. The proportion of field-collected larvae that were parasitized by C. aretas or fungi was assessed. Among the tested floral diets, buckwheat was superior for C. aretas, increasing its longevity by 1.4 times compared to water. Although buckwheat also increased longevity of A. comariana, its longevity and survival on buckwheat, borage, and strawberry was not significantly different, so buckwheat was chosen for field experiments. A. comariana densities in the 3 fields with sown buckwheat flower strips were 0.5, 4.0, and 8.3 larvae per m per row of strawberry respectively. Of the collected larvae, a total of 1%, 39%, and 65% were parasitized by C. aretas, respectively. The density of A. comariana and the proportion parasitized by C. aretas were highly significantly correlated. Distance from floral strips had no significant effect on either A. comariana larval density or on the proportion of individuals parasitized by C. aretas. Few other parasitoids emerged from collected larvae, and no larvae were infected by entomopathogenic fungi. Still, total A. comariana mortality was significantly affected by distance to flower strips, with the highest mortality near the flower strips. As no effect of buckwheat flower strips on C. aretas parasitism was found, the positive effect they had on A. comariana control stems from unknown mortality factors. As literature indicates that buckwheat for flower strips can augment a more complex suite of natural enemies, one such mortality factor could be a non-consumptive predator and/or parasitoid effect, but this requires further study. If confirmed, buckwheat may be utilized together with a selective food plant, once identified.

At3g08030 transcript: a molecular marker of seed ageing.

BACKGROUND AND AIMS: Prolonged storage generally reduces seed viability and vigour, although the rate of deterioration varies among species and environmental conditions. Here, we suggest a possible ageing molecular marker: At3g08030 mRNA. At3g08030 is a member of the DUF642 highly conserved family of cell-wall-associated proteins that is specific for spermatophytes. METHODS: At3g08030 expression was performed by RT-PCR and qRT-PCR analysis in seed samples differing in their rate of germination and final germination following a matrix priming and/or controlled deterioration (rapid ageing) treatment. KEY RESULTS: The At3g08030 gene transcript was present during the entire Arabidopsis thaliana plant life cycle and in seeds, during maturation, the ripening period and after germination. Matrix priming treatment increased the rate of germination of control seeds and seeds aged by controlled deterioration. Priming treatments also increased At3g08030 expression. To determine whether the orthologues of this gene are also age markers in other plant species, At3g08030 was cloned in two wild species, Ceiba aesculifolia and Wigandia urens. As in A. thaliana, the At3g08030 transcript was not present in aged seeds of the tested species but was present in recently shed seeds. A reduction in germination performance of the aged seeds under salt stress was determined by germination assays. CONCLUSIONS: At3g08030 mRNA detection in a dry seed lot has potential for use as a molecular marker for germination performance in a variety of plant species.

Reversal of the inhibitory effect of light and high temperature on germination of Phacelia tanacetifolia seeds by melatonin.

Possible role of melatonin in the germination of negatively photoblastic and thermosensitive seeds of Phacelia tanacetifolia Benth was studied. Final germination percentage (FGP) was determined in the presence or absence of light at various temperatures, ranging from 0 to 40°C. The highest FGP was determined as 48.7% and 92% at temperature of 15°C in the presence and absence of light, respectively. Seeds were primed with 1% KNO(3) containing various concentrations (0.3, 1, 6, 12, 30, 60, or 90 µM) of melatonin for 2 days at 15°C in darkness. Primed seeds were germinated at an inhibitory temperature of 30°C, and results were compared to those occurring at the optimum temperature of 15°C under both light and no light conditions. Melatonin incorporated into priming medium significantly reversed the inhibitory effects of light and high temperature. Germination was elevated from 2.5% to 52% of FGP for seeds primed in the presence of 6 µM melatonin in darkness at 30°C, while 1 µM melatonin had the highest FGP (21.0%) in the presence of light at 30°C. The highest FGP (47.5%) was obtained from seeds primed in the presence of 0.3 µM melatonin under the light condition at 15°C, while untreated seeds had 1.5% of FGP. The fastest seed germination was determined from seeds primed in the presence of 0.3 µM melatonin (G(50) = 0.56 days) at 15°C in darkness. The possible roles of melatonin in promoting germination parameters of photo- and thermosensitive seed germination are discussed.

Flowering catch crops used as forage plants for dairy cows: influence on fatty acids and tocopherols in milk.

The effect of several flowering dicotyledonous catch crop plants (dicots) on milk fat quality in cows was investigated to test the hypothesis that their phenolic compounds may inhibit ruminal biohydrogenation and thus enhance the transfer to milk of intact, plant-derived polyunsaturated fatty acids. Berseem clover (Trifolium alexandrinum), buckwheat (Fagopyrum esculentum), and phacelia (Phacelia tanacetifolia) were sown in mixture with ryegrass (Lolium multiflorum; intended biomass proportion of 0.2) on 1ha. For comparison, nonflowering chicory (Cichorium intybus, also sown in mixture with ryegrass) and pure ryegrass were cultivated. Realized biomass proportions (wet weight) were 91% for berseem clover, 69% for buckwheat, 54% for phacelia, and 51% for chicory. At the start of flowering (or from d 47 after sowing onward), cultures were harvested daily and fed for 20 d ad libitum to groups of 6 midlactation cows each. Additionally, 1 kg each of energy and protein concentrate and pure ryegrass hay were fed. Individual intake and milk yield of the cows were measured daily. Milk samples were obtained twice daily 5 d before and from 11 to 20 d after the start of treatment feeding. Feed samples were drawn twice a week from the fresh feeds. Apart from standard traits, feeds and milk were analyzed for fatty acids, tocopherols, and phenolic fractions. Only a few substantial treatment effects on intake and performance were observed. All diets based on dicots increased α-linolenic acid (ALA) concentrations in milk fat compared with the ryegrass diet even though the corresponding swards were not generally richer in ALA. The highest ALA concentration in milk fat (1.3 g/100g of fatty acids) occurred with the berseem clover diet. Transfer rate of ALA from feed to milk was highest with the buckwheat diet (0.09) and lowest with ryegrass (0.05). This was congruent with the differences in total extractable phenols, being high in the buckwheat sward (2.6% of dry matter) and low in the ryegrass sward (1.2% of dry matter). Intermediates of ALA biohydrogenation were lowest in the milk fat of the buckwheat group, indicating an inhibitory effect of this treatment, which provided the highest dietary levels of phenols. The α-tocopherol concentration in milk was higher with the buckwheat diet than with berseem clover and phacelia diets. The study provides evidence that the ALA concentration in milk fat could be enhanced by feeding flowering dicots; however, this was due to different modes of action.

Seasonal timing of first rain storms affects rare plant population dynamics.

A major challenge in forecasting the ecological consequences of climate change is understanding the relative importance of changes to mean conditions vs. changes to discrete climatic events, such as storms, frosts, or droughts. Here we show that the first major storm of the growing season strongly influences the population dynamics of three rare and endangered annual plant species in a coastal California (USA) ecosystem. In a field experiment we used moisture barriers and water addition to manipulate the timing and temperature associated with first major rains of the season. The three focal species showed two- to fivefold variation in per capita population growth rates between the different storm treatments, comparable to variation found in a prior experiment imposing eightfold differences in season-long precipitation. Variation in germination was a major demographic driver of how two of three species responded to the first rains. For one of these species, the timing of the storm was the most critical determinant of its germination, while the other showed enhanced germination with colder storm temperatures. The role of temperature was further supported by laboratory trials showing enhanced germination in cooler treatments. Our work suggests that, because of species-specific cues for demographic transitions such as germination, changes to discrete climate events may be as, if not more, important than changes to season-long variables.

Evidence of restoration cost in the annual gynodioecious Phacelia dubia.

A negative pleiotropic effect on fitness of nuclear sex-determining genes (cost of restoration) could explain nuclear-cytoplasmic gynodioecy but rarely has been demonstrated empirically. In a gynodioecious Phacelia dubia population, maternal lineages produce only hermaphroditic progenies irrespective of the pollen parent (N) or can segregate females (S). Natural progenies of N maternal plants had lower seed viability than that of S. Full-sib progenies of unrelated hermaphrodites from all possible matings between N and S lineages had similar pollen filling but differed in sporophyte performance, mainly at seed germination stage. A discrete multivariate analysis reveals that the performance of N(female symbol) x S(male symbol) progeny at early stages of development was significantly lower than that of the other three types of mating in agreement with the silent-cost-of-restoration hypothesis, affecting the sporophyte. The restoration cost and male sterility appear to be dominant and consequence of nuclear-cytoplasmic incompatibilities that may maintain nuclear-cytoplasmic polymorphism by frequency-dependent selection.

Role and regulation of cAMP in seed germination of Phacelia tanacetifolia.

Although adenosine 3',5'-cyclic monophosphate (cAMP) is known as a key second messenger in many living organisms, regulating a wide range of cellular responses, its biological function in higher plants is not well understood. In this study, the role and the regulation mechanism of cAMP in seed germination of Phacelia tanacetifolia Benth. were examined. The cAMP level of the seeds incubated under optimal conditions for germination showed a transient elevation before germination. When the seeds were exposed to light or supraoptimal temperature during incubation, elevation of cAMP levels as well as germination of the seeds were inhibited. Addition of membrane-permeable cAMP to the medium restored the germination rates of these seeds, suggesting that cAMP functions during germination. Treatment of the seeds with gibberellin (GA) was also effective to restore the elevation of cAMP levels and germination of the seeds. Uniconazole, a potent inhibitor of GA biosynthesis, blocked elevation of cAMP level under optimal conditions for germination. These results suggest that cAMP plays a role in the regulation of germination and that the cAMP level is regulated by GA in P. tanacetifolia seeds.

Florivory affects pollinator visitation and female fitness in Nemophila menziesii.

While herbivory has traditionally been studied as damage to leaves, florivory - herbivory to flowers prior to seed set - can also have large effects on plant fitness. Florivory can decrease fitness directly, either through the destruction of gametes or through alterations to plant physiology during fruit set, and can also change the appearance of a flower, deterring pollinators and reducing seed set. In order to distinguish between these hypotheses, it is necessary to both damage flowers and add pollen in excess to study the effects of damage on pollen limitation. Very few studies have used this technique over the lifetime of a plant. Here I describe a series of experiments showing the effects of natural and artificial damage on reproductive success in the annual plant Nemophila menziesii (Hydrophyllaceae, sensu lato). I show that natural and artificial petal damage decreased radial symmetry relative to controls and that both types of damage deterred pollinator activity. Both naturally damaged flowers and artificially damaged flowers in the field set fewer fruit or seed relative to undamaged control flowers. Finally, in an experiment crossing artificial petal damage with pollen addition, petal damage alone over the lifetime of this plant decreased female fitness, but only after a threshold of damage was reached. The fitness effect appeared to be direct because there was no detectable effect of pollen addition on the relationship between florivory and fitness. This result implies that both damaged and undamaged plants show similar amounts of pollen limitation and suggests that pollinator-mediated effects contributed little to the negative effects of florivory on female fitness. Florivores may thus be an under-appreciated agent of selection in certain plants, although more experimental manipulation of florivory is needed to determine if it is important over a range of taxa.

Preparation of 2H-furo[2,3-c]pyran-2-one derivatives and evaluation of their germination-promoting activity.

The butenolide, 3-methyl-2H-furo[2,3-c]pyran-2-one (1), has recently been identified as the germination stimulant present in smoke that promotes the germination of seeds from a wide range of plant species. In this paper, we describe the preparation of a number of analogues of 1 and compare their efficacy in promoting seed germination of three highly smoke-responsive plant species, Lactuca sativa L. cv. Grand Rapids (Asteraceae), Emmenanthe penduliflora Benth. (Hydrophyllaceae), and Solanum orbiculatum Poir. (Solanaceae). The results show that the methyl substituent at C-3 in 1 is important for germination-promoting activity while substitution at C-7 reduces activity. In contrast, bioactivity is mostly retained with analogues substituted at C-4 or C-5.

Structure of anthocyanin from the blue petals of Phacelia campanularia and its blue flower color development.

The dicaffeoyl anthocyanin, phacelianin, was isolated from blue petals of Phacelia campanularia. Its structure was determined to be 3-O-(6-O-(4'-O-(6-O-(4'-O-beta-d-glucopyranosyl-(E)-caffeoyl)-beta-d-glucopyranosyl)-(E)-caffeoyl)-beta-d-glucopyranosyl)-5-O-(6-O-malonyl-beta-d-glucopyranosyl)delphinidin. The CD of the blue petals of the phacelia showed a strong negative Cotton effect and that of the suspension of the colored protoplasts was the same, indicating that the chromophores of phacelianin may stack intermolecularly in an anti-clockwise stacking manner in the blue-colored vacuoles. In a weakly acidic aqueous solution, phacelianin displayed the same blue color and negative Cotton effect in CD as those of the petals. However, blue-black colored precipitates gradually formed without metal ions. A very small amount of Al(3+) or Fe(3+) may be required to stabilize the blue solution. Phacelianin may take both an inter- and intramolecular stacking form and shows the blue petal color by molecular association and the co-existence of a small amount of metal ions. We also isolated a major anthocyanin from the blue petals of Evolvulus pilosus and revised the structure identical to phacelianin.

Soil moisture and sex ratio in a plant with nuclear-cytoplasmic sex inheritance.

I investigated whether soil moisture affects relative fitness of females and hermaphrodites and sex ratio in a gynodioecious plant with nuclear-cytoplasmic sex inheritance. I contrast these results with those from species with strictly nuclear sex inheritance. I performed a manipulative watering experiment on seed fitness of the two sexes, and field studies measuring seed fitness and sex ratio as a function of soil moisture. In the dry site, watered hermaphrodites produced approximately twice as many seeds as unwatered hermaphrodites, with little treatment effect on female seed production. Over a natural soil moisture gradient, the ratio of female to hermaphrodite seed production was higher in dry than in wet sites. These data show that the seed fitness advantage of females is a function of soil moisture. Despite this, regression of soil moisture on the sex ratio of 23 populations was not significant. These results indicate a sex-dependent effect of soil moisture on resource allocation to seeds that does not translate into a strong effect on sex ratio. This is consistent with theory based on genomic conflict in which sex ratios are predicted to be only partly determined by fitness differences of the sexes.

Hybridization and regional sex ratios in Nemophila menziesii.

I tested whether a region of high female frequencies in the gynodioecious plant, Nemophila menziesii, may be due to hybridization between regionally distributed populations with different corolla colours. I crossed plants in the greenhouse from populations with different corolla colours and found that hybrid crosses yielded higher frequencies of females than within-colour crosses. In the field, I found that populations with high female frequencies had intermediate mean corolla colours and higher variance in corolla colour, two traits suggesting hybridization. Nemophila menziesii has nuclear-cytoplasmic sex inheritance, thus if populations with different corolla colours are fixed for different male-sterile cytoplasms and matching nuclear restorer alleles, hybridization between populations with different corolla colour should yield high frequencies of females. Two populations that are all hermaphroditic in the field segregated females in hybrid crosses suggesting that field populations may contain sex ratio distorters but appear undistorted, a prediction of genomic conflict theory.

Phenolic derivatives from Wigandia urens with weak activity against the chemokine receptor CCR5.

Three compounds, 2,3-dihydroxy-4-methoxy-6,6,9-trimethyl-6H-dibenzo[b,d]pyran (1), 8-methoxy-2-methyl-2-(4-methyl-3-pentenyl)-2H-1-benzopyran-6-ol (2) and 4-methoxy-3-(3-methyl-2-butenyl)-benzoic acid (3), have been isolated from Wigandia urens. The structures of compounds 1, 2 and 3 were determined from spectroscopic data and showed activity in a CCR5 assay with IC(50) values of 33, 46 and 26 muM respectively.

Isolation and structural identification of a germination inhibitor in fire-recruiters from the California chaparral.

A role for inhibitors as regulators of seed dormancy in fire-dependent annuals and their nondormant (fire-independent) congeners was examined in the family Hydrophyllaceae. From seeds of the fire-dependent species, Emmenanthe penduliflora, Phacelia minor, P. brachyloba, and P. grandiflora, extracts were obtained that were found to be largely self-inhibitory and potent inhibitors of seed germination in nondormant congeners. Lower activity was detected in the fire-independent species, P. tanacetifolia and P. campanularia. The inhibitory activity was associated with the new sucrose ester, 6-O-linoleyl-alpha-D-glucopyranosyl-beta-D-fructofuranoside.