program konferencji i abstrakty - conference programme and abstracts

Program Konferencji - Conference programme 

 

Dzień I – 1^st day, Poniedziałek - Monday, 25.09.2023

On line - in English          

Rozpoczęcie – Łączenie się z uczestnikami konferencji – Przywitanie

Commencement - Connecting with conference participants – Greetings  10.00 – 10.30

Wykłady zaproszonych prelegantów – ¬Lectures of invited speakers

1. Peter Nick

Haberlandt's legacy: how cytokinins helped to prove the cell theory           10.30

2. Michał Jasiński

Medicago truncatula ABCG40 is a cytokinin importer negatively regulating lateral root density and nodule number               

3. Renata Bączek-Kwinta

Participation of cytokinins in signal transduction triggered by cadmium and butenolides  

4. Julia Stachurska

The effect of deacclimation on metabolism of plants - special attention to hormonal management

5. Andrzej Kaźmierczak

Dual face of kinetin

6. dr Anna Król-Górniak

Smart cell cultivation

 

Zakończenie 1. dnia konferencji …………………………………………………15.00

End of the 1st day of the conference ………………………………………... 15.00

 

Dzień II -  2^nd day,  Wtorek - Thusday,  26.09.2023

Stacjonarnie – Stationary in Polish 10.00 – 15.30

Wydział Biologii i Ochrony Środowiska UŁ, Aula w budynku D       

Faculty of Biology and Environmental Protection ¬ University of Lodz, Hall in building D

 

Prezentacje sprzętu laboratoryjnego i odczynników firmy Witko ¬ Seminaria i warsztaty aplikacyjne:

Presentations of laboratory equipment and reagents from Witko:

 

Seminars and application workshops:

•              Preparatyka próbek roślinnych -  Preparation of plant samples

•              Dobra praktyka ważenia GWP - Good GWP Weighing Practice

•              Automatyzacja w laboratorium - Automation in the laboratory

•              Selektywna i ekologiczna ekstrakcja roślinna nadkrytycznym CO₂ - Selective and ecological plant extraction of supercritical CO^₂

•              Warsztaty aplikacyjne z przygotowania próbek metodą SPE na przykładzie oznaczania cytokinin     

                Application workshop on sample preparation by the SPE method on the example of cytokinin determination     

•              S-Biosystem – testowanie ekstraktów roślinnych na liniach komórkowych

                S-Biosystem –  testing plant extracts on cell lines

•              Microfluidics - stabilne nanoemulsje

                Microfluidics – stable nanoemulsions

 

Prezentowane metody ¬- Presented methods:

1.            ekstrakcja próbek – maceracja, aparat Soxhleta, SFE

               sample extraction – maceration, Soxhlet apparatus, SFE

2.            mikroskopowa (liczba komórek roślinnych)

               microscopic (number of plant cells)

3.            spektroskopowa (zawartość chlorofili, cukrów, białek, dysmutazy ponadtlenkowej, katalazy, peroksydazy askorbinianowej, reduktazy glutationowej i askorbinianu)

              spectroscopic (content of chlorophylls, sugars, proteins, superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase and ascorbate)

4.            SDS-PAGE i western blotting (analiza białek szoku termicznego)

                SDS-PAGE and western blotting (analysis of thermal shock proteins)

5.            AAS (analizy zawartość metali ciężkich)

                AAS (heavy metal content analysis)

6.            HPLC (analizy zawartości fitochelatyn i glutationu)

                HPLC (phytochelatin and glutathione content determinations)

7.            SPE + HPLC-MS (analiza jakościowa i ilościowa cytokinin)

                SPE + HPLC-MS (qualitative and quantitative cytokinin analysis)  

 

Prezentowany sprzęt - Presented equipments:

1.            Pipety automatyczne – Automatic pipettes

www.witko.com.pl/segmenty/pipety-jednokanalowe-regulowana-pojemnosc-acurasupsup-manual-825-835,5011

2.            Urządzenia do elektroforezy, wizualizacji Cleaver Scientific - Cleaver Scientific Electrophoresis and Visualization Equipment,

 https://www.cleaverscientific.com/

3.            Materiały zużywalne, plastiki ¬- Consumables, plastics

4.            Komory laminarne - Laminar chambers

 https://www.witko.com.pl/segmenty/komory-laminarne-safefast-classic,9681

5.            Komory PCR ¬- PCR chambers

www.faster-air.com/en/products/special-cabinets/PCR-FAST

6.            Wirówki -¬ Centrifuges

pl.ohaus.com/pl-PL/Frontier5000SeriesMicro-1

7.            Wagi ¬- Scale for weighing

 

https://pl.ohaus.com/pl-PL/produc

 

Abstrakty - Abstracts

 

Haberlandt's legacy: how cytokinins helped to prove the cell theory

Peter Nick¹*
¹ Molecular Cell Biology, Joseph Gottlieb Kölreuter Institute for Plant Sciences, Karlsruhe Institute for Technology, Fritz-Haber-Weg 4, 76131 Karlsruhe, Germany

* email of corresponding author: peter.nick@kit.edu

ORCID number: 0000-0002-0763-4175

The contribution describes the achievements of Haberlandt and the conceptual importance of cytokinins in the historical context. Starting with early thoughts by Aristoteles, the attempt to explain life without the need to resort to a deus ex machina has first led to the idea of abiogenesis, i.e., that life can be generated from abiotic components, stimulating the concept of life as a “field” that can incarnate in matter, when the conditions are favourable. This idea was later discarded stepwise, first by the experiments of Francesco Redi in the 17^th century repeating Aristoteles’ experiments, but adding negative controls and one of the first microscopes. Later, Spallanzani and Pasteur, independently demonstrated that also microbial life cannot emerge de-novo but derives from microbial life. The discovery of cells by Hooke was first not linked with this debate, only more than two centuries later, Schwann (an animal physiologist) and Schleiden (a botanist) coined the Cell Theory that is still valid today and accepted so generally that few are aware of it. A central statement of the Cell Theory is that life is cellular and that a cell harbours everything, what is needed for life. This bold prediction was difficult to confirm empirically, though.

Haberlandt was inspired by this prediction and developed plant-tissue culture (much earlier than this was attempted in animal cells). He succeeded to cultivate a couple of different plant cells and keep the cells alive for quite some time. He even succeeded to see cell expansion, but he failed, despite hard work, to induce any division. This failure induced him to propose chemical factors exchanged by cells into their neighbourhood to instill division. He even was able to demonstrate a “wound factor” that can do exactly this and proposed the term phytohormone in analogy to animal hormones. However, since he did not know the chemical nature of this factor, cytokinins, he was not able to integrate this into tissue culture and thus, failed tragically. While animal tissue culture started later, it advanced more swiftly and also succeeded to get cell division. However, it took another century, until the experimental proof was reached that a somatic cell can generate an embryo. This work, the first cloning of metazoan animals by Gurdon, and the induction of pluripotent stem cells by introduction of four transcription factors by Yamanaka was awarded by the Noble Prize in 2012. Much earlier, in the early 1950ies, Steward could demonstrate the central claim of the Cell Theory by regenerating an entire carrot from a somatic cell. The reason, why he succeeded, where Haberlandt failed, had been the discovery of cytokinins as hormones that can induce cell division.  

Somatic embryogenesis in plants has been developed into a standardised process enabling numerous biotechnological applications. It is mainly the breakthrough in regeneration that enabled, from the 1980ies, the development of Plant Genetic Engineering. Independently of the fact, how one judges the use of GMOs in agriculture, it was the discovery and handling of cytokinin that enabled this technology that meanwhile is used on around 35% of agricultural land worldwide. Animal Gene Tech does not even come near to this impact.

The carrot system has allowed to study the cellular and molecular mechanisms behind somatic embryogenesis triggered by cytokinins. This led to a surprising discovery. A crucial step is a formative cell division separating the embryo proper from a vacuolated cell that later undergoes Regulated Cell Death. Using a cell-type specific antibody and a magnet, it was able to deplete the regenerating carrot culture from those cells that are doomed to death. Surprisingly, this disrupts the development of the embryonic precursor. Using bioactivity-guided fractionation, it was later possible to identify a soluble factor, an arabinogalactan protein that is secreted by the suicidal cell that can induce embryogenesis. Thus, death is needed to get life. This aspect shows that cytokinins, the hormones of life, have a hidden side, where they act as hormones of death.

 

 

 

Medicago truncatula ABCG40 is a cytokinin importer negatively regulating lateral root density and nodule number

Tomasz Jamruszka¹, Joanna Banasiak^1,2, Aleksandra Pawela¹, Karolina Jarzyniak^1,2, Jian Xia³, Wanda Biała-Leonhard¹, Lenka Plačková⁴, Francesca Romana Iacobini³, Ondřej Novák⁴, Markus Geisler³, Michał Jasiński^1,2*

¹ Department of Plant Molecular Physiology, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Z. Noskowskiego 12/14, 61-704 Poznań, Poland

² Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Dojazd 11, 60-632 Poznań, Poland

³ Department of Biology, University of Fribourg, Chemin du Musée 10, CH-1700 Fribourg, Switzerland

⁴ Laboratory of Growth Regulators, Faculty of Science, Palacký University and Institute of Experimental Botany, The Czech Academy of Sciences, Šlechtitelů 27, CZ-78371, Olomouc, Czech Republic

* email of corresponding author: jasinski@ibch.poznan.pl

ORCID number: 0000-0002-8883-5998

Plant roots show high plasticity to meet the needs during fluctuations of nitrogen availability in the soil and can adapt both their physiology and morphology, accordingly. Numerous studies suggest a relevant role of cytokinin (CK) distribution in shaping of root systems. Nonetheless, our knowledge about an involvement of short-distance CK translocation in root mineral nutrition is still scarce and specific role of CK transporters in root morphology has yet to be established. We identified and characterized the Medicago truncatula full-size ATP-binding cassette (ABC) transporter belonging to the G subfamily, namely MtABCG40 as a CK importer. Its expression is root-specific and is induced by nitrogen deprivation and CKs. Our analyses indicate that MtABCG40 has a negative impact on lateral root density through decreased lateral root initiation and enhancement of primary root elongation. The mtabcg40 plants have an accelerated pace of cell division in the developing lateral root primordia and reduced the size of root apical meristem (RAM). The MtABCG40 action affects CK signaling and impacts the cellular auxin content. Moreover, in line with postulated resemblance to lateral roots, we also observed an inhibitory influence of this transporter on nodule number. We present data that demonstrate a full-size ABCG transporter with a novel function in legumes and CK transport.

This work was supported by the National Science Centre, Poland (Grants No. 2015/19/B/NZ9/03548) Key words: ABCG transporters, cytokinin, root morphology, nodulation, lateral root, root apical meristem.

 

 

Participation of cytokinins in signal transduction triggered by cadmium and butenolides

Renata Bączek-Kwinta*

University of Agriculture in Kraków, Faculty of Agriculture and Economics, Department of Plant Breeding, Physiology and Seed Science, Podłużna 3, 30-029, Kraków

*email of corresponding author: renata.baczek-kwinta@urk.edu.pl

ORCID number: 0000-0002-9079-17 1X

Cadmium (Cd) is one of the most widespread ant toxic environmental pollutants, and its detrimental effect to plants has been largely demonstrated. Considering its impact on cytokinins (CKs), it affects auxin/ cytokinin ratio in roots, which triggers aberrations in root apical meristem (RAM) maintenance. Cd interacts also with other phytohormones, hence its role as the metallohormone has been postulated.

Physiologically active butenolides comprise strigolactones (SLs) and karrikins (KARs). SLs were discovered first in Orobanchaceae family plants that parasite other plants’ roots. Nowadays it is known that the role of SLs is broader, including e.g. stimulation of seed germination and seedling development, alterations in root architecture and enhancement of stress response. Interestingly, such activity is also associated with KARs mode of action. KARs are the products of pyrolysis of simple carbohydrates and generated during combustion of plant material. So far, SLs have been considered phytohormones, while KARs, due to their external origin, plant growth regulators.

During the lecture, similarities, differences and cross-talks within Cd, SLs and KARs signals transduction and plant phenotypic responses dependent on CKs will be discussed.

 

The effect of deacclimation on metabolism of plants - special attention to hormonal management

Julia Stachurska¹*

¹ The Franciszek Górski Institute of Plant Physiology Polish Academy of Sciences, Niezapominajek 21, 30-239 Krakow

* email of corresponding author: j.stachurska@ifr-pan.edu.pl

ORCID number:0009-0000-1778-9413

Oilseed rape (Brassica napus ssp. oleifera L.) is an important source of vegetable oil for food and industries. The yield of the winter cultivars (compare to spring cultivars) is higher but part of their vegetation season is during winter, which elevates the risk of frost injuries. Winter species developed mechanisms that enable them to survive temperatures below 0 °C. Cold acclimation improves the ability of plants to survive frost through many characteristic biochemical and physiological adjustments. Nowadays, more frequent episodes of higher temperatures in late autumn occur due to climate change and this may interrupt the cold acclimation of plants. This phenomenon is called deacclimation and leads to a decrease in plants’ frost tolerance. Even though cold acclimation is a well-studied process, still little is known about the biochemical and physiological basis of deacclimation. 

The aim of the current work was to characterize the hormonal balance and changes in the expression of brassinosteroid receptor (BRI1) as a result of deacclimation in selected cultivars of oilseed rape (with different frost tolerance – Feliks and Rokas). Samples for analyses of hormonal balance and BRI1 were collected from oilseed rape plants non-acclimated (three weeks old plants grown at 17 °C before cold acclimation), cold-acclimated (at 4 °C day/night, three weeks) and deacclimated (at 16/9 °C day/night, one week). The tested hormones from groups of gibberellins, auxins, cytokinins, brassinosteroids and stress hormones like ABA (including their precursors, intermediates and conjugates) were analyzed using an Ultra-High-Performance Liquid Chromatography. Unambiguous results were obtained for ABA, which concentration increased in all cold-acclimated plants, while it strongly decreased after deacclimation. Deacclimation resulted also in an elevated level of the typical growth hormones (for example GA₃). For the first time changes in brassinosteroids (brassinolide and its precursor castasterone) were described in cold-acclimated and deacclimated oilseed rape. The concentration of brassinolide increased in less frost-tolerant cultivar Feliks after deacclimation. In more frost-tolerant cultivar Rokas there was noted only a statistically insignificant tendency. A shift of hormonal balance with an increase of the content of growth/development hormones and a decrease in protective ABA as a result of deacclimation may be one of the possible reasons for decreased frost tolerance in oilseed rape. Additionally, research on the accumulation of BRI1 revealed that the amount of this protein decreased in cold-acclimated plants (compared to non-acclimated plants), while after deacclimation it increased - significantly in cultivar Feliks (less frost-tolerant) but only slightly in cultivar Rokas (more frost-tolerant). Changes in hormonal homeostasis in cold-acclimated and deacclimated oilseed rape plants in the context of their frost-tolerance changes will be discussed. Studies made within project of National Science Center (2019/35/B/NZ9/02868).

 

 

Dual face of kinetin

Andrzej Kaźmierczak¹*, Peter Nick², Karel Doležal³, Andrzej Kornaś⁴,

¹Department of Cytophysiology, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska    141/143, 90-236 Lodz, Poland

²Botanical Institute, Karlsruhe Institute of Technology, Fritz-Haber-Weg 4, 76131 Karlsruhe, Germany

³Laboratory of Growth Regulators, Palacký University & Institute of Experimental Botany AS CR, Šlechtitelů 27,

783 71 Olomouc, The Czech Republic

⁴Institute of Biology, Pedagogical University of Cracow, Podchorążych 2, 30-084, Cracow, Poland

* email of corresponding author: andrzej.kazmierczak@biol.uni.lodz.pl

ORCID number: 0000-0001-6446-7337

Cell death (CD), along with the cell division cycle, is the most important process controlling the differentiation of plant and animal cells and prokaryotic organisms. CD may be induced by endogenous or exogenous factors and contributes to all the steps of plant development. One of the factors, present in animal and plant organisms, which applied exogenously induced CD, is kinetin (Kin).

It is hypothesized that Kin, known as plant hormone of the cytokinin’s (CKs) group controlling cell cycle, inducing division both in the root cortex of faba bean (Vicia faba spp. minor) and in suspension of Nicotiana tabacum BY-2 line lead to mitotic catastrophe, which is manifested by death of about 45% and 60% of cells, respectively. This suggestion can be confirmed by the number of results on the nuclear level. The arrest of the cell cycle, induction of endoreplication, chromatin condensation and chromatin fragmentation were observed in both faba bean and BY-2 models. Moreover, formation of micronuclei and/or apoptotic-like (pseudoapoptotic) bodies in faba bean was observed, which are mentioned as one of the morphological exemplification of mitotic catastrophe in animals where the microtubular breakdown can have an important role (https://doi.org/10.1007/s00709-022-01814-6).  

Extensive studies in faba bean with 6-(2-hydroxy-3-methylbenzylamino)purine (PI-55) and  adenine (Ad), 5′-amine-5′-deoxyadenosine (Ado) and N-(2-chloro-4-piridylo)-N′-phenylurea (CPPU), the blocker of  CK receptors and inhibitors the activities of enzymes of CK metabolism,
i.e., hosphoribosyltransferase, kinases, and oxidases, respectively, as well as with ethylene glycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetra acetic acid (EGTA), lanthanum chloride (LaCl₃), ruthenium red (RRed) and cyclosporine A (CS-A), the chelator of extracellular calcium ions (Ca²⁺), blockers of  plasma-, endoplasmic reticulum-membrane Ca²⁺ ion channels and mitochondria-Ca²⁺ ions transition pores (PTP), respectively, indicated that Kin is converted to appropriate ribotides (5′-monophosphate ribonucleotides). The next studies 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), to cis-zeatin riboside (cZR) and to cis-zeatin riboside 5’-monophosphate (cZR5’MP), which
is indicated to be a ligand of cytokinin-dependent receptors inducing CD (https://doi.org/10.1038/s41598-021-03103-3).

It seems that crosstalk of phytohormones, protein kinase and Ca²⁺ ions pathways is responsible for transduce the cZ-dependent signal to induced kinetin-programmed cell death (Kin-PCD).