The (Plant) Cell – Vessel for Life Tomasz Nodzyński Advances and Challenges in Modern Biology 2021.10.04 The (Plant) Cell – Vessel for Life Can U see a cell..? Seeing is believing There was no concept a “cell” What was believed before… Spontaneous Generation In the early 1600’s and before, living organisms arising from the environment like dust and dirt. Disproven in 1864 Streptococcus pyogenes Swan-neck bottle used by Pasteur Drawing of cork by Robert Hooke, discoverer of “cells” He coined the tem „cells” Cells were first observed in plants. Photograph of cork cells Photo credit: ©David B. Fankhauser, Ph.D Little rooms ~ Cells 1665 1678: ut tensio, sic vis "as the extension, so the force" or „ the extension is proportional to the force"1635 -1703 1643 - 1727 Isaac Newton Diaries Diarrhoea Antony van Leeuwenhoek Dierkens Sperm cells observation helped to abolish the spontaneous generation theory Scanning line by line concept. epidermis of orchid spot seen also during the early stage of pollen formation Nucleus - Opaque spot in cells termed areola. The term cell nucleus was used by Robert Brown for the first time in 1831. Brown sensed that this spot was a key component of cells and called them "nucleus" - a term which is still being used today. Matthias Jakob Schleiden (botanist) - German botanist and co-founder of the cell theory, along with Theodor Schwann (zoologist/med.) and Rudolf Virchow. He declared that the cell is the basic building block of all plant matter. Cell wall→ nuclei Botanists of his day who limited themselves to merely naming and describing plants. Contemporary of Brown 1804-1881 The Small and Big Resolution for a human eye (naked eye) is between 100 and 200 um (about the diameter of a human hair) then the majority of bacteria cannot be seen with the naked eye. Mycoplasmas (about 0.25um) Green algae Stentor Protists Size: 2 mm in length Habitat: Freshwater Year Discovered: 1831 Gromia Sphaerica (ameba) Size: 3 cm in width Habitat: Ocean floor Year Discovered: 2008 Discovery Location: Arabian Sea Acetabularia is a genus of green algae Size: 10 cm in height Habitat: Shallow, subtropical waters Year Discovered: 1930’s Nucleus as keep for organismal features/information. Acetabularia Caulerpa Taxifolia (Aquarium Strain) Size: 3 meters in length Habitat: The Mediterranean Sea Year Successfully Bred: 1980 All large single cell organisms are in water environment What a Cell needs? PM as barrier – for what? Why we need a barrier? https://en.wikipedia.org/wiki/Lipid_bilayer Lipid mono-layer – lipid properties Benjamin Franklin Why we need a barrier? Electrocardiography life the ability to grow, …. The (Plant) Cell – Vessel for Life Growth of Membrane Jack Szostak IF a proto-cell becomes more complex – what would be good to have…? What is complexity? Preservation of information. life the ability to grow, change, etc., that separates plants and animals from things like water or rocks The (Plant) Cell – Vessel for Life Mendel’s observation of peas revealed the laws of inheritance Hynčice is a little Silesian village, administratively part of Vražné municipality, located. Gregor Johann Mendel (Řehoř Jan Mendel; 20 July 1822 – 6 January 1884) was a Germanspeaking Moravian-Silesian scientist and Augustinian friar and abbot of St. Thomas' Abbey in Brno who gained posthumous fame as the founder of the modern science of genetics. What was the key approach of Mendel? Establishment of homozygous lines He became a friar because it enabled him to obtain an education without having to pay for it himself. After he was elevated as abbot in 1868, his scientific work largely ended, as Mendel became consumed with his increased administrative responsibilities.. life the ability to grow, change …~ Evolve The (Plant) Cell – Vessel for Life Darwin, c. 1854, when he was preparing On the Origin of Species for publication. 1809 - 1882 Darwin's finches life the ability to grow, change …~ Evolve/adapt to environment Life: Self-sustaining chemical system capable of Darwinian evolution Journey to the Microcosmos Life and Death Preservation of information. Inside-cell Sub-structures to help Cell biology (subcellular structures) Nodzyński et al 2013 Intracellular trafficking Nodzyński, T. et al. (2012). Endocytic Trafficking of PIN Proteins and Auxin Transport. In Endocytosis in Plants, J. Šamaj, ed (Springer Berlin Heidelberg), pp. 165–183. Vacuolar mutants have problems with hypocotyl bending Vacuole role in plant turgor Complex cell biology phenotypes remain a quantification challenge.. Can Informatics help..? Storage Vacuole role in Germination Nodzyński et al 2013 MS-/no sugar Green Fluorescence Protein Ca2+ Actin Tubulin Subcellular structure markers Analysis of protein localisation Cell identity markers luciferin + O2 → oxyluciferin + light Multiple Cells ~ tissues Cell conglomerates. Antony van Leeuwenhoek 17th century that lens grinding by Antony van Leeuwenhoek provided the resolution needed to make major discoveries. In the first half of the 18th century botany was beginning to move beyond descriptive science into experimental science. Microscopic section through one-yearold ash tree Anatomy – internal structure of plants Looking for patterns, tissue organisation.. What is the significance of those patterns? epidermis + cortex vs endodermis The Casparian strip. Casparian strip deposition is not fully understood. BITS BIGGER PICTURE (Plant Biology) How to coordinate the BITS? How to coordinate the BITS. Coordinating signals. Vibrio harveyi – single cell communication Signalling discovered in the luminescent marine bacteria V. fischeri and Vibrio harveyi. In the early 1970s, researchers observed that supernatants from stationary phase cultures could be added to cells at low density and trigger light production, the signalling was species-specific and dependent on cell density rather than the nutritional status of the cells. Chemistry (identification of signals) Germinating shoot in of canary grass and oats Chemistry - the tropic signal is a chemical Chemistry (identification of signals) Germinating shoot in of canary grass and oats Chemistry - the tropic signal is a chemical Chemistry - the tropic signal is a chemical Cell to Cell translocation? indole-3-acetic acid (IAA) Auxin Transport Inhibited auxin transport AUX1 – Expression and Localization AUX1 proteinAUX1::GUS PIN1/AUX1 pin2Col-0 PIN2 – Root Gravitropism PIN2 protein • Embryo development • Organ initiation and positioning • Vascular tissue differentiation • Shoot and root elongation • Growth responses to light and gravity • Apical hook formation AUXIN mediates embryos Ln* Solving crystal structures of PM proteins remains a challenge, and even more observing that structure (significance) in action. -All defects in pin loss-off-function mutants are in auxin transport-dependent processes and can be phenocopied by auxin transport inhibitors - Local auxin distribution (gradients) are affected in pins - Polar PIN localization correlation with direction of auxin flow (Nodzyński et al., 2016) PIN gene family PIN Protein Topology PINs in Arabidopsis Root gravitropismApical organogenesis Root merist. Embryo Tropisms ER ER? ER Intracellular/Vesicular trafficking contributes to PIN polarity the PM. Vesicular trafficking delivers also membrane. Nodzyński, T. et al. (2012). Endocytic Trafficking of PIN Proteins and Auxin Transport. In Endocytosis in Plants, J. Šamaj, ed (Springer Berlin Heidelberg), pp. 165–183. Adopted from Gälweileret al., 1998 Mechanistic model for the generation of membrane curvature. (i) Lipid composition is changed by P4-ATPases, LPAT and PLA2; (ii) amphipathic helices of Arf family small GTPases are inserted into the cytoplasmic leaflet of the membrane; (iii) the resultant curvature is sensed and stabilized by BAR domain protein Membrane bound Mechanistic model for the generation of membrane curvature. (B) Helical-wheel representation of the Nterminal amphipathic helix of Arf1 (amino acids 2-13); (C) Structure of the BAR domain dimer of arfaptin; (D) The complex regulation of proteins involved in generating membrane curvature. Arfs are usually myristoylated on a glycine residue at position 2 of the N‐terminal amphipathic helix. Osmotic conditions influence turgor How the preserve membrane integrity during osmotic changes endoexo Regulate membrane delivery Do Osmotic conditions influence early steps of endocytic trafficking? MS+ NaCl[100] MQ FM4-64[2] 10’ remove deliver Osmotic conditions influence early steps of endocytic trafficking MS+ NaCl[100] MQ FM4-64[2] 10’ (Z-stack) remove deliver Preserve membrane integrity during osmotic changes Endo (remove)Exo (deliver) Regulate membrane delivery More flexible trafficking better plant survival. Zwiewka et al., 2015 Role of intracellular trafficking in the context of stress/plant adaptation is not fully understood. Thank you for your attention! CEITEC @CEITEC_Brno www.ceitec.eu Tomasz Nodzyński nodzynski@ceitec.muni.cz Building E26 Where to find us E26 building (indicated by red square) Other courses to consider Developmental and Cell Biology of Plants (S2008) Tomasz Nodzyński Marta Zwiewka MONDAY START @ 14:00 01/03/2022 Lecture will have a flexible on-line formula Given by Course is in Spring semester Exact starting dates might be subject to change, check IS system for updates. S2011 Hormones in plant development Given by Helene Boisivon Course is in Spring semester Exact starting dates might be subject to change, check IS system for updates.