Shoot bending promotes flower bud formation by miRNA‐mediated regulation in apple (Malus domestica Borkh.)

Flower induction in apple (Malus domestica Borkh.) trees plays an important life cycle role, but young trees produce fewer and inferior quality flower buds. Therefore, shoot bending has become an important cultural practice, significantly promoting the capacity to develop more flower buds during the growing seasons. Additionally, microRNAs (miRNAs) play essential roles in plant growth, flower induction and stress responses. In this study, we identified miRNAs potentially involved in the regulation of bud growth, and flower induction and development, as well as in the response to shoot bending. Of the 195 miRNAs identified, 137 were novel miRNAs. The miRNA expression profiles revealed that the expression levels of 68 and 27 known miRNAs were down‐regulated and up‐regulated, respectively, in response to shoot bending, and that the 31 differentially expressed novel miRNAs between them formed five major clusters. Additionally, a complex regulatory network associated with auxin, cytokinin, abscisic acid (ABA) and gibberellic acid (GA) plays important roles in cell division, bud growth and flower induction, in which related miRNAs and targets mediated regulation. Among them, miR396, 160, 393, and their targets associated with AUX, miR159, 319, 164, and their targets associated with ABA and GA, and flowering‐related miRNAs and genes, regulate bud growth and flower bud formation in response to shoot bending. Meanwhile, the flowering genes had significantly higher expression levels during shoot bending, suggesting that they are involved in this regulatory process. This study provides a framework for the future analysis of miRNAs associated with multiple hormones and their roles in the regulation of bud growth, and flower induction and formation in response to shoot bending in apple trees.     

Antimicrobial,antibiofilm and antitumor activities of essential oil of Agastache rugosa from Xinjiang,China

In the study, we evaluated chemical composition and antimicrobial, antibiofilm, and antitumor activities of essential oils from dried leaf essential oil of leaf and flower of Agastache rugosa for the first time. Essential oil of leaf and flower was evaluated with GC and GC–MS methods, and the essential oil of flower revealed the presence of 21 components, whose major compounds were pulegone (34.1%), estragole (29.5%), and p-Menthan-3-one (19.2%). 26 components from essential oil of leaf were identified, the major compounds were p-Menthan-3-one (48.8%) and estragole (20.8%). At the same time, essential oil of leaf, there is a very effective antimicrobial activity with MIC ranging from 9.4 to 42 μg ml⁻¹ and potential antibiofilm, antitumor activities for essential oils of flower and leaf essential oil of leaf. The study highlighted the diversity in two different parts of A. rugosa grown in Xinjiang region and other places, which have different active constituents. Our results showed that this native plant may be a good candidate for further biological and pharmacological investigations.     

Variation in morphological characters,chemical composition,and anthocyanin content of different Chrysanthemum morifolium cultivars from Iran

The morphological traits, essential oil composition, and anthocyanin content of 17 Iranian C. morifolium cultivars were evaluated. Based on Shannon diversity index (I), such morphological characters as flower head diameter, ray floret, shape of apex, and main color of ray floret revealed more variability than the other traits. Significant genotypic variations were also observed in essential oil content (ranging from 0.1 to 0.56% (w/w)) and composition. Gas chromatography-mass spectrometry (GC-MS) analysis of the essential oil was conducted to identify 47 compounds in which camphor (0–44.6%), chrysanthenone (0–48.96%), chrysanthenyl acetate (0–16.22%), verbenol (0–11.78%), (+)-5-epi-neointermedeol (0–25.41%), and aromadendrene (0–14.98%) were the main constituents. Results of cluster analysis of essential oil compositions divided the cultivars into four groups. Hybridization among cultivars of divergent clusters led to heterotic effects for flower quality and phytochemical characters. The relationships established between the components revealed high correlations of anthocyanin with the two major aroma compounds of verbenol and chrysanthenyl acetate while they also provided a clue to the association between aroma compounds and color pigmentation. Moreover, a correlation was established between chrysanthenone and flower head diameter (r = −0.502). Based on the results obtained, it might be suggested that smaller flowers are capable of accumulating higher amounts of chrysanthenone. Clearly, the associations between morphological traits and essential oil components provide new insights for improved breeding programs in chrysanthemum.     

Chemical Composition and Biological Activities of the Essential Oil from Leaves and Flowers of Pulicaria incisa sub. candolleana (Family Asteraceae)

The composition of the essential oil isolated from leaves and flowers of Pulicaria incisa sub. candolleana E. Gamal ‐Eldin , growing in Egypt, was analysed by GC and GC‐MS. Forty‐nine and 68 compounds were identified from the oils of the leaves and flowers accounting for 86.69 and 84.29%, respectively of the total detected constituents. Both leaves and flowers oils were characterized by the high content of carvotanacetone with 66.01, 50.87 and chrysanthenone 13.26, 24.3%, respectively. The cytotoxic activity of both essential oils was evaluated against hepatocellular carcinoma cell line HEPG‐2, using MTT assay and vinblastine as a reference drug. Leaf oil showed higher activity with IC₅₀ 11.4 μg/ml compared with 37.4 μg/ml for flower oil. The antimicrobial activity of both oils was evaluated using agar well diffusion method towards two representatives for each of Gram positive and Gram negative bacteria as well as four representatives for fungi. The minimum inhibitory concentration of both essential oils against bacterial and fungal strains was obtained in the range of 0.49 – 15.63 μg/ml.     

Leaf blade structure of Verbesina macrophylla (Cass.) F. S. Blake (Asteraceae): ontogeny,duct secretion mechanism and essential oil composition

• Secretory structures are common in Asteraceae, where they exhibit a high degree of morphological diversity. The species Verbesina macrophylla, popularly known as assa‐peixe, is native to Brazil where it is widely used for medicinal purposes. Despite its potential medical importance, there have been no studies of the anatomy of this species, especially its secretory structures and secreted compounds. This study examined leaves of V. macrophylla with emphasis on secretory structures and secreted secondary metabolites.
• Development of secretory ducts and the mechanism of secretion production are described for V. macrophylla using ultrastructure, yield and chemical composition of its essential oils.
• Verbesina macrophylla has a hypostomatic leaf blade with dorsiventral mesophyll and secretory ducts associated with vascular bundles of schizogenous origin. Histochemistry identified the presence of lipids, terpenes, alkaloids and mucopolysaccharides. Ultrastructure suggests that the secretion released into the duct lumen is produced in plastids of transfer cells, parenchymal sheath cells and stored in vacuoles in these cells and duct epithelial cells. The essential oil content was 0.8%, and its major components were germacrene D, germacrene D‐4‐ol, β‐caryophyllene, bicyclogermacrene and α‐cadinol.
• Secretory ducts of V. macrophylla are squizogenous. Substances identified in tissues suggest that both secretions stored in the ducts and in adjacent parenchyma cells are involved in chemical defence. The essential oil is rich in sesquiterpenes, with germacrene D and its derivatives being notable components.

     

Diversity of Essential Oil-Secretory Cells and Oil Composition in Flowers and Buds of Magnolia sirindhorniae and Its Biological Activities

Magnolia sirindhorniae Noot. & Chalermglin produces fragrant flowers. The volatile oil secretary cells, quantity and quality as well as antioxidant and antimicrobial activities of the oils extracted from buds and flowers, have been investigated. The distribution of essential oil secretory cell in bud and flower revealed that the density and size of the oil cells were significantly higher in flowers compared to buds. In different floral parts, carpel has a higher oil cell density followed by gynophore and tepal. The histochemical analysis revealed the essential oil is synthesized in oil secretory cells. The volatile oil yield was 0.25 % in the buds and 0.50 % in flowers. GC/FID and GC/MS analysis identified 33 compounds contributing 83.2–83.5 % of the total essential oil composition. Linalool is the main constituent contributing 58.9 % and 51.0 % in the buds and flowers oils, respectively. The essential oil extracted from the flowers showed higher antimicrobial efficacy against Klebsiella pneumoniae and Staphylococcus aureus. Similarly, the essential oil isolated from the flowers depicts higher free radical scavenging, and antioxidant activity compared to buds’ oil.     

Assessment of reproductive characteristics of Jatropha curcas L. in south Florida

Jatropha curcas L. (jatropha) is a species identified for biofuel production because of the high quality of the oil produced by its seeds. However, jatropha is undomesticated and little information is available about its reproductive characteristics. Breeding and genetic improvement programs are much needed for the jatropha development as a bioenergy crop. Information about floral display and mode of reproduction are considered essential for breeding programs. In this study, the total number of female flowers, male to female flower ratio, fruit set, in vitro pollen germination, and the formation of fruits by apomixis, self‐pollination, and natural pollination were evaluated in 17 jatropha accessions planted in South Florida. The total number of female flowers per inflorescence and male to female flower ration ranged from 2.8 to 9.1 and 9.9:1 to 55.4 : 1, respectively. During summer 2011, high fruit setting average was observed (75.5%). In vitro pollen germination varied from 64.6% during spring 2011 to 51.6% during fall 2011. The fruit set observed was from 10.1% to 64.0% through natural pollination and apomixes, respectively. Characteristics such as fruit fresh weight, number of seeds per fruit, seed dry weight, and oil content were influenced by mode of reproduction.     

Chemical Composition and Allelopathic Potential of Essential Oils from Citharexylum spinosum L. Grown in Tunisia

Citharexylum spinosum L. (Verbenaceae) also known as Citharexylum quadrangulare Jacq . or Citharexylum fruticosum L. is an exotic tree introduced many years ago in Tunisia, specially used as a street and park ornamental tree. Essential oils (EOs) were obtained by hydrodistillation of the different parts (roots, stems, leaves, flowers and fruits; drupes) collected from trees grown in the area of Monastir (Tunisia). In total, 84 compounds, representing 90.1 – 98.4% of the whole oil composition, were identified by GC‐FID and GC/MS analyses. The root EO was distinguished by its high content in monoterpene hydrocarbons (α‐phellandrene; 30.8%) whereas that obtained from stems was dominated by sesquiterpene hydrocarbons (cuparene; 16.4%). The leaf oil was rich in an apocarotenoid derivative (hexahydrofarnesylacetone; 26%) and an aliphatic hydrocarbon (nonadecane; 14.5%). Flowers oil was rich in esters (2‐phenylethyl benzoate; 33.5%). Finally, drupes oil was rich in oxygenated sesquiterpenes (β‐eudesmol; 33.1%). Flowers oil showed a significant phytotoxic effect against lettuce seeds germination, it induces a total inhibition when tested at 1 mg/ml. Root and shoot elongation seemed to be more affected than germination. The inhibition of the shoot length varied from 3.6% to 100% and that of the root from 16.1% to 100%. The highest inhibition of 100% was detected for flower oil tested at 1 mg/ml. Our in vitro studies suggest a possible and new alternative use of C. spinosum EOs in herbicidal formulations, further experiments involving field conditions are necessary to confirm its herbicidal potential.     

Chemical composition and chemometric analysis of essential oils variation of <Emphasis Type="Italic">Bidens tripartita</Emphasis> L<Emphasis Type="Italic">.</Emphasis> during vegetation stages

The variation in the content and composition of essential oils of Bidens tripartita L. grown in Lithuania has been studied using supercritical CO₂ extraction followed by GC–MS. Extraction of essential oils was performed at two different pressures (9.1 and 15.1 MPa). Plant material has been harvested at different phenological stages (from June to September). A total of 26 different compounds were identified in the essential oils. The highest content and diversity of compounds was determined during the full-flowering stage. Identified substances were mainly monoterpene hydrocarbons and sesquiterpene hydrocarbons. The main components were α-pinene, p-cymene, trans-β-ocimene, and β-elemene. Chemometric analysis, including principal component and hierarchical cluster analyses, showed the dependence of the essential oils qualitative as well as quantitative composition on the phenological stage the plant was collected at. It also revealed that there was no correlation between age of the plant and quantity of the identified compounds.     

Biological activities of the essential oils and methanol extract of tow cultivated mint species (<Emphasis Type="Italic">Mentha longifolia</Emphasis> and <Emphasis Type="Italic">Mentha pulegium</Emphasis>) used in the Tunisian folkloric medicine

The composition of the essential oils and methanolic extracts of two cultivated mint species (M. longifolia and M. pulegium), as well as the in vitro antimicrobial and antioxidant activities of the essential oil and methanol extract of Mentha longifolia and Mentha pulegium were compared. GC-MS analysis of the essential oil identified 41 compounds constituting 96.66 and 96.13% of the total oil from M. longifolia and M. pulegium, respectively. The later oils were rich on pulegone (47.15 and 61.11%, respectively). Moreover, 1,8 cineole (11.54%), menthone (10.7%), α-pinene (3.57%), α-terpineol (3.17%) and d-cadinene (3.53%) were only present in M. longifolia oil, while isomenthone (17.02%), and piperitone (2.63%), were characteristic of M. pulegium oil. Shoot extract of the two species showed significantly different contents in total polyphenols (89.1 and 37.41 mg GAE/g DW), flavonoids (63.93 and 33.83 mg CE/g DW) and tannins (1.47 and 3.07 mg CE/g DW), respectively in M. longifolia and M. pulegium. The essential oils showed strong antimicrobial activity against all 16 microorganisms tested, whereas the methanol extracts were inactive. Moreover, the essential oils showed higher antioxidant activity than the methanolic extracts against the DPPH and superoxide radical scavenging. In fact, antioxidant activities of the oils were the same for both M. longifolia and M. pulegium against DPPH (IC₅₀ = 9 and 10 μg/ml, respectively) and 2-fold and 4-fold higher than shoot extracts (IC₅₀ = 20 and 48 μg/ml, respectively). Moreover, both oils showed the same antioxidative abilities as compared to the positive control (butylated hydroxytoluene). In the same way, the capacity to inhibit superoxide anion was very significant for the two oils (0.1 μg/ml for M. longifolia and 0.11 μg/ml for M. pulegium).     

Variation in Endogenous Gibberellins,Abscisic Acid,and Carbohydrate Content During the Growth Cycle of Colored <Emphasis Type="Italic">Zantedeschia</Emphasis> spp., a Tuberous Geophyte

Phenologic changes and variation in the level of endogenous gibberellins (GAs), abscisic acid (ABA), carbohydrate content, and α-amylase activity were examined in colored Zantedeschia spp. cv. Cala Gold. These changes were examined in the primary bud tissues and in the attached tuber tissue during the growth cycle. Dormant tubers were dry-stored at 20°C for 3 months, planted in a phytotron, and grown under 22/16 ± 1°C. Plant development was monitored under continued irrigation until leaf senescence and tuber dormancy. GAs and ABA were extracted from the primary bud tissues, fractionated by HPLC, and analyzed using GC-SIM. Starch, glucose, soluble protein, and α-amylase activity were monitored in the tuber tissue attached to the primary bud. Endogenous changes in GAs and ABA in the primary bud were correlated with endogenous changes in carbohydrate content and α-amylase activity in the attached tuber tissue. These correlations were observed during the rest and the growth periods and were associated with developmental changes in the plant, that is, bud dormancy relaxation, bud growth, and inflorescence differentiation. ABA content decreased and a transient pulse of GA was measured in the primary bud concomitantly with the onset of shoot elongation in dry tubers during storage, before planting. The sharp increase of GAs in the bud preceded inflorescence differentiation as observed in dissected apices by about 15 days, as well as the increase in α-amylase activity in the attached tuber tissue. A steep decrease in starch level was measured in the tuber after planting, concomitantly with massive plant growth. These findings suggest a possible involvement of gibberellin in the initiation of α-amylase activity during dormancy relaxation in colored Zantedeschia and in the autonomous induction of flowering.     

Roots volatiles and fatty acids of coriander (<Emphasis Type="Italic">Coriandrum sativum</Emphasis> L.) grown in saline medium

An hydroponic culture was conducted to investigate the effect of saline stress on the essential oil and fatty acid composition of Tunisian coriander (Coriandrum sativum L.) roots. Ten days old coriander seedlings were treated during 3 weeks with different NaCl concentrations (0, 25, 50 and 75 mM). Roots volatile components and fatty acids were analyzed. The essential oil yield was 0.06% in the control, on the basis of dry matter weight, and did not changed at low concentration (25 mM), while it increased significantly with increasing NaCl concentrations to reach 0.12 and 0.21% at 50 and 75 mM NaCl, respectively. The major volatile component was (E)-2-dodecenal with 52% of total essential oil constituents, followed by decanal, dodecanal, (E)-2-tridecenal and (E)-2-dodecenal. Further, the amount of these compounds was affected differently by the NaCl level. Total fatty acid amount of coriander roots increased significantly only with 50 and 75 mM NaCl. Three major fatty acids: linoleic (43%), oleic (25.5%) and palmitic (21.6%) were identified. Linoleic acid amount remains unchanged at 25 mM, while it increased with raising NaCl concentrations. However, oleic acid amount decreased only at 25 mM and no effect was observed at 50 and 75 mM. Fatty acid percentages were differently affected by salt. The oleic/linoleic ratio was reduced with raising NaCl concentrations.     

Chemical Composition of Ballota macedonica Vandas and Ballota nigra L. ssp. foetida (Vis.) Hayek Essential Oils – The Chemotaxonomic Approach

The essential oils isolated from fresh aerial parts of Ballota macedonica (two populations) and Ballota nigra ssp. foetida were analyzed by GC and GC/MS. Eighty five components were identified in total; 60 components in B. macedonica oil (population from the Former Yugoslav Republic of Macedonia), 34 components in B. macedonica oil (population from the Republic of Serbia), and 33 components in the oil of B. nigra ssp. foetida accounting for 93.9%, 98.4%, and 95.8% of the total oils, respectively. The most abundant components in B. macedonica oils were carotol (13.7 – 52.1%), germacrene D (8.6 – 24.6%), and (E)‐caryophyllene (6.5 – 16.5%), while B. nigra ssp. foetida oil was dominated by (E)‐phytol (56.9%), germacrene D (10.0%), and (E)‐caryophyllene (4.7%). Multivariate statistical analyses (agglomerative hierarchical cluster analysis and principal component analysis) were used to compare and discuss relationships among Ballota species examined so far based on their volatile profiles. The chemical compositions of B. macedonica essential oils are reported for the first time.     

Changes in IAA,Phenolic Compounds,Peroxidase, IAA Oxidase,and Polyphenol Oxidase in Relation to Flower Formation in Crocus sativus

Changes in levels of IAA, phenolic compounds, peroxidase, polyphenol oxidase, and IAA oxidase activities in the corm and the apical bud of Crocus sativusL. during bud growth and development, with special emphasis on the flowering stage, were studied. In the bud, flower formation was accompanied by enhanced activities of peroxidase, polyphenol oxidase, IAA oxidase, and higher contents of phenolic compounds as well as lower levels of IAA. In the corm, during the flower formation, these enzymes showed an opposite behavior. Moreover, the contents of phenolics and IAA in the corm tissues during flower formation and growth were higher than at the other developmental stages. It may be concluded that the transition of saffron plants to flowering is correlated with peroxidase, polyphenol oxidase, and IAA oxidase. Furthermore, these enzymes might exert their roles in the regulation of flowering through their participation in IAA catabolism. The hypothesis of regulation of bud development by an interaction between phenolics and the enzymes involved in IAA catabolism is discussed.     

Essential Oil Composition of Phagnalon sordidum (L.) from Corsica,Chemical Variability and Antimicrobial Activity

The chemical composition of Phagnalon sordidum (L.) essential oil was investigated for the first time using gas chromatography and chromatography/mass spectrometry. Seventy‐six compounds, which accounted for 87.9% of the total amount, were identified in a collective essential oil of P. sordidum from Corsica. The main essential oil components were (E)‐β‐caryophyllene (14.4%), β‐pinene (11.0%), thymol (9.0%), and hexadecanoic acid (5.3%). The chemical compositions of essential oils from 19 Corsican locations were investigated. The study of the chemical variability using statistical analysis allowed identifying direct correlation between the three populations of P. sordidum widespread in Corsica and the essential oil compositions they produce. The in vitro antimicrobial activity of P. sordidum essential oil was evaluated and it exhibited a notable activity on a large panel of clinically significant microorganisms.     

Tocopherol,Carotene, Phenolic Contents and Antibacterial Properties of Rose Essential Oil,Hydrosol and Absolute

The antioxidant and antibacterial activities, and total phenolic contents of Rosa damascena Mill. flower extracts (absolute, essential oil and hydrosol) were investigated. The chemical compositions of these extracts were analysed by GC-MS. Phenylethyl alcohol (78.38%) was found to be the main constituent of rose absolute, while citrenellol and geraniol were the major compounds (>55%) of rose essential oil and hydrosol. Tocopherol and carotene levels were determined by high performance liquid chromatography (HPLC) analysis. The levels of beta carotene (422.3±35.6 ppm), alpha tocopherol (2397.1±72.5 ppm) and gamma tocopherol (343.1±28.4 ppm) of rose absolute were found to be higher than that of essential oil and hydrosol. Their total phenolic contents were also evaluated. The total phenolic content of the tested extracts varied from 5.2 to 2134.3 GAE/mg L⁻¹. Rose absolute and essential oil contained high levels of phenolics and demonstrated strong antibacterial activity against Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Bacillus subtilis (ATCC 6633), Staphylococcus aureus (ATCC 6538), Chromobacterium violaceum (ATCC 12472) and Erwinia carotovora (ATCC 39048) strains.     

Chemical Composition and Interpopulation Variability of Essential Oils of Taxus baccata L. from Serbia

The composition of the essential oil of the twigs and needles of Taxus baccata L. from three natural populations in Serbia has been determined by GC/MS analysis. Of the 91 detected compounds, 87 were identified. The most abundant compound classes were aliphatic alcohols, terpenes, aliphatic hydrocarbons, and aliphatic aldehydes, which together comprised ca. 86.92% of the total oil composition. The dominant constituents were oct‐1‐en‐3‐ol (23.48%), (3Z)‐hex‐3‐en‐1‐ol (11.46%; aliphatic alcohols), and myrtenol (11.38%; oxygenated monoterpene). The PCA of 22 selected compounds revealed differentiations of populations based on geographic distribution. The CA showed that Populations I and II from the Dinaric Alps were similar, and that Population III from the Balkan mountain system was distinct. This was the first investigation of interpopulation variability of T. baccata populations based on essential oil composition. The results of this study were compared with those of previous studies concerning volatile compounds produced by Taxus species. The results indicate that the essential oil content of T. baccata populations from this study is unique, mostly resembling the population from southeast Serbia.     

Essential oil composition of <Emphasis Type="Italic">Taraxacum officinale</Emphasis>

The gas chromatography–mass spectrometry (GC–MS) analysis of essential oil obtained by hydrodistillation from the flower of Taraxacum officinale L. revealed the presence of 25 compounds with 1,3-dimethylbenzene, 1,2-dimethylbenzene, 1-ethyl-3-methylbenzene, heneicosane and tricosane as the main components.     

Micropropagation of mature <Emphasis Type="Italic">Crataegus aronia</Emphasis> L., a medicinal and ornamental plant with rootstock potential for pome fruit

The effects of culture media and cytokinin types on micropropagation of mature Crataegus aronia L. were investigated. Using single-axillary bud explants, the growth of cultures on MS, WPM, DKW and NRM containing 4.44 μM benzyladenine (BA) plus 0.05 μM indole-3-butyric acid (IBA), and on NRM containing thidiazuron, meta-Topolin (mT) or BA at 1.25, 2.5, 5.0 or 7.5 μM plus 0.05 μM IBA were compared. The culture medium had significant effects on shoot number and length. In comparison with MS, DKW and WPM, shoot production was greater on NRM (5.7 shoots per explant). Shoot production on MS, DKW and WPM (4.2, 4.2 and 4.1, respectively) were statistically similar to each other. Thidiazuron was detrimental to shoot formation and caused formation of rosette shoots and/or large callus to form on explants. In the presence of mT, only some of the explants developed into shoots. Benzyladenine was the only cytokinin that promoted both shoot proliferation and shoot elongation. Higher shoot numbers were obtained at 5.0 and 7.5 μM BA compared to lower concentrations of BA. Over 80% of microshoots rooted and rooted shoots were successfully acclimatized to ex vitro conditions.