A number of scientific reports published to date contain data on endogenous levels of various phytohormones in potato (Solanum tuberosum L.) but a complete cytokinin profile of potato tissues, that would include data on all particular molecular forms of cytokinin, has still been missing. In this work, endogenous levels of all analytically detectable isoprenoid cytokinins, as well as the auxin indole-3-acetic acid (IAA), and abscisic acid (ABA) have been determined in shoots and roots of 30 day old in vitro grown potato (cv. Désirée). The results presented here are generally similar to other data reported for in vitro grown potato plants, whereas greenhouse-grown plants typically contain lower levels of ABA, possibly indicating that in vitro grown potato is exposed to chronic stress. Cytokinin N-glucosides, particularly N7-glucosides, are the dominant cytokinin forms in both shoots and roots of potato, whereas nucleobases, as the bioactive forms of cytokinins, comprise a low proportion of cytokinin levels in tissues of potato. Differences in phytohormone composition between shoots and roots of potato suggest specific patterns of transport and/or differences in tissue-specific metabolism of plant hormones. These results represent a contribution to understanding the hormonomics of potato, a crop species of extraordinary economic importance.
- MeSH
- cytokininy analýza metabolismus MeSH
- fyziologický stres MeSH
- kořeny rostlin metabolismus MeSH
- kyselina abscisová analýza metabolismus MeSH
- kyseliny indoloctové analýza metabolismus MeSH
- regulátory růstu rostlin analýza metabolismus MeSH
- Solanum tuberosum růst a vývoj metabolismus MeSH
- tandemová hmotnostní spektrometrie MeSH
- výhonky rostlin metabolismus MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Hormonal dynamics after Plasmodiophora brassicae infection were compared in two Brassica napus cultivars-more resistant SY Alister and more sensitive Hornet, in order to elucidate responses associated with efficient defense. Both cultivars responded to infection by the early transient elevation of active cytokinins (predominantly cis-zeatin) and auxin indole-3-acetic acid (IAA) in leaves and roots, which was longer in Hornet. Moderate IAA levels in Hornet roots coincided with a high expression of biosynthetic gene nitrilaseNIT1 (contrary to TAA1, YUC8, YUC9). Alister had a higher basal level of salicylic acid (SA), and it stimulated its production (via the expression of isochorismate synthase (ICS1)) in roots earlier than Hornet. Gall formation stimulated cytokinin, auxin, and SA levels-with a maximum 22 days after inoculation (dai). SA marker gene PR1 expression was the most profound at the time point where gall formation began, in leaves, roots, and especially in galls. Jasmonic acid (JA) was higher in Hornet than in Alister during the whole experiment. To investigate SA and JA function, SA was applied before infection, and twice (before infection and 15 dai), and JA at 15 dai. Double SA application diminished gall formation in Alister, and JA promoted gall formation in both cultivars. Activation of SA/JA pathways reflects the main differences in clubroot resistance.
- MeSH
- aminohydrolasy genetika MeSH
- Brassica napus růst a vývoj metabolismus parazitologie MeSH
- cyklopentany analýza MeSH
- cytokininy analýza MeSH
- intramolekulární transferasy genetika MeSH
- kořeny rostlin růst a vývoj metabolismus parazitologie MeSH
- kyseliny indoloctové analýza MeSH
- listy rostlin růst a vývoj metabolismus parazitologie MeSH
- nemoci rostlin parazitologie MeSH
- odolnost vůči nemocem MeSH
- oxylipiny analýza MeSH
- Plasmodiophorida patogenita MeSH
- regulace genové exprese u rostlin MeSH
- regulátory růstu rostlin analýza MeSH
- rostlinné proteiny genetika MeSH
- vývojová regulace genové exprese MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
BACKGROUND AND AIMS: The metabolism of cytokinins (CKs) and auxins in vascular plants is relatively well understood, but data concerning their metabolic pathways in non-vascular plants are still rather rare. With the aim of filling this gap, 20 representatives of taxonomically major lineages of cyanobacteria and algae from Cyanophyceae, Xanthophyceae, Eustigmatophyceae, Porphyridiophyceae, Chlorophyceae, Ulvophyceae, Trebouxiophyceae, Zygnematophyceae and Klebsormidiophyceae were analysed for endogenous profiles of CKs and auxins and some of them were used for studies of the metabolic fate of exogenously applied radiolabelled CK, [(3)H]trans-zeatin (transZ) and auxin ([(3)H]indole-3-acetic acid (IAA)), and the dynamics of endogenous CK and auxin pools during algal growth and cell division. METHODS: Quantification of phytohormone levels was performed by high-performance or ultrahigh-performance liquid chromatography-electrospray tandem mass spectrometry (HPLC-MS/MS, UHPLC-MS/MS). The dynamics of exogenously applied [(3)H]transZ and [(3)H]IAA in cell cultures were monitored by HPLC with on-line radioactivity detection. KEY RESULTS: The comprehensive screen of selected cyanobacteria and algae for endogenous CKs revealed a predominance of bioactive and phosphate CK forms while O- and N-glucosides evidently did not contribute greatly to the total CK pool. The abundance of cis-zeatin-type CKs and occurrence of CK 2-methylthio derivatives pointed to the tRNA pathway as a substantial source of CKs. The importance of the tRNA biosynthetic pathway was proved by the detection of tRNA-bound CKs during the course of Scenedesmus obliquus growth. Among auxins, free IAA and its oxidation catabolite 2-oxindole-3-acetic acid represented the prevailing endogenous forms. After treatment with [(3)H]IAA, IAA-aspartate and indole-3-acetyl-1-glucosyl ester were detected as major auxin metabolites. Moreover, different dynamics of endogenous CKs and auxin profiles during S. obliquus culture clearly demonstrated diverse roles of both phytohormones in algal growth and cell division. CONCLUSIONS: Our data suggest the existence and functioning of a complex network of metabolic pathways and activity control of CKs and auxins in cyanobacteria and algae that apparently differ from those in vascular plants.
- MeSH
- Chlorophyta chemie metabolismus fyziologie MeSH
- cytokininy analýza metabolismus MeSH
- fylogeneze MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací metody MeSH
- homeostáza fyziologie MeSH
- kyseliny indoloctové analýza metabolismus MeSH
- regulátory růstu rostlin analýza metabolismus MeSH
- sinice chemie metabolismus fyziologie MeSH
- Streptophyta chemie metabolismus fyziologie MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
Parallel determination of auxin and cytokinin levels within plant organs and tissues represents an invaluable tool for studies of their physiological effects and mutual interactions. Thanks to their different chemical structures, auxins, cytokinins and their metabolites are often determined separately, using specialized procedures of sample purification, extraction, and quantification. However, recent progress in the sensitivity of analytical methods of liquid chromatography coupled to mass spectrometry (LC-MS) allows parallel analysis of multiple compounds. Here we describe a method that is based on single step purification protocol followed by LC-MS separation and detection for parallel analysis of auxins, cytokinins and their metabolites in various plant tissues and cell cultures.
- MeSH
- chromatografie kapalinová MeSH
- cytokininy analýza chemie izolace a purifikace MeSH
- kyseliny indoloctové analýza chemie izolace a purifikace MeSH
- metabolomika metody MeSH
- regulátory růstu rostlin analýza MeSH
- rostlinné extrakty analýza chemie izolace a purifikace MeSH
- tandemová hmotnostní spektrometrie MeSH
- Publikační typ
- časopisecké články MeSH
KEY MESSAGE: The current evidence of regulatory effect of smoke-water (SW) and karrikinolide (KAR(1)) on the concentrations of endogenous cytokinins in plants partly explain the basis for their growth stimulatory activity. Karrikinolide (KAR1) which is derived from smoke-water (SW) is involved in some physiological aspects in the life-cycle of plants. This suggests a potential influence on the endogenous pool (quantity and quality) of phytohormones such as cytokinins (CKs). In the current study, the effect of SW (1:500; 1:1000; 1:1500 v/v dilutions) and KAR1 (10(-7); 10(-8); 10(-9) M) applied during micropropagation of Eucomis autumnalis subspecies autumnalis on the ex vitro growth and CKs after 4 months post-flask duration was evaluated. The interactions of SW and KAR(1) with benzyladenine (BA), α-naphthaleneacetic acid (NAA) or BA+NAA were also assessed. Plants treated with SW (1:500) and KAR1 (10(-8) M) demonstrated superior growth in terms of the rooting, leaf and bulb sizes and fresh biomass than the control and plants treated with BA and BA+NAA. However, plant growth was generally inhibited with either SW (1:500) or KAR1 (10(-8) M) and BA when compared to BA (alone) treatment. Relative to NAA treatment, the presence of KAR(1) (10(-7) M) with NAA significantly increased the leaf area and fresh biomass. Both SW and KAR1-treated plants accumulated more total CKs, mainly isoprenoid-type than the control and NAA-treated plants. The highest CK content was also accumulated in SW (1:500) with BA+NAA treatments. Similar stimulatory effects were observed with increasing concentrations of KAR(1) and BA. The current findings establish that SW and KAR1 exert significant influence on the endogenous CK pools. However, the better growth of plants treated with SW and KAR1 treatments was not exclusively related to the endogenous CKs.
- MeSH
- aklimatizace * MeSH
- Asparagaceae fyziologie růst a vývoj účinky léků MeSH
- biomasa MeSH
- cytokininy analýza metabolismus MeSH
- furany farmakologie MeSH
- kořeny rostlin účinky léků růst a vývoj fyziologie MeSH
- kouř MeSH
- kyseliny naftalenoctové metabolismus MeSH
- léčivé rostliny MeSH
- lipnicovité MeSH
- listy rostlin účinky léků růst a vývoj fyziologie MeSH
- pyrany farmakologie MeSH
- regulátory růstu rostlin analýza metabolismus MeSH
- voda chemie MeSH
- výhonky rostlin účinky léků růst a vývoj fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Plant beneficial microbes mediate biocontrol of diseases by interfering with pathogens or via strengthening the host. Although phytohormones, including cytokinins, are known to regulate plant development and physiology as well as plant immunity, their production by microorganisms has not been considered as a biocontrol mechanism. Here we identify the ability of Pseudomonas fluorescens G20-18 to efficiently control P. syringae infection in Arabidopsis, allowing maintenance of tissue integrity and ultimately biomass yield. Microbial cytokinin production was identified as a key determinant for this biocontrol effect on the hemibiotrophic bacterial pathogen. While cytokinin-deficient loss-of-function mutants of G20-18 exhibit impaired biocontrol, functional complementation with cytokinin biosynthetic genes restores cytokinin-mediated biocontrol, which is correlated with differential cytokinin levels in planta. Arabidopsis mutant analyses revealed the necessity of functional plant cytokinin perception and salicylic acid-dependent defence signalling for this biocontrol mechanism. These results demonstrate microbial cytokinin production as a novel microbe-based, hormone-mediated concept of biocontrol. This mechanism provides a basis to potentially develop novel, integrated plant protection strategies combining promotion of growth, a favourable physiological status and activation of fine-tuned direct defence and abiotic stress resilience.
- MeSH
- Arabidopsis mikrobiologie MeSH
- cytokininy analýza biosyntéza farmakologie MeSH
- kyselina salicylová farmakologie MeSH
- listy rostlin mikrobiologie MeSH
- nemoci rostlin mikrobiologie MeSH
- Pseudomonas fluorescens metabolismus MeSH
- Pseudomonas syringae účinky léků růst a vývoj patogenita MeSH
- regulátory růstu rostlin farmakologie MeSH
- tandemová hmotnostní spektrometrie MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Bryophytes represent a very diverse group of non-vascular plants such as mosses, liverworts and hornworts and the oldest extant lineage of land plants. Determination of endogenous phytohormone profiles in bryophytes can provide substantial information about early land plant evolution. In this study, we screened thirty bryophyte species including six liverworts and twenty-four mosses for their phytohormone profiles in order to relate the hormonome with phylogeny in the plant kingdom. METHODOLOGY: Samples belonging to nine orders (Pelliales, Jungermanniales, Porellales, Sphagnales, Tetraphidales, Polytrichales, Dicranales, Bryales, Hypnales) were collected in Central and Northern Bohemia. The phytohormone content was analysed with a high performance liquid chromatography electrospray tandem-mass spectrometry (HPLC-ESI-MS/MS). PRINCIPAL FINDINGS: As revealed for growth hormones, some common traits such as weak conjugation of both cytokinins and auxins, intensive production of cisZ-type cytokinins and strong oxidative degradation of auxins with abundance of a major primary catabolite 2-oxindole-3-acetic acid were pronounced in all bryophytes. Whereas apparent dissimilarities in growth hormones profiles between liverworts and mosses were evident, no obvious trends in stress hormone levels (abscisic acid, jasmonic acid, salicylic acid) were found with respect to the phylogeny. CONCLUSION: The apparent differences in conjugation and/or degradation strategies of growth hormones between liverworts and mosses might potentially show a hidden link between vascular plants and liverworts. On the other hand, the complement of stress hormones in bryophytes probably correlate rather with prevailing environmental conditions and plant survival strategy than with plant evolution.
- MeSH
- Bryophyta klasifikace metabolismus MeSH
- cyklopentany analýza metabolismus MeSH
- cytokininy analýza metabolismus MeSH
- fylogeneze MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- kyselina abscisová analýza metabolismus MeSH
- kyselina salicylová analýza metabolismus MeSH
- kyseliny indoloctové analýza metabolismus MeSH
- oxylipiny analýza metabolismus MeSH
- regulátory růstu rostlin analýza metabolismus MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Celiac is a chronic hereditary intestinal disorder caused by autoimmune stimulation and injury of intestinal mucosa and different studies have reported an increase of inflammatory cytokines production in these patients. This study aimed to investigate the serum level of inflammatory cytokines and their correlations with the severity of histological variations among celiac patients. Material and methods: By a cross sectional study all celiac patients attending to gastrointestinal clinics in Ilam city during Oct. 2011- MAR. 2013 were investigated. Demographic data was collected using a validate questionnaire and patient's interview and each patient was taken a blood sample for laboratory investigation of tTG-IgA antibody via ELISA test and the serum level of hs-CRP, IL-6 and TNF-a was measured. All patients underwent endoscopic evaluation and biopsy samples were evaluated according to Marsh classification and correlation between severity of histological variations and serum levels of inflammatory cytokines was measured. Results: Totally 274 patients including 90 male and 184 female with a mean age of 28.8±14.4 years were evaluated. The mean serum levels of hs-CRP, IL-6 and TNF-a were 7.8±3.1 mg/l, 2.6±0.9 pg/ml and 6.9±4.6 pg/ml respectively (p=0.599, p=0.629 and p= 0.651). Hs-CRP showed a significant relationship with the severity of mucosal damage upon class I of Marsh classification (p=0.01); however, IL-6 showed a significant level among patients with Marsh I, II and IIIC (p=0.000, p=0.02 and p=0.000) but not in those with Marsh class IIIA and IIIB. Conclusion: There was a positive relationship between mean serum levels of IL-6 and TNF-a with increasing the Marsh classification from class I to class IIIC and the highest rate of IL-6 and TNF-a was shown in Marsh IIIC but the serum level of hs-CRP was not increased with Marsh class increasing. It was concluded that as the Marsh classification increased from class I to class IIIC, and/or the severity of pathologic variations was increased, the mean serum levels of inflammatory markers were increased too.
- Klíčová slova
- predikce celiakie, hs-CRP, Marsh klasifikace, sérové hladiny zánětlivých cytokinů,
- MeSH
- biopsie MeSH
- celiakie * diagnóza dietoterapie genetika imunologie MeSH
- cytokininy * analýza škodlivé účinky MeSH
- dospělí MeSH
- histologie srovnávací MeSH
- interleukin-6 analýza krev MeSH
- klinické laboratorní techniky metody využití MeSH
- lidé MeSH
- mladiství MeSH
- průřezové studie klasifikace metody statistika a číselné údaje využití MeSH
- sérologické testy * metody využití MeSH
- TNF-alfa analýza krev MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
We have developed a N6-dimethylallyladenine (cytokinin) dehydrogenase-based microbiosensor for real-time determination of the family of hormones known as cytokinins. Cytokinin dehydrogenase from Zea mays (ZmCKX1) was immobilised concurrently with electrodeposition of a silica gel film on the surface of a Pt microelectrode, which was further functionalized by free electron mediator 2,6-dichlorophenolindophenol (DCPIP) in supporting electrolyte to give a bioactive film capable of selective oxidative cleavage of the N6- side chain of cytokinins. The rapid electron shuffling between freely diffusible DCPIP and the FAD redox group in ZmCKX1 endowed the microbiosensor with a fast response time of less than 10 s. The immobilised ZmCKX1 retained a high affinity for its preferred substrate N6-(Δ2-isopentenyl) adenine (iP), and gave the miniaturized biosensor a large linear dynamic range from 10 nM to 10 µM, a detection limit of 3.9 nM and a high sensitivity to iP of 603.3 µAmM-1cm-2 (n = 4, R2 = 0.9999). Excellent selectivity was displayed for several other aliphatic cytokinins and their ribosides, including N6-(Δ2-isopentenyl) adenine, N6-(Δ2-isopentenyl) adenosine, cis-zeatin, trans-zeatin and trans-zeatin riboside. Aromatic cytokinins and metabolites such as cytokinin glucosides were generally poor substrates. The microbiosensors exhibited excellent stability in terms of pH and long-term storage and have been used successfully to determine low nanomolar cytokinin concentrations in tomato xylem sap exudates.
- MeSH
- biosenzitivní techniky metody MeSH
- cytokininy analýza metabolismus MeSH
- enzymy imobilizované chemie metabolismus MeSH
- isopentenyladenosin analogy a deriváty analýza metabolismus MeSH
- kukuřice setá enzymologie MeSH
- limita detekce MeSH
- oxidoreduktasy chemie metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Phytohormones are key regulators in various physiological processes of plant growth and development. Their chemical analyses, together with their genomics and proteomics, are an integral part of plant development studies. The information on hormone levels is often valuable for biologists dealing with any hormone-regulated processes. Plant tissue is a complex multicomponent mixture containing phytohormones in minute quantities (pg or ng/g fresh weight) along with many other related compounds with similar structures and/or physicochemical properties. Therefore, their analysis requires rapid, sensitive and sufficiently selective analytical methods. The significance of solid-phase extraction for purification and hyphenated techniques such as GC-MS, LC-MS and CE-MS for routine analyses of phytohormones is discussed.
- MeSH
- Brassica MeSH
- chemické techniky analytické MeSH
- chromatografie kapalinová MeSH
- cyklopentany analýza MeSH
- cytokininy analýza MeSH
- elektroforéza kapilární MeSH
- financování organizované MeSH
- fyziologie rostlin MeSH
- gibereliny analýza MeSH
- kyselina abscisová analýza MeSH
- kyseliny indoloctové analýza MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- rostliny MeSH
- steroidy MeSH
