1. Plant Responses to Signals A weed is a plant that has mastered every survival skill except for learning how to grow in rows. - anonymous

2. External Signals External signals are used by plant cells to alter their physiology, morphology and development, –physical environment, –chemical environment, –biological environment, sometimes other plants, Plants receive signals at the cell level, and have no well defined sensory organs, Except for gravity, all other signals are constantly variable.

3. Internal Signals Signals can be processed by growing and non-growing cells, Signals are transduced into biologically meaningful results through numerous and co- ordinated pathways, –changes in ion flux, –regulation of metabolic pathways, –regulation of gene expression, –changes in the cytoskeleton.

4. Signal Transduction models Linear Model Signal Response Network Model Signals Signal Response

5. Reception Cell surface receptors, –hydrophilic molecules such as peptides and carbohydrates don’t readily cross the membrane, are perceived on the cell surface, Amphiphilic and hydrophobic molecules may pass through the membrane to receptors, –steroid hormones for example, Light may be perceived at the cell surface, or in the cell.

6. Plant Receptors i.e. light responses Where does one look? –no clearly differentiated organs (i.e., eyes, ears, etc.), –sensitive tissues, however, no clearly differentiated cells, –lots of responses. Germination (+/-) Stem length (-) Leaf expansion (+) Flowering (+/-) Phototropism (+/-) Stomatal opening (+) Chloroplast development (+) Pigment synthesis (+) De-Etiolation (italics)

7. De-etiolation greening Etiolated Etiolated growth habit, –long stem, –unexpanded closed leaves, etioplasts vs chloroplasts, lack of chlorophyll, –apical hook, –short root, Photomorphogenesis, –Inhibited stem growth, –Expanded leaves, –Pigmentation. –Root development. De-etiolated Nature Reviews Molecular Cell Biology 3; 85-93

8. Phytochrome plant photoreceptor 1920’s, –researchers observed chlorophyll deficient mutants (albinos) that underwent de- etiolation when given physiologically active light, 1950’s, –phytochrome discovered, Molecular switch, signal transducer.

9. Phytochrome Proteins with tetra-pyrrole chromophores, and transmission kinase domains, Phytochrome gene family contains at least five members, Gene family members serve different functions. chromophore kinase domains N C or

10. The Experiments

11. cGMP is a second messenger, Calcium is a second messenger. Signal red light Response greening Phytochrome Signal Transduction hypothesis Phytochrome is a greening receptor, See Fig. 39.2 for general signal transduction.

12. Phytochrome Signal Transduction for real Active phytochrome (Pfr) is transported to the nucleus, … binds and inactivates transcriptional repressor (PIF3), phytochrome (Pfr) …enhances the expression of MYB, red light activates phytocrome, active phytochrome (Pfr) is transported to the nucleus, CCA1 expression, in turn, enhances greening proteins. …MYB transcription factor is expressed, in turn activates CCA1 transcription, … CCA1 enhances the expression of CAB (chlorophyll A/B) proteins.

13. Nature Reviews Molecular Cell Biology 3; 85-93...thousands of genes,...response. …hundreds of transcription factors, phytochromes

14. Plant Responses to Signals II Phytohormones No one can look at the plants growing on a bank or on the borders of a thick wood, and doubt that the young stems and leaves place themselves so that the leaves may be well illuminated...they are extremely heliotropic; and this probably serves...as a guide (for) the buried seeds through fissures in the ground or through overlying masses of vegetation, into the light and air. - Charles Darwin “The Power of Movement in Plants” (1880) Charles Darwin was a Plant Physiologist, Phototropism, Introduction to the plant hormone Auxin.

15. Phytohormones …a plant product that is able to stimulate physiological responses at very low concentrations, –either in the tissue in which it is synthesised, –or in other regions of the plant to which it is transported,...do not operate in isolation from one another, but often act in co-ordination to produce subtle responses, …affect gene expression, enzyme activity and membrane function.

16. Charles Darwin and his son Francis localized the location of perception for blue light phototropism. Tropism: a growth response in plants that results in curvature toward, or away from a stimulus. Phototropism

17. Peter Boysen-Jensen demonstrated that a diffusable substance was involved. Positive Phototropism: growth toward a light stimulus

18. Peter Went demonstrated that the diffusible substance resulted in cell expansion. Went isolated the active compound. Natural auxins... IAA

19. Bioassays Bioassay: identification (or quantitation) of a biologically active substance by measuring the effect the substance has on living material. Oat Coleoptile (2 cm) auxin Oat Coleoptile (> 2 cm)

20. Auxin Transport polar Auxin moves basipetally (from apex to base). Synthesized in the SAM

21. Auxin …induces apical dominance, basipetal movement of auxin inhibits axillary buds, SAM intact axillary buds are released. SAM removed “Leader”

22. Apical Dominance basipetal movement of auxin inhibits axillary buds, SAM intact axillary buds are released, SAM removed

23. Auxins …induce vascular differentiation …xylem differentiation occurs around the wound,...following the path of auxin diffusion. …xylem differentiates between hydathodes and leaf vasculature...following the path of basipetal auxin transport. hydathode

24. Auxins …induces lateral and adventitious root formation, Rooting compounds. [ auxin ] Honeysuckle cut stems

25. Auxins …promotes fruit development, Strawberry seeds removed + auxin seed seeds removed Normal …embryo produces auxin that stimulates fruit development.

26. Auxin …promotes cell expansion, Auxin activates the H + -ATPase. Acidification of the cell wall activates expansin (cell wall proteins). See Fig. 39.11

27. Acid Growth Hypothesis Time course of action. Mode of action (hypothesis) AUXIN BINDING PROTEIN ABP + Auxin …activates the pump. See Fig. 39.11a

28. Auxin Binding Protein auxin receptor? over-express ABP1 gene add auxin Auxin receptor has not conclusively been identified, –a protein that is found to bind auxin, (Auxin Binding Protein, ABP) has been proposed as a candidate receptor, A common test for receptors, is to over- express (express more than the normal amount of protein), and test to see if the signal is amplified, ABP passes this test.

29. Auxin Binding Protein auxin receptor? wild typeabp mutant ABP controls cell division and elongation in embryogenesis. dead http://genesdev.cshlp.org/content/15/7/902

30. TIR1 auxin receptor …a ffects transcriptional control U U U TIR1, F-box portion of SCF TIR1 ubiquitin ligase complex, represents an auxin receptor CUL1 ASK RBX E2 CSN TIR1 I II III IV (Dharmasiri et al. (2005) Nature 435: 441-445; Kepinski and Leyser (2005) Nature 435: 446-451) AuxRE GENE “X” III IV III IV (Harper et al. (2000) Plant Cell 12: 757-770; Tatematsu et al. (2004) Plant Cell 16: 379-393) (Liscum & Reed (2002) Plant Mol. Biol. 49: 387-400; Esmon et al. (2005) Int. J. Dev. Biol. 49: 665-674 To know: TIR1 represents a new signal transduction paradigm. IAA

31. Auxins …promote gene expression, Genes Products 5 - 50 minutes

32. Concept Map Auxin Discovery Receptor Transport Function(s)