Jan Kopcewicz

Nicolaus Copernicus University | umk · Chair of Plant Physiology and Biotechnology

The timing of the transition from vegetative to reproductive development is of great fundamental and practical interest. Physiological event is a subject to strict and precise control. Among the environmental factors, the most important is photoperiod defined as the following changes in the light and darkness duration in the daily cycle, while the...

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 abu...

This paper presents a comprehensive study on the mechanisms of tree responses to pathogens. We describe natural resistance concerning the presence of different types of barriers protecting plants from invasion and also give them strength after a pathogen attack. We put emphasis on induced resistance functioning both locally and systemically and inv...

The involvement of gibberellins (GAs) in the control of flower induction in the short-day plant Ipomoea nil has been investigated. To clarify the molecular basis of this process, we identified the full-length cDNAs of the InGA20ox3 and InGA2ox1 genes, which encode enzymes responsible for GA biosynthesis and catabolism, respectively. We studied the...

qPCR validation of differential expression patterns of selected unigenes. The relative quantification of the gene expression level was determined with qPCR assay using ACTIN as a reference gene. The values presented on top of each bar indicate the expression level derived from the RNA-Seq data.

Annotation of L. luteus DEGs from an FAB vs. FNAB library comparison by KEGG database. (A) Distribution of DEGs into KEGG biological categories. The numbers of KEGG pathways belonging to a category are shown in brackets. (B) Classification of DEGs into the KEGG “Metabolism” category. The numbers of DEGs belonging to a pathway are shown.

Annotation of L. luteus DEGs from an FPAB vs. FPNAB library comparison by KEGG database. (A) Distribution of DEGs into KEGG biological categories. The numbers of KEGG pathways belonging to a category are shown in brackets. (B) Classification of DEGs into the KEGG “Metabolism” category. The numbers of DEGs belonging to a pathway are shown.

Statistical analysis of differentially expressed gene in compared libraries of L. luteus.

Differential expression patterns of cell wall-related genes in library comparison of FAB vs. FNAB, FPAB vs. FPNAB, PAB vs. PNAB.

Annotation of L. luteus DEGs from comparison of PAB vs. PNAB libraries by KEGG database. (A) Distribution of DEGs into KEGG biological categories. The numbers of KEGG pathways belonging to a category are shown in brackets. (B) Classification of DEGs into the KEGG “Metabolism” category. The numbers of DEGs belonging to a pathway are shown in bracket...

Differential expression patterns of plant hormone signaling-related genes in library comparison of PAB vs. PNAB.

List and annotation of differentially expressed genes (DEGs) between FAB and FNAB with PPDE < 0.05.

The enriched pathways (KEGG) of DEGs from library comparison of FPAB vs. FPNAB.

Quality filtering and statistics of raw reads obtained for eight transcriptome libraries of L. luteus.

List and annotation of differentially expressed genes (DEGs) between FPAB and FPNAB with PPDE < 0.05.

List and annotation of common DEGs identified in all three library comparisons of FAB vs. FNAB, FPAB vs. FPNAB, PAB vs. PNAB with FPKM (Fragment Per Kilobase of exon per Million fragments mapped) in each of the libraries.

The enriched pathways (KEGG) of DEGs from library comparison of PAB vs. PNAB.

List of primers used for the RT-qPCR validation and the RT-qPCR validation results.

Differential expression patterns of plant hormone signaling-related genes in library comparison of FAB vs. FNAB.

Differential expression patterns of plant hormone signaling-related genes in library comparison of FPAB vs. FPNAB.

List and annotation of differentially expressed genes (DEGs) between PAB and PNAB with PPDE < 0.05.

The enriched pathways (KEGG) of DEGs from library comparison of FAB vs. FNAB.

Ent-kaurene oxidase (EKO) catalyze three sequential oxidations in the early steps of gibberellin biosynthesis pathway. In this research, a cDNA sequence of InEKO1 gene in the model short-day plant Ipomoea nil was identified. Our studies revealed that inductive conditions for flowering caused an increase in the transcriptional activity of examined g...

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 abscission of certain organs from the plant is part of the fulfilment of its developmental programs. The separation process occurs in a specialized abscission zone usually formed at the base of detached organ. The changing level of phytohormones, particularly ethylene, is the element responsible for coordinating anatomical and physiological tra...

Interactions among jasmonates and auxin in the photoperiodic flower induction of a short-day plant Ipomoea nil were examined. Therefore, we measured changes in jasmonic acid (JA) and jasmonic acid methyl ester (JAMe) levels in the cotyledons of I. nil during the inductive night, as well as the effects of indole-3-acetic acid (IAA) on their content....

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 an...

Jasmonic acid methyl ester (JAMe) has been recently shown to play a crucial role in many physiological processes. In this paper, we focused on cotyledon senescence in Ipomoea nil and revealed that JAMe and darkness are the main factors stimulating the process examined. What is more, we showed that mefenamic acid (a jasmonate biosynthesis inhibitor)...

Allene oxide synthase (AOS) encodes the first enzyme in the lipoxygenase pathway, which is responsible for jasmonic acid (JA) formation. In this study we report the molecular cloning and characterization of InAOS from Ipomoea nil. The full-length gene is composed of 1662 bp and encodes for 519 amino acids. The predicted InAOS contains PLN02648 moti...

Auxin affects almost all of the growth and development processes in plants. The auxin signal transduction pathway involves a number of proteins, among which the key elements are: TAAR auxin receptors (TIR1 and AFB1-3), AUX/IAA auxin response repressors and Auxin Response Factor (ARF) transcription factors. The activity of genes encoding some compon...

The great agronomic potential of Lupinus luteus, species widely cultivated in many European countries as well as Australia, is strongly affected by premature and excessive generative organ abscission, mainly flowers. The unwanted process takes place in a specialized group of cells, called abscission zone (AZ). During their development they become c...

Yellow lupine like the rest belonging to the family Fabaceae (Fabaceae L.), has important practical significance. Development of lupines flowers and fruit and are crucial for the productivity of these plants. Phytohormone auxin (IAA) plays an important role in the regulation of plant ontogeny, including apical dominance, the fruit maturation and ag...

Yellow lupine like the rest belonging to the family Fabaceae (Fabaceae L.), has important practical significance. Development of lupines flowers and fruit and are crucial for the productivity of these plants. Phytohormone auxin (IAA) plays an important role in the regulation of plant ontogeny, including apical dominance, the fruit maturation and ag...

Twenty days-long stratification of Scots pine seeds results in the seeds germinating in similar high percentage in darkness as in light. The period of stratification enough to overcome the "barrier" of light is connected with simultaneous increase of gibberellin contents, especially with the appearance of the new groups of gibberellins qualitativel...

Increasing the number of flowers and pods set, as well as reducing the intensity of their abortion, is of crucial importance for the yielding of leguminous plants. This study examined the effects of the type of soil used and mineral fertilization applied on the generative development of the traditional and self-completing (restricted branching) cul...

Yellow lupin (Lupinus luteus L.) is an important legume crop characterized by high content of seed proteins. The crucial stage for lupin productivity, the formation and development of flowers and seeds, is often associated with the abscission of flowers. Plant development processes are determined by a number of proteins and phytohormones, whose lev...

Interactions between indole-3-acetic acid (IAA), abscisic acid (ABA), and ethylene (ET) in the photoperiodic flower induction of a short-day (SD) plant Pharbitis nil were investigated. It was shown that both IAA and ABA applied just before and during the first half of the 16-h-long inductive night inhibited flower induction in P. nil. Ethylene is a...

It has been previously suggested that 1-O-IAGluc growth stimulation occurs as the effect of its hydrolysis into a free IAA. In present experiments castanospermine, a known β-glucosidase inhibitor, was included. 1-O-IAGluc in the presence of castanospermine stimulated growth of corn coleoptiles segments even stronger then free IAA. So, it seems that...

The endogenous content of IAA in the cotyledons of Pharbitis nil is low before and during the first half of the inductive 16-h-long dark period. From the 8th to the 12th hour the level of IAA increased and then again was going down at the end of a dark period. Exogenous IAA applied to the cotyledones before and during the first half of the inductiv...

The study examined the influence of light and auxin on the transcription level of PnACO3, a gene involved in ethylene production, in relation to the inhibitory effect of ethylene on flower induction in the short-day plant Pharbitis nil (=Ipomoea nil). Exogenous auxin was shown to increase the level of PnACO3 mRNA, with the effect depending on the e...

Plant hormones auxin have been implicated in almost every aspects of plant development. In the past few years, it was shown that TIR1 (TRANSPORT INHIBITOR RESPONSE1) and some close homologs act as auxin receptors in Arabidopsis thaliana. TIR1 is an integral component of the SKP1/CULLIN/F-BOX PROTEIN (SCF)TIR1 complex that mediates the degradation o...

The formation and development of flowers and seeds of lupines are crucial for the productivity of the plants. This stage of development, however, is associated with undesirable phenomenon, which is the flowers abscission. The physiological studies and field tests show that to the moment the seeds reach full maturity, on plant stay only pods on the...

Phytohormone auxin is a key player during pattern formation, organogenesis, and various physiological processes. Auxin signaling and plant development depend on a family of partially redundant F-box receptors of the TAAR (TRANSPORT INHIBITOR RESPONSE 1 / AUXIN SIGNALING F-BOX ROTEIN 2 AUXIN RECEPTOR) clade. They act as a part of the ubiquitin ligas...

MicroRNAs regulate gene expression by guiding the cleavage or attenuating the translation of target mRNAs. In Arabidopsis thaliana, the subset of class II TCP genes (plant-specific group of transcription factors) contains an miR319-binding site. One of them, AtTCP4, regulates negatively leaf growth and positively leaf senescence. In addition, miR31...

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...

The plant hormone auxin regulates a broad spectrum of developmental processes. Recent advances have been made in the understanding of auxin response pathways, primarily by the characterization of genes encoding Auxin Response Factors (ARF) in Arabidopsis. In addition, expression of some of ARFs is regulated by microRNAs (miRNAs). In A. thaliana, AR...

The plant hormone auxin regulates a broad spectrum of developmental processes. Recent advances have been made in the understanding of auxin response pathways, primarily by the characterization of genes encoding Auxin Response Factors (ARF) in Arabidopsis. In addition, expression of some of ARFs is regulated by microRNAs (miRNAs). In A. thaliana, AR...

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 precur...

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 jasmo...

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 ro...

Little work was done so far with phytochrome from Pharbitis nil. Purification of phyA from this plant has been exceptionally difficult. Labile phytochrome was presented in too small amount to obtain either absorption spectra or enough protein to produce antibodies. Monoclonal antibodies mAP5, MAC 50, 52, 198 recognized Pharbitis nil labile phytochr...

In this study, the first ACC oxidase (PnACO1) cDNA from model short-day plant Pharbitis nil was isolated. The expression pattern of PnACO1 was studied under different conditions (photoperiod and auxin), an adequate balance of which determines P. nil flowering. It was shown that the gene was transcribed in all the examined organs of the 5-day-old se...

Streszczenie: Gibereliny (GA), jako jedne z podstawowych fitohormonów, kontrolują tak ważne procesy wzrostu i rozwoju roślin jak kiełkowanie nasion, wydłużanie łodyg i indukcję kwitnienia. Z ponad stu trzydziestu różnych GA zidentyfikowanych u roślin, grzybów i bakterii tylko nieliczne – GA1, GA3, GA4, GA5, GA6, GA7 – wykazują aktywność biologiczną...

Gibberellins (GAs), as one of the most important phytohormones, control different aspect of plant growth and development including seed germination, stem elongation and flower induction. Among more than a hundred and thirty GAs identified from plants, fungi and bacteria only a small number of them, such as GA(1), GA(3), GA(4), GA(5), GA(6) and GA(7...

Application of gibberellic acid (GA3) on the cotyledons of 5-d-old Pharbitis nil reversed the inhibitory effect of both abscisic acid (ABA) and ethylene on flowering. Application of GA3 slightly decreased ethylene production and did not affect the endogenous ABA content in the cotyledons during the night. However, it reversed the stimulating effect...

In this study using biochemical approaches we identified two calcium-dependent protein kinases (CDPKs) named PnCDPK52 and PnCDPK56 in soluble protein extracts from seedlings of Pharbitis nil. Both enzymes phosphorylated the specific substrate histone III-S in the presence of Ca2+ and cross-reacted with antibodies against the CDPK. PnCDPKs exhibited...

Interactions between methyl jasmonate (JA-Me) and ethylene in the photoperiodic flower induction of short-day plant Pharbitis nil were investigated. Both JA-Me and gaseous ethylene applied during the inductive long night caused a decrease in the number of flower buds generated by P. nil. Application of ethylene did not affected niether the level of...

Flowering of plants is controlled by hormones among which both stimulators and inhibitors are present. The role of abscisic acid (ABA) in flower induction of the short day plant Pharbitis nil was shown in our experiments through exogenous applications and endogenous level determination of the hormone in cotyledons of seedlings grown under special l...

Precise control of flowering time and generative organs development is essential for successful plants reproduction. Recent years showed that microRNA is one of the factors involved in regulation of generative development in plants. MicroRNA are 21 bp length regulatory molecules. They are involved in target genes silencing by inactivating they expr...

Precise control of flowering time and generative organs development is essential for successful plants reproduction. Recent years showed that microRNA is one of the factors involved in regulation of generative development in plants. MicroRNA are 21 bp length regulatory molecules. They are involved in target genes silencing by inactivating they expr...

Calcium-dependent protein kinases (CDPKs) are central calcium signal decoders. Here we report the isolation and characterization of the Japanese Morning Glory’s (Pharbitis nil) CDPK gene, termed PnCDPK1. The full-length cDNA of 1943bp contains an open reading frame for PnCDPK1 consisting of 514 amino acid residues and a calculated molecular mass of...

In this work we studied interactions between auxin and ethylene in the inhibition of photoperiodic flower induction in SDP Pharbitis nil. To this end, PnACS2 was cloned and its expression was studied under various photoperiodic conditions, as well as under the influence of auxin. The full-length PnACS2 cDNA consists of 1,841bp and encodes for 472 a...

Guanylyl cyclases (GCs) are enzymes involved in the biosynthesis of cyclic guanosine monophosphate (cGMP). Here we report the cloning and characterization of a new guanylyl cyclase, designated PnGC-1, from Pharbitis nil. This gene encodes a protein of 286 amino acids, with a calculated molecular mass of 32kDa. The predicted amino acid sequence cont...

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...

The ZEITLUPE (ZTL) protein is involved in the control of circadian period, hypocotyl elongation and flowering time in Arabidopsis thaliana. The aim of the present work was the identification of the InZTL gene and localization of its mRNA in the model short-day plant Ipomoea nil. The deduced InZTL protein of 622 amino acid residues contained a LOV d...

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...

Phototropin 1 (phot1) is a blue-light Ser/Thr receptor kinase that contains two LOV domains. It is a plasma membrane-associated protein that mediates phototropism, blue-light induced chloroplast movement, and stomatal opening. The aim of the present work was to analyze the intracellular localization of phot1 protein in Ipomoea nil seedlings. In cot...

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 ord...

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 t...

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 t...

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...

It has been shown that both IAA and ethylene application inhibit flower induction in the short-day plant Pharbitis nil. However application of IAA has elevated ethylene production in this plant, as well. Strong enhancement of ethylene production is also correlated with the night-break effect, which completely inhibits flowering. In order to determi...

It is known that the level of cGMP is modulated in plant cells in response to a number of stimuli but intracellular events dependent on cGMP metabolism are not clear. Guanylyl cyclases (GCs) are enzymes which are responsible for synthesis of cGMP in eukaryotic and prokaryotic cells. To collect evidence for the participation of cGMP in light signal...

It was revealed that cGMP is involved in the control of photoperiodic flower induction. Further insight into the signalling function of cGMP is likely to be obtained by analysis of its effectors. Therefore, in the present study, we used various agents that cause changes in cGMP-dependent kinase (PKG) activity and examined their effects on the activ...

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 negat...

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 enc...

The effect of the exogenous application of polyamines on the flowering induction of the short-day plant Pharbtis nil was investigated. Putrescine, spermidine and spermine applied on the cotyledons of 4-day seedlings had no significant effect on the flowering of this plant under conditions of full induction caused by a 16-hour-long inductive night....

Cyclic adenosine diphosphate ribose (cADPR) is a potent endogenous calcium-mobilizing agent synthesized from NAD+ by ADP-ribosyl cyclases described for several animal cells. Pharmacological studies suggest that cADPR is an endogenous modulator of Ca2+-induced Ca2+ release channels. There is also information about the sub-micromolar concentration of...