Abiotic Environmental Interactions - Abiotic Stress -

1. Abiotic Environmental Interactions - Abiotic Stress - Stress is usually defined as an external factor that exerts a disadvantageous influence on the plant. These can be environmental or abiotic factors that produce stress in plants, although biotic factors such as weeds, pathogens, and insect predation can also produce stress. In most cases, stress is measured in relation to plant survival, crop yield, growth (biomass production), or the primary assimilation processes (CO2 and mineral uptake), which are related to overall growth. Taiz, Zeiger (Plant Physiology)

2. Abiotic and Biotic environmental stress factors

3. Stress: live and let die?

4. Why bother about plant stress? As human population increases, agriculture must feed more people while competing with urban development for premium arable land If record yields can be assumed to represent plant growth under ideal conditions, then the losses associated with biotic and abiotic stresses can reduce the average productivity by 65-87%, depending on the crop

5. Resistance and Acclimation Resistanceiscomposed of : –  Avoidance: mechanisms that preventexposure to stress –Tolerance: mechanisms that permit the plant to withstand the stress Resistance: Saguaro: succulent photo-synthetic stem (highy drought tolerant) Honey mesquite: deep roots (drought avoiding) Mohave desert star: wet-season life cycle Acclimation: Spinach: osmotic adjustment Black spruce: freezing tolerance

6. Adaptation and Acclimation Acclimation of Several Cereals to cold Adaptation (to stress): An inherited level of stress resistance acquired by a process of selection over many generations. Contrary with acclimation Electrolytleakage Acclimation (hardening): The increase in plant stress tolerance due to exposure to prior stress. May involve gene expression. Contrary with adaptation Summeroat No acclimation Winter oat Withacclimation Winter rye

7. Gene Expression Patterns often Change in Response to AbioticStress Stress-induced changes in metabolism and development canoften be attributed to altered patterns of gene expression

8. Reactive oxygen species production as is a common, first response to many stresses Oxidative stress results from conditions that promote formation of ROS, which damage or kill cells (ROS)

9. Reactive Oxygen Species (ROS) These reactive molecules are highly destructive to lipids, nucleic acids and proteins Except H2O2, ROS have a very short half life Plants scavenge and dispose of these reactive molecules by use of antioxidant defense systems present in several subcellular compartments When these defenses failto halt the self-propagating auto-oxydation reactions associated with ROS, cell death ultimately ensues

10. Formation of ROS by Electron Transfer v Singlet-Oxygen Hydrogenperoxide MolecularOxygen Hydroxyl radical Superoxide Water

11. EnzymaticScavenger Reactions thatEliminate ROS • OS: oxidative stress • HS: heat stress • WS: water deficiency stress • CS: cold stress • Salz: salt stress • HL: high light stress Non Enzymatic Antioxydant Defensesystems •-tocopherol (vitamin E) •ascorbate (Vitamin C) • reduced glutathione (GSH) • polyamines (cabbage, broccoli, cauliflower) •flavonoids

12. Disfunctionof the Photosynthetic Transport Chainprovides a major source of electrons for ROSformation

13. ROS Formation during Photosynthesis If light absorption and CO2-fixation are in balance: minimal ROS-formation

14. Electron Transfer from Reduced PSI to O2is an electron sink creatingsuperoxide ions

15. Photoinhibition of PSII leads to singlet oxygenbuil up

16. CO2-Limitation: Enhancement of Photorespiration and H2O2-Formation

17. Water Deficit Stress • Periods of little or no rainfall can lead to a meteorological condition called drought •Transient or prolonged drought conditions reduce the amount of water available for plant growth (saline habitats or low temperatures can have the same effect) •Water deficit stress is measured by the water potential of the cell interior an cell exterior w=water potential = s + p s = solute potential (number of particles dissolved in water) p = pressure potential (physical forces exerted on water by environment)

18. Resurrection plants Plants that “completely” dry out (no physiological activity detectable) and have the ability to green and resume physiological activity after watering are called resurrection plants. Craterostigmaplantagineum Top: dried out plants are extremely fragile and crumble when touched Bottom: same plant, 24 hours after watering The Rose of Jericho (AsteriscuspygmaeusandAnastaticahierochuntia) is not, though often claimed, a resurrection plant. These plants die when they dry out; after watering, they simply swell and release the seeds which germinate and form a new plant.

19. Resurrection plants as a model for water-deficit stress adaptation Dehydration –> Activation of “desiccation-related” genes –> (1) Alterations in metabolism and –> (2) Production of “protective” proteins • Alterations in metabolism: • accumulation of protective solutes such as sucrose, trehalose, and proline that stabilize proteins and cellular membranes, • production of antioxidant compounds (such as galloylquinic acids), • (c) biochemical alterations in membrane and cell wall composition. • (2) Production of “protective” proteinssuch as “dehydrins” and “expansins” that help preserve the structural integrity of intracellular organelles and the cell walls.

20. Role of abscisicacid (ABA) in stomataclosure and water retention -ABA H2O +ABA In the absence of ABA, the phosphatase PP2Cis free to inhibit autophosphorylation of SnRk kinases ABA receptor (PYR/RCAR) onycloned in 2010! ABA enablesPYR/RCAR proteins to bind and sequester PP2C Other ABA receptorlikelyexist. This relieves inhibition of SnRk, whichbecomesautoactivated and phosphorylateABF transcription factors

21. Freezing Stress Cold temperatures can cause a type of water deficit stress: as ice formation is initiated in the intercellular spaces, cellular water moves down the water potential gradient, across the plasma membrane, and toward the extracellular ice. Therefore, a water deficit develops within the cell in response to freezing. Injury symptoms become apparent after transfer to normal temperatures: • - cellular dehydration • - alteration of membrane structure • - plasma membrane destabilization (lipid-lipid demixing) • - loss of interactions between cell wall and plasma membrane • - acidification of cytoplasm (vacuole rupture?) Candidate sensingproteinsmay monitor changes in membrane fluidity

22. The Miracle of the Blue Orchid Membrane lipids maintain fluid phase (liquid crystalline) at normal, warm temperatures, which ensures maintenance of cellular function. When temperature decreases, however, lipids with high melting temperatures begin to solidify (gel phase) and become phase-separated within the membrane, resulting in the membranes becoming leaky and/or dysfunctional, including the membrane of the vacuole (tonoplast) Intracellular water and solutes are lost and membrane-associated reactions such as carrier-mediated transport, enzyme-mediated processes, and receptor function are inactivated. In Prague, during the cold winter of 1875/76, Herrmann Müller placed the flowers of the orchid Calanthetriplicataon the window sill over night. On the next morning he took the frozen flowers back into the room. After de-freezing, the flowers turned blue. What happened? A chromogenoussecondary metabolite, the colorless indican, came into contact with glycosidase after the disruption of the tonoplast and was converted to indoxyl (yellow) which was then oxidized to indigo (blue).

23. Cold acclimation (or freezing tolerance) The ability to survive temperatures below freezing is genotype-specific. Among the genotypes unable to withstand temperatures belowfreezing are many important crop plants, including corn, tomato, and rice. Other plants are able to survive temperatures below freezing; somecan survive temperatures lower than -40°C. Freezing tolerance develops in a process known as cold acclimation, a response to low, but non freezing temperatures that occur beforefreezing. Arabidopsis, after exposure to temperatures in the range of 1-5°C for one to five days, can survive temperatures of -8-12°C. - alteration of membrane composition (increase in phospholipids) - accumulation of compatible solutes (membrane protection) - improved water retention at the membrane surface - change in plasma membrane protein composition (phospholipases, synaptotagmin 1  membrane trafficking)

24. Flooding and Oxygen Deficit Plants are obligate aerobes: oxygen is the terminal electron acceptor in the mitochrondrial electron transfer chain During flooding, too much water blocks the entry of O2 intothe soil so that roots and other organs cannot carry out respiration Plant speciesgenerallycanbeclassified as wetland, flood-tolerant, or flood sensitive, according to theirability to withstandperiods of oxygendeficit Wetlandplants have specificanatomicaladaptations: - aerenchymafacilitate O2 transport - lenticels in periderm for gas exchange - pneumatophores: shallowrootswithnegativegeotropy

25. Wetland plants are fullyadapted : Pneumatophore: shallow roots that grow with negative geotrophy out of the aquatic environment prevent oxygen deficit stress for instance in mangroves Thickenedroothypodermis to reduce O2loss Adventitiousrootsfrom the hypocotyl or stem, lenticells (openings in the periderm) Zea mays: root cortex under aerobic (A) and anaerobic (B) conditions Aerenchyma (continuous, columnarintracellularspaces) faciliate transport of O2 fromaerial structures to submergedroots

26. Phenotypic plasticity alsohelpssome plants to adapt to flooding The (genetically) same plant at a wet site Agropyronsmithii at a dry site root length

27. Phenotypic plasticity alsohelpssome plants to adapt to flooding Oryza sativa L. var. Indica: growth response of seedlings to flooding Rice reduces root growth and increasecoleoptile growth or internodalgrowth

28. Flood Tolerant Plants Flood tolerant plants can endure anoxia temporarily but not for prolonged periods. Like wetland species, these plants generate ATP through anaerobic metabolism during short term flooding. In most cases, root elongation is inhibited, overall rates of protein synthesis diminish, and patterns of gene expression are altered Flood sensitive plants They show injury response to anoxia, causing death within 24 hrs Generally sensing of oxygen deprivation is very poorly understood

29. Tropospheric Ozone is linked to Oxidative Stress in Plants Stratospheric ozone (O3) is beneficial because it shields the earth from UV irradiation But tropospheric ozone is harmful to life because it is a highly reactive oxidant… …and is produced by human activity… … which also destroysStratospheric ozone mostly by emission of CFCs(ozone hole) The negative effects of ozone on plants include: -decreased photosynthesis rates -leaf injury -reduced growth of shoots and roots -accelerated senescence -reduced crop yield BUT: ozone exposure induces pathogen resistance

30. Ozone causes Oxidative Damages to Biomolecules

31. Plants vary Greatly in their Ability to Survive in High-Ozone Environments Their resistance to ozone utilizes either avoidance or tolerancemechanisms: - Avoidance by physically excluding the pollutant by closing the stomata - Tolerance via biochemical responses that induce or activate the antioxidant defense system and DNA repair mechanisms Ozone-damagedoatleaves acclimation

32. Heat Stress Heat stress may arise under numerous temporal and developmental circumstances: –  in leaves when transpiration is insufficent (water limitation, high temperature) or with closed stomata and high irradiance –  in germinating seedlings when the soil is warmed by the sun –  in organs with reduced capacity for transpiration (e.g. fruits) –  in general from high temperature or irradiation Plants exposed to excess heat exhibit a characteristic set of cellular and metabolic responses including: –  damage of cellular structures, including organelles and the cytoskeleton, and impairment of membrane function –  decrease in the synthesis of “normal” proteins, denaturation of proteins –  transcription and translation of a new set of proteins known as heat shock proteins (HSPs) This response is observed when plants are exposed to temperatures at least 5°C above their optimal growing conditions

33. Plants can Acclimate to Heat Stress Plants can acquire thermotolerance if subjected to a nonlethal (permissive) high temperature for a few hours before encountering heat shock conditions . An acclimated plant can survive exposure to a temperature that would otherwise be lethal (there is, of course, a limit to how much heat a plant can withstand) . 45°C 28°C 40°C, 2hrs -> 45°C

34. Heat shock proteins • A and C: silver staining • B and D: fluorography of newly synthesised proteins Fluorographyis a methodused to visualize substances present in gels, blots, or otherbiochemicalseparations. Radioactivelylabeledsubstances emit radiation that excites a moleculeknown as a fluor or scintillatorthatispresent in the gel. When the excitedmolecule relaxes to itsground state, itemits a photon of visible or ultraviolet light thatisdetected by photographic film. The developed film indicateswhich bands in the gel containradioactivelylabeledmaterial (Waterborget al., 1994). Activelysynthesizedproteins are labelledwithS35incorporatedintomethionin.

35. 5 Classes of HSPs are Defined According to Size •HSP100 – contain 2 conserved ATP domains. 3 subfamilies: ClpB is heat inducible; members of this familiy are required for thermotolerance in plants and yeast – The HSP100 familiy of chaperones may function in disaggregation rather than prevent protein aggregation and misfolding during exposure to high temperatures •HSP90 – are found in bacteria and in the cytosolic, nuclear, and endoplasmic reticulum compartments of eukaryotic cells, where they may function as molecular chaperones •HSP70 – are essential for normal cell function. Some members are expressed constitutively; others are induced by heat or cold. Localised to the nucleolus during heat stress, HSP70 is redistributed to the cytoplasm during stress recovery

36. 5 Classes of HSPs are Defined According to Size •HSP60 – are thought to function as molecular chaperones. HSP60 proteins are abundant even at normal temperatures. Their major role is thought to involve protein assembly. In vitro, HSP60 proteins prevent other proteins from aggregating at physiologically relevant temperatures and are important in protein refolding as temperatures increase •smHSPs – Plants contain 5-6 classes of smHSPs (2 in the cytosol, 1 in the chloroplast, 1 in the ER, 1 in the mitochrondrion, and possibly another in a membrane compartment that has not been defined), whereas other eukaryotes have only one single class of smHSP – HSP18.1 from pea has been demonstrated to prevent protein aggregation in response to high temperature in vitro

37. HeatShock Transcription Factor (HSF) •  The heat shock transcription factor (HSF) is expressed constitutivelybut must be activatedvia trimerizationduring heat stress to recognise its DNA target, the heat shock element (HSE). •  The HSE is made up of 5-bp repeats in alternating orientations withthe consensus nGAAn. An HSF-regulated promotor maycontain5 to 7 of theserepeats close to the TATA box

38. 5) HSF - phosphorylation 6) Interaction of phosphoryl. HSF with HSP70 7) Release of HSF from DNA 8) Conversion to non DNA – binding monomer 1) Monomeric form of heat shock factor (HSF) + HSP70 2) Trimeric form upon heat shock 3) Binding of HSF to heat shock promoter 4) HSP – gene transcription