ABSTRACT
The main purpose of this study was to demonstrate that the course of anther development, including post-meiotic maturation, dehiscence and senescence, is ensured by the interdependencies between jasmonic acid (JA) and indole-3-acetic acid (IAA) in yellow lupin (Lupinus luteus L.). The concentration of JA peaked during anther dehiscence when IAA level was low, whereas the inverse relationship was specific to anther senescence. Cellular and tissue localization of JA and IAA, in conjunction with broad expression profile for genes involved in biosynthesis, signalling, response, and homeostasis under different conditions, allowed to complete and define the role of studied phytohormones during late anther development, as well as predict events triggered by them. The development/degeneration of septum and anther wall cells, dehydration of epidermis, and rupture of stomium may involve JA signalling, while the formation of secondary thickening in endothecial cell walls is rather JA independent. The IAA is involved in programmed cell death (PCD)-associated processes during anther senescence but does not exclude its participation in the anther dehiscence processes, mainly related to cell disintegration and degeneration. A detailed understanding of these multistage processes, especially at the level of phytohormonal interplay, can contribute to the effective control of male fertility, potentially revolutionizing the breeding of L. luteus.
Subject(s)
Cyclopentanes , Flowers , Gene Expression Regulation, Plant , Indoleacetic Acids , Lupinus , Oxylipins , Plant Growth Regulators , Indoleacetic Acids/metabolism , Cyclopentanes/metabolism , Oxylipins/metabolism , Lupinus/metabolism , Lupinus/growth & development , Lupinus/drug effects , Flowers/metabolism , Flowers/growth & development , Plant Growth Regulators/metabolism , Gene Expression Regulation, Plant/drug effects , Signal TransductionABSTRACT
The abscission of plant organs is a phytohormone-controlled process. Our study provides new insight into the involvement of gibberellic acid (GA3) in the functioning of the flower abscission zone (AZ) in yellow lupine (Lupinus luteus L.). Physiological studies demonstrated that GA3 stimulated flower abortion. Additionally, this phytohormone was abundantly presented in the AZ cells of naturally abscised flowers, especially in vascular bundles. Interesting interactions among GA3 and other modulators of flower separation were also investigated. GA3 accumulated after treatment with the ethylene (ET) precursor 1-aminocyclopropane-1-carboxylic acid (ACC). Abscisic acid (ABA) treatment did not cause such an effect. Furthermore, the expression of the newly identified LlGA20ox1 and LlGA2ox1 genes encoding 2-oxoglutarate-dependent dioxygenases fluctuated after ACC or ABA treatment which confirmed the existence of regulatory crosstalk. GA3 appears to cooperate with the ET precursor in the regulation of AZ function in L. luteus flowers; however, the presented mechanism is ABA-independent.
Subject(s)
Abscisic Acid/pharmacology , Flowers/metabolism , Gibberellins/pharmacology , Lupinus/metabolism , Flowers/drug effects , Gene Expression Regulation, Plant/drug effects , Lupinus/drug effects , Plant Proteins/genetics , Plant Proteins/metabolismABSTRACT
Yellow lupine (Lupinus luteus L., Taper c.), a member of the legume family (Fabaceae L.), has an enormous practical importance. Its excessive flower and pod abscission represents an economic drawback, as proper flower and seed formation and development is crucial for the plant's productivity. Generative organ detachment takes place at the basis of the pedicels, within a specialized group of cells collectively known as the abscission zone (AZ). During plant growth these cells become competent to respond to specific signals that trigger separation and lead to the abolition of cell wall adhesion. Little is known about the molecular network controlling the yellow lupine organ abscission. The aim of our study was to establish the divergences and similarities in transcriptional networks in the pods, flowers and flower pedicels abscised or maintained on the plant, and to identify genes playing key roles in generative organ abscission in yellow lupine. Based on de novo transcriptome assembly, we identified 166,473 unigenes representing 219,514 assembled unique transcripts from flowers, flower pedicels and pods undergoing abscission and from control organs. Comparison of the cDNA libraries from dropped and control organs helped in identifying 1,343, 2,933 and 1,491 differentially expressed genes (DEGs) in the flowers, flower pedicels and pods, respectively. In DEG analyses, we focused on genes involved in phytohormonal regulation, cell wall functioning and metabolic pathways. Our results indicate that auxin, ethylene and gibberellins are some of the main factors engaged in generative organ abscission. Identified 28 DEGs common for all library comparisons are involved in cell wall functioning, protein metabolism, water homeostasis and stress response. Interestingly, among the common DEGs we also found an miR169 precursor, which is the first evidence of micro RNA engaged in abscission. A KEGG pathway enrichment analysis revealed that the identified DEGs were predominantly involved in carbohydrate and amino acid metabolism, but some other pathways were also targeted. This study represents the first comprehensive transcriptome-based characterization of organ abscission in L. luteus and provides a valuable data source not only for understanding the abscission signaling pathway in yellow lupine, but also for further research aimed at improving crop yields.
ABSTRACT
Flower abscission is a highly regulated developmental process activated in response to exogenous (e.g. changing environmental conditions) and endogenous stimuli (e.g. phytohormones). Ethylene (ET) and abscisic acid (ABA) are very effective stimulators of flower abortion in Lupinus luteus, which is a widely cultivated species in Poland, Australia and Mediterranean countries. In this paper, we show that artificial activation of abscission by flower removal caused an accumulation of ABA in the abscission zone (AZ). Moreover, the blocking of that phytohormone's biosynthesis by NDGA (nordihydroguaiaretic acid) decreased the number of abscised flowers. However, the application of NBD - an inhibitor of ET action - reversed the stimulatory effect of ABA on flower abscission, indicating that ABA itself is not sufficient to turn on the organ separation. Our analysis revealed that exogenous ABA significantly accelerated the transcriptional activity of the ET biosynthesis genes ACC synthase (LlACS) and oxidase (LlACO), and moreover, strongly increased the level of 1-aminocyclopropane-1-carboxylic acid (ACC) - ET precursor, which was specifically localized within AZ cells. We cannot exclude the possibility that ABA mediates flower abscission processes by enhancing the ET biosynthesis rate. The findings of our study will contribute to the overall basic knowledge on the phytohormone-regulated generative organs abscission in L. luteus.
Subject(s)
Abscisic Acid/pharmacology , Biosynthetic Pathways/drug effects , Ethylenes/biosynthesis , Flowers/physiology , Lupinus/physiology , Amino Acids, Cyclic/metabolism , Biosynthetic Pathways/genetics , Flowers/drug effects , Fluorescent Antibody Technique , Gene Expression Regulation, Plant/drug effects , Lupinus/drug effects , Lupinus/genetics , Masoprocol/pharmacology , Plant Proteins/genetics , Plant Proteins/metabolism , Transcription, Genetic/drug effectsABSTRACT
The BLADE-ON-PETIOLE (BOP) genes have been recently shown to play an essential role in many physiological processes, including embryogenesis, meristem determinacy, leaf patterning and nodule development. In our research we used Lupinus luteus, a plant with great agronomic potential due to its high protein content and nitrogen fixation ability. In this work, LlBOP in L. luteus was identified for the first time and its expression during nodule development was analyzed. The high expression levels of LlBOP and LlLbI (LEGHEMOGLOBIN), essential to nitrogen-fixing symbiosis, were noted in the developing root nodules and were correlated with the occurrence of leghemoglobin. All of these data indicate that LlBOP is an important regulator of root nodule formation and functioning in L. luteus.
Subject(s)
Gene Expression Regulation, Plant , Genes, Plant , Lupinus/growth & development , Lupinus/genetics , Root Nodules, Plant/growth & development , Root Nodules, Plant/genetics , Amino Acid Sequence , Base Sequence , Cloning, Molecular , DNA, Complementary/genetics , DNA, Complementary/isolation & purification , Gene Expression Regulation, Developmental , Molecular Sequence Data , Plant Proteins/chemistry , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Root Nodulation/geneticsABSTRACT
The plant hormone auxin plays a critical role in regulating plant growth and development. Recent advances have been made that having improved our understanding of auxin response pathways, primarily by characterizing the genes encoding auxin response factors (ARFs) in Arabidopsis. In addition, the expression of some ARFs is regulated by microRNAs (miRNAs). In Arabidopsis thaliana, ARF6 and ARF8 are targeted by miR167, whereas ARF10, ARF16 and ARF17 are targeted by miR160. Nevertheless, little is known about any possible interactions between miRNAs and the auxin signaling pathway during plant development. In this study, we isolated the miR167 target gene InARF8 cDNA from the cotyledons of the short day plant (SDP) Ipomoea nil (named also Pharbitis nil). Additionally, the In-miR167 precursor was identified from the I. nil EST database and analyses of InARF8 mRNA, In-pre-miR167 and mature miR167 accumulation in the plant's vegetative and generative organs were performed. The identified cDNA of InARF8 contains a miR167 complementary sequence and shows significant similarity to ARF8 cDNAs of other plant species. The predicted amino acid sequence of InARF8 includes all of the characteristic domains for ARF family transcription factors (B3 DNA-binding domain, AUX/IAA-CTD and a glutamine-rich region). Quantitative RT-PCR reactions and in situ hybridization indicated that InARF8 was expressed primarily in the shoot apices, leaf primordia and hypocotyls of I. nil seedlings, as well as in flower pistils and petals. The InARF8 transcript level increased consistently during the entire period of pistil development, whereas in the stamens, the greatest transcriptional activity occurred only during the intensive elongation phase. Additionally, an expression analysis of both the precursor In-pre-miR167 molecules identified and mature miRNA was performed. We observed that, in most of the organs examined, the InARF8 expression pattern was opposite to that of MIR167, indicating that the gene's activity was regulated by mRNA cleavage. Our findings suggested that InARF8 and InMIR167 participated in the development of young tissues, especially the shoot apices and flower elements. The main function of MIR167 appears to be to regulate InARF8 organ localization.
Subject(s)
Gene Expression Regulation, Plant , Ipomoea nil/genetics , Ipomoea nil/metabolism , MicroRNAs/genetics , Indoleacetic Acids/metabolism , Ipomoea nil/growth & development , Transcription Factors/geneticsABSTRACT
Abscisic acid is one of the plant hormones that determines normal growth and development, i.e. seeds ripening and germination, stomata opening and closure, flowering and stress responses. An appropriate level of endogenous ABA plays a key role in the regulation of most of these processes. Its content in a particular tissue is a balance between the rate of its biosynthesis, oxidative degradation and formation of inactive derivatives (mainly ester). The progress on ABA metabolism was relatively slow in the past. Application of modern molecular biology methods let the most of genes encoding enzymes involved in the regulation of ABA metabolism be identified and contributed to the understanding of its action.
Subject(s)
Abscisic Acid/genetics , Abscisic Acid/metabolism , Plant Growth Regulators/genetics , Plant Growth Regulators/metabolism , Abscisic Acid/biosynthesis , Germination , Lipid Metabolism , Metabolism , Mutation , Plant Growth Regulators/biosynthesis , Seeds/metabolism , Signal TransductionABSTRACT
Gibberellins (GA), as one of the most important phytohormones, control different aspect of plant growth and development such as seed germination, stem elongation and floral induction. Although identified more than a hundred and thirty GA, only a small number of them are biological active. Many non-bioactive GA are present in plant tissues as precursors or deactivated metabolites. Biochemical and genetic approaches have led to the recognition most of the genes that encode GA biosynthesis and deactivation enzymes, and conducted investigation has helped us to better understand GA functions in plants. Many enzymes involved in GA metabolism are multifunctional and therefore fewer enzymes than might be expected are required to created the various gibberellins structures. In this review, we summarized current knowledge on the GA biosynthesis and deactivation pathways in plants and showed precise characteristic of genes and encoding protein which are involved in gibberellins metabolism.
Subject(s)
Gibberellins/biosynthesis , Gibberellins/chemistry , Plant Growth Regulators/biosynthesis , Plant Growth Regulators/chemistry , Plants/metabolism , Models, Molecular , Plants/geneticsABSTRACT
Jasmonates are plant hormones involved in many growth and development processes. They also participate in plant defense responses. Current progress in the study on biosynthesis and signaling of jasmonates has contributed to the understanding of the mechanisms regulating concentration of these hormones in the cell. Sustaining a proper level of jasmonates allow the plant to respond appropriately to changing conditions. It is possible due to the large number of enzymes and genes involved in biosynthesis of these hormones as well as multilevel control of their expression.
Subject(s)
Cyclopentanes/metabolism , Oxylipins/metabolism , Plant Growth Regulators/biosynthesis , Plants/metabolism , Adaptation, Physiological , Gene Expression Regulation, Plant , Lipoxygenase/classification , Lipoxygenase/metabolism , Phylogeny , Plants/geneticsABSTRACT
Signaling pathways, and specifically the signaling pathway of calcium, have been widely implicated in the regulation of a variety of signals in plants. Calcium-dependent protein kinases (CDPKs) are essential sensor-transducers of calcium signaling pathways, the functional characterization of which is of great interest because they play important roles during growth and in response to a wide range of environmental and developmental stimuli. Here, we report the first evidence of transient and specific elevation of PnCDPK1 transcript level and enzyme activity following conversion of a leaf bud to a flower bud, as well as participation of PnCDPK1 in evocation and flower morphogenesis in Pharbitis nil. Fluorescence microscopy immunolocalization and biochemical analysis confirmed the presence of CDPK in shoot apexes. The protein level was low in leaves, vegetative apexes and increased significantly in apexes after a flowering long-induction night. In the vegetative apex, a very weak PnCDPK1 protein signal was accumulated prominently in the zone of the ground meristem and in external layers of tissues of the cortex. After the dark treatment, the signal in cells of the ground meristem was still present, but a significantly stronger signal appeared in epidermal cells, cortex tissue, and leaf primordium. At the onset of flower meristem development, the PnCDPK1 level diverged significantly. PnCDPK1 mRNA, protein level and enzyme activity were very low at the beginning of flower bud development and gradually increased in later stages, reaching the highest level in a fully open flower. Analysis of flower organs revealed that PnCDPK1 was accumulated mainly in petals and sepals rather than in pistils and stamens. Our results clearly indicate that PnCDPK1 is developmentally regulated and may be an important component in the signal transduction pathways for flower morphogenesis. Findings from this research are important for further dissecting mechanisms of flowering and functions of CDPKs in flowering plants.
Subject(s)
Flowers/enzymology , Gene Expression Regulation, Developmental , Ipomoea nil/enzymology , Protein Kinases/metabolism , Signal Transduction , Flowers/genetics , Flowers/growth & development , Flowers/physiology , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Plant , Ipomoea nil/genetics , Ipomoea nil/growth & development , Ipomoea nil/physiology , Light , Meristem/enzymology , Meristem/genetics , Meristem/growth & development , Meristem/metabolism , Photoperiod , Plant Leaves/enzymology , Plant Leaves/genetics , Plant Leaves/growth & development , Plant Leaves/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Shoots/enzymology , Plant Shoots/genetics , Plant Shoots/growth & development , Plant Shoots/metabolism , Protein Kinases/genetics , RNA, Messenger/genetics , RNA, Plant/geneticsABSTRACT
Significant progress which was made during last few years in research of jasmonic acid signaling pathway yielded surprising information about chemical form of a signaling molecule of the hormone, which turned out to be its conjugate with amino acid isoleucine. Function of jasmonic acid receptor is more and more frequently attributed to COI1 protein which is structurally and functionally similar to the auxin receptor TIR1. Signal perception takes place in the nucleus and leads to the activation of SCF(COI1) ubiquitine ligase and consequently to proteolysis of transcription repressors, the JAZ proteins. Reduced pool of these negative regulators enables to activate the transcription factors (i. e. ERF1, WRKY70, MYC2), as well as expression of genes involved in defense responses of plants (i. e. PDF1.2, VSP1, CHI-B). Jasmonic acid signal transduction pathway, is also subjected to complicated regulations, including both positive, and negative feedbacks, which enable plants react adequately to variable environmental conditions.
Subject(s)
Cyclopentanes/metabolism , Oxylipins/metabolism , Plant Physiological Phenomena , Arabidopsis Proteins/metabolism , Gene Expression Regulation, Plant/physiology , Indoleacetic Acids/chemistry , Indoleacetic Acids/metabolism , Nuclear Proteins/metabolism , Plant Proteins/chemistry , Plant Proteins/metabolism , Signal Transduction/physiology , Transcription Factors/metabolismABSTRACT
The miR172 gene is involved in the regulation of flowering time and floral organ identity in Arabidopsis thaliana through regulation of APETALA2 (AP2)-like genes' activity. AP2 plays critical roles in establishing meristem and organ identity during floral development. Additionally, the AP2-like genes including TARGET OF EAT1 (TOE1), TOE2, SMZ, SNZ are involved in the timing of flowering in Arabidopsis thaliana. In our study, a full-length cDNA encoding InAP2-like transcription factor was isolated from cotyledons of morning glory (Ipomoea nil named also Pharbitis nil), a model short day plant. The identified sequence shows significant similarity to the cDNA of TOE1 from Arabidopsis thaliana and contains nucleotides complementary to miR172. Semi-quantitative RT-PCR analysis and in situ hybridization showed that the accumulation of InAP2-like transcripts was high, especially in cotyledons of 5-d-old seedlings. During the 16h-long inductive night, an increase in the expression of InAP2-like and a decrease in the accumulation of miR172 were observed. Auxin and ethylene treatment, as well as a "night-break", which completely eliminated flowering induction of Ipomoea nil, caused a decrease in the InAP2-like mRNAs levels in cotyledons of Ipomoea nil. These results suggest the potential involvement of miR172 and InAP2-like in the mechanism of flowering induction in Ipomoea nil.
Subject(s)
Flowers , Ipomoea/growth & development , MicroRNAs/genetics , Photoperiod , Base Sequence , DNA Primers , DNA, Complementary , Ipomoea/genetics , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
The light- and indole-3-acetic acid (IAA)-regulated 1-aminocyclopropane-1-carboxylic acid (ACC) synthase gene (PnACS) from Pharbitis nil was isolated. Here, it was shown that the gene was expressed in cotyledons, petioles, hypocotyls, root and shoot apexes both in light- and dark-grown seedlings. The highest expression level of PnACS was found in the roots. IAA applied to the cotyledons of P. nil seedlings caused a clear increase of PnACS messenger accumulation in all the organs examined. In this case, the most IAA-responsive were the hypocotyls. Our studies revealed that the PnACS transcript level in the cotyledons exhibited diurnal oscillations under both long-day (LD) and short-day (SD) conditions. IAA applied at the beginning of inductive darkness caused a dramatic increase in the expression of PnACS, suggesting that the inhibitory effect of IAA on P. nil flowering may result from its stimulatory effect on ethylene production.
Subject(s)
Flowers/enzymology , Gene Expression Regulation, Plant/drug effects , Indoleacetic Acids/pharmacology , Ipomoea nil/enzymology , Ipomoea nil/genetics , Light , Lyases/genetics , Amino Acid Sequence , Base Sequence , Cotyledon/drug effects , Cotyledon/enzymology , Cotyledon/genetics , Cotyledon/radiation effects , DNA, Complementary/isolation & purification , Flowers/drug effects , Flowers/radiation effects , Gene Expression Profiling , Gene Expression Regulation, Enzymologic/drug effects , Gene Expression Regulation, Enzymologic/radiation effects , Gene Expression Regulation, Plant/radiation effects , Ipomoea nil/drug effects , Ipomoea nil/radiation effects , Lyases/chemistry , Lyases/metabolism , Molecular Sequence Data , Organ Specificity/drug effects , Organ Specificity/radiation effects , Photoperiod , Phylogeny , RNA, Messenger/genetics , RNA, Messenger/metabolism , Seedlings/drug effects , Seedlings/enzymology , Seedlings/genetics , Seedlings/radiation effectsABSTRACT
Cyclic GMP acts as a chemical switch in plant cells to modulate cellular reactions. However, its metabolism has not been extensively explored and is still poorly understood. Previous experiments suggest that an endogenous cGMP system could participate in the mechanism of phytochrome controlled photoperiodic flower induction in Pharbitis nil. In order to gain further information on the role of cGMP, we have begun to study the enzyme of cGMP synthesis. In this article, the presence of the enzyme with guanylyl cyclase (GC) activity in soluble protein fractions of P. nil is reported. A large portion of the enzymatic activity is present in the cotyledons, where enzyme activity amounted to 0.45 pmol cGMP/min/mg protein. The enzyme exhibited a K(m) 0.5mM for GTP. A plot of 1/v versus 1/[GTP] was linear and V(max) was 0.74 pmol cGMP/min/mg protein. It was shown that the anti-sGC antibody recognise a 40 kDa protein. Moreover, the NO-donor, sodium nitroprusside (SNP) and YC-1, as a NO-independent stimulator, enhanced enzyme activity. The NS 2028 (a potent GC inhibitor) treatments provoked a 3-fold reduction of the enzyme activity in comparison to the untreated fractions. Furthermore, the influence of light on GC activity was analysed. It was noted that cGMP level increased in cool white light, and darkness inhibited enzyme activity. Exposure to blue light acts to stimulate cGMP formation, whereas in red light a rapid decrease in GC activity was observed that returned to the high level when far-red light was applied after the red light treatment. The results presented in this work strongly argue that an enzyme with guanylyl cyclase activity is present in P. nil organs and its activity is controlled by light via the photoreceptors-dependent pathways.
Subject(s)
Guanylate Cyclase/metabolism , Ipomoea nil/enzymology , Seedlings/enzymology , Cotyledon/drug effects , Cotyledon/enzymology , Cyclic AMP/metabolism , Cyclic GMP/metabolism , Guanosine Triphosphate/metabolism , Guanylate Cyclase/isolation & purification , Guanylate Cyclase/radiation effects , Hypocotyl/enzymology , Ipomoea nil/radiation effects , Kinetics , Light , Plant Roots/enzymology , Seedlings/radiation effects , Seeds/enzymology , ThermodynamicsABSTRACT
Ethylene is involved in the regulation of many growth and developmental processes in plants. Signaling pathways of the hormone are activated by five receptors, which are localized in membranes of endoplasmic reticulum and are similar to bacterial two-component histidine kinases. In the air, ethylene receptors activate CTR1 protein, which is a negative regulator (repressor) of nuclear protein--EIN2. In turn, EIN2 is an activator of transcriptional factors cascade responsible for the regulation of the expression of ethylene response genes. The level of EIN3, as well as other elements of ethylene signal transduction pathway, is subjected to complicated regulations on transcriptional and posttranslational levels, in which other internal and environmental factors are involved.
Subject(s)
Endoplasmic Reticulum/physiology , Ethylenes/metabolism , Gene Expression Regulation, Plant , Plant Growth Regulators/metabolism , Signal Transduction , Arabidopsis Proteins/physiology , DNA-Binding Proteins , Gene Expression , Nuclear Proteins/physiology , Protein Kinases/physiology , Receptors, Cell Surface/physiology , Transcription Factors/physiologyABSTRACT
Hormones are included in the essential elements that control the induction of flowering. Ethylene is thought to be a strong inhibitor of flowering in short day plants (SDPs), whereas the involvement of abscisic acid (ABA) in the regulation of flowering of plants is not well understood. The dual role of ABA in the photoperiodic flower induction of the SDP Pharbitis nil and the interaction between ABA and ethylene were examined in the present experiments. Application of ABA on the cotyledons during the inductive 16-h-long night inhibited flowering. However, ABA application on the cotyledons or the shoot apices during the subinductive 12-h-long night resulted in slight stimulation of flowering. Application of ABA also resulted in enhanced ethylene production. Whereas nordihydroguaiaretic acid (NDGA) - an ABA biosynthesis inhibitor - applied on the cotyledons of 5-d-old seedlings during the inductive night inhibited both the formation of axillary and of terminal flower buds, application of 2-aminoethoxyvinylglycine (AVG) and 2,5-norbornadiene (NBD) - inhibitors of ethylene action - reversed the inhibitory effect of ABA on flowering. ABA levels in the cotyledons of seedlings exposed to a 16-h-long inductive night markedly increased. Such an effect was not observed when the inductive night was interrupted with a 15-min-long red light pulse or when seedlings were treated at the same time with gaseous ethylene during the dark period. Lower levels of ABA were observed in seedlings treated with NDGA during the inductive night. These results may suggest that ABA plays an important role in the photoperiodic induction of flowering in P. nil seedlings, and that the inhibitory effect of ethylene on P. nil flowering inhibition may depend on its influence on the ABA level. A reversal of the inhibitory effect of ethylene on flower induction through a simultaneous treatment of induced seedlings with both ethylene and ABA strongly supports this hypothesis.
Subject(s)
Abscisic Acid/pharmacology , Ethylenes/pharmacology , Flowers/drug effects , Flowers/physiology , Ipomoea nil/drug effects , Ipomoea nil/physiology , Abscisic Acid/biosynthesis , Cotyledon/drug effects , Cotyledon/metabolism , Cotyledon/radiation effects , Ethylenes/biosynthesis , Flowers/radiation effects , Ipomoea nil/radiation effects , Light , Masoprocol/pharmacology , Photoperiod , Seedlings/drug effects , Seedlings/radiation effectsABSTRACT
Light is one of the most important environmental factors influencing the induction of flowering in plants. Light is absorbed by specific photoreceptors--the phytochromes and cryptochromes system--which fulfil a sensory and a regulatory function in the process. The absorption of light by phytochromes initiates a cascade of related biochemical events in responsive cells, and subsequently changes plant growth and development. Induction of flowering is controlled by several paths. One is triggered by the guanosine-3':5'-cyclic monophosphate (cGMP) level. Thus, the aim of our study was to investigate the role of cGMP in phytochrome-controlled flowering. It is best to conduct such research on short-day plants because the photoperiodic reactions of only these plants are totally unequivocal. The most commonly used plant is the model short-day plant Pharbitis nil. The seedlings of P. nil were cultivated under special photoperiodic conditions: 72-h-long darkness, 24-h-long white light with low intensity and 24-h-long inductive night. Such light conditions cause a degradation of the light-labile phytochrome. Far red (FR) treatment before night causes inactivation of the remaining light-stable phytochrome. During the 24-h-long inductive darkness period, the total amount of cGMP in cotyledons underwent fluctuations, with maxima at the 4th, 8th and 14th hours. When plants were treated with FR before the long night, fluctuations were not observed. A red light pulse given after FR treatment could reverse the effect induced by FR, and the oscillation in the cGMP level was observed again. Because the intracellular level of cGMP is controlled by the opposite action of guanylyl cyclases (GCs) and phosphodiesterases (PDEs), we first tested whether accumulation of the nucleotide in P. nil tissue may be changed after treatment with a GC stimulator or PDE inhibitor. Accumulation of the nucleotide in P. nil cotyledons treated with a stimulator of cGMP synthesis (sodium nitroprusside) was markedly (approximately 80%) higher. It was highest in the presence of dipyridamole, whereas 3-isobutyl-1-methylxanthine did not significantly affect cGMP level. These results show that the analysed compounds were able to penetrate the cotyledons' tissue, and that they influenced enzyme activity and cGMP accumulation. FR light applied at the end of the 24-h-long white light period inhibited flowering. Exogenous cGMP added on cotyledons could reverse the effect of FR, especially when the compound was applied in the first half of the long night. Flowering was also promoted by exogenous application of guanylyl cyclase activator and phosphodiesterase inhibitors, and in particular dipyridamole. The results obtained suggest that an endogenous cGMP system could participate in the mechanism of a phytochrome-controlled flowering in P. nil.
Subject(s)
Cyclic GMP/metabolism , Flowers/physiology , Ipomoea nil/metabolism , Phytochrome/metabolism , 1-Methyl-3-isobutylxanthine/pharmacology , Cotyledon/drug effects , Cotyledon/metabolism , Cotyledon/radiation effects , Cyclic GMP/pharmacology , Dipyridamole/pharmacology , Flowers/drug effects , Flowers/radiation effects , Ipomoea nil/drug effects , Ipomoea nil/radiation effects , Light , Nitroprusside/pharmacology , PhotoperiodABSTRACT
Ethylene is one of the plant hormones that controls growth and development. There are many responses regulated via ethylene in response to exogenous stimuli. Research on ethylene biosynthesis and the signalling pathway enabled us to understand the mechanism of the regulation of these responses. Different temporal and spatial expression of genes encoding enzymes involved in ethylene biosynthesis is of great importance for the regulation of ethylene responses. Also, post-translational regulation of the enzymes seems to be a key regulatory mechanism for the control of their activity. Because of versatile regulation of its production, ethylene can control plant development at many levels.
Subject(s)
Amino Acid Oxidoreductases/metabolism , Ethylenes/biosynthesis , Gene Expression Regulation, Plant/physiology , Plant Growth Regulators/biosynthesis , Plant Physiological Phenomena , Signal Transduction , Amino Acid Oxidoreductases/genetics , Arabidopsis , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Circadian Rhythm , Phylogeny , Plant Proteins/metabolism , Protein Processing, Post-Translational/physiology , Sequence Alignment , Sequence Homology, Amino Acid , Transcription, GeneticABSTRACT
Calcium signals play an important role in many aspects of plant growth and development, including plant response to biotic and abiotic stress. The stimulus characteristic intracellular Ca2+ signals are generated in plant cells by a variety of stimuli, including changes in environmental conditions, interaction with microbes and growth and development processes. Cytoplasmatic calcium brings about responses by interacting with target proteins, like calcium-dependent kinases. In plant there are at least five classes of protein kinases (CDPK, CRK, CCaMK, CaMK and SnRK3), which activity is regulated by calcium ions. In this article the structure, regulation and function of calcium stimulated protein kinases are briefly reviewed.
Subject(s)
Calcium/metabolism , Plants/metabolism , Protein Kinases/metabolism , Signal Transduction , Plants/enzymologyABSTRACT
The involvement of cGMP in the regulation of the flowering of Pharbitis nil was investigated through exogenous applications of cGMP and chemicals that are able to change the cGMP level and analyses of endogenous cGMP level. Exogenous applications of cGMP and 8-pCPT-cGMP (a cyclic GMP non hydrolyzed analog) to P. nil plants, which were exposed to a 12-h-long subinductive night, significantly increased flowering response. NS-2028 (guanylyl cyclase inhibitor) inhibited flowering when that compound was applied during a 16-h-long inductive night, whereas SNP (guanylyl cyclase activator) increased the flowering when plants were subjected to a 12-h-long subinductive night. The inhibitors of cyclic nucleotides phosphodiesterase (isobutyl-methylxanthine and dipyridamole), which increase the cytosolic cGMP level, promoted the flowering and allowed the length of the dark period necessary for induction of flowering to be reduced. The endogenous cGMP level was also measured after the treatment of P. nil seedlings with those chemicals. Results have clearly shown that compounds that were used in physiological experiments modulated endogenous cGMP level. There was a significant difference in the cyclic GMP level between 16-h-long night conditions and a long night with a night-break. During a long inductive night the oscillation of cGMP was observed with four main peaks in 4, 7, 11, 14 h, whereas a 10 min flash of red light in the middle of the night was able to modify these rhythmical changes in the second half of the long night. These results have shown that there are oscillations in the concentration of cGMP in the night and the biosynthesis and/or deactivation of cGMP is affected by light treatment and therefore it may be involved in the regulation of photoinduction processes in cotyledons. From these combined results, we propose a hypothesis that cGMP is involved in the control of photoperiodic flower induction in Pharbitis nil.