Reduced height (Rht) genes have revolutionised wheat cultivation, but they can compromise freezing tolerance, and only a few alleles are in use. Thus, evaluating the role of other Rht alleles in stress responses is crucial. Far-red supplementation of white light (W+FR) can induce pre-hardening in cereals at 15°C. However, the relevant effect of blue light enrichment (W+B) is poorly described. This study investigates the influence of W+FR or W+B exposure in young winter wheat leaves of a tall (wild-type, rht12) and a dwarf, gibberellin-deficient (near-isogenic line, Rht12) genotype in cv. Maris Huntsman background over 10 days at 15°C. The main objectives were to investigate the relationship between light quality, gibberellin homeostasis, and freezing tolerance. Key parameters such as frost injury, hormonal pools and the expression of relevant genes were examined. Results provided evidence about the involvement of Rht alleles in the basal freezing tolerance of wheat leaves from the side of gibberellin availability. It was revealed that W+FR and W+B treatments partially rescued the freezing-sensitive phenotype of Rht12 leaves, suggesting a potential compensatory mechanism. Analysis of gibberellic acid (GA) metabolism indicated differential responses to light treatments between the Rht12 and wild-type leaves, with implications for freezing tolerance. Moreover, alterations in hormone levels, including jasmonic acid (JA) and salicylic acid (SA), were observed, highlighting the complex interplay between light signalling and hormonal regulation in wheat. Overall, these findings suggest that manipulating light responses may offer a strategy to enhance freezing tolerance in gibberellin-deficient dwarf wheat genotypes.

• MeSH
• červené světlo MeSH
• cyklopentany MeSH
• genotyp MeSH
• gibereliny metabolismus   MeSH
• listy rostlin * účinky záření   fyziologie   metabolismus   MeSH
• oxylipiny MeSH
• pšenice * fyziologie   účinky záření   genetika   metabolismus   MeSH
• regulace genové exprese u rostlin účinky záření   MeSH
• regulátory růstu rostlin * metabolismus   MeSH
• světlo * MeSH
• zmrazování MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cyklopentany MeSH
• gibereliny MeSH
• jasmonic acid MeSH Prohlížeč
• oxylipiny MeSH
• regulátory růstu rostlin * MeSH

Two papers with quite different objectives established protocols that proved pivotal for future work on the role of gibberellins in seed germination. In their paper published in 1967, Russell Jones and Joseph Varner (Planta 72: 155-161) developed a bioassay based on induction of α-amylase activity in barley embryo-less half-seeds that was specific for bioactive gibberellins. The induction of α-amylase in the aleurone of barley and other cereals was to become the experimental system of choice to study gibberellin signalling. However, despite much progress in identifying the molecular events linking gibberellin action and α-amylase gene expression, in many cases their role in the process is still unclear. In 1987, Steven Groot and Cees Karssen (Planta 171:525-531) showed that germination of tomato seeds was limited by the ability of the radicle to penetrate the surrounding layers, with the endosperm forming the major barrier. They used a modified needle attached to a tensiometer to measure the force required to break through the endosperm. While in wild-type seeds, a factor from the embryo, assumed to be gibberellin, promoted breakdown of the endosperm, gibberellin-deficient seeds required an external supply of the hormone to weaken the endosperm or for it to be mechanically disrupted for germination to occur. The paradigm of seed germination being physically restricted by surrounding layers and the role of gibberellin in weakening these tissues has been confirmed in many eudicot species. Gibberellin signalling induces the production of cell-wall loosening enzymes in the micropylar endosperm adjacent to the radicle, but it is unclear whether or not this is a direct response. In both eudicot and monocot systems, there is still much to learn about the role of gibberellin signalling in germination.

• Klíčová slova
• Aleurone, Bioassays, Endosperm, Germination, Gibberellin action, α-amylase,
• MeSH
• alfa-amylasy * metabolismus   genetika   MeSH
• endosperm * fyziologie   metabolismus   MeSH
• gibereliny * metabolismus   MeSH
• ječmen (rod) enzymologie   genetika   MeSH
• klíčení fyziologie   MeSH
• regulace genové exprese u rostlin MeSH
• regulátory růstu rostlin metabolismus   MeSH
• semena rostlinná * fyziologie   enzymologie   MeSH
• vegetační klid * fyziologie   MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• alfa-amylasy * MeSH
• gibereliny * MeSH
• regulátory růstu rostlin MeSH

Plants have evolved several strategies to cope with the ever-changing environment. One example of this is given by seed germination, which must occur when environmental conditions are suitable for plant life. In the model system Arabidopsis thaliana seed germination is induced by light; however, in nature, seeds of several plant species can germinate regardless of this stimulus. While the molecular mechanisms underlying light-induced seed germination are well understood, those governing germination in the dark are still vague, mostly due to the lack of suitable model systems. Here, we employ Cardamine hirsuta, a close relative of Arabidopsis, as a powerful model system to uncover the molecular mechanisms underlying light-independent germination. By comparing Cardamine and Arabidopsis, we show that maintenance of the pro-germination hormone gibberellin (GA) levels prompt Cardamine seeds to germinate under both dark and light conditions. Using genetic and molecular biology experiments, we show that the Cardamine DOF transcriptional repressor DOF AFFECTING GERMINATION 1 (ChDAG1), homologous to the Arabidopsis transcription factor DAG1, is involved in this process functioning to mitigate GA levels by negatively regulating GA biosynthetic genes ChGA3OX1 and ChGA3OX2, independently of light conditions. We also demonstrate that this mechanism is likely conserved in other Brassicaceae species capable of germinating in dark conditions, such as Lepidium sativum and Camelina sativa. Our data support Cardamine as a new model system suitable for studying light-independent germination studies. Exploiting this system, we have also resolved a long-standing question about the mechanisms controlling light-independent germination in plants, opening new frontiers for future research.

• Klíčová slova
• Cardamine hirsuta, DOF AFFECTING GERMINATION1, gibberellins, light, seed germination,
• MeSH
• Arabidopsis genetika   metabolismus   MeSH
• Cardamine * genetika   metabolismus   fyziologie   účinky záření   růst a vývoj   MeSH
• gibereliny * metabolismus   MeSH
• klíčení * účinky záření   genetika   fyziologie   MeSH
• proteiny huseníčku metabolismus   genetika   MeSH
• regulace genové exprese u rostlin MeSH
• semena rostlinná * růst a vývoj   genetika   účinky záření   metabolismus   MeSH
• světlo MeSH
• transkripční faktory metabolismus   genetika   MeSH
• zpětná vazba fyziologická MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• gibereliny * MeSH
• proteiny huseníčku MeSH
• transkripční faktory MeSH

BACKGROUND: Semi-dwarfing alleles are used widely in cereals to confer improved lodging resistance and assimilate partitioning. The most widely deployed semi-dwarfing alleles in rice and barley encode the gibberellin (GA)-biosynthetic enzyme GA 20-OXIDASE2 (GA20OX2). The hexaploid wheat genome carries three homoeologous copies of GA20OX2, and because of functional redundancy, loss-of-function alleles of a single homoeologue would not be selected in wheat breeding programmes. Instead, approximately 70% of wheat cultivars carry gain-of-function mutations in REDUCED HEIGHT 1 (RHT1) genes that encode negative growth regulators and are degraded in response to GA. Semi-dwarf Rht-B1b or Rht-D1b alleles encode proteins that are insensitive to GA-mediated degradation. However, because RHT1 is expressed ubiquitously these alleles have pleiotropic effects that confer undesirable traits in some environments. RESULTS: We have applied reverse genetics to combine loss-of-function alleles in all three homoeologues of wheat GA20OX2 and its paralogue GA20OX1 and evaluated their performance in three years of field trials. ga20ox1 mutants exhibited a mild height reduction (approximately 3%) suggesting GA20OX1 plays a minor role in stem elongation in wheat. ga20ox2 mutants have reduced GA1 content and are 12-32% shorter than their wild-type segregants, comparable to the effect of the Rht-D1b 'Green Revolution' allele. The ga20ox2 mutants showed no significant negative effects on yield components in the spring wheat variety 'Cadenza'. CONCLUSIONS: Our study demonstrates that chemical mutagenesis can expand genetic variation in polyploid crops to uncover novel alleles despite the difficulty in identifying appropriate mutations for some target genes and the negative effects of background mutations. Field experiments demonstrate that mutations in GA20OX2 reduce height in wheat, but it will be necessary to evaluate the effect of these alleles in different genetic backgrounds and environments to determine their value in wheat breeding as alternative semi-dwarfing alleles.

• Klíčová slova
• Dwarfing alleles, Gibberellin, Green revolution, TILLING, Wheat,
• MeSH
• alely MeSH
• fenotyp * MeSH
• gibereliny metabolismus   MeSH
• mutace MeSH
• oxygenasy se smíšenou funkcí genetika   metabolismus   MeSH
• pšenice * genetika   MeSH
• rostlinné geny MeSH
• rostlinné proteiny * genetika   metabolismus   MeSH
• rýže (rod) genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• gibberellin, 2-oxoglutarate-oxygen oxidoreductase (20-hydroxylating, oxidizing) MeSH Prohlížeč
• gibereliny MeSH
• oxygenasy se smíšenou funkcí MeSH
• rostlinné proteiny * MeSH

In situ separation and visualization of synthetic and naturally occurring isomers from heterogeneous plant tissues, especially when they share similar molecular structures, are a challenging task. In this study, we combined the ion mobility separation with desorption electrospray ionization mass spectrometry imaging (DESI-IM-MSI) to achieve a direct separation and visualization of two synthetic auxin derivatives, auxinole and its structural isomer 4pTb-MeIAA, as well as endogenous auxins from Arabidopsis samples. Distinct distribution of these synthetic isomers and endogenous auxins in Arabidopsis primary roots and hypocotyls was achieved in the same imaging analysis from both individually treated and cotreated samples. We also observed putative metabolites of synthetic auxin derivatives, i.e. auxinole amino acid conjugates and hydrolysed 4pTb-MeIAA product - 4pTb-IAA, based on their unique drifting ion intensity patterns. Furthermore, DESI-IM-MSI-revealed abundance of endogenous auxins and synthetic isomers was validated by liquid chromatography-mass spectrometry (LC-MS). Our results demonstrate that DESI-IM-MSI could be used as a robust technique for detecting endogenous and exogenous isomers and provide a spatiotemporal evaluation of hormonomics profiles in plants.

• Klíčová slova
• Auxin, Desorption electrospray ionization, Ion mobility, Isomer, Mass spectrometry imaging, Metabolite,
• MeSH
• Arabidopsis * MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• isomerie MeSH
• kyseliny indoloctové analýza   MeSH
• molekulární struktura MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kyseliny indoloctové MeSH

The field of plant hormonomics focuses on the qualitative and quantitative analysis of the hormone complement in plant samples, akin to other omics sciences. Plant hormones, alongside primary and secondary metabolites, govern vital processes throughout a plant's lifecycle. While active hormones have received significant attention, studying all related compounds provides valuable insights into internal processes. Conventional single-class plant hormone analysis employs thorough sample purification, short analysis and triple quadrupole tandem mass spectrometry. Conversely, comprehensive hormonomics analysis necessitates minimal purification, robust and efficient separation and better-performing mass spectrometry instruments. This review summarizes the current status of plant hormone analysis methods, focusing on sample preparation, advances in chromatographic separation and mass spectrometric detection, including a discussion on internal standard selection and the potential of derivatization. Moreover, current approaches for assessing the spatiotemporal distribution are evaluated. The review touches on the legitimacy of the term plant hormonomics by exploring the current status of methods and outlining possible future trends.

• Klíčová slova
• Hormonomics, Internal standard, Liquid chromatography, Mass spectrometry, Matrix effect, Metabolomics, Omics, Plant hormone, Solid phase extraction,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Studies of vitality/mortality of cortex cells, as well as of the concentrations of ethylene (ETH), gibberellins (GAs), indolic compounds/auxins (ICs/AUXs) and cytokinins (CKs), were undertaken to explain the hormonal background of kinetin (Kin)-regulated cell death (RCD), which is induced in the cortex of the apical parts of roots of faba bean (Vicia faba ssp. minor) seedlings. Quantification was carried out with fluorescence microscopy, ETH sensors, spectrophotometry and ultrahigh-performance liquid chromatography tandem mass spectrometry (UHPLC‒MS/MS). The results indicated that Kin was metabolized to the transport form, i.e., kinetin-9-glucoside (Kin9G) and kinetin riboside (KinR). KinR was then converted to cis-zeatin (cZ) in apical parts of roots with meristems, to cis-zeatin riboside (cZR) in apical parts of roots without meristems and finally to cis-zeatin riboside 5'-monophosphate (cZR5'MP), which is indicated to be a ligand of cytokinin-dependent receptors inducing CD. The process may be enhanced by an increase in the amount of dihydrozeatin riboside (DHZR) as a byproduct of the pathway of zeatin metabolism. It seems that crosstalk of ETH, ICs/AUXs, GAs and CKs with the cZR5'MP, the cis-zeatin-dependent pathway, but not the trans-zeatin-dependent pathway, is responsible for Kin-RCD, indicating that the process is very specific and offers a useful model for studies of CD hallmarks in plants.

• MeSH
• buněčná smrt MeSH
• cytokininy metabolismus   MeSH
• kinetin farmakologie   MeSH
• kyseliny indoloctové MeSH
• semenáček metabolismus   MeSH
• tandemová hmotnostní spektrometrie MeSH
• Vicia faba * metabolismus   MeSH
• zeatin metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokininy MeSH
• kinetin MeSH
• kyseliny indoloctové MeSH
• zeatin MeSH

We showed that wild pea seeds contained a more diverse combination of bioactive GAs and had higher ABA content than domesticated peas. Although the role of abscisic acid (ABA) and gibberellins (GAs) interplay has been extensively studied in Arabidopsis and cereals models, comparatively little is known about the effect of domestication on the level of phytohormones in legume seeds. In legumes, as in other crops, seed dormancy has been largely or entirely removed during domestication. In this study, we have measured the endogenous levels of ABA and GAs comparatively between wild and domesticated pea seeds during their development. We have shown that wild seeds contained more ABA than domesticated ones, which could be important for preparing the seeds for the period of dormancy. ABA was catabolised particularly by an 8´-hydroxylation pathway, and dihydrophaseic acid was the main catabolite in seed coats as well as embryos. Besides, the seed coats of wild and pigmented cultivated genotypes were characterised by a broader spectrum of bioactive GAs compared to non-pigmented domesticated seeds. GAs in both seed coat and embryo were synthesized mainly by a 13-hydroxylation pathway, with GA29 being the most abundant in the seed coat and GA20 in the embryos. Measuring seed water content and water loss indicated domesticated pea seeds´ desiccation was slower than that of wild pea seeds. Altogether, we showed that pea domestication led to a change in bioactive GA composition and a lower ABA content during seed development.

• Klíčová slova
• Desiccation, Legume, Maturation, Phytohormones, Pigmentation, Seed-coat,
• MeSH
• Arabidopsis * genetika   MeSH
• domestikace MeSH
• gibereliny metabolismus   MeSH
• hrách setý genetika   metabolismus   MeSH
• klíčení MeSH
• kyselina abscisová * metabolismus   MeSH
• semena rostlinná MeSH
• vegetační klid genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• gibereliny MeSH
• kyselina abscisová * MeSH

The view on the role of light during seed germination stems mainly from studies with Arabidopsis (Arabidopsis thaliana), where light is required to initiate this process. In contrast, white light is a strong inhibitor of germination in other plants, exemplified by accessions of Aethionema arabicum, another member of Brassicaceae. Their seeds respond to light with gene expression changes of key regulators converse to that of Arabidopsis, resulting in opposite hormone regulation and prevention of germination. However, the photoreceptors involved in this process in A. arabicum remain unknown. Here, we screened a mutant collection of A. arabicum and identified koy-1, a mutant that lost light inhibition of germination due to a deletion in the promoter of HEME OXYGENASE 1, the gene for a key enzyme in the biosynthesis of the phytochrome chromophore. koy-1 seeds were unresponsive to red- and far-red light and hyposensitive under white light. Comparison of hormone and gene expression between wild type and koy-1 revealed that very low light fluence stimulates germination, while high irradiance of red and far-red light is inhibitory, indicating a dual role of phytochromes in light-regulated seed germination. The mutation also affects the ratio between the 2 fruit morphs of A. arabicum, suggesting that light reception via phytochromes can fine-tune several parameters of propagation in adaptation to conditions in the habitat.

• MeSH
• Arabidopsis * metabolismus   MeSH
• Brassicaceae * genetika   MeSH
• fytochrom * genetika   metabolismus   MeSH
• hormony metabolismus   MeSH
• klíčení genetika   MeSH
• proteiny huseníčku * genetika   metabolismus   MeSH
• semena rostlinná genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• fytochrom * MeSH
• hormony MeSH
• proteiny huseníčku * MeSH

Dormancy and heteromorphism are innate seed properties that control germination timing through adaptation to the prevailing environment. The degree of variation in dormancy depth within a seed population differs considerably depending on the genotype and maternal environment. Dormancy is therefore a key trait of annual weeds to time seedling emergence across seasons. Seed heteromorphism, the production of distinct seed morphs (in color, mass or other morphological characteristics) on the same individual plant, is considered to be a bet-hedging strategy in unpredictable environments. Heteromorphic species evolved independently in several plant families and the distinct seed morphs provide an additional degree of variation. Here we conducted a comparative morphological and molecular analysis of the dimorphic seeds (black and brown) of the Amaranthaceae weed Chenopodium album. Freshly harvested black and brown seeds differed in their dormancy and germination responses to ambient temperature. The black seed morph of seedlot #1 was dormant and 2/3rd of the seed population had non-deep physiological dormancy which was released by after-ripening (AR) or gibberellin (GA) treatment. The deeper dormancy of the remaining 1/3rd non-germinating seeds required in addition ethylene and nitrate for its release. The black seeds of seedlot #2 and the brown seed morphs of both seedlots were non-dormant with 2/3rd of the seeds germinating in the fresh mature state. The dimorphic seeds and seedlots differed in testa (outer seed coat) thickness in that thick testas of black seeds of seedlot #1 conferred coat-imposed dormancy. The dimorphic seeds and seedlots differed in their abscisic acid (ABA) and GA contents in the dry state and during imbibition in that GA biosynthesis was highest in brown seeds and ABA degradation was faster in seedlot #2. Chenopodium genes for GA and ABA metabolism were identified and their distinct transcript expression patterns were quantified in dry and imbibed C. album seeds. Phylogenetic analyses of the Amaranthaceae sequences revealed a high proportion of expanded gene families within the Chenopodium genus. The identified hormonal, molecular and morphological mechanisms and dormancy variation of the dimorphic seeds of C. album and other Amaranthaceae are compared and discussed as adaptations to variable and stressful environments.

• Klíčová slova
• abscisic acid, coat-imposed dormancy, gibberellins, hormone metabolism, seed coat properties, seed heteromorphism, thermal time modelling, weed seed bank,
• Publikační typ
• časopisecké články MeSH