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1.
Plants (Basel) ; 10(10)2021 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-34685902

RESUMO

Yukon Draba (Draba yukonensis) is a small, short-lived perennial mustard species that is endemic to southwestern Yukon in Canada. This plant has been categorized as a species of Special Concern. It faces the threat of habitat loss due to natural and man-made causes and a population that is unevenly distributed to a few large and several small subpopulations in the area. It will therefore be judicious to undertake investigations on the conservation of this species to save it from further deterioration which may lead to its extinction. In this study, a protocol was developed for in vitro propagation and cryopreservation of Yukon Draba. The micropropagation protocol was optimized using shoot tips which enabled clonal propagation and in vitro storage of the species. Shoots grew best in the medium containing MS basal salts and had the highest multiplication with the addition of 2 µM 6-benzylaminopurine or 5 µM Kinetin with 3% sucrose. The addition of 10 µM Indole Butyric Acid (IBA) produced the highest number of adventitious roots on the shoots and the longest root length was observed at 2 µM IBA. The rooted plantlets were transferred to greenhouse and the highest survival (87.5%) was observed for the plantlets treated with a lower concentration of IBA (2 µM). Cryopreservation protocol was developed using the droplet-vitrification method for in vitro shoot tips. Two-week-old shoots had the highest survival and regrowth following exposure to plant vitrification solution 3 (PVS3) for 30 min, prior to direct immersion of the droplets into the liquid nitrogen. The optimized protocols for the micropropagation and cryopreservation may be useful for the long-term germplasm conservation and reintroduction of this species in its natural habitat.

2.
Plants (Basel) ; 10(9)2021 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-34579422

RESUMO

Cryopreservation is considered an ideal strategy for the long-term preservation of plant genetic resources. Significant progress was achieved over the past several decades, resulting in the successful cryopreservation of the genetic resources of diverse plant species. Cryopreservation procedures often employ in vitro culture techniques and require the precise control of several steps, such as the excision of explants, preculture, osmo- and cryoprotection, dehydration, freeze-thaw cycle, unloading, and post-culture for the recovery of plants. These processes create a stressful environment and cause reactive oxygen species (ROS)-induced oxidative stress, which is detrimental to the growth and regeneration of tissues and plants from cryopreserved tissues. ROS-induced oxidative stresses were documented to induce (epi)genetic and somatic variations. Therefore, the development of true-to-type regenerants of the source germplasm is of primary concern in the application of plant cryopreservation technology. The present article provides a comprehensive assessment of epigenetic and genetic integrity, metabolic stability, and field performance of cryopreserved plants developed in the past decade. Potential areas and the directions of future research in plant cryopreservation are also proposed.

3.
Int J Mol Sci ; 22(16)2021 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-34445105

RESUMO

In grafted plants, the movement of long-distance signals from rootstocks can modulate the development and function of the scion. To understand the mechanisms by which tolerant rootstocks improve scion responses to osmotic stress (OS) conditions, mRNA transport of osmotic responsive genes (ORGs) was evaluated in a tomato/potato heterograft system. In this system, Solanum tuberosum was used as a rootstock and Solanum lycopersicum as a scion. We detected changes in the gene expression levels of 13 out of the 21 ORGs tested in the osmotically stressed plants; of these, only NPR1 transcripts were transported across the graft union under both normal and OS conditions. Importantly, OS increased the abundance of StNPR1 transcripts in the tomato scion. To examine mRNA mobility in transgrafted plants, StNPR1 and StDREB1 genes representing the mobile and non-mobile transcripts, respectively, were overexpressed in tobacco (Nicotiana tabacum). The evaluation of transgenic tobacco plants indicated that overexpression of these genes enhanced the growth and improved the physiological status of transgenic plants growing under OS conditions induced by NaCl, mannitol and polyethylene glycol (PEG). We also found that transgenic tobacco rootstocks increased the OS tolerance of the WT-scion. Indeed, WT scions on transgenic rootstocks had higher ORGs transcript levels than their counterparts on non-transgenic rootstocks. However, neither StNPR1 nor StDREB1 transcripts were transported from the transgenic rootstock to the wild-type (WT) tobacco scion, suggesting that other long-distance signals downstream these transgenes could have moved across the graft union leading to OS tolerance. Overall, our results signify the importance of StNPR1 and StDREB1 as two anticipated candidates for the development of stress-resilient crops through transgrafting technology.


Assuntos
Lycopersicon esculentum/genética , Osmose/fisiologia , Pressão Osmótica/fisiologia , Solanum tuberosum/genética , Tabaco/genética , Raízes de Plantas/genética , Plantas Geneticamente Modificadas/genética , Transgenes/genética
4.
Int J Mol Sci ; 22(15)2021 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-34361028

RESUMO

The growth and productivity of several apple rootstocks have been evaluated in various previous studies. However, limited information is available on their tolerance to osmotic stress. In the present study, the physiological and molecular responses as well as abscisic acid (ABA) levels were assessed in six apple rootstocks (M26, V3, G41, G935, B9 and B118) osmotically stressed with polyethylene glycol (PEG, 30%) application under greenhouse conditions. Our results showed that V3, G41, G935 and B9 had higher relative water content (RWC), and lower electrolyte leakage (EL) under stress conditions compared to M26 and B118. Additionally, water use efficiency (WUE) was higher in V3, G41 and B9 than M26, which might be partially due to the lower transpiration rate in these tolerant rootstocks. V3, G41 and B9 rootstocks also displayed high endogenous ABA levels which was combined with a reduction in stomatal conductance and decreased water loss. At the transcriptional level, genes involved in ABA-dependent and ABA-independent pathways, e.g., SnRK, DREB, ERD and MYC2, showed higher expression in V3, G41, G935 and B9 rootstocks compared to M26 in response to stress. In contrast, WRKY29 was down-regulated in response to stress in the tolerant rootstocks, and its expression was negatively correlated with ABA content and stomatal closure. Overall, the findings of this study showed that B9, V3 and G41 displayed better osmotic stress tolerance followed by G935 then M26 and B118 rootstocks.


Assuntos
Regulação da Expressão Gênica de Plantas , Malus/genética , Pressão Osmótica , Proteínas de Plantas/genética , Ácido Abscísico/metabolismo , Malus/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo
5.
Int J Mol Sci ; 22(16)2021 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-34445535

RESUMO

Apples (Malus domestica Borkh) are prone to preharvest fruit drop, which is more pronounced in 'Honeycrisp'. Hexanal is known to improve fruit retention in several economically important crops. The effects of hexanal on the fruit retention of 'Honeycrisp' apples were assessed using physiological, biochemical, and transcriptomic approaches. Fruit retention and fruit firmness were significantly improved by hexanal, while sugars and fresh weight did not show a significant change in response to hexanal treatment. At commercial maturity, abscisic acid and melatonin levels were significantly lower in the treated fruit abscission zone (FAZ) compared to control. At this stage, a total of 726 differentially expressed genes (DEGs) were identified between treated and control FAZ. Functional classification of the DEGs showed that hexanal downregulated ethylene biosynthesis genes, such as S-adenosylmethionine synthase (SAM2) and 1-aminocyclopropane-1-carboxylic acid oxidases (ACO3, ACO4, and ACO4-like), while it upregulated the receptor genes ETR2 and ERS1. Genes related to ABA biosynthesis (FDPS and CLE25) were also downregulated. On the contrary, key genes involved in gibberellic acid biosynthesis (GA20OX-like and KO) were upregulated. Further, hexanal downregulated the expression of genes related to cell wall degrading enzymes, such as polygalacturonase (PG1), glucanases (endo-ß-1,4-glucanase), and expansins (EXPA1-like, EXPA6, EXPA8, EXPA10-like, EXPA16-like). Our findings reveal that hexanal reduced the sensitivity of FAZ cells to ethylene and ABA. Simultaneously, hexanal maintained the cell wall integrity of FAZ cells by regulating genes involved in cell wall modifications. Thus, delayed fruit abscission by hexanal is most likely achieved by minimizing ABA through an ethylene-dependent mechanism.


Assuntos
Ácido Abscísico/metabolismo , Aldeídos/farmacologia , Parede Celular/metabolismo , Frutas/crescimento & desenvolvimento , Malus/crescimento & desenvolvimento , Melatonina/metabolismo , Proteínas de Plantas/metabolismo , Frutas/efeitos dos fármacos , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Malus/efeitos dos fármacos , Malus/metabolismo , Proteínas de Plantas/genética
6.
Sci Rep ; 10(1): 18706, 2020 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-33127982

RESUMO

An optimized empirical pseudopotential method (EPM) in conjunction with virtual crystal approximation (VCA) and the compositional disorder effect is used for simulation to extract the electronic material parameters of wurtzite nitride alloys to ensure excellent agreement with the experiments. The proposed direct bandgap results of group-III nitride alloys are also compared with the different density functional theories (DFT) based theoretical results. The model developed in current work, significantly improves the accuracy of calculated band gaps as compared to the ab-initio method based results. The physics of carrier transport in binary and ternary nitride materials is investigated with the help of in-house developed Monte Carlo algorithms for solution of Boltzmann transport equation (BTE) including nonlinear scattering mechanisms. Carrier-carrier scattering mechanisms defined through Coulomb-, piezoelectric-, ionized impurity-, surface roughness-scattering with acoustic and intervalley scatterings, all have been given due consideration in present model. The direct and indirect energy bandgap results have been calibrated with the experimental data and use of symmetric and asymmetric form factors associated with respective materials. The electron mobility results of each binary nitride material have been compared and contrasted with experimental results under appropriate conditions and good agreement has been found between simulated and experimental results.

7.
Eng Life Sci ; 20(3-4): 126-132, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32874176

RESUMO

Plant tissue culture techniques have been used to propagate horticultural crops at a commercial scale for more than three decades. However, due to the high cost it is generally only used for high value crops. To increase production efficiency and make micropropagation viable for a wider range of species, new approaches to address key steps of the process with high labor inputs need to be evaluated. For this study, a two-piece scaffold system was designed, prototyped using 3D printing, and tested to physically hold plants upright thereby facilitating liquid based rooting. This system was evaluated with Malus domestica, Betula lenta, and Musa sp. using static liquid culture as well as rocker based temporary immersion system and compared to rooting in semi-solid based medium as is commonly practiced. Significantly, earlier rooting was observed in all three species in liquid when compared to semi-solid culture system, and plants cultured in liquid on the rocker generally performed better than those in static liquid. In addition to quicker, more uniform rooting, reducing labor requirements, and preventing root damage. This newly designed system is simple, easy to use, will help to improve efficiency, and reduce the cost of micropropagation.

8.
Biomolecules ; 10(9)2020 08 28.
Artigo em Inglês | MEDLINE | ID: mdl-32872300

RESUMO

Thidiazuron (TDZ) is a diphenylurea synthetic herbicide and plant growth regulator used to defoliate cotton crops and to induce regeneration of recalcitrant species in plant tissue culture. In vitro cultures of African violet thin petiole sections are an ideal model system for studies of TDZ-induced morphogenesis. TDZ induces de novo shoot organogenesis at low concentrations and somatic embryogenesis at higher concentrations of exposure. We used an untargeted metabolomics approach to identify metabolites in control and TDZ-treated tissues. Statistical analysis including metabolite clustering, pattern and pathway tools, logical algorithms, synthetic biotransformations and hormonomics identified TDZ-induced changes in metabolism. A total of 18,602 putative metabolites with extracted masses and predicted formulae were identified with 1412 features that were found only in TDZ-treated tissues and 312 that increased in response to TDZ. The monomer of TDZ was not detected intact in the tissues but putative oligomers were found in the database and we hypothesize that these may form by a Diels-Alder reaction. Accumulation oligomers in the tissue may act as a reservoir, slowly releasing the active TDZ monomer over time. Cleavage of the amide bridge released TDZ-metabolites into the tissues including organic nitrogen and sulfur containing compounds. Metabolomics data analysis generated six novel hypotheses that can be summarized as an overall increase in uptake of sugars from the culture media, increase in primary metabolism, redirection of terpene metabolism and mediation of stress metabolism via indoleamine and phenylpropanoid metabolism. Further research into the specific mechanisms hypothesized is likely to unravel the mode of action of TDZ and to provide new insights into the control of plant morphogenesis.


Assuntos
Lamiaceae/fisiologia , Compostos de Fenilureia/farmacologia , Reguladores de Crescimento de Plantas/farmacologia , Tiadiazóis/farmacologia , Metabolômica , Morfogênese , Desenvolvimento Vegetal/efeitos dos fármacos , Reguladores de Crescimento de Plantas/fisiologia , Técnicas de Cultura de Tecidos
9.
PLoS One ; 15(4): e0231741, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32298393

RESUMO

Hill's thistle (Cirsium hillii (Canby) Fernald) is a perennial plant endemic to the Great Lakes region of North America. Hill's thistle is listed as threatened in Ontario and Canada where it is found in globally rare alvar habitats. The main objective of this study was ex-situ conservation of Hill's thistle using in vitro culture techniques and reintroduction of micropropagated plants back to their natural habitat in Bruce Peninsula National Park, Ontario, Canada. Two out of twenty-nine available seeds were successfully germinated under in vitro condition. An efficient micropropagation protocol was optimized with 100% survival during acclimatization of plantlets in the greenhouse. Three hundred micropropagated plants were reintroduced to twelve different sites within Bruce Peninsula National Park in June and July 2017. Plants were monitored for survival, rosette growth, and flowering on all sites from 2017-2019. After four months of planting, 67 to 99% of the plants were alive in different sites and 90 to 99% of them survived over winter. In the following years, shoot regeneration and flowering were observed on most sites. This study further confirms the benefit of plant tissue culture techniques to ensure revival of Hill's thistle ecological biodiversity through the reintroduction of micropropagated plants. This approach consisting of the components of conservation, propagation, and reintroduction (CPR) may potentially serve as a model for saving and enriching other species at risk.


Assuntos
Cirsium/crescimento & desenvolvimento , Conservação dos Recursos Naturais/métodos , Espécies em Perigo de Extinção , Sementes/crescimento & desenvolvimento , Aclimatação , Biodiversidade , Ecossistema , Flores/crescimento & desenvolvimento , Germinação , Great Lakes Region , Herbivoria , Técnicas In Vitro , América do Norte , Ontário , Brotos de Planta/crescimento & desenvolvimento , Estações do Ano
10.
Metabolomics ; 16(5): 62, 2020 04 25.
Artigo em Inglês | MEDLINE | ID: mdl-32335734

RESUMO

INTRODUCTION: Plants respond to changes in their environments through hormonal activation of a physiological cascade that redirects metabolic resources and growth. In filberts (Corylus sp.), chelated iron promotes the growth of new shoots but the mechanism(s) are not understood. OBJECTIVES: To use untargeted metabolomics and hormonomics approaches to generate novel hypotheses for the morphoregulatory role of ferric ethylenediamine-N,N'-di-(ortho-hydroxyphenyl) acetic acid (Fe-EDDHA) in filbert shoot organogenesis in vitro. METHODS: Data were generated using previously optimized standardized untargeted metabolomics protocols with time of flight mass spectrometry. Multivariate statistical tools (principal component and partial least squares discriminant analysis) did not detect significant differences. Discovery tools Significance Analysis of Microarrays (SAM), multiple linear regression analysis, Bayesian analysis, logical algorithms, machine learning, synthetic biotransformations, targeted hormonomics, and online resources including MetaboAnalyst were used. RESULTS: Starch/sucrose metabolism and shikimate pathway metabolites were increased. Dose dependent decreases were found in polyphenol metabolism, specifically ellagic acid and its methylated derivative 3,4,3'-tri-O-methylellagic acid. Hormonomics analysis revealed significant differences in phytohormones and their conjugates. FeEDDHA treatment reduced indole-3-acetic acid, abscisic acid, salicylic acid, jasmonic acid conjugates (JA-Trp, JA-Ile, OH-JA) and dihydrozeatinglucoside in regenerating explants. Serotonin (5HT) was decreased in FeEDDHA-treated regenerating tissues while the related metabolite melatonin was increased. Eight phenolic conjugates of 5HT and eight catabolites were affected by FeEDDHA indicating that metabolism to sequester, deactivate and metabolize 5HT was induced by Fe(III). Tryptophan was metabolized through kynurenine but not anthranilate. CONCLUSION: Seven novel hypotheses were generated to guide future studies to understand the regulatory control(s) of shoot organogenesis.


Assuntos
Corylus/metabolismo , Metabolômica , Brotos de Planta/metabolismo , Corylus/química , Etilenodiaminas/química , Etilenodiaminas/metabolismo , Compostos Férricos/química , Compostos Férricos/metabolismo , Análise Multivariada , Brotos de Planta/química
11.
Planta ; 249(6): 1863-1874, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30859306

RESUMO

MAIN CONCLUSION: This DNA fingerprinting test confirmed 195 unique Corylus sp. accessions that were used to build a reference database for identity verification of unknown hazelnut trees from three locations in Ontario. Hazelnut is one of the most profitable tree nuts worldwide. Development of a hazelnut industry in Ontario is urgently required, but economically important cultivars must be genetically verified first in order to meet industry standards. Traditional methods for cultivar identification are largely trait-based and unreliable. In this study, a multiplexed fingerprinting test was modified to allow for hazelnut cultivar discrimination at the DNA level. Fourteen highly polymorphic SSR markers covering the 11 linkage groups of Corylus genome were PCR amplified in multiplex using fluorescent-labelled primers. PCR conditions and primer physical properties were optimized to generate a clear signal for each locus. The 14 SSRs were used to fingerprint 195 unique Corylus accessions collected from the USDA-NCGR. Fragment sizes were subjected to a UPGMA clustering analysis which separated Corylus accessions based on species and geographic origin. For validation purposes, hazelnut leaves from three locations in Ontario were collected for identity verification using this DNA fingerprinting test. As a result, 33.3% of the unknown trees were duplicates of seven distinct genotypes and a small percentage (8.3%) of these were identical to reference Corylus hybrids. These results reflect common mislabelling issues and genotype duplications that can prevent a uniform plant propagation system. Implementation of this test together with the addition of more unique accessions to the reference database will help verification of trueness-to-type of economically important cultivars for the hazelnut industry.


Assuntos
Corylus/genética , Impressões Digitais de DNA , Bases de Dados de Ácidos Nucleicos , Genoma de Planta/genética , Ligação Genética , Genótipo , Técnicas de Genotipagem , Repetições de Microssatélites/genética , Reação em Cadeia da Polimerase Multiplex , Fenótipo , Filogenia
12.
J Pineal Res ; 66(1): e12527, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30267543

RESUMO

Melatonin and serotonin are important phytochemicals enabling plants to redirect growth in response to environmental stresses. Despite much research on their biosynthetic routes, localization of their biosynthetic enzymes and recent identification of a phytomelatonin receptor, localization of the molecules themselves has to date not been possible. Elucidation of their locations in living tissues can provide an effective tool to facilitate indolamine research across systems including both plants and animals. In this study, we employed a novel technique, quantum dot nanoparticles, to directly visualize melatonin and serotonin in axenic roots. Melatonin was absorbed through epidermal cells, travelled laterally, and accumulated in endodermal and rapidly dividing pericycle cells. Serotonin was absorbed by cells proximal to the crown with rapid polar movement toward the root tip. Thermal stress disrupted localization and dispersed melatonin and serotonin across cells. These data demonstrate the natural movement of melatonin and serotonin in roots directing cell growth and suggest that plants have a mechanism to disperse the indolamines throughout tissues as antioxidants in response to environmental stresses.


Assuntos
Hypericum/metabolismo , Melatonina/metabolismo , Serotonina/metabolismo , Regulação da Expressão Gênica de Plantas , Pontos Quânticos , Estresse Fisiológico
13.
J Pineal Res ; 64(2)2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29149453

RESUMO

Melatonin and serotonin are important signaling and stress mitigating molecules that play important roles across growth and development in plants. Despite many well-documented responses, a systematic investigation of the entire metabolic pathway (tryptophan, tryptamine, and N-acetylserotonin) does not exist, leaving many open questions. The objective of this study was to determine the responses of Hypericum perforatum (L.) to melatonin, serotonin, and their metabolic precursors. Two well-characterized germplasm lines (#4 and 112) created by mutation and a haploid breeding program were compared to wild type to identify specific responses. Germplasm line 4 has lower regenerative and photosynthetic capacity than either wild type or line 112, and there are documented significant differences in the chemistry and physiology of lines 4 and 112. Supplementation of the culture media with tryptophan, tryptamine, N-acetylserotonin, serotonin, or melatonin partially reversed the regenerative recalcitrance and growth impairment of the germplasm lines. Quantification of phytohormones revealed crosstalk between the indoleamines and related phytohormones including cytokinin, salicylic acid, and abscisic acid. We hypothesize that melatonin and serotonin function in coordination with their metabolites in a cascade of phytochemical responses including multiple pathways and phytohormone networks to direct morphogenesis and protect photosynthesis in H. perforatum.


Assuntos
Hypericum/crescimento & desenvolvimento , Hypericum/metabolismo , Melatonina/metabolismo , Desenvolvimento Vegetal/fisiologia , Serotonina/metabolismo , Hypericum/efeitos dos fármacos , Melatonina/farmacologia , Desenvolvimento Vegetal/efeitos dos fármacos , Reguladores de Crescimento de Plantas/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Sementes/crescimento & desenvolvimento , Serotonina/farmacologia , Transdução de Sinais/fisiologia
14.
Funct Plant Biol ; 45(2): 58-69, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32291021

RESUMO

Melatonin is an indoleamine neurotransmitter that has recently become well established as an important multi-functional signalling molecule in plants. These signals have been found to induce several important physiological responses that may be interpreted as behaviours. The diverse processes in which melatonin has been implicated in plants have expanded far beyond the traditional roles for which it has been implicated in mammals, which include sleep, tropisms and reproduction. These functions, however, appear to also be important melatonin mediated processes in plants, though the mechanisms underlying these functions have yet to be fully elucidated. Mediation or redirection of plant physiological processes induced by melatonin can be summarised as a series of behaviours including, among others: herbivore defence, avoidance of undesirable circumstances or attraction to opportune conditions, problem solving and response to environmental stimulus. As the mechanisms of melatonin action are elucidated, its involvement in plant growth, development and behaviour is likely to expand beyond the aspects discussed in this review and hold promise for applications in diverse fundamental and applied plant sciences including conservation, cryopreservation, morphogenesis, industrial agriculture and natural health products.

15.
Plant Signal Behav ; 12(5): e1296997, 2017 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-28448744

RESUMO

American Elms were devastated by an outbreak of Dutch Elm Disease is caused by the fungus Ophiostoma novo-ulmi Brasier that originated in Asia and arrived in the early 1900s. In spite of decades of study, the specific mechanisms and disease resistance in some trees is not well understood. the fungus is spread by several species of bark beetles in the genus Scolytus, during their dispersal and feeding. Our objective was to understand elm responses to beetle feeding in the absence of the fungus to identify potential resistance mechanisms. A colony of Scolytus multistriatus was established from wild-caught beetles and beetles were co-incubated with susceptible or resistant American elm varieties in a controlled environment chamber. Beetles burrowed into the auxillary meristems of the young elm shoots. The trees responded to the beetle damage by a series of spikes in the concentration of plant growth regulating compounds, melatonin, serotonin, and jasmonic acid. Spikes in melatonin and serotonin represented a 7,000-fold increase over resting levels. Spikes in jasmonic acid were about 10-fold higher than resting levels with one very large spike observed. Differences were noted between susceptible and resistant elms that provide new understanding of plant defenses.


Assuntos
Besouros/patogenicidade , Ulmus/metabolismo , Ulmus/parasitologia , Animais , Ciclopentanos/metabolismo , Melatonina/metabolismo , Oxilipinas/metabolismo , Doenças das Plantas/parasitologia , Serotonina/metabolismo , Transdução de Sinais
16.
Plant Methods ; 13: 6, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28115979

RESUMO

BACKGROUND: Due to the complex process of designing and manufacturing new plant tissue culture vessels through conventional means there have been limited efforts to innovate improved designs. Further, development and availability of low cost, energy efficient LEDs of various spectra has made it a promising light source for plant growth in controlled environments. However, direct replacement of conventional lighting sources with LEDs does not address problems with uniformity, spectral control, or the challenges in conducting statistically valid experiments to assess the effects of light. Prototyping using 3D printing and LED based light sources could help overcome these limitations and lead to improved culture systems. RESULTS: A modular culture vessel design in which the fluence rate and spectrum of light are independently controlled was designed, prototyped using 3D printing, and evaluated for plant growth. This design is compatible with semi-solid and liquid based culture systems. Observations on morphology, chlorophyll content, and chlorophyll fluorescence based stress parameters from in vitro plants cultured under different light spectra with similar overall fluence rate indicated different responses in Nicotiana tabacum and Artemisia annua plantlets. This experiment validates the utility of 3D printing to design and test functional vessels and demonstrated that optimal light spectra for in vitro plant growth is species-specific. CONCLUSIONS: 3D printing was successfully used to prototype novel culture vessels with independently controlled variable fluence rate/spectra LED lighting. This system addresses several limitations associated with current lighting systems, providing more uniform lighting and allowing proper replication/randomization for experimental plant biology while increasing energy efficiency. A complete procedure including the design and prototyping of a culture vessel using 3D printing, commercial scale injection molding of the prototype, and conducting a properly replicated experiment are discussed. This open source design has the scope for further improvement and adaptation and demonstrates the power of 3D printing to improve the design of culture systems.

17.
J Clin Sleep Med ; 13(2): 275-281, 2017 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-27855744

RESUMO

STUDY OBJECTIVES: Melatonin is an important neurohormone, which mediates circadian rhythms and the sleep cycle. As such, it is a popular and readily available supplement for the treatment and prevention of sleep-related disorders including insomnia and jet lag. This study quantified melatonin in 30 commercial supplements, comprising different brands and forms and screened supplements for the presence of serotonin. METHODS: A total of 31 supplements were analyzed by ultraperformance liquid chromatography with electrochemical detection for quantification of melatonin and serotonin. Presence of serotonin was confirmed through analysis by ultraperformance liquid chromatography with mass spectrometry detection. RESULTS: Melatonin content was found to range from -83% to +478% of the labelled content. Additionally, lot-to-lot variable within a particular product varied by as much as 465%. This variability did not appear to be correlated with manufacturer or product type. Furthermore, serotonin (5-hydroxytryptamine), a related indoleamine and controlled substance used in the treatment of several neurological disorders, was identified in eight of the supplements at levels of 1 to 75 µg. CONCLUSIONS: Melatonin content did not meet label within a 10% margin of the label claim in more than 71% of supplements and an additional 26% were found to contain serotonin. It is important that clinicians and patients have confidence in the quality of supplements used in the treatment of sleep disorders. To address this, manufacturers require increased controls to ensure melatonin supplements meet both their label claim, and also are free from contaminants, such as serotonin. COMMENTARY: A commentary on this article appears in this issue on page 163.


Assuntos
Produtos Biológicos/análise , Suplementos Nutricionais/análise , Melatonina/análise , Serotonina/análise , Produtos Biológicos/química , Cromatografia Líquida , Técnicas Eletroquímicas , Espectrometria de Massas , Melatonina/química , Serotonina/química
18.
Front Plant Sci ; 7: 1721, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27899931

RESUMO

Despite growing evidence of the importance of melatonin and serotonin in the plant life, there is still much debate over the stability of melatonin, with extraction and analysis methods varying greatly from lab to lab with respect to time, temperature, light levels, extraction solvents, and mechanical disruption. The variability in methodology has created conflicting results that confound the comparison of studies to determine the role of melatonin in plant physiology. We here describe a fully validated method for the quantification of melatonin, serotonin and their biosynthetic precursors: tryptophan, tryptamine and N-acetylserotonin by liquid chromatography single quadrupole mass spectrometry (LC-MS) in diverse plant species and tissues. This method can be performed on a simple and inexpensive platform, and is both rapid and simple to implement. The method has excellent reproducibility and acceptable sensitivity with percent relative standard deviation (%RSD) in all matrices between 1 and 10% and recovery values of 82-113% for all analytes. Instrument detection limits were 24.4 ng/mL, 6.10 ng/mL, 1.52 ng/mL, 6.10 ng/mL, and 95.3 pg/mL, for serotonin, tryptophan, tryptamine, N-acetylserotonin and melatonin respectively. Method detection limits were 1.62 µg/g, 0.407 µg/g, 0.101 µg/g, 0.407 µg/g, and 6.17 ng/g respectively. The optimized method was then utilized to examine the issue of variable stability of melatonin in plant tissue culture systems. Media composition (Murashige and Skoog, Driver and Kuniyuki walnut or Lloyd and McCown's woody plant medium) and light (16 h photoperiod or dark) were found to have no effect on melatonin or serotonin content. A Youden trial suggested temperature as a major factor leading to degradation of melatonin. Both melatonin and serotonin appeared to be stable across the first 10 days in media, melatonin losses reached a mean minimum degradation at 28 days of approximately 90%; serotonin reached a mean minimum value of approximately 60% at 28 days. These results suggest that melatonin and serotonin show considerable stability in plant systems and these indoleamines and related compounds can be used for investigations that span over 3 weeks.

19.
Biotechnol Adv ; 34(8): 1347-1361, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27742596

RESUMO

Serotonin is an ancient indoleamine that was presumably part of the life cycle of the first prokaryotic life forms on Earth millions of years ago where it functioned as a powerful antioxidant to combat the increasingly oxygen rich atmosphere. First identified as a neurotransmitter signaling molecule in mammals, it is ubiquitous across all forms of life. Serotonin was discovered in plants many years after its discovery in mammals; however, it has now been confirmed in almost all plant families, where it plays important roles in plant growth and development, including functions in energy acquisition, seasonal cycles, modulation of reproductive development, control of root and shoot organogenesis, maintenance of plant tissues, delay of senescence, and responses to biotic and abiotic stresses. Despite its widespread presence and activity, there are many questions which remain unanswered about the role of serotonin in plants including the mode of signaling and receptor identity as well as the mechanisms of action of this important molecule. This review provides an overview of the role of serotonin in plant life and their ability to adapt.


Assuntos
Desenvolvimento Vegetal/fisiologia , Serotonina , Plantas/embriologia , Sementes/crescimento & desenvolvimento , Sementes/fisiologia , Serotonina/metabolismo , Serotonina/fisiologia , Transdução de Sinais/fisiologia
20.
Plant Cell Rep ; 35(1): 91-102, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26400684

RESUMO

KEY MESSAGE: Essential oils have growth regulating properties comparable to the well-documented methyl jasmonate and may be involved in localized and/or airborne plant communication. Aromatic plants employ large amounts of resources to produce essential oils. Some essential oils are known to contain compounds with plant growth regulating activities. However, the potential capacity of essential oils as airborne molecules able to modulate plant growth/development has remained uninvestigated. Here, we demonstrate that essential oils from eight taxonomically diverse plants applied in their airborne state inhibited auxin-induced elongation of Pisum sativum hypocotyls and Avena sativa coleoptiles. This response was also observed using five monoterpenes commonly found in essential oils as well as isoprene, the basic building block of terpenes. Upon transfer to ambient conditions, A. sativa coleoptiles resumed elongation, demonstrating an antagonistic relationship rather than toxicity. Inclusion of essential oils, monoterpenes, or isoprene into the headspace of culture vessels induced abnormal cellular growth along hypocotyls of Arabidopsis thaliana. These responses were also elicited by methyl jasmonate (MeJA); however, where methyl jasmonate inhibited root growth essential oils did not. Gene expression studies in A. thaliana also demonstrated differences between the MeJA and isoprenoid responses. This series of experiments clearly demonstrate that essential oils and their isoprenoid components interact with endogenous plant growth regulators when applied directly or as volatile components in the headspace. The similarities between isoprenoid and MeJA responses suggest that they may act in plant defence signalling. While further studies are needed to determine the ecological and evolutionary significance, the results of this study and the specialized anatomy associated with aromatic plants suggest that essential oils may act as airborne signalling molecules.


Assuntos
Arabidopsis/efeitos dos fármacos , Avena/efeitos dos fármacos , Cuminum/química , Óleos Voláteis/farmacologia , Ervilhas/efeitos dos fármacos , Óleos Vegetais/farmacologia , Acetatos/farmacologia , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Avena/genética , Avena/crescimento & desenvolvimento , Butadienos/farmacologia , Ciclopentanos/farmacologia , Regulação da Expressão Gênica de Plantas , Hemiterpenos/farmacologia , Hipocótilo/efeitos dos fármacos , Hipocótilo/genética , Hipocótilo/crescimento & desenvolvimento , Ácidos Indolacéticos/metabolismo , Oxilipinas/farmacologia , Ervilhas/genética , Ervilhas/crescimento & desenvolvimento , Pentanos/farmacologia , Reguladores de Crescimento de Plantas/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Transdução de Sinais/efeitos dos fármacos , Terpenos/farmacologia
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