Unbalanced Expression of Structural Genes in Carotenoid Pathway Contributes to the Flower Color Formation of the Osmanthus Cultivar 'Yanzhi Hong'.

Carotenoids are important natural pigments that are responsible for the fruit and flower colors of many plants. The composition and content of carotenoid can greatly influence the color phenotype of plants. However, the regulatory mechanism underling the divergent behaviors of carotenoid accumulation, especially in flower, remains unclear. In this study, a new cultivar Osmanthus fragrans 'Yanzhi Hong' was used to study the regulation of carotenoid pigmentation in flower. Liquid chromatograph-mass spectrometer (LC-MS) analysis showed that ß-carotene, phytoene, lycopene, γ-carotene, and lutein were the top five pigments enriched in the petals of 'Yanzhi Hong'. Through transcriptome analysis, we found that the expression of the structural genes in carotenoid pathway was imbalanced: most of the structural genes responsible for lycopene biosynthesis were highly expressed throughout the flower developmental stages, while those for lycopene metabolism kept at a relatively lower level. The downregulation of LYCE, especially at the late developmental stages, suppressed the conversion from lycopene to α-carotene but promoted the accumulation of ß-carotene, which had great effect on the carotenoid composition of 'Yanzhi Hong'. Ethylene response factor (ERF), WRKY, basic helix-loop-helix (bHLH), v-myb avian myeloblastosis viral oncogene homolog (MYB), N-Acetylcysteine (NAC), auxin response factor (ARF), and other transcription factors (TFs) have participated in the flower color regulation of 'Yanzhi Hong', which formed co-expression networks with the structural genes and functioned in multiple links of the carotenoid pathway. The results suggested that the cyclization of lycopene is a key link in determining flower color. The modification of the related TFs will break the expression balance between the upstream and downstream genes and greatly influence the carotenoid profile in flowers, which can be further used for creating colorful plant germplasms.

Analysis of the Aging-Related AP2/ERF Transcription Factor Gene Family in Osmanthus fragrans.

Ethylene-Responsive Factor (ERF) is a key element found in the middle and lower reaches of the ethylene signal transduction pathway. It is widely distributed in plants and plays important roles in plant growth and development, hormone signal transduction, and various stress processes. Although there is research on AP/ERF family members, research on AP2/ERF in Osmanthus fragrans is lacking. Thus, in this work, AP2/ERF in O. fragrans was extensively and comprehensively analyzed. A total of 298 genes encoding OfAP2/ERF proteins with complete AP2/ERF domains were identified. Based on the number of AP2/ERF domains and the similarity among amino acid sequences between AP2/ERF proteins from A. thaliana and O. fragrans, the 298 putative OfAP2/ERF proteins were divided into four different families, including AP2 (45), ERF (247), RAV (5), and SOLOIST (1). In addition, the exon-intron structure characteristics of these putative OfAP2/ERF genes and the conserved protein motifs of their encoded OfAP2/ERF proteins were analyzed, and the results were found to be consistent with those of the population classification. A tissue-specific analysis showed the spatiotemporal expression of OfAP2/ERF in the stems and leaves of O. fragrans at different developmental stages. Specifically, 21 genes were not expressed in any tissue, while high levels of expression were found for 25 OfAP2/ERF genes in several tissues, 60 genes in the roots, 34 genes in the stems, 37 genes in young leaves, 34 genes in old leaves, 32 genes in the early flowering stage, 18 genes in the full flowering stage, and 37 genes in the late flowering stage. Quantitative RT-PCR experiments showed that OfERF110a and OfERF110b had the highest expression levels at the full-bloom stage (S4), and this gradually decreased with the senescence of petals. The expression of OfERF119c decreased first and then increased, while the expression levels of OfERF4c and OfERF5a increased constantly. This indicated that these genes may play roles in flower senescence and the ethylene response. In the subsequent subcellular localization experiments, we found that ERF1-4 was localized in the nucleus, indicating that it was expressed in the nucleus. In yeast self-activation experiments, we found that OfERF112, OfERF228, and OfERF23 had self-activation activity. Overall, these results suggest that OfERFs may have the function of regulating petal senescence in O. fragrans.

Characterization of YABBY transcription factors in Osmanthus fragrans and functional analysis of OfYABBY12 in floral scent formation and leaf morphology.

BACKGROUND: The plant-specific YABBY transcription factor family plays important roles in plant growth and development, particularly leaf growth, floral organ formation, and secondary metabolite synthesis. RESULTS: Here, we identified a total of 13 OfYABBY genes from the Osmanthus fragrans genome. These 13 OfYABBY genes were divided into five subfamilies through phylogenetic analysis, and genes in the same subfamily showed similar gene structures and conserved protein motifs. Gene duplication promoted the expansion of the OfYABBY family in O. fragrans. Tissue-specific expression analysis showed that the OfYABBY family was mainly expressed in O. fragrans leaves and floral organs. To better understand the role of OfYABBY genes in plant growth and development, OfYABBY12 was selected for heterologous stable overexpression in tobacco, and OfYABBY12-overexpressing tobacco leaves released significantly fewer volatile organic compounds than wild-type tobacco leaves. Overexpression of OfYABBY12 led to the downregulation of NtCCD1/4 and decreased ß-ionone biosynthesis. Correspondingly, a dual-luciferase assay showed that OfYABBY12 negatively regulated the expression of OfCCD4, which promotes ß-ionone synthesis. Furthermore, tobacco leaves overexpressing OfYABBY12 were curled and wrinkled and had significantly reduced leaf thickness and leaf inclusions and significantly extended flower pistils (styles). CONCLUSION: Overall, the results suggest that the OfYABBY gene family may influence the biosynthesis of the floral scent (especially ß-ionone) in O. fragrans and may regulate leaf morphogenesis and lateral organs.

The homomorphic self-incompatibility system in Oleaceae is controlled by a hemizygous genomic region expressing a gibberellin pathway gene.

In flowering plants, outcrossing is commonly ensured by self-incompatibility (SI) systems. These can be homomorphic (typically with many different allelic specificities) or can accompany flower heteromorphism (mostly with just two specificities and corresponding floral types). The SI system of the Oleaceae family is unusual, with the long-term maintenance of only two specificities but often without flower morphology differences. To elucidate the genomic architecture and molecular basis of this SI system, we obtained chromosome-scale genome assemblies of Phillyrea angustifolia individuals and related them to a genetic map. The S-locus region proved to have a segregating 543-kb indel unique to one specificity, suggesting a hemizygous region, as observed in all distylous systems so far studied at the genomic level. Only one of the predicted genes in this indel region is found in the olive tree, Olea europaea, genome, also within a segregating indel. We describe complete association between the presence/absence of this gene and the SI types determined for individuals of seven distantly related Oleaceae species. This gene is predicted to be involved in catabolism of the gibberellic acid (GA) hormone, and experimental manipulation of GA levels in developing buds modified the male and female SI responses of the two specificities in different ways. Our results provide a unique example of a homomorphic SI system, where a single conserved gibberellin-related gene in a hemizygous indel underlies the long-term maintenance of two groups of reproductive compatibility.

SWATH-MS based quantitive proteomics reveal regulatory metabolism and networks of androdioecy breeding system in Osmanthus fragrans.

BACKGROUND: The fragrant flower plant Osmanthus fragrans has an extremely rare androdioecious breeding system displaying the occurrence of males and hermaphrodites in a single population, which occupies a crucial intermediate stage in the evolutionary transition between hermaphroditism and dioecy. However, the molecular mechanism of androdioecy plant is very limited and still largely unknown. RESULTS: Here, we used SWATH-MS-based quantitative approach to study the proteome changes between male and hermaphroditic O. fragrans pistils. A total of 428 proteins of diverse functions were determined to show significant abundance changes including 210 up-regulated and 218 down-regulated proteins in male compared to hermaphroditic pistils. Functional categorization revealed that the differentially expressed proteins (DEPs) primarily distributed in the carbohydrate metabolism, secondary metabolism as well as signaling cascades. Further experimental analysis showed the substantial carbohydrates accumulation associated with promoted net photosynthetic rate and water use efficiency were observed in purplish red pedicel of hermaphroditic flower compared with green pedicel of male flower, implicating glucose metabolism serves as nutritional modulator for the differentiation of male and hermaphroditic flower. Meanwhile, the entire upregulation of secondary metabolism including flavonoids, isoprenoids and lignins seem to protect and maintain the male function in male flowers, well explaining important feature of androdioecy that aborted pistil of a male flower still has a male function. Furthermore, nine selected DEPs were validated via gene expression analysis, suggesting an extra layer of post-transcriptional regulation occurs during O. fragrans floral development. CONCLUSION: Taken together, our findings represent the first SWATH-MS-based proteomic report in androdioecy plant O. fragrans, which reveal carbohydrate metabolism, secondary metabolism and post-transcriptional regulation contributing to the androdioecy breeding system and ultimately extend our understanding on genetic basis as well as the industrialization development of O. fragrans.

Photoprotective Role of Photosynthetic and Non-Photosynthetic Pigments in Phillyrea latifolia: Is Their "Antioxidant" Function Prominent in Leaves Exposed to Severe Summer Drought?

Carotenoids and phenylpropanoids play a dual role of limiting and countering photooxidative stress. We hypothesize that their "antioxidant" function is prominent in plants exposed to summer drought, when climatic conditions exacerbate the light stress. To test this, we conducted a field study on Phillyrea latifolia, a Mediterranean evergreen shrub, carrying out daily physiological and biochemical analyses in spring and summer. We also investigated the functional role of the major phenylpropanoids in different leaf tissues. Summer leaves underwent the most severe drought stress concomitantly with a reduction in radiation use efficiency upon being exposed to intense photooxidative stress, particularly during the central hours of the day. In parallel, a significant daily variation in both carotenoids and phenylpropanoids was observed. Our data suggest that the morning-to-midday increase in zeaxanthin derived from the hydroxylation of ß-carotene to sustain non-photochemical quenching and limit lipid peroxidation in thylakoid membranes. We observed substantial spring-to-summer and morning-to-midday increases in quercetin and luteolin derivatives, mostly in the leaf mesophyll. These findings highlight their importance as antioxidants, countering the drought-induced photooxidative stress. We concluded that seasonal and daily changes in photosynthetic and non-photosynthetic pigments may allow P. latifolia leaves to avoid irreversible photodamage and to cope successfully with the Mediterranean harsh climate.

Identification of the Female-Produced Sex Pheromone of the Dotted White Geometrid Naxa seriaria (Lepidoptera: Geometridae).

The dotted white geometrid moth, Naxa seriaria Motschulsky (Lepidoptera: Geometridae), is a pest of Oleaceae in Korea, Japan, and China. In this study, we identified (3Z,6Z,9Z,12Z,15Z)-heneicosapentaene (C-21 pentaene) as the only compound in extracts of the pheromone glands of female N. seriaria causing a response from receptors on the antennae of males in analyses by gas chromatography with electroantennographic detection. The synthetic sex pheromone elicited dose-dependent electrophysiological responses from antennae of male N. seriaria. In field tests, more male moths were captured in traps baited with synthetic C-21 pentaene than in unbaited traps, and increasing the loading of C-21 pentaene in the lure increased catches of male moths. Significantly more male N. seriaria moths were caught in delta traps than in bucket traps. Based on these results, C-21 pentaene is proposed to be the major, if not the only, component of the sex pheromone of N. seriaria.

Transcriptomic analysis of flower opening response to relatively low temperatures in Osmanthus fragrans.

BACKGROUND: Sweet osmanthus (Osmanthus fragrans Lour.) is one of the top ten traditional ornamental flowers in China. The flowering time of once-flowering cultivars in O. fragrans is greatly affected by the relatively low temperature, but there are few reports on its molecular mechanism to date. A hypothesis had been raised that genes related with flower opening might be up-regulated in response to relatively low temperature in O. fragrans. Thus, our work was aimed to explore the underlying molecular mechanism of flower opening regulated by relatively low temperature in O. fragrans. RESULTS: The cell size of adaxial and abaxial petal epidermal cells and ultrastructural morphology of petal cells at different developmental stages were observed. The cell size of adaxial and abaxial petal epidermal cells increased gradually with the process of flower opening. Then the transcriptomic sequencing was employed to analyze the differentially expressed genes (DEGs) under different number of days' treatments with relatively low temperatures (19 °C) or 23 °C. Analysis of DEGs in Gene Ontology analysis showed that "metabolic process", "cellular process", "binding", "catalytic activity", "cell", "cell part", "membrane", "membrane part", "single-organism process", and "organelle" were highly enriched. In KEGG analysis, "metabolic pathways", "biosynthesis of secondary metabolites", "plant-pathogen interaction", "starch and sucrose metabolism", and "plant hormone signal transduction" were the top five pathways containing the greatest number of DEGs. The DEGs involved in cell wall metabolism, phytohormone signal transduction pathways, and eight kinds of transcription factors were analyzed in depth. CONCLUSIONS: Several unigenes involved in cell wall metabolism, phytohormone signal transduction pathway, and transcription factors with highly variable expression levels between different temperature treatments may be involved in petal cell expansion during flower opening process in response to the relatively low temperature. These results could improve our understanding of the molecular mechanism of relatively-low-temperature-regulated flower opening of O. fragrans, provide practical information for the prediction and regulation of flowering time in O. fragrans, and ultimately pave the way for genetic modification in O. fragrans.

Genome-Wide Identification, Classification, and Expression Profiling Reveals R2R3-MYB Transcription Factors Related to Monoterpenoid Biosynthesis in Osmanthus fragrans.

Osmanthus fragrans is widely grown for the purpose of urban greening and the pleasant aroma emitted from its flowers. The floral scent is determined by several monoterpenoid volatiles, such as linalool and its oxides, which are a few of the most common volatiles and the main components of the essential oils in most sweet osmanthus cultivars. In addition, the relative contents of cis- and trans-linalool oxide (furan) may affect the aromas and quality of the essential oils. MYB proteins represent the largest family of transcription factors in plants and participate in regulating secondary metabolites. Several cis-elements, especially AC-rich regions, are known to be bound by 2R-MYBs and could be found in the promoter of the enzyme genes in the terpenoid metabolic pathway. However, there has to date been no investigation into the 2R-MYB family genes involved in regulating terpenoid biosynthesis in O. fragrans. Here, 243 non-redundant 2R-MYB proteins were grouped into 33 clusters based on the phylogeny and exon-intron distribution. These genes were unevenly distributed on 23 chromosomes. Ka/Ks analysis showed that the major mode of 2R-MYB gene evolution was purifying selection. Expression analysis indicated that 2R-MYB genes in O. fragrans exhibited varied expression patterns. A total of 35 OfMYBs representing the highest per kilobase per million mapped reads in the flower were selected for quantitative real-time PCR analysis. The correlation analysis between the expression level and the contents of fragrant compounds at different flowering stages suggested that OfMYB19/20 exhibited remarkably positive correlation with the accumulation of cis-linalool oxides. OfMYB51/65/88/121/137/144 showed significantly negative correlations with one or more linalool oxides. Characterization of these proteins revealed that OfMYB19 and OfMYB137 were localized in the nuclei, but did not show transcriptional activation in the yeast system, which suggested that they may be bound to other transcription factors to exert regulatory functions. These findings provide useful information for further functional investigation of the 2R-MYBs and offer a foundation for clarifying the 2R-MYB transcription factors involved in the molecular mechanism of the regulation of monoterpenoid biosynthesis in Osmanthus fragrans.

Biochemical and Comparative Transcriptome Analyses Reveal Key Genes Involved in Major Metabolic Regulation Related to Colored Leaf Formation in Osmanthus fragrans 'Yinbi Shuanghui' during Development.

Osmanthus fragrans 'Yinbi Shuanghui' not only has a beautiful shape and fresh floral fragrance, but also rich leaf colors that change, making the tree useful for landscaping. In order to study the mechanisms of color formation in O. fragrans 'Yinbi Shuanghui' leaves, we analyzed the colored and green leaves at different developmental stages in terms of leaf pigment content, cell structure, and transcriptome data. We found that the chlorophyll content in the colored leaves was lower than that of green leaves throughout development. By analyzing the structure of chloroplasts, the colored leaves demonstrated more stromal lamellae and low numbers of granum thylakoid. However, there was a large number of plastoglobuli. Using transcriptome sequencing, we demonstrated that the expression of differentially expressed genes (DEGs) involved in chlorophyll degradation was upregulated, i.e., heme oxygennase-1 (HO1), pheophorbide a oxidase (PAO), and chlorophyllase-2 (CLH2), affecting the synthesis of chlorophyll in colored leaves. The stay-green gene (SGR) was upregulated in colored leaves. Genes involved in carotenoid synthesis, i.e., phytoene synthase 1 (PSY1) and 1-Deoxyxylulose-5-phosphate synthase (DXS), were downregulated in colored leaves, impeding the synthesis of carotenoids. In the later stage of leaf development, the downregulated expression of Golden2-Like (GLK) inhibited chloroplast development in colored leaves. Using weighted gene co-expression network analysis (WGCNA) to investigate the correlation between physiological indicators and DEGs, we chose the modules with the highest degree of relevance to chlorophyll degradation and carotenoid metabolism. A total of five genes (HSFA2, NFYC9, TCP20, WRKY3, and WRKY4) were identified as hub genes. These analyses provide new insights into color formation mechanisms in O. fragrans 'Yinbi Shuanghui' leaves at the transcriptional level.

Fraxitoxin, a New Isochromanone Isolated from Diplodia fraxini.

A new isochromanone, named fraxitoxin, was isolated together with (-)-mellein and tyrosol from liquid cultures of Diplodia fraxini, a pathogen involved in the etiology of canker and dieback disease of Fraxinus spp. in Europe. It was characterized as 5-methoxy-3-methylisochroman-1-one using spectroscopic methods (essentially NMR and HR-EI-MS). Its absolute configuration (R) at C(3) was assigned by electronic circular dichroism (ECD) measurements and calculations. Phytotoxic activity of the compound was evaluated on ash, cork and holm oak leaves at concentration of 1 mg/ml by the leaf-puncture assay. Interestingly, fraxitoxin caused necrotic lesions only on ash leaves.

Identification and validation of reference genes for gene expression studies in sweet osmanthus (Osmanthus fragrans) based on transcriptomic sequence data.

Accurate normalized data is a primary requisite for quantifying gene expression using RT-qPCR technology. Despite this importance, however, suitable reference genes in Osmanthus fragrans are not available. In this study, seven potential candidate reference genes (OfL25-1, OfL25-10, OfRP2, OfTUA, OfTUB3, OfUBQ2 and Of18S) were evaluated to determine which one would be the most reliable reference genes. The expression levels of the candidate reference genes were analysed by RT-qPCR in flower, leaf, pedicel, blossom bud tissues, as well as in floral organs at different developmental stages.GeNormand NormFinderwere used to statistically analyse transcript variation.Results indicated that OfRP2 and OfL25-10 were the optimal reference genes for use in RT-qPCR when analysing different stages of floral development; while OfTUB3 and OfL25-1 were optimal across tissues. The selected reference genes were used to examineOfMYB1 expression. The results appeared to be useful for future gene expression analyses aiming to characterize developmental stages and tissues of O. fragrans.

Larval Survival and Growth of Emerald Ash Borer (Coleoptera: Buprestidae) on White Ash and White Fringetree Saplings Under Well-Watered and Water-Deficit Conditions.

Emerald ash borer (Agrilus planipennis Fairmaire) was recently found on a novel host in North America, white fringetree (Chionanthus virginicus L.) (Oleaceae). In this study, we artificially infested 4-yr-old, naïve white fringetree and white ash (Fraxinus americana L.) saplings under well-watered and water-deficit conditions with emerald ash borer eggs. We used physiological and phenotypical approaches to investigate both plant response to emerald ash borer and insect development at 21, 36, and 61 d postinfestation. Photosynthesis was reduced in both tree species by larval feeding, but not by water deficits. Emerald ash borer larvae established and survived successfully on white ash. Both establishment and survival were lower on white fringetree than on white ash. Larvae were larger, and had reached higher instars at all three time points on white ash than on white fringetrees. Larvae grew faster in white ash under water-deficit conditions; however, water-deficit conditions negatively impacted survival of larvae at 61 d postinfestation in white fringetrees, although head size did not differ among surviving larvae. White ash showed higher callus formation in well-watered trees, but no impact on larval survival was observed. In white fringetree, callus formation was not affected by water treatment, and was inversely related to larval survival. The higher rate of mortality and slow growth rate of larvae in white fringetree as compared to white ash suggest that populations of emerald ash borer may be sustained by white fringetree, but may grow more slowly than in white ash.

High Stocking Density Controls Phillyrea Angustifolia in Mediterranean Grasslands.

Extensive grazing applied in the form of low instantaneous pressure over a long period is a widespread management practice in protected areas. However this kind of stocking method does not always achieve the expected results, in particular because it fails to limit colonization by woody plants.This is the case in the relict xero-halophytic grasslands of the northern Mediterranean coastal region, subjected to widespread colonization by the shrub Phillyrea angustifolia despite the presence of extensive grazing. In this study, we investigated, for an equal annual stocking rate, the respective impact of high stocking density applied over a short period (mob grazing) and low stocking density applied over a long period on both P. angustifolia and herbaceous cover, using an in situ experimental design run for 7 years. Only mob grazing was effective both in controlling the establishment and increasing the mortality of P. angustifolia individuals. We did not find any difference after the 7 years of experimentation between the two stocking methods with regard to the herbaceous community parameters tested: species richness, diversity, evenness, contribution of annual characteristic species. By contrast, the exclusion of domestic grazing led to a strong reduction of these values.The use of mob grazing may be well suited for meeting conservation goals such as maintaining open habitats in these grasslands.

Distribution, Predictors, and Impacts of Emerald Ash Borer (Agrilus planipennis) (Coleoptera: Buprestidae) Infestation of White Fringetree (Chionanthus virginicus).

Emerald ash borer, Agrilus planipennis (Fairmaire), is an invasive pest of ash trees (Fraxinus spp.) in North America that was recently found infesting white fringetree (Chionanthus virginicus L.). Initial reports of the infestation of white fringetree by emerald ash borer occurred in southwestern Ohio and Chicago, IL. We examined white fringetrees at additional sites in Illinois, Indiana, Ohio, and Pennsylvania in Summer and Fall 2015 and Winter 2016 for emerald ash borer infestation. Our aim was to examine white fringetrees at a limited number of sites with emerald ash borer infestation and to relate tree size, crown dieback, epicormic sprouting, tree sex, and adjacency to ash or white fringetrees with the likelihood of beetle infestation. A higher proportion of infested trees exhibited epicormic sprouting and the likelihood that a tree was infested increased with increasing crown dieback, variables that may be both predictors and responses to attack. The proportion of trees infested with emerald ash borer increased with increasing tree size. Signs consistent with emerald ash borer infestation were found in 26% of 178 white fringetrees, with at least one host infested at each site in all states. Infestation rates of white fringetrees increased with the density of white fringetrees at each site. The Chicago Botanic Garden site had a significantly lower infestation (3.7%) than other sites, which may be due to proactive management of ash. Overall, these data indicate white fringetree has been utilized by emerald ash borer throughout their overlapping ranges in the United States in ornamental settings likely due to ecological fitting.

Effects of Mulching on Soil Properties and Growth of Tea Olive (Osmanthus fragrans).

Different mulches have variable effects on soil physical properties and plant growth. This study aimed to compare the effects of mulching with inorganic (round gravel, RG), organic (wood chips, WC), and living (manila turf grass, MG) materials on soil properties at 0-5-cm and 5-10-cm depths, as well as on the growth and physiological features of Osmanthus fragrans L. 'Rixianggui' plants. Soil samples were collected at three different time points from field plots of O. fragrans plants treated with the different mulching treatments. Moisture at both soil depths was significantly higher after mulching with RG and WC than that in the unmulched control (CK) treatment. Mulching did not affect soil bulk density, pH, or total nitrogen content, but consistently improved soil organic matter. The available nitrogen in the soil increased after RG and WC treatments, but decreased after MG treatment during the experimental period. Mulching improved plant growth by increasing root activity, soluble sugar, and chlorophyll a content, as well as by providing suitable moisture conditions and nutrients in the root zone. Plant height and trunk diameter were remarkably increased after mulching, especially with RG and WC. However, while MG improved plant growth at the beginning of the treatment, the 'Rixianggui' plants later showed no improvement in growth. This was probably because MG competed with the plants for water and available nitrogen in the soil. Thus, our findings suggest that RG and WC, but not MG, improved the soil environment and the growth of 'Rixianggui' plants. Considering the effect of mulching on soil properties and plant growth and physiology, round gravel and wood chips appear to be a better choice than manila turf grass in 'Rixianggui' nurseries. Further studies are required to determine the effects of mulch quality and mulch-layer thickness on shoot and root growths.

Fundamental Host Range of Leptoypha hospita (Hemiptera: Tingidae), a Potential Biological Control Agent of Chinese Privet.

Chinese privet, Ligustrum sinense Lour., is an invasive shrub within riparian areas of the southeastern United States. Biological control is considered the most suitable management option for Chinese privet. The potential host range of the lace bug, Leptoypha hospita Drake et Poor, was evaluated on the basis of adult feeding and oviposition, combined oviposition-nymphal development no-choice tests, nymphal development no-choice tests, multiple generation comparison on Forestiera pubescens Nutt. and L. sinense no-choice tests, and multiple-choice tests with 45 plant species in 13 families. No-choice tests showed that the host range of L. hospita was restricted to the tribe Oleeae. In adult feeding and oviposition no-choice tests, the bug fed and oviposited significantly more on Chinese privet than all other test plant species except for three native Forestiera spp., two nonnative Syringa spp., and another exotic Ligustrum sp. Among those, only F. pubescens supported complete development in numbers comparable to Chinese privet. However, when reared for multiple generations lace bugs reared on F. pubescens were smaller and had lower fecundity than those reared on L. sinense, suggesting F. pubescens is not an optimal host. In multiple-choice tests, L. hospita displayed a strong preference for feeding and ovipositing on Chinese privet over other test plant species, with the exception of the closely related nonnative Syringa spp. and its congenic species Ligustrum vulgare. The results of this study suggest that the risk to nontarget plant species in North America is minimal, and L. hospita would be a promising candidate for Chinese privet biological control.

Contrasting impacts of continuous moderate drought and episodic severe droughts on the aboveground-biomass increment and litterfall of three coexisting Mediterranean woody species.

Climate change is predicted to increase the aridity in the Mediterranean Basin and severely affect forest productivity and composition. The responses of forests to different timescales of drought, however, are still poorly understood because extreme and persistent moderate droughts can produce nonlinear responses in plants. We conducted a rainfall-manipulation experiment in a Mediterranean forest dominated by Quercus ilex, Phillyrea latifolia, and Arbutus unedo in the Prades Mountains in southern Catalonia from 1999 to 2014. The experimental drought significantly decreased forest aboveground-biomass increment (ABI), tended to increase the litterfall, and decreased aboveground net primary production throughout the 15 years of the study. The responses to the experimental drought were highly species-specific. A. unedo suffered a significant reduction in ABI, Q. ilex experienced a decrease during the early experiment (1999-2003) and in the extreme droughts of 2005-2006 and 2011-2012, and P. latifolia was unaffected by the treatment. The drought treatment significantly increased branch litterfall, especially in the extremely dry year of 2011, and also increased overall leaf litterfall. The drought treatment reduced the fruit production of Q. ilex, which affected seedling recruitment. The ABIs of all species were highly correlated with SPEI in early spring, whereas the branch litterfalls were better correlated with summer SPEIs and the leaf and fruit litterfalls were better correlated with autumn SPEIs. These species-specific responses indicated that the dominant species (Q. ilex) could be partially replaced by the drought-resistant species (P. latifolia). However, the results of this long-term study also suggest that the effect of drought treatment has been dampened over time, probably due to a combination of demographic compensation, morphological and physiological acclimation, and epigenetic changes. However, the structure of community (e.g., species composition, dominance, and stand density) may be reordered when a certain drought threshold is reached.

Analysis of aroma-active compounds in three sweet osmanthus (Osmanthus fragrans) cultivars by GC-olfactometry and GC-MS.

OBJECTIVE: Aroma is the core factor in aromatherapy. Sensory evaluation of aromas differed among three sweet osmanthus (Osmanthus fragrans) cultivar groups. The purpose of this study was to investigate the aroma-active compounds responsible for these differences. METHODS: Gas chromatography-olfactometry (GC-O) and GC-mass spectrometry (GC-MS) were used to analyze the aroma-active compounds and volatiles of creamy-white ('Houban Yingui', HBYG), yellow ('Liuye Jingui', LYJG), and orange ('Gecheng Dangui', GCDG) cultivars. RESULTS: Seventeen aroma-active compounds were detected among 54 volatiles. trans-ß-Ocimene, trans-ß-ionone, and linalool, which were major volatiles, were identified as aroma-active, while cis-3-hexenyl butanoate, γ-terpinene, and hexyl butanoate were also aroma-active compounds, although their contents were low. Analysis of the odors was based on the sum of the modified frequency (MF) values of aroma-active compounds in different odor groups. HBYG contained more herb odors, contributed by cis-ß-ocimene and trans-ß-ocimene, while LYJG had more woody/violet/fruity odors released by trans-ß-ionone, α-ionone, and hexyl butanoate. In GCDG, the more floral odors were the result of cis-linalool oxide, trans-linalool oxide, and linalool. CONCLUSIONS: Aroma-active compounds were not necessarily only the major volatiles: some volatiles with low content also contributed to aroma. The aroma differences among the three cultivars resulted from variation in the content of different odor groups and in the intensities of aroma-active compounds.

Dampening effects of long-term experimental drought on growth and mortality rates of a Holm oak forest.

Forests respond to increasing intensities and frequencies of drought by reducing growth and with higher tree mortality rates. Little is known, however, about the long-term consequences of generally drier conditions and more frequent extreme droughts. A Holm oak forest was exposed to experimental rainfall manipulation for 13 years to study the effect of increasing drought on growth and mortality of the dominant species Quercus ilex, Phillyrea latifolia, and Arbutus unedo. The drought treatment reduced stem growth of A. unedo (-66.5%) and Q. ilex (-17.5%), whereas P. latifolia remained unaffected. Higher stem mortality rates were noticeable in Q. ilex (+42.3%), but not in the other two species. Stem growth was a function of the drought index of early spring in the three species. Stem mortality rates depended on the drought index of winter and spring for Q. ilex and in spring and summer for P. latifolia, but showed no relation to climate in A. unedo. Following a long and intense drought (2005-2006), stem growth of Q. ilex and P. latifolia increased, whereas it decreased in A. unedo. Q. ilex also enhanced its survival after this period. Furthermore, the effect of drought treatment on stem growth in Q. ilex and A. unedo was attenuated as the study progressed. These results highlight the different vulnerabilities of Mediterranean species to more frequent and intense droughts, which may lead to partial species substitution and changes in forest structure and thus in carbon uptake. The response to drought, however, changed over time. Decreased intra- and interspecific competition after extreme events with high mortality, together with probable morphological and physiological acclimation to drought during the study period, may, at least in the short term, buffer forests against drier conditions. The long-term effects of drought consequently deserve more attention, because the ecosystemic responses are unlikely to be stable over time.Nontechnical summaryIn this study, we evaluate the effect of long-term (13 years) experimental drought on growth and mortality rates of three forest Mediterranean species, and their response to the different intensities and durations of natural drought. We provide evidence for species-specific responses to drought, what may eventually lead to a partial community shift favoring the more drought-resistant species. However, we also report a dampening of the treatment effect on the two drought-sensitive species, which may indicate a potential adaptation to drier conditions at the ecosystem or population level. These results are thus relevant to account for the stabilizing processes that would alter the initial response of ecosystem to drought through changes in plant physiology, morphology, and demography compensation.