The Plant Growth Inhibitor Nagilactone Does Not Work Directly in a Stabilized Podocarpus nagi Forest

Nagilactones isolated from Podocarpus nagi are known for poisonous physiological activities to organisms. To clarify the allelopathic potential of nagilactone to wild plants, the seasonal changes in nagilactone contents in throughfall, leaf litter, and soil were monitored monthly in a podocarp forest at Mt. Mikasa, Nara City, central Japan. The average annual nagilactone concentrations in throughfall, leaf litter, and surface soil at 0–10 cm depth were 4.7 × 10^–12 g/ml, 3.8 × 10^–4 g/g and 3.1 × 10^–7 g/g, respectively. The nagilactone flux from canopy leaves to the forest floor via throughfall was 5.2 × 10^–2 g/ha/yr, which was far smaller than that via leaf litterfall: 765.3 g/ha/year. The disappearance of nagilactones from leaf litter and the nagilactone accumulation in the Ao layer and mineral soil were also studied. Nagilactone in leaf litter rapidly disappeared and its relative disappearance rate was 5.8/yr. Nagilactone concentration in mineral soil also decreased rapidly with an increase in soil depth and could not be detected in soil at 40–100 cm depth. The total mass of nagilactones was 177.3 g/ha in the Ao layer and 105.0 g/ha in mineral soil. These quantities in nagilactone dynamics give a very rapid turnover time of 0.37 year (4.4 months) in a soil nagilactone pool, implying the possibility of the reabsorption of nagilactone by P. nagi, while the allelopathic effects of nagilactone to other plants in the podocarp forest are questionable.     

Sequestration of host plant-derived compounds by geometrid moth, Milionia basalis, toxic to a predatory stink bug, Eocanthecona furcellata

A predatory stink bug, Eocanthecona furcellata, died after feeding on Milionia basalis larvae. The compounds toxic to E. furcellata were isolated from the hemolymph of M. basalis larvae and identified as inumakilactone A, nagilactone C, and nagilactone C glucoside. The concentrations of inumakilactone A, nagilactone C, and nagilactone C glucoside in the hemolymph of the final instar larvae were 130, 50, and 770 g/ml, respectively. Nagilactone C showed the highest insecticidal activity against second-instar nymphs of E. furcellata, while nagilactone C glucoside showed the lowest, one twentieth of that of nagilactone C. When mixed compounds were given at the same concentrations as those in hemolymph of M. basalis, all nymphs of E. furcellata died with in three days. Inumakilactone A and nagilactone C were found to be in the leaves of podocarp, Podocarpus macrophyllus, the only host plant of M. basalis, at concentrations of 13 and 175 g/g fresh weight, respectively. However, no nagilactone C glucoside was detected in the leaves of this species. These results suggested that M. basalis may transform nagilactone C to its glucoside.     

Do defoliation and subsequent phytochemical responses reduce future herbivory on oak trees?

Perennial plants are thought to respond to partial or complete defoliation by producing new foliage that is less susceptible to herbivores because of induction of allelochemicals. Here, I tested this hypothesis by manually removing primary foliage from branches ofQuercus emoryi (Fagaceae) at two different times in the season and monitoring changes in protein and tannin levels and the amount of herbivory relative to control branches. New, secondary leaves had 2.5 × greater hydrolyzable tannin content than mature foliage of control branches. Condensed tannins, which constitute a relatively low fraction of leaf mass, were lower, while protein content was temporarily greater, in new secondary leaves relative to mature leaves. Despite large increases in hydrolyzable tannins, herbivory levels were greater on refoliated branches than on control branches. New foliage is susceptible to herbivory regardless of when it is produced in the season, possibly because lower toughness and higher water content override any induced or developmentally related changes in allelochemistry. My results do not support the hypothesis that postherbivore changes in phytochemistry protect perennial plants from future herbivory, at least within a growing season.     

Persistence of nonprotein seed amino acid s-(β-carboxyethyl)-cysteine in young leaves ofCalliandra rubescens Ecological implications

The insecticidal nonprotein amino acid S-(-carboxyethyl)-cysteine (S-CEC) is the major free amino acid in the seeds of severalCalliandra spp. where it accounts for up to 2.9 % of dry weight. Lesser amounts of other related S-containing amino acids and an array of nonprotein imino acids derived from pipecolic acid are other constituents. High concentrations of imino acids, which also show insecticidal activity, are maintained in the mature leaves, but sulfur compounds are lacking. In this study the disappearance of S-CEC from the germinating seeds and young seedlings ofC. rubescens was monitored over time. After 10 weeks, S-CEC continues to be found in high concentrations in the stems and new leaves. As young leaves mature, sulfur compounds quickly decrease in concentration. Traces of S-CEC are found in new leaves of plants up to nine months after germination. Whether high concentration of S-CEC in young leaves is due to transportation from the seed or de novo synthesis is unclear. The ecological implications are discussed.     

Multichemical resistance of the coniferPodocarpus gracilior (Podocarpaceae) to insect attack

Podocarpus gracilior is resistant in nature to insect attack. Apparently, the resistance ofP. gracilior is due to a multichemical defense mechanism. Chemicals identified as potential components of the multichemical defense are four norditerpenedilactones, including nagilactones, C, D, and F, which cause insect feeding deterrent activity ultimately coupled to an insecticidal activity, and podolide, an insecticide; two nonlethal growth-inhibiting biflavones, podocarpusflavone A and 7,4-dimethylamentoflavone; and the ecdysis-inhibiting phytoecdysone, ponasterone A.     

Neonate Silkworm (<Emphasis Type="Italic">Bombyx mori</Emphasis>) Larvae Are Attracted to Mulberry (<Emphasis Type="Italic">Morus alba</Emphasis>) Leaves with Conspecific Feeding Damage

The silkworm Bombyx mori is a molecular genetic model for the Lepidoptera. Its odorant receptor genes have been described, and preliminary studies suggest that several are expressed specifically in the larval caterpillar stage. This study was undertaken to identify olfactory behaviors specific to the larvae. A two-choice leaf disk bioassay with naive neonate larvae was used to evaluate the attractiveness of three types of mulberry leaf (Morus alba): newly flushed leaves from branch tips, mature leaves, and mature leaves with feeding damage caused by conspecific larvae. Mature leaves with feeding damage were the most attractive, newly flushed leaves were moderately favored, and undamaged mature leaves were the least attractive. Volatile odors collected from whole mulberry leaves by using solid-phase microextraction fibers were analyzed by gas chromatography-mass spectrometry. The volatile profile of newly flushed leaves and mature leaves damaged by conspecific larvae was more complex compared to undamaged mature leaves. By comparing the volatile makeup of each leaf type, a list of 22 candidate odors responsible for attracting the neonate larvae was generated; α-farnesene was particularly notable as a herbivore-induced volatile. These odors will be used in future in vitro studies to determine whether they activate larval-specific odorant receptors. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Interactive Influence of Leaf Age,Light Intensity,and Girdling on Green Ash Foliar Chemistry and Emerald Ash Borer Development

Biotic and abiotic environmental factors affect plant nutritional quality and defensive compounds that confer plant resistance to herbivory. Influence of leaf age, light availability, and girdling on foliar nutrition and defense of green ash (Fraxinus pennsylvanica Marsh) was examined in this study. Longevity of the emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), adults reared on green ash foliage subjected to these factors was assayed. Mature leaves generally were more nutritious with greater amino acids and a greater ratio of protein to non-structural carbohydrate (P:C) than young leaves, in particular when trees were grown in shade. On the other hand, mature leaves had lower amounts of trypsin and chymotrypsin inhibitors, and total phenolics compared to young leaves. Lower defense of mature leaves alone, or along with higher nutritional quality may lead to increased survival and longevity of emerald ash borer feeding on mature leaves. Sunlight reduced amino acids and P:C ratio, irrespective of leaf age and girdling, and elevated total protein of young foliage, but not protein of mature leaves. Sunlight also dramatically increased all investigated defensive compounds of young, but not mature leaves. Girdling reduced green ash foliar nutrition, especially, of young leaves grown in shade and of mature leaves grown in sun. However emerald ash borer performance did not differ when fed leaves from trees grown in sun or shade, or from girdled or control trees. One explanation is that emerald ash borer reared on lower nutritional quality food may compensate for nutrient deficiency by increasing its consumption rate. The strong interactions among leaf age, light intensity, and girdling on nutrition and defense highlight the need for caution when interpreting data without considering possible interactions.     

Electrophysiological Responses of Female and Male Hypsipyla grandella (Zeller) to Swietenia macrophylla Essential Oils

The mahogany shoot borer, Hypsipyla grandella, is one of the most economically important Neotropical forest pest insects because it attacks valuable timber of the Meliaceae. Despite detailed information on the life cycle of H. grandella, factors influencing attack by the shoot borer are not well known. Electroantennogram studies of borer antennae indicate that both female and male moths elicited significant antennal responses to essential oil of the terminal shoots, mature and senescent leaves, in three concentrations. -Himachalene, germacrene D, germacrene A, cadina-1,4-diene, hexadecanoic acid, and ethyl hexadecanoate appeared in all essential oils from Swietenia macrophylla. Germacrene-D and -himachalene were also present in the active essential oil of Cedrela odorata and Toona ciliata. These findings suggest that these compounds play a role in attracting H. grandella to oviposit on the leaves of these plants. The GC-EAD of essential oil from S. macrophylla mature leaves to female H. grandella antennae led to the identification of -caryophyllene as the main constituent responsible for the antennae response.     

Depletion of Host-Derived Cyanide in the Gut of the Eastern Tent Caterpillar, Malacosoma americanum

Using a colorimetric procedure, we assessed the HCN-p of black cherry leaves (Prunus serotina) ingested by the eastern tent caterpillar, Malacosoma americanum, and the cyanide content of the bolus as it passed thorough the caterpillar's digestive tract and into the detritus pool. The mean HCN-p of leaves in our study area was 1902 ± 174 (SE) ppm. Young leaves found at the tips of growing branches, which the caterpillars preferred, had a significantly higher HCN-p (3032 ± 258 ppm) than older leaves found at the middle (1542 ± 243 ppm) or base of the shoot (1131 ± 159 ppm). Following a bout of overnight feeding on young leaves, the cyanide content of the foregut and midgut boluses of early sixth-instar caterpillars averaged 631 ± 161 ppm, and 14 ± 3 ppm, respectively, indicating that host-derived cyanide is rapidly depleted as the bolus transits the gut. Some cyanide, however, remains. In three studies, the mean cyanide content of fresh fecal pellets ranged from approximately 20 to 38 ppm, while the dried, compacted pellets ranged from 63 to 85 ppm. Food in the foreguts of mature caterpillars dispersing over the ground in search of pupation sites had 417 ± 99 ppm cyanide. The potential impact of this egested and caterpillar-transported cyanide on the consumer and detritivore communities is discussed.     

Systemic induction of feeding deterrents in cotton plants by feeding ofSpodoptera SPP. Larvae

Cotton,Gossypium hirsutum L., has been shown to exhibit systemic induced resistance to arthropods under certain conditions. We conducted experiments to determine the effects of previous feeding ofSpodoptera exigua Hübner andSpodoptera littoralis (Boisd.) larvae on feeding behavior, growth, and survival of larvae subsequently feeding on cotton. In one feeding choice test,S. exigua larvae preferred young leaves from undamaged control plants to undamaged young leaves from a previously damaged plant. Feeding deterrence was noticeable after only 6 hr of initial feeding damage by larvae, and there was almost complete deterrence after 30 and 54 hr of continuous feeding. In a second feeding choice test,S. littoralis larvae fed more on mature leaves from undamaged control plants than on undamaged mature leaves from previously damaged plants. In no-choice tests, third instars ofS. littoralis fed undamaged young leaves from damaged plants did not gain weight and died by the seventh day, whereas larvae fed young leaves from undamaged control plants gained weight and pupated within 11 days. Sixth instars ofS. littoralis fed either old damaged leaves, old undamaged leaves, or young undamaged leaves all from previously damaged plants gained weight slowly and took more than 12 days to pupate, whereas larvae fed young leaves from undamaged plants gained weight rapidly and pupated within five days of the beginning of the experiment.     

Protein-precipitating capacity of tannins inShorea (Dipterocarpaceae) seedling leaves

The protein-precipitating capacities of tanniferous extracts from immature and mature leaves of threeShorea spp. (Dipterocarpaceae) seedlings were measured by an adaptation of Goldstein and Swain's -glucosidase precipitation assay. Protein precipitation by the extracts was not correlated with total phenolics (Folin-Denis assay) or proanthocyanidin content (BuOH-HCl assay) as measured in an earlier study. Extracts ofS. maxwelliana mature leaves had much lower protein-precipitating capacity than those ofS. acuminata andS. leprosula, but fewer insect species feed on and cause less damage to the foliage ofS. maxwelliana compared with the other species' foliage. Immature leaf extracts ofS. leprosula andS. acuminata had substantial protein-precipitating capacities which in the latter species exceeded that of its mature leaf extracts. Leaf extracts precipitated less protein when initial protein concentration was reduced, although not limiting, but no effect or the reverse effect occurred with quebracho tannin and tannic acid. Problems in the characterization of foliage astringency and the interpretation of its role as a potential antiherbivore defense are discussed.     

Influence of Light and Photosynthesis on Alkaloid Concentration in Larkspur

Concentrations of toxic norditerpenoid alkaloids vary greatly in tall larkspur (Delphinium barbeyi) and may be influenced by environmental stress. We evaluated the effect of shade, darkness, and inhibition of photosynthesis on toxic alkaloid concentration. In plants treated with metribuzin to inhibit photosynthesis, alkaloid concentration increased, but dry weight of the plants decreased as growth ceased, leaving absolute alkaloid content similar to that of control plants. Short-term shade (70% reduction in sunlight for three days), dark treatments from leaves collected at night, and aluminum foil covered leaves all increased alkaloid concentration in comparison to untreated control plants. It appears that absolute amounts of alkaloids remained the same, but the mass of stressed plants declined as nonstructural carbohydrates were depleted, thus increasing the relative concentration of alkaloids. We conclude that norditerpenoid alkaloids in larkspur do not respond to short-term light stress. Alkaloid concentration was lower in larkspur plants growing beneath forest canopy and in potted plants in a long-term shade study (70% reduction in sun light for 21 days) than plants growing in open sunlight. Long-term shade may have reduced synthesis of norditerpenoid alkaloids, particularly in the earlier developmental stages of the plant. Shade stress or photosynthesis inhibition apparently did not increase norditerpenoid alkaloid synthesis, which contrasts with the carbon/nutrient balance theory of plant defense.     

Volatile Compounds on the Leaf Surface of Intact and Regrowth Tarbush (<Emphasis Type="Italic">Flourensia cernua</Emphasis> DC) Canopies

Shrub expansion into desert grasslands is a serious problem resulting in loss of forage and rangeland productivity. Flourensia cernua DC (tarbush) is one such shrub contributing to the decline of Chihuahuan Desert grasslands. Our previous research has shown tarbush consumption by sheep and goats to be negatively related to leaf surface concentration of individual terpenes and epicuticular wax. Concentrations of compounds such as terpenes often change with plant age and phenology. Our objective was to examine the effect of altering the vegetative state of tarbush on volatile chemicals. Ninety tarbush plants were randomly selected, and all biomass within 10 cm of the soil surface was removed from 45 plants during winter dormancy. Leaves were collected the following summer during active growth from the canopy of intact controls and resprouts. Leaf surface volatiles were analyzed by gas chromatography-mass spectroscopy and subjected to univariate analysis of variance and stepwise discriminate analysis. Of the 87 compounds present on tarbush leaves, 35 were greater in canopy samples and 16 were greater in regrowth samples based on univariate analysis (P < 0.05). Mean concentration of total volatiles on canopy leaves tended to be less (P = 0.062) than that of regrowth (3,642 vs. 4,684 μg/g DM). Nine compounds in the discriminant analysis (α-muurolene, iso-borneol, unknown#6, p-cymen-8-ol, unknown#7, sabinene, β-caryophyllene, δ-cadinene, and α-copaene) explained 95% of the variation between canopy and regrowth samples. Lower cumulative concentration of volatile compounds in canopy than regrowth samples suggests repsrouts may be less vulnerable to herbivory than intact tarbush.     

Allelochemic control of biomass allocation in interacting shrub species

Aqueous leachates derived from canopy phyllodes of invasiveAcacia cyclops affected growth of a range of shrub species independently of nutrient input effects. All plants showed a sublethal phytotoxic response. Root mass was generally less adversely affected than shoot mass and, while decreasing significantly in response to the 10% concentration, showed no such response to the 1% solution. Root-shoot biomass ratios increased, except inEuphorbia burmannii, which may recognize intrinsic root architecture limitations on extensive exploitation of toxin-free soil. Application of surface plant litter from underA. cyclops canopies stimulated the production of basal stems inProtasparagus capensis andEriocephalus racemosus but was insufficient to significantly reduce root-shoot ratios. Plant growth inhibition was maximized by canopy leachate compounded by surface litter effects inAnthospermum spathulatum. The net effect of leachate at high concentration on biomass allocation in certain shrub species may help explain their patterns of association and disassociation withA. cyclops.     

Psoralens in senescing leaves ofRuta graveolens

Concentrations of three furanocoumarins, psoralen, xanthotoxin, and bergapten, were measured on the surface and within mature whole leaves of two groups ofRuta graveolens L. late autumn plants, 2 and 6 years old, which contained green, yellow, and dry yellow leaves. Upper green leaves contained higher concentrations of these coumarins than lower green leaves, green leaves contained several times as much as yellow leaves, and dry leaves contained even smaller amounts than yellow ones. The dry yellow leaves contained only a very small percentage of furanocoumarins on the surface, suggesting that extrusion to the surface of yellow leaves was slower or had stopped, while loss from the surface continued. The loss of psoralen was the most dramatic in and on the dry leaves. Bergapten's ratio to the other cournarins increased during senescence. Xanthotoxin was always the predominant furanocoumarin in this species.     

Decomposition and nutrient release from pruning residues of two indigenous agroforestry species during the wet and dry seasons

The decomposition of leaves from Cordia africana Lam. and Albizia gummifera G. F.Gmel was investigated during the wet and dry seasons at Wondo Genet (Ethiopia). Litterbags of leaves were buried in soils under farmland and shaded-coffee agroforestry systems. Residual matter was recovered after 4, 8, 12, and 16 weeks and analysed for nitrogen (N), phosphorus (P), potassium (K), cellulose, lignin, soluble polyphenol and condensed tannin content. Mass-loss and release of N, polyphenols and condensed tannins were greater from Albizia leaves than from Cordia leaves, suggesting that a high polyphenol content does not necessarily retard decomposition. The rates of mass loss and release of the majority of leaf constituents were considerably faster during the wet season than during the dry season. Lignin decomposition, however, proceeded more rapidly during the dry season, and no significant seasonal differences were observed for polyphenol decomposition. The decomposition kinetics of most leaf components during the wet season were best described by a single-exponential model, but a quadratic model provided the best fit during the dry season. Initial leaf chemistry and season were important decomposition factors, while land-use effects were negligible. However, land-use effects showed distinct seasonal differences, with leaf litter decomposing more rapidly in soil under shaded-coffee than under farmland management, especially during the wet season. This study also demonstrated that polyphenol content does not show the predictive effects it has been attributed to have and that other constituents, such as condensed tannins, would be better suited for this purpose.     

Comparison of furanocoumarin concentrations of greenhouse-grownRuta chalepensis with outdoor plants later transferred to a greenhouse

Ruta chalepensis contained concentrations of furanocoumarins 25–50% of those found inR. graveolens both in the whole leaf and on its surface. On the leaf surface of plants grown all year indoors in a greenhouse, they increased steadily between November 1 and December 14 on mature upper and lower leaves. New growth upper leaves on December 14 contained less than mature upper leaves. Plants transferred from outdoors to the greenhouse showed decreased concentrations after the first two weeks, followed by recovery both in the whole leaf and on the leaf surface. Proportions of xanthotoxin and bergapten to psoralen changed during the experiment. On the leaf surfaces and in the whole upper leaves of the indoor plants, the proportions were often similar, but in the transferred plants, in most cases, psoralen was less than bergapten or xanthotoxin in the upper leaves and markedly less in the lower leaves. Implications of these findings for possible effects of environmental changes on secondary plant metabolism are discussed.     

Growth Inhibitory Effects of Nutgrass (Cyperus rotundus) on Rice (Oryza sativa) Seedlings

Growth inhibitory effects of aqueous extracts and leachates of leaves and tubers of Cyperus rotundus L. were investigated by using rice (Oryza sativa L.) seedling as a bioassay material. Both the extracts and leachates of Cyperus were inhibitory to the growth of rice seedlings. Growth inhibition was more pronounced in the presence of aqueous extracts than the leachates. The extract and leachate of leaves had higher total phenolic contents than those of the tubers. Soil amendment with fresh leaves of Cyperus reduced plant height, leaf area, and root and shoot weight of rice seedlings. Total phenolic content was higher in soil amended with fresh Cyperus leaves than the unamended control soil. Nineteen compounds were tentatively identified from the aqueous extracts of leaves and tubers by ethyl acetate extraction followed by gas chromatography–mass spectroscopy (GC-MS). Dicarboxylic, phenolic, and fatty acids were the major compounds. Our results suggest that Cyperus may affect the growth and establishment of rice seedlings after sowing or transplanting, especially when Cyperus plants are mixed in soil during land preparation by ploughing in rain-fed rice culture.     

Light-induced variation in phenolic levels in foliage of rain-forest plants

Levels of phenolic secondary metabolites in the leaves of four west African rain-forest plants,Acacia pennata, Cynometra leonensis, Diopyros thomasii, andTrema guineensis, were correlated with incident light intensity at both the inter- and intraindividual level. Enhanced phenolic levels under high light intensity appeared to be due to production of both polyphenolics (condensed and hydrolyzable tannins) and simple phenolics. InTrema guineensis, where it is possible to separate leaves in terms of both their age and the light incident upon them, condensed tannin production progressed differently during the development of sun and shade leaves, suggesting continuing production of new oligomers in the former but not in the latter. The results of this study suggest that the production of phenolics in relation to variation in incident light is a finely tuned process, which must be explained in terms of plant physiology and intermediate metabolism rather than in terms of resource allocation or a direct response to herbivory.     

Effects of rosemary extracts and major constituents on lipid oxidation and soybean lipoxygenase activity

Rosemary (Rosemarinus officinalis L.) leaves were extracted with three different solvents, namely hexane, acetone and methanol. A reverse-phase high-performance liquid chromatography system in combination with a mass detector was used to quantitate the content of carnosol, carnosic acid and ursolic acid in the rosemary extracts. All rosemary extracts showed strong inhibitory effects on lipid oxidation and soybean lipoxygenase activity.