Coevolution of the checkerspot butterfly Euphydryas chalcedona and its larval food plant Diplacus aurantiacus: larval response to protein and leaf resin

Summary Prediapause larvae of the checkerspot butterfly Euphydryas chalcedona were raised from hatch until entrance into diapause on artificial diets. The proportions of protein and host plant leaf resin differed among the diets. Larval size growth rates and mortality were monitored and overall rates and efficiencies of food use were computed.Larval survivorship, growth rate and size of larvae at idapause were significantly enhanced by increasing dietary protein content, particularly over the range found in leaves of the host plant. In contrast, an increasing dietary content of Diplacus aurantiacus leaf resin significantly depressed larval surviviorship, growth rates and size of larvae at diapause. A simple dosedependent interaction was observed between the effects of dietary leaf resin and protein on larval success. Dietary content of leaf resin and protein significantly influenced some measures of food utilization efficiency (ECI and ECD), but not others (AD and NUE).The negative interaction between the effects of dietary leaf resin and protein content suggests the leaf resin phenolic compounds reduce the availability of protein to the larvae. The results for efficiency indices of larval food use are potentially in conflict with this interpretation.The influence of host plant leaf resin and protein on larval success, coupled with the relation between photosynthesis and leaf nitrogen content, are consistent with the hypothesis that productivity can be enhanced by herbivore deterrence resulting from leaf resin production.     

Effect of moisture stress on composition and yield in leaf resin of Hymenaea courbaril

Previous controlled environment studies of the effect of photoperiod, light intensity and temperature have shown little phenotypic plasticity in leaf resin composition of Hymenaea courbaril which ranges throughout New World lowland tropical ecosystems. The present control chamber study also showed negligible variation in resin composition as a result of moisture status. Although some change in composition occurred under dry treatment conditions, leaf development was retarded and the variation is consistent with the progressive change in composition with leaf development from very young to fully expanded leaves. Yield (mg resin/g leaf tissue), not measured in previous studies, also showed little variation between wet and dry treatments. It is suggested that these indications of low plasticity in both composition and yield in Hymenaea leaf resins may be significant in herbivore interactions.     

大孔树脂分离纯化辣椒叶总黄酮的工艺研究

目的:筛选适合分离纯化辣椒叶总黄酮的一种大孔树脂,同时用响应面法进行优化得到最佳纯化工艺。方法:采用热回流法提取辣椒叶总黄酮,以吸附率和解吸率为考察指标,考察6种不同型号的大孔树脂(HPD100、HPD450、HPD600、HPD826、D101、AB-8)对辣椒叶总黄酮的吸附能力与解吸能力,确定最佳树脂。通过动态吸附解吸实验考察此树脂对辣椒叶总黄酮的最佳分离纯化工艺。结果:通过对辣椒叶总黄酮吸附分离性能的分析显示HPD600为最佳树脂,最优工艺为:上样浓度为10 mg/mL,上样量为10 mL,洗脱体积为4 BV,洗脱液流速为4 mL/min,洗脱液pH为7,依次用水、10%、30%乙醇冲洗树脂柱,50%乙醇为洗脱液。纯化后的黄酮纯度435.4 mg/g。结论:该方法简便,操作简单,对辣椒叶总黄酮的纯化效果较好。     

Integrated antiherbivore, antidesiccant and ultraviolet screening properties of creosotebush resin

The surface leaf resin of creosotebush, Larrea spp. (Zygophyllaceae) exhibits antiherbivore, antidesiccant and UV screening properties. Composed mainly of monomeric phenolic aglycones, the resin absorbs most radiation of wavelength shorter than 410 nm, reduces the rate of evaporative water loss across cellulose membrane and exhibits tannin-like protein-complexing and digestibility-reducing properties. Leaf-chewing insects prefer mature leaves which have a lower resin content than young leaves. A phenoloxidaes system present in the leaves probably enhances the digestibility-reducing action of resin on in vitro proteolysis and growth-rates of Astroma quadrilobatum (Orthoptera: Proscopiidae) are negatively correlated with resin-phenoloxidase activity as measured by macerated leaf oxygen demand. Non-ideal behaviour of the resin is suggestive of cooperative action between components.     

Sesquiterpenes in leaf pocket resins of Copaifera species

Four sesquiterpene leaf resin components were isolated and identified from Copaifera leaf resin. Additional GC and mass spectrometric evidence support the close similarity of Copaifera leaf pocket resin composition with that of the related genus, Hymenaea.     

不同工艺提取老鹰茶总黄酮含量的比较

比较不同工艺提取富集的老鹰茶中总黄酮含量,分别采用传统乙醇提取-正丁醇萃取法和水提取-大孔树脂法提取老鹰茶总黄酮,并用紫外分光光度法,以芦丁为标准品,270nm下测定总黄酮含量。水提取-大孔树脂法富集所得固体中黄酮含量较高,可达55.86%,回收率为105.6%,RSD=1.64%。水提取-大孔树脂富集的工艺操作简单,所得固体黄酮含量高,更适于老鹰茶总黄酮的提取,对进一步的工业化生产开发有较好的参考价值。     

The response of leaf resin to artificial herbivory in Eriodictyon californicum

Herbivore damage induces the production of defensive chemicals in many plant species. However, defensive chemicals sometimes have functions other than protection from herbivores, and these chemicals may respond to hervibory differentialy than those which are primarily defensive. A well-studied group of leaf constituents with multiple functions is the phenolic leaf resins, which protect leaves from herbivores, UV radiation, and possibly excess water loss. The effects of herbivory on phenolic leaf resins were investigated in the chapparral shrub Eriodictyon californicum by artificially damaging leaves during the growing season and subsequently measuring resin content and composition. We found that (i) resin content declined in leaves damaged by artificial herbivory; (ii) resin declined (at the P < 0.1 level) in undamaged leaves present on the same plant as those heavily damaged; and (iii) resin composition, as measured by the ratio of the major components, was unaffected by artificial herbivory.     

Sesquiterpene Deterrence of a Leaf-Tying Lepidopteran, Stenoma ferrocanella, on Hymenaea stigonocarpa in Central Brazil

Study of two populations of Hymenaea stigonocarpa in the central Brazilian cerrado (savanna/woodland) shows attack by a leaf-tying lepidopteran, Steoma ferrocanella, occurred after inferred peak concentration (% of leaf dry wt) of sesquiterpene leaf resins during leaf development. Also a highly statistically significant relationship existed between leaf-tier attack and level of caryophyllene in one population and γ-muurolene in the other population. The data from one population are consistent with laboratory experiments demonstrating highly significant dose-dependent effects of caryophyllene in Hymenaea leaf resin on mortaility of the lepidopteren Spodoptera exigua. Results from both populations support other evidence that changes in the amount of single terpenes in the mixture constituting the resin may have significant antiherbivore effects and, concomitantly, that herbivory may be a source of some of the leaf resin variation.     

The seasonal dynamics of leaf resin,nitrogen, and herbivore damage in Eriodictyon californicum and their parallels in Diplacus aurantiacus

Summary The chaparral shrub Eriodictyon californicum produces a flavonoid leaf resin with a chemically similar composition to that previously reported for the sympatric shrub Diplacus aurantiacus. We determined the phenology, resin content, and nitrogen content of Eriodictyon leaves and the leaf area lost to herbivores. Nitrogen content and resin content were both negatively correlated with leaf age at each sampling date, but nitrogen decreased during the growing season while resin increased. The fraction of leaf area lost during the growing season averaged less than 7% and was highest on the oldest leaves. The seasonal pattern of resin production in Eriodictyon corresponds to that in Diplacus, indicating that the similar environments of Eriodyctyon and Diplacus have led to convergent leaf resins. This convergence in these two plants has implications for chemicals of similar function in other chaparral shrubs.     

Vegetative growth and leaf resin composition in Hymenaea courbaril under photoperiodic extremes

Seedlings from five geographic populations of the widespread Hymenaea courbaril were grown for one year under two extremes of photoperiod. The short photoperiod treatment consisted of 8 hr of unsupplemented greenhouse daylight and the long photoperiod treatment of greenhouse daylight at the natural photoperiod of Santa Cruz, California (37° N) plus a 3-hr night interruption (0.25 mW/cm²). Growth was measured by total stem length and leaf resin sesquiterpene composition was quantitatively analyzed by GLC. Photoperiod had no significant effect on leaf resin chemistry but the longer photoperiod markedly promoted vegetative growth. Large population differences in overall vegetative growth and quantitative leaf resin composition were also evident. Growth patterns indicated that the low intensity night break had little or no effect on growth.     

Flavonoid aglycones from Eriodictyon californicum resin and their implications for herbivory and UV screening

The flavones luteolin and chrysoeriol and five methylated derivatives of the flavonone eriodictyol were identified in the external leaf resin of Eriodictyon californicum. These flavonoids comprise approx. 80% of the mass of the resin. The flavonones can be classified into 7-hydroxy- or 7-methoxy-compounds, and these two groups of compounds can be separately quantified in a crude resin extract using UV spectroscopy in the presence of sodium methoxide. The different properties of these flavonoids as herbivore deterrents and UV screens may explain the complexity of the leaf resin mixture.     

The impact of plasticity and maternal effect on the evolution of leaf resin production in Diplacus aurantiacus

Our previous quantitative genetic study of leaf resin production in Diplacus aurantiacus revealed large environmental and maternal effects on variation in resin production, which suggests the possibility of a genotype×environment interaction for this trait when plants grow in heterogeneous environments. Our objectives in this study were to observe the genetic variation in plasticity of resin production under field and chamber conditions, compare phenotypic correlations of resin content with growth traits under these two environmental conditions, and distinguish the possible basis of the maternal effect on resin production using parents and half-sib progeny. A significant genotype×environment interaction (P<0.0001) in leaf resin production was found, which suggests a potential for the evolution of plasticity of these secondary metabolites under heterogeneous environments. The phenotypic correlation between resin content and growth rate also exhibited plasticity. In addition, the resin content of dam half-sib families grown in the chamber had a closer relationship with their maternal parents in the field (r=0.65, P=0.059) than in the chamber (r=0.39, P=0.34), suggesting an environmentally based maternal effect on the secondary chemicals. We suggest that the maternal environmental effect may act as a contributor to plasticity of resin production and, while it may not diminish the appearance of the genotype×environment interaction, the heritable variation of plasticity of resin production may be confounded.     

Multitrophic interactions among fungal endophytes,bees, and <Emphasis Type="Italic">Baccharis dracunculifolia</Emphasis>: resin tapering for propolis production leads to endophyte infection

The tropics are known for their high diversity of plants, animals, and biotic interactions, but the role of the speciose endophytic fungi in these interactions has been mostly neglected. We report a unique interaction among plant sex, bees, and endophytes on the dioecious shrub, Baccharis dracunculifolia (Asteraceae). We assessed whether there was an association between resin collection by bees and fungal endophytes considering the host plant sex. We hypothesized that resin collection by the Africanized honey bee, Apis mellifera L. (Apidae) could favor the entry of endophytes in B. dracunculifolia. Specifically, we tested the hypotheses that (1) bees damage the leaf buds of female and male plant at different proportions; (2) damage on leaf buds increases the richness of endophytic fungi; (3) endophyte richness differs between female and male plants; and (4) in vitro growth of endophytes depends on the sex of the plant individual from which the resin was extracted. Endophyte richness and proportion of leaf bud damage did not vary between the plant sexes. However, species similarity of endophytes between female and male plants was 0.33. Undamaged leaf buds did not show culturable endophytes, with all fungi exclusively found in damaged leaf buds. Endophyte composition changed with the plant sex. The endophytes exclusively found in female plants did not develop in the presence of male resin extract. These findings highlight that resin collection by A. mellifera for propolis production favors the entry of endophytic fungi in B. dracunculifolia. Additionally, endophyte composition and growth are influenced by plant sex.     

The role of leaf resin in the interaction between Eriodictyon californicum (Hydrophyllaceae) and its herbivore,Trirhabda diducta (Chrysomelidae)

Summary The chaparral shrub Eriodictyon californicum secretes a phenolic leaf resin composed of flavonoid aglycones. We used leaves with artificially altered resin contents to test the effects of resin on the feeding, growth, and oviposition of the specialist herbivore Trirhabda diducta. In addition, we compared Trirhabda feeding and growth on young foliage with that on foliage from the preceding year. Our results show that the Eriodictyon leaf resin affects Trirhabda larvae and adults similarly, having no significant effect on growth rates or on nutrient utilization at up to 5X the resin levels normally encountered by larvae in the field. Both Trirhabda larvae and adults respond to high resin concentrations by increasing their consumption rates, with concomitant decreases in digestibility and the efficiency of conversion of ingested food to biomass. Low-resin foliage is preferred by larvae for feeding and by adults for oviposition. Larvae feeding on leaves of the current season have higher growth efficiencies, consumption, and growth compared to larvae feeding on leaves from the preceding year.     

Distribution, Development and Structure of Resin Ducts in Guayule (Parthenium argentatum Gray)

The distribution, development and structure of resin ducts inguayule (Parthenium argentatum Gray), the second best sourceof natural rubber, have been studied. Resin ducts are widelydistributed in stem, root, leaf, petiole and peduncle. The ductsin the primary tissues are initiated schizogenously and theirfurther development is schizolysigenous. The ducts in the cortexof the root do not have a well-defined epithelium. Ducts developedfrom the vascular cambium are initiated and develop schizogenously.Both resin and rubber are produced in the epithelial cells ofresin ducts. While resin is secreted into the duct lumen, rubberis stored within these cells. Epithelial cells store more thanneighbouring parenchyma cells. Guayule, rubber, resin, ducts, epithelial cells     

THE EVOLUTION OF CARBON ALLOCATION TO PLANT SECONDARY METABOLITES: A GENETIC ANALYSIS OF COST IN DIPLACUS AURANTIACUS

Diplacus aurantiacus contains large amounts of a leaf phenolic resin, an important deterrent to a leaf-eating caterpillar, Euphydryas chalcedona. The resin can also retard water loss during drought. Furthermore, the leaf resin content differs among plants and populations. This study investigates the existence of heritable variation (h²) in resin production and tests for a genetic correlation (r_G) between carbon allocation to secondary metabolites and growth rate, as well as with three other vegetative traits. Nine dam and 10 sire plants were chosen randomly at a field site and used to generate 78 full-sib families (19 half-sib families) by crossing all males to all females in a factorial design. Heritability was estimated in two ways, and genetic correlations were estimated by three methods. We found: (1) the heritability of resin production estimated by the regression of offspring on sires was significantly greater than zero ; (2) the maternal variance in resin content was significantly greater than zero (21.3% of total phenotypic variance); (3) significant negative genetic correlation between resin content and growth rate was observed from two of three methods and was consistent with the phenotypic correlation; and (4) the cost of resin could be assessed quantitatively. The genetic cost of 1 mg in resin is equivalent to 25 mg of dry shoot-biomass growth, but the phenotypic cost is only 2.1 mg. This study indicates that carbon allocation to these secondary metabolites may respond to natural selection, and the phenotypic cost of resin production has a genetic basis in D. aurantiacus. This trade-off suggests that once selection occurs, increased phenolic resin production may result in decreased growth, or vice versa.     

MORPHOLOGY,SEASONAL VARIATION,AND FUNCTION OF RESIN GLANDS ON BUDS AND LEAVES OF POPULUS DELTOIDES (SALICACEAE)

When buds form in summer or early fall, modified stipules act as bud scales and their adaxial epidermis secretes a resin that fills the bud. This secretory layer collapses in the dormant bud. Immature leaves, stipules, and leaf primordia occupy the center of the bud; all lack functional resin glands. In spring, stipules of emerging leaves develop an adaxial palisadelike secretory epidermis that becomes more ridged longitudinally in successive stipules. Marginal teeth of the first leaves to emerge are covered with trichomes and lack a secretory epidermis. In successive leaves the teeth become glandular and secrete resin as the lamina unrolls. Later in the season, marginal leaf glands account for much of the resin. Unspecialized hydathodes or extrafloral nectaries occur proximal to each glandular tip. Guttation of water or nectar occurs here through stomata located above a vein ending. On the basis of field observations and a laboratory feeding experiment, the resin seems to function mainly as an insect repellent. It may also reduce water loss from young leaves.     

The effect of solar UV-B radiation on terpenes and biomass production in Grindelia chiloensis (Asteraceae), a woody perennial of Patagonia,Argentina

UV-B radiation is absorbed effectively by nucleic acids and other sensitive targets, potentially causing harmful photochemical effects. Protection against UV-B radiation may be afforded by flavonoids and other phenolics, which absorb strongly in the UV region, but little is known about the role played by other compounds, such as terpenes. Grindelia chiloensis, native of Patagonia (Argentina), can accumulate as much as 25% resin (terpenes) in its leaves. The present investigation was carried out to test the effect of solar UV-B radiation on the allocation of photoassimilates to biomass and terpenes. Exposure to UV-B radiation reduced whole plant biomass, plant height and leaf area, and increased leaf thickness and resin accumulation in Grindelia chiloensis. Higher absorbance was found for refined resin in the UV-B waveband from plants grown under solar UV-B radiation than plants without UV-B radiation. These chemical and structural changes could protect the plant from UV radiation, and may help elucidate the importance of epicuticular resins for a species as G. chiloensis native to an environment with maximum daily integrated values of solar UV-B irradiance.     

Ecological significance of the chemistry of the leaf resin of elytropappus rhinocerotis

Elytropappus rhinocerotis (Asteraceae) is a dominant, fire-adapted species of renosterveld shrub communities in the Cape region of South Africa. The leaves are covered by a thick lipophilic resin comprising up to 20% of their dry weight. Approximately 80% of the leaf resin is composed of phenolic and acidic products with the methoxylated flavones cirsimaritin, hispidulin, eupafolin and the flavonol quercetin the main components. Seven benzoic and cinnamic acid derivatives are also present in minor amounts. About 20% of the resin consisted of aliphatic constituents. Bioassays indicated allelopathic effects of the resin on the seed germination of Lactuca and Raphanus. The predominance of lipophilic flavonoids and phenolics suggests a dual role of repelling herbivores and reducing cuticular transpiration. Calorimetric measurements indicated no significant contribution of the non-volatile resin to the flammability of E. rhinocerotis.     

EVOLUTIONARY IMPLICATIONS OF LEAF RESIN POCKET PATTERNS IN THE TROPICAL TREE HYMENAEA (CAESALPINIOIDEAE: LEGUMINOSAE)

ABSTRACT: Measurement of the size (volume), distribution, and density of resin pockets are compared in margin and center areas of leaves in 11 species (including six varieties of two widespread species) of Hymenaea. Resin composition and yield (mg/g leaf tissue) were analyzed by gas-liquid chromatography. Resin pocket patterns in certain species are highly distinctive; also a relationship seems to exist between patterns of closely related species, with variation having occurred in species or populations of widespread species which have radiated into different kinds of habitats. Thus these patterns may provide useful taxonomic characters, but we suggest that some of the patterns (and thus composition and yield of resin of different portions of the leaf) have evolved under selective pressures of herbivorous insects.