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
The methodology described here represents an improved strategy for analysis of a broad range of stress-related plant hormones including jasmonates, salicylic acid, abscisic acid, and auxin metabolites. The method conditions are optimized in order to reduce the background effect of complicated plant matrix, allow effective preconcentration and thus perform highly sensitive profiling of multiple plant hormones by ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS).
- MeSH
- cyklopentany analýza MeSH
- kyselina abscisová analýza MeSH
- kyselina salicylová analýza MeSH
- kyseliny indoloctové analýza MeSH
- oxylipiny analýza MeSH
- regulátory růstu rostlin analýza MeSH
- tandemová hmotnostní spektrometrie metody MeSH
- vysokoúčinná kapalinová chromatografie metody 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
Stress-induced changes in phytohormone metabolite profiles have rapid effects on plant metabolic activity and growth. The jasmonates (JAs) are a group of fatty acid-derived stress response regulators with roles in numerous developmental processes. To elucidate their dual regulatory effects, which overlap with those of other important defence-signalling plant hormones such as salicylic acid (SA), abscisic acid (ABA) and indole-3-acetic acid (IAA), we have developed a highly efficient single-step clean-up procedure for their enrichment from complex plant matrices that enables their sensitive quantitative analysis using hyphenated mass spectrometry technique. The rapid extraction of minute quantities of plant material (less than 20mg fresh weight, FW) into cold 10% methanol followed by one-step reversed-phase polymer-based solid phase extraction significantly reduced matrix effects and increased the recovery of labile JA analytes. This extraction and purification protocol was paired with a highly sensitive and validated ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method and used to simultaneously profile sixteen stress-induced phytohormones in minute plant material samples, including endogenous JA, several of its biosynthetic precursors and derivatives, as well as SA, ABA and IAA.
- MeSH
- chromatografie kapalinová MeSH
- cyklopentany analýza chemie MeSH
- extrakce na pevné fázi MeSH
- fyziologický stres MeSH
- kyselina abscisová analýza chemie MeSH
- kyselina salicylová analýza chemie MeSH
- kyseliny indoloctové analýza MeSH
- listy rostlin chemie MeSH
- mastné kyseliny analýza chemie MeSH
- molekulární struktura MeSH
- oxylipiny analýza chemie MeSH
- regulátory růstu rostlin chemie izolace a purifikace fyziologie MeSH
- tandemová hmotnostní spektrometrie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
