Volatile composition and classification of Lilium flower aroma types and identification, polymorphisms, and alternative splicing of their monoterpene synthase genes

Abstract Lily is a well-known ornamental plant with a diversity of fragrant types. Basic information on lily floral scent compounds has been obtained for only a few accessions, and little is known about Lilium aroma types, the terpene synthase genes that may play roles in the production of key volatiles, or the range of monoterpenes that these genes produce. In this study, 41 cultivars were analyzed for volatile emissions, and a total of 46 individual volatile compounds were identified, 16 for the first time in lilies. Lily accessions were classified into six groups according to the composition of major scent components: faint-scented, cool, fruity, musky, fruity-honey, and lily. Monoterpenes were one of the main groups of volatiles identified, and attention was focused on terpene synthase (TPS) genes, which encode enzymes that catalyze the last steps in monoterpene synthesis. Thirty-two candidate monoterpene synthase cDNAs were obtained from 66 lily cultivars, and 64 SNPs were identified. Two InDels were also shown to result from variable splicing, and sequence analysis suggested that different transcripts arose from the same gene. All identified nucleotide substitution sites were highly correlated with the amounts of myrcene emitted, and InDel site 230 was highly correlated with the emission of all major monoterpenoid components, especially (E)-β-ocimene. Heterologous expression of five cDNAs cloned from faint-scented and strong-scented lilies showed that their corresponding enzymes could convert geranyl diphosphate to (E)-β-ocimene, α-pinene, and limonene. The findings from this study provide a major resource for the assessment of lily scent volatiles and will be helpful in breeding of improved volatile components.

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Composition and Biosynthesis of Scent Compounds from Sterile Flowers of an Ornamental Plant Clematis florida cv. ‘Kaiser’

Molecules ◽

10.3390/molecules25071711 ◽

2020 ◽

Vol 25 (7) ◽

pp. 1711

Author(s):

Yifan Jiang ◽

Renjuan Qian ◽

Wanbo Zhang ◽

Guo Wei ◽

Xiaohua Ma ◽

...

Keyword(s):

Floral Scent ◽

Ornamental Plant ◽

Terpene Synthase ◽

Enzyme Assays ◽

Total Volatile ◽

Terpene Biosynthesis ◽

Geranyl Diphosphate ◽

Multiple Products ◽

Key Genes

Clematis florida is a popular ornamental vine species known for diverse colors and shapes of its flowers but not for scent. Here we investigated the composition and biosynthesis of floral scent in ‘Kaiser’, a fragrant cultivar of C. florida that has sterile flowers. Volatile profiling revealed that flowers of ‘Kaiser’ emit more than 20 compounds, with monoterpenes being most abundant. Among the three floral organs, namely sepals, transformed-petals, and ovaries, ovaries had the highest rates of total volatile emission. To determine the molecular mechanism underlying floral scent biosynthesis in ‘Kaiser’, we sequenced a flower transcriptome and searched the transcriptome for terpene synthase genes (TPSs), which are key genes for terpene biosynthesis. Among the TPS genes identified, three were putative intact full-length genes and were designated CfTPS1, CfTPS2, and CfTPS3. Phylogenetic analysis placed CfTPS1, CfTPS2, and CfTPS3 to the TPS-g, TPS-b, and TPS-a subfamily, respectively. Through in vitro enzyme assays with Escherichia coli-expressed recombinant proteins, both CfTPS1 and CfTPS2 were demonstrated to catalyze the conversion of geranyl diphosphate to linalool, the most abundant constituent of C. florida floral scent. In addition, CfTPS1 and CfTPS2 produced the sesquiterpene nerolidol from (E,E)-farnesyl diphosphate. CfTPS3 showed sesquiterpene synthase activity and produced multiple products in vitro. All three CfTPS genes showed higher levels of expression in sepals than those in transformed-petals and ovaries. Our results show that despite being sterile, the flowers of ‘Kaiser’ have normal mechanisms for floral scent biosynthesis that make the flowers fragrant.

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Assessing Genetic Variability of Pelargonium Species Using PCR-based TRAP Markers

HortScience ◽

10.21273/hortsci.39.4.812d ◽

2004 ◽

Vol 39 (4) ◽

pp. 812D-812

Author(s):

Wai-Foong Hong ◽

Chang-Qing Bai ◽

Michael Broe ◽

Jinguo Hu ◽

Charles Krause ◽

...

Keyword(s):

Cluster Analysis ◽

Genetic Variation ◽

Genetic Variability ◽

Morphological Traits ◽

Molecular Data ◽

Ornamental Plant ◽

Target Region ◽

Marker System ◽

Highly Correlated

Pelargonium is one of the important flower crops in USA. It is a priority genus for conservation at the USDA Ornamental Plant Germplasm Center (OPGC). It belongs to Geraniaceae family and comprises of about 280 species. To understand the genetic variation of the Pelargonium collection at OPGC, the PCR-based TRAP (target region amplified polymorphism) marker system which was newly developed in sunflower was used in this study. Twelve sets of primers were used to fingerprint 46 accessions representing 21 commercial P. hortorum, 17 scented geraniums and 8 other unidentified Pelargonium taxa. About 150 DNA bands could be detected in each primer and accession combination. Cluster analysis showed that molecular data was highly correlated with the phenotypes. Cultivars with similar morphological traits were clustered together. These results demonstrated that the TRAP system is a useful technique for the characterization and classification of Pelargonium collections.

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Volatiles of the Inflorescences of the Madeiran Orchids, Goodyera macrophylla Lowe and Gennaria diphylla (Link) Parl. and Their Role in Pollination

Natural Product Communications ◽

10.1177/1934578x0700200414 ◽

2007 ◽

Vol 2 (4) ◽

pp. 1934578X0700200

Author(s):

Francisco M. Fernandes ◽

A. Cristina Figueiredo ◽

José G. Barroso ◽

Luís G. Pedro ◽

Christopher C. Wilcock ◽

...

Keyword(s):

Volatile Compounds ◽

Floral Scent ◽

Volatile Components ◽

Rare Endemic ◽

First Time ◽

Headspace Sorption ◽

Type Apparatus

Goodyera macrophylla, considered a rare endemic, and Gennaria diphylla are two native terrestrial Orchidaceae species that can be found mainly in Madeiran Laurisilva. The volatile compounds contributing to the floral scent of these two Madeiran orchids were analysed and compared for the first time. The volatiles, isolated by distillation-extraction in a Likens-Nickerson-type apparatus, and those extracted by headspace sorption, were analysed by GC and GC-MS. The main volatile components isolated by distillation-extraction were γ-terpinene (13%) and n-nonanal (11%) in G. macrophylla and n-octacosane (19%) and n-heptacosane (13%) in G. diphylla. The main volatile components isolated by headspace sorption were p-cymene (21%), α-pinene (15%) and linalool (14%) in G. macrophylla and cis-arbusculone (28%) and lilac alcohol (26%) in G. diphylla. The importance of the volatiles in the pollination of these orchids is discussed.

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СОЦИАЛЬНАЯ КОНФЛИКТОЛОГИЯ: ТЕОРЕТИКО-ИСТОРИЧЕСКИЙ АСПЕКТ

Konfliktologia ◽

10.31312/2310-6085-2020-15-2-68-80 ◽

2020 ◽

Vol 15 (2) ◽

pp. 68

Author(s):

А. Н. Сухов

Keyword(s):

Theory And Practice ◽

Important Task ◽

Social Conflicts ◽

Psychological Approach ◽

Structure Dynamics ◽

History Of ◽

The Subject ◽

First Time

This given article reveals the topicality not only of destructive, but also of constructive, as well as hybrid conflicts. Practically it has been done for the first time. It also describes the history of the formation of both foreign and domestic social conflictology. At the same time, the chronology of the development of the latter is restored and presented objectively, in full, taking into account the contribution of those researchers who actually stood at its origins. The article deals with the essence of the socio-psychological approach to understanding conflicts. The subject of social conflictology includes the regularities of their occurrence and manifestation at various levels, spheres and conditions, including normal, complicated and extreme ones. Social conflictology includes the theory and practice of diagnosing, resolving, and resolving social conflicts. It analyzes the difficulties that occur in defining the concept, structure, dynamics, and classification of social conflicts. Therefore, it is no accident that the most important task is to create a full-fledged theory of social conflicts. Without this, it is impossible to talk about effective settlement and resolution of social conflicts. Social conflictology is an integral part of conflictology. There is still a lot of work to be done, both in theory and in application, for its complete design. At present, there is an urgent need to develop conflict-related competence not only of professionals, but also for various groups of the population.

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Morphological and Ultrastructural Characterization of Hemocytes in an Insect Model, the Hematophagous Dipetalogaster maxima (Hemiptera: Reduviidae)

Insects ◽

10.3390/insects12070640 ◽

2021 ◽

Vol 12 (7) ◽

pp. 640

Author(s):

Natalia R. Moyetta ◽

Fabián O. Ramos ◽

Jimena Leyria ◽

Lilián E. Canavoso ◽

Leonardo L. Fruttero

Keyword(s):

Cell Biology ◽

Cell Types ◽

Transmission Electron ◽

Ultrastructural Characterization ◽

Current Classification ◽

Contrast Microscopy ◽

First Time ◽

Controversial Aspect

Hemocytes, the cells present in the hemolymph of insects and other invertebrates, perform several physiological functions, including innate immunity. The current classification of hemocyte types is based mostly on morphological features; however, divergences have emerged among specialists in triatomines, the insect vectors of Chagas’ disease (Hemiptera: Reduviidae). Here, we have combined technical approaches in order to characterize the hemocytes from fifth instar nymphs of the triatomine Dipetalogaster maxima. Moreover, in this work we describe, for the first time, the ultrastructural features of D. maxima hemocytes. Using phase contrast microscopy of fresh preparations, five hemocyte populations were identified and further characterized by immunofluorescence, flow cytometry and transmission electron microscopy. The plasmatocytes and the granulocytes were the most abundant cell types, although prohemocytes, adipohemocytes and oenocytes were also found. This work sheds light on a controversial aspect of triatomine cell biology and physiology setting the basis for future in-depth studies directed to address hemocyte classification using non-microscopy-based markers.

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Kinetic studies and homology modeling of a dual-substrate linalool/nerolidol synthase from Plectranthus amboinicus

Scientific Reports ◽

10.1038/s41598-021-96524-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Nur Suhanawati Ashaari ◽

Mohd Hairul Ab. Rahim ◽

Suriana Sabri ◽

Kok Song Lai ◽

Adelene Ai-Lian Song ◽

...

Keyword(s):

Active Site ◽

Catalytic Efficiency ◽

Kinetic Studies ◽

Homology Model ◽

Biochemical Properties ◽

Terpene Synthase ◽

Monoterpene Synthase ◽

Terminal Domain ◽

Catalytic Active Site ◽

Plectranthus Amboinicus

AbstractLinalool and nerolidol are terpene alcohols that occur naturally in many aromatic plants and are commonly used in food and cosmetic industries as flavors and fragrances. In plants, linalool and nerolidol are biosynthesized as a result of respective linalool synthase and nerolidol synthase, or a single linalool/nerolidol synthase. In our previous work, we have isolated a linalool/nerolidol synthase (designated as PamTps1) from a local herbal plant, Plectranthus amboinicus, and successfully demonstrated the production of linalool and nerolidol in an Escherichia coli system. In this work, the biochemical properties of PamTps1 were analyzed, and its 3D homology model with the docking positions of its substrates, geranyl pyrophosphate (C10) and farnesyl pyrophosphate (C15) in the active site were constructed. PamTps1 exhibited the highest enzymatic activity at an optimal pH and temperature of 6.5 and 30 °C, respectively, and in the presence of 20 mM magnesium as a cofactor. The Michaelis–Menten constant (Km) and catalytic efficiency (kcat/Km) values of 16.72 ± 1.32 µM and 9.57 × 10–3 µM−1 s−1, respectively, showed that PamTps1 had a higher binding affinity and specificity for GPP instead of FPP as expected for a monoterpene synthase. The PamTps1 exhibits feature of a class I terpene synthase fold that made up of α-helices architecture with N-terminal domain and catalytic C-terminal domain. Nine aromatic residues (W268, Y272, Y299, F371, Y378, Y379, F447, Y517 and Y523) outlined the hydrophobic walls of the active site cavity, whilst residues from the RRx8W motif, RxR motif, H-α1 and J-K loops formed the active site lid that shielded the highly reactive carbocationic intermediates from the solvents. The dual substrates use by PamTps1 was hypothesized to be possible due to the architecture and residues lining the catalytic site that can accommodate larger substrate (FPP) as demonstrated by the protein modelling and docking analysis. This model serves as a first glimpse into the structural insights of the PamTps1 catalytic active site as a multi-substrate linalool/nerolidol synthase.

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Measuring Dysgraphia: A Graded-Difficulty Spelling Test

Behavioural Neurology ◽

10.1155/1994/659593 ◽

1994 ◽

Vol 7 (3-4) ◽

pp. 107-116 ◽

Cited By ~ 65

Author(s):

D. M. Baxter ◽

E. K. Warrington

Keyword(s):

Left Hemisphere ◽

Right Hemisphere ◽

Language Disorders ◽

Lower Quartile ◽

Verbal Skills ◽

Spelling Test ◽

Spelling Disorder ◽

Highly Correlated ◽

Orthopaedic Injuries

In this paper we describe the construction of a graded-difficulty spelling test for adults consisting of two alternative forms each containing 30 words (GDST, Forms A and B). The spelling test, together with background tests of verbal and non-verbal skills, was administered to 100 control patients with orthopaedic injuries. The two forms of the spelling test were highly correlated (0.92). Spelling was highly correlated with reading (0.75, 0.77) and moderately correlated with vocabulary (0.57) and naming (0.39, 0.40). There was no correlation between spelling skills and non-verbal reasoning. The test was validated in a group of 26 patients with left hemisphere and 20 patients with right hemisphere lesions. Spelling was shown to be lateralized to the left hemisphere and there appeared to be a shift in scores of the left hemisphere group towards the lower quartile, with 65% of the left hemisphere group falling within this band. The most severe spelling impairments were invariably associated with other language disorders but a number of dissociations were documented at spelling levels falling between the 5th and 25th percentile band. Two patients with left hemisphere lesions (8%) were identified as having selective dysgraphias. The lack of overlap between the anatomical sites of the two patients with specific lexical dysgraphia argues against a single site for this type of dysgraphia and argues for further refinement of this classification of spelling disorder.

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A comparison of methodological approaches to the subfamilial classification of the Naididae (Oligochaeta)

Canadian Journal of Zoology ◽

10.1139/z87-108 ◽

1987 ◽

Vol 65 (3) ◽

pp. 691-707 ◽

Cited By ~ 16

Author(s):

A. F. L. Nemec ◽

R. O. Brinkhurst

Keyword(s):

Data Matrix ◽

Jaccard Coefficient ◽

Data Set ◽

Parsimony Method ◽

Fit Statistics ◽

Reconstruction Methods ◽

Linkage Methods ◽

The Family ◽

First Time

A data matrix of 23 generic or subgeneric taxa versus 24 characters and a shorter matrix of 15 characters were analyzed by means of ordination, cluster analyses, parsimony, and compatibility methods (the last two of which are phylogenetic tree reconstruction methods) and the results were compared inter alia and with traditional methods. Various measures of fit for evaluating the parsimony methods were employed. There were few compatible characters in the data set, and much homoplasy, but most analyses separated a group based on Stylaria from the rest of the family, which could then be separated into four groups, recognized here for the first time as tribes (Naidini, Derini, Pristinini, and Chaetogastrini). There was less consistency of results within these groups. Modern methods produced results that do not conflict with traditional groupings. The Jaccard coefficient minimizes the significance of symplesiomorphy and complete linkage avoids chaining effects and corresponds to actual similarities, unlike single or average linkage methods, respectively. Ordination complements cluster analysis. The Wagner parsimony method was superior to the less flexible Camin–Sokal approach and produced better measure of fit statistics. All of the aforementioned methods contain areas susceptible to subjective decisions but, nevertheless, they lead to a complete disclosure of both the methods used and the assumptions made, and facilitate objective hypothesis testing rather than the presentation of conflicting phylogenies based on the different, undisclosed premises of manual approaches.

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Bearing Severity Fault Evaluation Using Contour Maps—Case Study

Applied Sciences ◽

10.3390/app11146452 ◽

2021 ◽

Vol 11 (14) ◽

pp. 6452

Author(s):

César Ricardo Soto-Ocampo ◽

Juan David Cano-Moreno ◽

José Manuel Mera ◽

Joaquín Maroto

Keyword(s):

Health Indicators ◽

Bearing Faults ◽

Contour Maps ◽

Initial Stage ◽

Critical Components ◽

Bearing Damage ◽

First Time ◽

Current Operating

Increasing industrial competitiveness has led to an increased global interest in condition monitoring. In this sector, rotating machinery plays an important role, where the bearing is one of the most critical components. Many vibration-based signal treatments are already being used to identify features associated with bearing faults. The information embedded in such features are employed in the construction of health indicators, which allow for evaluation of the current operating status of the machine. In this work, the use of contour maps to represent the diagnosis map of a bearing, used as a health map, is presented for the first time. The results show that the proposed method is promising, allowing for the satisfactory detection and evaluation of the severity of bearing damage. In this initial stage of the research, our results suggest that this method can improve the classification of bearing faults and, therefore, optimise maintenance processes.

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Prognostic impact of β-2-microglobulin expression in colorectal cancers stratified by mismatch repair status

Journal of Clinical Pathology ◽

10.1136/jclinpath-2012-200742 ◽

2012 ◽

Vol 65 (11) ◽

pp. 996-1002 ◽

Cited By ~ 21

Author(s):

Viktor Hendrik Koelzer ◽

Kristi Baker ◽

Daniela Kassahn ◽

Daniel Baumhoer ◽

Inti Zlobec

Keyword(s):

Protein Expression ◽

Mismatch Repair ◽

Immune Cell ◽

Prognostic Significance ◽

Clinicopathological Features ◽

Prognostic Impact ◽

Mismatch Repair Status ◽

First Time ◽

Intra Class Correlation Coefficient

Backgroundβ-2-microglobulin (B2M) is essential for antigen presentation, yet may also possess proto-oncogenic properties.AimTo determine the prognostic impact of B2M in patients with mismatch repair (MMR) proficient and deficient colorectal cancer (CRC) and to investigate whether this effect on outcome is dependent on the local immune response. MethodsB2M protein expression and tumour-infiltrating immune cells (CD3, CD16, CD163, CD20, CD4, CD45RO, CD56, CD68, CD8, FoxP3, GranzymeB, iNOS, mast cell tryptase, MUM1, PD1, TIA-1) were evaluated in a well characterised tissue microarray of 408 CRCs. The predictive value for clinicopathological features and the prognostic significance of B2M expression were analysed, stratified by MMR status and the immunohistological characteristics of immune cell infiltrates. ResultsInterobserver agreement for B2M staining was high (intra-class correlation coefficient=0.91). Complete B2M loss was more frequent in MMR-deficient (19.4%) compared to MMR-proficient (7.1%) tumours (p<0.001). In MMR-deficient cases, B2M loss predicted rare local recurrence (p=0.034), infrequent nodal-positivity (p=0.035), absence of distant metastasis (p=0.048; sensitivity=100%) and a trend towards favourable survival (p=0.124) independent of immune infiltrates. No associations between B2M and clinicopathological features were observed in MMR-proficient cases.ConclusionsOur data show for the first time that absence of B2M protein expression identifies MMR-deficient cancers with a favourable clinical course and absence of metastatic disease. Validation of B2M protein expression for sub-classification of MMR-deficient CRC is recommended for future clinical trials.

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