The chromosome-scale genome of Phoebe bournei reveals contrasting fates of terpene synthase (TPS)-a and TPS-b subfamilies.

Terpenoids, including aromatic volatile monoterpenoids and sesquiterpenoids, function in defense against pathogens and herbivores. Phoebe trees are remarkable for their scented wood and decay resistance. Unlike other Lauraceae species investigated to date, Phoebe species predominantly accumulate sesquiterpenoids instead of monoterpenoids. Limited genomic data restrict the elucidation of terpenoid variation and functions. Here, we present a chromosome-scale genome assembly of a Lauraceae tree, Phoebe bournei, and identify 72 full-length terpene synthase (TPS) genes. Genome-level comparison shows pervasive lineage-specific duplication and contraction of TPS subfamilies, which have contributed to the extreme terpenoid variation within Lauraceae species. Although the TPS-a and TPS-b subfamilies were both expanded via tandem duplication in P. bournei, more TPS-a copies were retained and constitutively expressed, whereas more TPS-b copies were lost. The TPS-a genes on chromosome 8 functionally diverged to synthesize eight highly accumulated sesquiterpenes in P. bournei. The essential oil of P. bournei and its main component, ß-caryophyllene, exhibited antifungal activities against the three most widespread canker pathogens of trees. The TPS-a and TPS-b subfamilies have experienced contrasting fates over the evolution of P. bournei. The abundant sesquiterpenoids produced by TPS-a proteins contribute to the excellent pathogen resistance of P. bournei trees. Overall, this study sheds light on the evolution and adaptation of terpenoids in Lauraceae and provides valuable resources for boosting plant immunity against pathogens in various trees and crops.

Kaempferol-3-O-α-(3,4-di-E-p-coumaroyl)-rhamnopyranoside from Nectandra oppositifolia releases Ca2+ from intracellular pools of Trypanosoma cruzi affecting the bioenergetics system.

Phytochemical analysis of EtOH extract from leaves of Nectandra oppositifolia afforded three flavonoids: kaempferol (1), kaempferol-3-O-α-rhamnopyranoside (2) and kaempferol-3-O-α-(3,4-di-E-p-coumaroyl)-rhamnopyranoside (3), which were characterized by NMR and ESI-HRMS. When tested against the protozoan parasite Trypanosoma cruzi, the etiologic agent of Chagas disease, flavonoids 1 and 3 were effective to kill the trypomastigotes with IC50 values of 32.0 and 6.7 µM, respectively, while flavonoid 2 was inactive. Isolated flavonoids 1-3 were also tested in mammalian fibroblasts and showed CC50 values of 24.8, 48.7 and 153.1 µM, respectively. Chemically, these results suggested that the free aglycone plays an important role in the bioactivity while the presence of p-coumaroyl unities linked in the rhamnoside unity is important to enhance the antitrypanosomal activity and reduce the mammalian cytotoxicity. The mechanism of cellular death was investigated for the most potent flavonoid 3 in the trypomastigotes using fluorescent and luminescent-based assays. It indicated that this compound induced neither permeabilization of the plasma membrane nor depolarization of the membrane electric potential. However, early time incubation (20 min) with flavonoid 3 resulted in a constant elevation of the Ca2+ levels inside the parasite. This effect was followed by a mitochondrial imbalance, leading to a hyperpolarization and depolarization of the mitochondrial membrane potential, with reduction of the ATP levels. During this time, the levels of reactive oxygen species levels (ROS) were unaltered. The leakage of Ca2+ from the intracellular pools can affect the bioenergetics system of T. cruzi, leading to the parasite death. Therefore, flavonoid 3 can be a useful tool for future studies against T. cruzi parasites.

Interaction of isolinderanolide E obtained from Nectandra oppositifolia with biomembrane models.

A long-tail lactone, named isolinderanolide E, was obtained from Nectandra oppositifolia and incorporated in Langmuir monolayers of dipalmitoyl-phosphoethanolamine (DPPE) as a model of microbial membranes. The compound was dissolved in chloroform and mixed with DPPE to provide mixed solutions spread on the air-water interface. After solvent evaporation, mixed monolayers were formed, and surface pressure-area isotherms, dilatational rheology, Brewster angle microscopy (BAM), and infrared spectroscopy were employed to characterize the prodrug-membrane interactions. Isolinderanolide E expanded DPPE monolayers, denoting repulsive interactions. At 30 mN/m, the monolayer presented higher viscoelastic and in-plane elasticity parameters and an increased ratio of all-trans/gauche conformers of the alkyl chains, confirming molecular order. Morphology of the monolayer was analyzed by BAM, which revealed a more homogeneous distribution of Isolinderanolide E along the DPPE monolayer than the prodrug directly spread at the interface, which tends to aggregate. A molecular model proposing the molecular orientation of the amphiphilic drug is presented and explained by the distortion of the alkyl chains as well as by viscoelastic changes. In conclusion, the prodrug changes the thermodynamic, rheological, morphological, and structural properties of the DPPE monolayer, which may be essential to understand, at the molecular level, the action of bioactives in selected membrane models.

Alkaloids of the Lauraceae.

This chapter presents an overview of the chemistry and pharmacology of the alkaloids found in species of the Lauraceae family. The occurrence of alkaloids from Lauraceae species as well as their chemical structures is summarized in informative and easy-to-understand tables. Within the Lauraceae family, the genera Ocotea (195), Litsea (180), Cryptocarya (133), and Neolitsea (110) have led to the greater number of publications regarding alkaloids content. Valuable and comprehensive information about the structure of these alkaloids is provided. The alkaloids of the aporphine type, found in 22 of the 23 genera, represent the predominant group in this family. Many of the isolated alkaloids exhibit unique structures. From plants of this family, 22 different types of skeletons have been isolated, among them only the purine alkaloids are classified as pseudoalkaloids, and the types phenethylamines, phenethylcinnamides, and phthalidoisoquinoline are classified as protoalkaloids. The chapter is presented as a contribution for the scientific community, mainly to enable the search for alkaloids in species belonging to the Lauraceae family.

Biosynthetic investigation of γ-lactones in Sextonia rubra wood using in situ TOF-SIMS MS/MS imaging to localize and characterize biosynthetic intermediates.

Molecular analysis by parallel tandem mass spectrometry (MS/MS) imaging contributes to the in situ characterization of biosynthetic intermediates which is crucial for deciphering the metabolic pathways in living organisms. We report the first use of TOF-SIMS MS/MS imaging for the cellular localization and characterization of biosynthetic intermediates of bioactive γ-lactones rubrynolide and rubrenolide in the Amazonian tree Sextonia rubra (Lauraceae). Five γ-lactones, including previously reported rubrynolide and rubrenolide, were isolated using a conventional approach and their structural characterization and localization at a lateral resolution of ~400 nm was later achieved using TOF-SIMS MS/MS imaging analysis. 2D/3D MS imaging at subcellular level reveals that putative biosynthetic γ-lactones intermediates are localized in the same cell types (ray parenchyma cells and oil cells) as rubrynolide and rubrenolide. Consequently, a revised metabolic pathway of rubrynolide was proposed, which involves the reaction between 2-hydroxysuccinic acid and 3-oxotetradecanoic acid, contrary to previous studies suggesting a single polyketide precursor. Our results provide insights into plant metabolite production in wood tissues and, overall, demonstrate that combining high spatial resolution TOF-SIMS imaging and MS/MS structural characterization offers new opportunities for studying molecular and cellular biochemistry in plants.

Transcriptome analysis reveals a composite molecular map linked to unique seed oil profile of Neocinnamomum caudatum (Nees) Merr.

BACKGROUND: Neocinnamomum caudatum (Nees) Merr., a biodiesel tree species in the subtropical areas of South China, India and Burma, is distinctive from other species in Lauraceae family and its seed oil is rich in linoleic acid (18:2) and stearic acid (18:0). However, there is little genetic information about this species so far. In this study, a transcriptomic analysis on developing seeds of N. caudatum was conducted in an attempt to discern the molecular mechanisms involving the control of the fatty acid (FA) and triacylglycerol (TAG) biosynthesis. RESULTS: Transcriptome analysis revealed 239,703 unigenes with an average length of 436 bp and 137 putative biomarkers that are related to FA formation and TAG biosynthesis. The expression patterns of genes encoding ß-ketoacyl-acyl carrier protein synthase I (KASI), ß- ketoacyl-acyl carrier protein synthase II (KASII), stearoyl-ACP desaturase (SAD), fatty acid desaturase 2 (FAD2), fatty acid desaturase 8 (FAD8) and acyl-ACP thioesterase A/B (FATA/B) were further validated by qRT-PCR. These genes displayed a very similar expression pattern in two distinct assays. Moreover, sequence analysis of different FATBs from diverse plant species revealed that NcFATB is structurally different from its counterpart in other species in producing medium-chain saturated FAs. Concertedly, heterologous expression of NcFATB in E. coli BL21 (DE3) strain showed that this corresponding expressed protein, NcFATB, prefers long-chain saturated fatty acyl-ACP over medium-chain fatty acyl-ACP as substrate. CONCLUSIONS: Transcriptome analysis of developing N. caudatum seeds revealed a composite molecular map linked to the FA formation and oil biosynthesis in this biodiesel tree species. The substrate preference of NcFATB for long-chain saturated FAs is likely to contribute to its unique seed oil profile rich in stearic acid. Our findings demonstrate that in the tree species of Lauraceae family, the FATB enzymes producing long-chain FAs are structurally distinct from those producing medium-chain FAs, thereby suggesting that the FATB genes may serve as a biomarker for the classification of tree species of Lauraceae family.

Gelsemium poisoning mediated by the non-toxic plant Cassytha filiformis parasitizing Gelsemium elegans.

INTRODUCTION: Gelsemium poisoning is caused by consumption of the deadly Gelsemium species such as Gelsemium elegans, leading to significant gastrointestinal, neurological and cardio-respiratory toxicities. In 2011 (Cluster 1) and 2012 (Cluster 2), the authors encountered two clusters of gelsemium poisoning after consumption of the non-toxic parasitic plant Cassytha filiformis. The current study aims to examine the mechanism of gelsemium poisoning mediated by a benign parasitic plant. METHODS: Qualitative analysis of toxic gelsemium alkaloids using liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed on the herbal and urine samples from both clusters to confirm exposure. Morphological examination, qualitative analysis of aporphine alkaloids using liquid chromatography-ion trap-time of flight mass spectrometry (LC-IT-TOF/MS) and Sanger sequencing were performed on the plant sample from Cluster 2 to confirm its identity. A field study was conducted in local countryside and C. filiformis was collected for histological, LC-MS/MS and LC-IT-TOF/MS analyses to study its interaction with G. elegans. RESULTS: Gelsemium alkaloids that are not naturally present in C. filiformis were detected in the patients' herbal and urine samples. Misidentification and contamination with G. elegans during the preparation process were excluded by morphological examination of the plant sample from Cluster 2. Its identity as C. filiformis was verified with LC-IT-TOF/MS and molecular analyses. Histological, LC-MS/MS and LC-IT-TOF/MS analyses of C. filiformis collected during the field study confirmed that its haustoria penetrated the vascular bundles of G. elegans and absorbed its gelsemium toxins. CONCLUSIONS: The non-toxic plant C. filiformis absorbed toxic gelsemium alkaloids from its host, G. elegans, and led to gelsemium poisoning in our patients. Our study provides new insights into the toxicology of such plants. Benign parasitic plants may lead to potentially life-threatening poisoning if it parasitizes toxic hosts and absorbs their phytotoxins. The public awareness of risks associated with the use of these medicinal parasitic plants should be raised.

In depth investigation of the metabolism of Nectandra megapotamica chemotypes.

Plants produce a wide range of secondary metabolites. Within a single species, chemotypes can be distinguished by the differences in the composition of the secondary metabolites. Herein, we evaluated Nectandra megapotamica (Spreng.) chemotypes and the balance of different classes of metabolites to verify how significant differences in plant metabolism are regarding chemotypes. We collected N. megapotamica leaves from eight adult plants in two Brazilian states. The essential oils and ethanol/water extracts were analyzed by GC-MS and LC-DAD-MS, respectively. Histochemical tests were performed, as well as chemical analyses of leaves from adaxial and abaxial foliar surfaces of N. megapotamica, and the stereochemistry of α-bisabolol was determined. Two different chemotypes, based on volatile compounds, were identified, distinguished by the presence of isospathulenol, α-bisabolol, ß-bisabolene, and (E)-nerolidol for chemotype A, and bicyclogermacrene and elemicin for chemotype B. A stereochemical analysis of chemotype A extract revealed (+)-α-bisabolol enantiomer. Histochemical tests of chemotypes showed similar results and suggested the presence of essential oil in idioblasts stained with the dye NADI. The analyses of chemotype A leaves by GC-MS revealed similar compositions for abaxial and adaxial surfaces, such pattern was also observed for chemotype B. Medium and high polarity metabolites showed high chemical similarities between the chemotypes, highlighting the presence of proanthocyanidins and glycosylated flavonoids (O- and C-glycosides). Thus, N. megapotamica produced distinct volatile chemotypes with highly conserved medium to high polarity compounds. Such results suggest that phenolic derivatives have a basal physiological function, while genetic or environmental differences lead to differentiation in volatile profiles of N. megapotamica.

The floral transcriptome of Machilus yunnanensis, a tree in the magnoliid family Lauraceae.

Machilus yunnanensis Lecomte is an important Chinese timber species in the family Lauraceae, subclass Magnoliidae. To understand the genetic and molecular basis of flowering in M. yunnanensis, we applied RNA-seq for transcriptome analysis of young and mature flowers and compared these with the transcriptome of leaves. From approximately 26 Gb Illumina clean reads, we obtained a total of 85 823 high-quality transcripts using Trinity software. Comparisons of gene expression in young and mature flowers showed that 37 877 genes were differentially expressed. Among those genes, 121 and 111 genes are implicated in the flowering network and in plant hormone signaling transduction, respectively. Moreover, 141 homologs of these genes displayed similar expression patterns during flower development in M. yunnanensis and Arabidopsis thaliana. Using cluster analysis, we further compared the gene expression profiles in the different pathways that control flowering and phytohormone signaling, and found similar gene expression patterns in the temperature, gibberellin, brassinosteroid, and auxin mediated flowering pathways. Our study provides a clear gene network for flowering and related plant hormone regulating crosstalk in M. yunnanensis and shows that the temporal regulatory programs in the development of flower organs of Machilus are similar to those in the very different flowers of Arabidopsis.

Tandem Mass Spectrometry Imaging and in Situ Characterization of Bioactive Wood Metabolites in Amazonian Tree Species Sextonia rubra.

Driven by a necessity for confident molecular identification at high spatial resolution, a new time-of-flight secondary ion mass spectrometry (TOF-SIMS) tandem mass spectrometry (tandem MS) imaging instrument has been recently developed. In this paper, the superior MS/MS spectrometry and imaging capability of this new tool is shown for natural product study. For the first time, via in situ analysis of the bioactive metabolites rubrynolide and rubrenolide in Amazonian tree species Sextonia rubra (Lauraceae), we were able both to analyze and to image by tandem MS the molecular products of natural biosynthesis. Despite the low abundance of the metabolites in the wood sample(s), efficient MS/MS analysis of these γ-lactone compounds was achieved, providing high confidence in the identification and localization. In addition, tandem MS imaging minimized the mass interferences and revealed specific localization of these metabolites primarily in the ray parenchyma cells but also in certain oil cells and, further, revealed the presence of previously unidentified γ-lactone, paving the way for future studies in biosynthesis.

Antinociceptive activity of Riparin II from Aniba riparia: Further elucidation of the possible mechanisms.

Riparin II (RipII) has an anti-inflammatory activity potentially due its ability to decrease TNF-α and IL-1ß production and its histamine antagonism. The objective of this study was to evaluate the role of RipII in the pain process and the possible antinociceptive mechanisms involved, using classic models of nociception. Male Swiss mice were used in the assays. Determinate the acute toxicity according to the OECD 425 test guideline. The models used were the acetic acid-, formalin-, hot plate and glutamate-induced nociception. For evaluation of antinociceptive effect, the involvement of TRPV1, TRPA1, TRPM8, ASICS, Bradykinin, PKC and PKA were performed using the paw licking using agonists. The acute toxicity study did not detect any clinical signs or changes in behavior or mortality. RipII, administered orally (25 and 50 mg/kg) caused a reduction of nociception induced by acetic acid, formalin (on the second phase) and glutamate. In the investigation of antinociceptive mechanism, we used capsaicin (2.2 µg/paw), cinnamaldehyde (10 nmol/paw), menthol (1.2 µmol/paw), ASICS (2% acetic acid, pH 1.98) and bradykinin (10 µg/paw). The results showed that TRPV1, TRPA1, TRPM8, ASICS and bradykinin play a role in the antinociceptive effect of RipII. The results also showed that PKA is involved too. These data demonstrate that RipII has a low or not toxicity and produced an important antinociceptive effect through mechanisms that probably involve an interaction, at least in part, TRPV1, TRPA1, TRPM8, ASICS, bradykinin and PKA participate in the RipII's antinociceptive effect.

Photodamage attenuating potential of Nectandra hihua against UVB-induced oxidative stress in L929 fibroblasts.

The total phenolic content (TP) and antioxidant activity of the ethanolic extract and fractions that were obtained from the leaves of Nectandra hihua were assessed using different methods. The ethanolic extract (EE) and ethyl acetate fraction (EAF) had the best antioxidant capacity, which was comparable to butylated hydroxytoluene and quercetin (ABTS+ 2.55 ±â€¯0.06, 3.54 ±â€¯0.03 mmol TE/g; DPPH IC50 10.27 ±â€¯0.05, 9.88 ±â€¯0.02 µg/mL; FRAP 2.17 ±â€¯0.08, 2.38 ±â€¯0.04 mmol TE/g; ORAC 5.16 ±â€¯0.08, 5.35 ±â€¯0.07 mmol TE/g; TP 568.05 ±â€¯18.15, 397.20 ±â€¯17.88 mg GAE/g, respectively). The cytoprotective effect, reactive oxygen species (ROS) and lipid peroxidation inhibitions on L929 fibroblasts irradiated with UVB (600 mJ/cm2) in pre- and post-treatments with EE and EAF were determined. These plant materials demonstrated high ROS scavenging activity and lipid peroxidation inhibition on L929 fibroblasts in both treatments, especially with pre-treatment (EE 38.47 ±â€¯1.95% and EAF 40.20 ±â€¯4.5% inhibition of ROS production, and EE 39.03 ±â€¯3.33% and EAF 41.67 ±â€¯7.6% of lipid peroxidation inhibition), indicating the best cytoprotection with pre-treatment (13.52 ±â€¯1.66% and 13.34 ±â€¯2.61% increases in cell viability). The antioxidant flavonoids quercitrin, avicularin, juglalin, afzelin and astragalin were isolated from EAF. The results obtained indicate that EE and EAF present photodamage attenuating potential against UVB-induced oxidative stress and can be useful as a starting point for developing dermatological products to prevent oxidative skin damage.

Effects of elevated O3 on physiological and biochemical responses in three kinds of trees native to subtropical forest in China during non-growing period.

Numerous studies have documented the negative effects of ozone (O3) on tree species in growing season, however, little is done in non-growing season. Three evergreen tree species, Phoebe bournei (Hemsl.) Yang (P. bournei), Machilus pauhoi Kanehira (M. pauhoi) and Taxus chinensis (Pilger) Rehd (T. chinensis), were exposed to non-filtered air, 100 nmol mol-1 O3 air (E1) and 150 nmol mol-1 O3 air (E2) in open-top chambers in subtropical China. In the entire period of experiment, O3 fumigation decreased net photosynthesis rate (Pn) through stomatal limitation during the transition period from growing to non-growing season (TGN), and through non-stomatal limitation during the period of non-growing season (NGS) in all species tested. Meanwhile, O3 fumigation reduced and delayed the resilience of Pn in all species tested during the transition period from non-growing to growing season (TNG). O3 fumigation significantly decreased chlorophyll contents during NGS, whereas no obvious injury symptoms were observed till the end of experiment. O3 fumigation induced increases in levels of malondialdehyde, superoxide dismutase, total phenolics and reduced ascorbic acid, and changes in four plant endogenous hormones as well in all species tested during NGS. During NGS, E1 and E2 reduced Pn by an average of 80.11% in P. bournei, 94.56% in M. pauhoi and 12.57% in T. chinensis, indicating that the O3 sensitivity was in an order of M. pauhoi > P. bournei > T. chinensis. Overall, O3 fumigation inhibited carbon fixation in all species tested during NGS. Furthermore, O3-induced physiological activities also consumed the dry matter. All these suggested that elevated O3, which is likely to come true during NGS in the future, will adversely affect the accumulation of dry matter and the resilience of Pn during TNG in evergreen tree species, and further inhibit their growth and development in the upcoming growing season.

The Fungus Raffaelea lauricola Modifies Behavior of Its Symbiont and Vector, the Redbay Ambrosia Beetle (Xyleborus Glabratus), by Altering Host Plant Volatile Production.

The redbay ambrosia beetle Xyleborus glabratus is the vector of the symbiotic fungus, Raffaelea lauricola that causes laurel wilt, a highly lethal disease to members of the Lauraceae family. Pioneer X. glabratus beetles infect live trees with R. lauricola, and only when tree health starts declining more X. glabratus are attracted to the infected tree. Until now this sequence of events was not well understood. In this study, we investigated the temporal patterns of host volatiles and phytohormone production and vector attraction in relation to laurel wilt symptomology. Following inoculations with R. lauricola, volatile collections and behavioral tests were performed at different time points. Three days after infection (DAI), we found significant repellency of X. glabratus by leaf odors of infected swamp bay Persea palustris as compared with controls. However, at 10 and 20 DAI, X. glabratus were more attracted to leaf odors from infected than non-infected host plants. GC-MS analysis revealed an increase in methyl salicylate (MeSA) 3 DAI, whereas an increase of sesquiterpenes and leaf aldehydes was observed 10 and 20 DAI in leaf volatiles. MeSA was the only behaviorally active repellent of X. glabratus in laboratory bioassays. In contrast, X. glabratus did not prefer infected wood over healthy wood, and there was no associated significant difference in their volatile profiles. Analyses of phytohormone profiles revealed an initial increase in the amount of salicylic acid (SA) in leaf tissues following fungal infection, suggesting that the SA pathway was activated by R. lauricola infection, and this activation caused increased release of MeSA. Overall, our findings provide a better understanding of X. glabratus ecology and underline chemical interactions with its symbiotic fungus. Our work also demonstrates how the laurel wilt pathosystem alters host defenses to impact vector behavior and suggests manipulation of host odor by the fungus that attract more vectors.

The Genus Neolitsea of Lauraceae: A Phytochemical and Biological Progress.

As one of the biggest genera in Lauraceae, Neolitsea owns a great application potential in industrial and traditional medical fields. Modern phytochemical and biological investigations revealed its structural diversity and diverse activities. This review summarizes the most recent progress on this regards, hopefully to provide a reference for full use and development of the Neolitsea resource.

Biological evaluation of secondary metabolites from the root of Machilus obovatifolia.

Bioassay-guided fractionation of the root of Machilus obovatifolia led to the isolation of four new lignans, epihenricine B (1), threo-(7'R,8'R) and threo-(7'S,8'S)-methylmachilusol D (2 and 3), and isofragransol A (4), along with 23 known compounds. The compounds were obtained as isomeric mixtures (i.e., 2/3 and 4/20, resp.). The structures were elucidated by spectral analyses. Among the isolates, 1, licarin A (12), guaiacin (14), (±)-syringaresinol (21), and (-)-epicatechin (23) showed ABTS (=2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) cation radical-scavenging activity, with SC50 values of 11.7±0.5, 12.3±1.1, 11.0±0.1, 10.6±0.3, and 9.5±0.2 µM in 20 min, respectively. In addition, kachirachirol B (17) showed cytotoxicity against the NCI-H460 cell line with an IC50 value of 3.1 µg/ml.

Responses of the photosynthetic apparatus to winter conditions in broadleaved evergreen trees growing in warm temperate regions of Japan.

Photosynthetic characteristics of two broadleaved evergreen trees, Quercus myrsinaefolia and Machilus thunbergii, were compared in autumn and winter. The irradiance was similar in both seasons, but the air temperature was lower in winter. Under the winter conditions, net photosynthesis under natural sunlight (Anet) in both species dropped to 4 µmol CO2 m(-2) s(-1), and the quantum yield of photosystem II (PSII) photochemistry in dark-adapted leaves (Fv/Fm) also dropped to 0.60. In both species the maximum carboxylation rates of Rubisco (V(cmax)) decreased, and the amount of Rubisco increased in winter. A decline in chlorophyll (Chl) concentration and an increase in the Chl a/b ratio in winter resulted in a reduction in the size of the light-harvesting antennae. From measurements of Chl a fluorescence parameters, both the relative fraction and the energy flux rates of thermal dissipation through other non-photochemical processes were markedly elevated in winter. The results indicate that the photosynthetic apparatus in broadleaved evergreen species in warm temperate regions responds to winter through regulatory mechanisms involving the downregulation of light-harvesting and photosynthesis coupled with increased photoprotective thermal energy dissipation to minimize photodamage in winter. These mechanisms aid a quick restart of photosynthesis without the development of new leaves in the following spring.

Flavonoids from Machilus japonica stems and their inhibitory effects on LDL oxidation.

Stems of Machilus japonica were extracted with 80% aqueous methanol (MeOH) and the concentrated extract was successively extracted with ethyl acetate (EtOAc), normal butanol (n-BuOH), and water. Six flavonoids were isolated from the EtOAc fraction: (+)-taxifolin, afzelin, (-)-epicatechin, 5,3'-di-O-methyl-(-)-epicatechin, 5,7,3'-tri-O-methyl-(-)-epicatechin, and 5,7-di-O-methyl-3',4'-methylenedioxyflavan-3-ol. The chemical structures were identified using spectroscopic data including NMR, mass spectrometry and infrared spectroscopy. This is the first report of isolation of these six compounds from M. japonica. The compounds were evaluated for their diphenyl picryl hydrazinyl scavenging activity and inhibitory effects on low-density lipoprotein oxidation. Compounds 1 and 3-6 exhibited DPPH antioxidant activity equivalent with that of ascorbic acid, with half maximal inhibitory concentration (IC50) values of 0.16, 0.21, 0.17, 0.15 and 0.07 mM, respectively. The activity of compound 1 was similar to the positive control butylated hydroxytoluene, which had an IC50 value of 1.9 µM, while compounds 3 and 5 showed little activity. Compounds 1, 3, and 5 exhibited LDL antioxidant activity with IC50 values of 2.8, 7.1, and 4.6 µM, respectively.

Eucalyptol is an attractant of the Redbay ambrosia beetle, Xyleborus glabratus.

The redbay ambrosia beetle, Xyleborus glabratus, is an invasive wood-boring beetle that has become established in the southeastern United States. The beetle transmits the causal pathogen of lethal laurel wilt to susceptible host trees, which include redbay, an important forest community species, and avocado, a valuable food crop. By examining odors of redbay wood, we developed an artificial lure that captured X. glabratus in redbay forests. Eucalyptol was a critical component of the blend for beetle attraction, and eucalyptol alone in large quantities attracted X. glabratus. Furthermore, eucalyptol stimulated boring by X. glabratus into paper arenas. The results suggest that eucalyptol contributes to host selection behavior of X. glabratus and may be useful for management of this pathogen vector.

Leaf-derived cecidomyiid galls are sinks in Machilus thunbergii (Lauraceae) leaves.

Three relevant hypotheses - nutrition, environment and the enemies hypothesis - often invoked to explore source and sink relationships between galls and their host plants are still under dispute. In this research, chlorophyll fluorescence, gas exchange capacity, stomatal conductance, total carbon and nitrogen, total soluble sugars and starches, and scanning and transmission electron microscopy of two types of galls were used to investigate source-sink relationships. Compared with host leaves, these galls demonstrated slightly lower chlorophyll fluorescence; however, gas exchange capacity and stomatal conductance were not detected at all. Scanning electron micrographs demonstrated that the abaxial epidermis of host leaves contain normal amounts of stomata, whereas no stomata were observed on the exterior and interior surfaces of both types of galls. In addition, gall inner surfaces were covered with many kinds of fungal hyphae. Gall total carbon (C) and nitrogen (N) levels were lower but the C/N ratio was higher in galls than host leaves. Both types of galls accumulated higher total soluble sugars and starches than host leaves. Transmission electron micrographs also revealed that both types of galls contain plastoglobuli and giant starch granules during gall development. Results strongly indicate that leaf-derived cecidomyiid galls are sinks in Machilus thunbergii leaves. However, it is perplexing how larvae cycle and balance CO(2) and O(2) in gall growth chambers without stomata.