Latiyah Timothy

816012983
Bioc 2162: Circulatory and Secretory
Tutorial #5: Plant Hormones

1. Similarities and Differences between Plant and Mammalian Hormones:

 While plant hormones and mammalian hormones are naturally occurring organic substances,
that exert a profound influence on physiological processes at low concentrations.
 The site of synthesis in plant hormones, unlike animal hormones, are not localized and seem
to be much more diffused and cannot be localized to specific tissues.
 For plant hormones, action at a distance is not an essential characteristic.
 It is also still being debated whether or not plant hormones act in a concentration dependant
manner.
 Some scientists argue that plant cells respond to hormone concentration; whereas others argue
that it is changing sensitivity of the target cells to the hormone. Evidence exists for both
arguments.
 Plant hormones have a multiplicity of effects. Each group of plant hormones is known to
influence a wide variety of developmental events and each of these events can be influenced
by more than one hormone group.

2. The evidence in which cell growth induced by IAA is initiated by a decrease in cell wall pH.

 The Acid-Growth Hypothesis proposed by Rayle and Cleveland and Hagar that auxin causes
the acidification of the cell wall by the pumping of protons via a plasma membrane V-type
ATPase.
 The lower pH activates wall-loosening enzymes which loosen the wall and yields turgor
pressure.
 This is evidenced by Avena coleoptiles in which the pH of the cell wall solution is lowered
from 5.7 to 4.7 within 10 minutes of auxin application.
 The lag period is consistent between the lag period of auxin addition and the beginning of
growth.
 Auxin-stimulated proton pumping is an energy-dependent process and can be inhibited by
both inhibitors of auxin-induced growth and metabolic inhibitors.
 If neutral buffers were added to the cell wall space of the coleoptile sections, auxin-induced
growth is almost completely inhibited.
 Agents besides auxin that can cause proton excretion can have a similar effect to auxin and
can promote growth.
 The problem with the acid-growth hypothesis is the probability of the existence if the docking
protein by which the ABP1 receptor interacts with to bind auxin and stimulate the V-type
ATPase.
3. The molecular process by which IAA can lead to the activation of a plasma membrane V-type
ATPase.

 ABP1 is the suggested receptor for auxin. It is a 43kDa glycoprotein dimer of 22kDa subunits
with a high affinity for auxin. Its role as the auxin receptor is evidenced indirectly with the
use of anti-ABP antibodies which inhibit auxin-induced coleoptile elongation and
hyperpolarization of the plasma membrane.
 However, there are some issues, ABP1 has no membrane-spanning domains. It is also located
in the lumen of the ER and not the plasma membrane. ABP1 also has a KDEL sequence.
 The targeting of the ABP1 protein has been refined. It is suggested that ABP1 interacts C-
terminal Binding Protein 1(CBP1) and this is evidenced by the synthetic peptide
corresponding to the C-terminus of Zm-ABP1.
 When ABP1 wants to go outside the cell, it binds to CBP1 which masks the KDEL sequence
on ABP1 and allows it to target the plasma membrane.
 This allows ABP1 to enter the exit site of the ER. It goes off to the Cis, moves through the
Golgi and gets to the TGN.
 Then both ABP1 and CBP1 fuse through the secretory pathway to the plasma membrane and
ABP1 is displayed to the outside.
 ABP1 binds to a hypothetical docking protein. Binding to auxin then occurs.
 By some downstream signalling, it turns on the V-type ATPase which then releases protons to
acidify the cell wall.
 At the same time, a potassium channel is activated which allows and efflux of potassium into
the cell and activates an aquaporin and allows water to come in and increase the turgor
pressure. This mechanism is still unknown.
 Auxin also activates a membrane-bound enzyme called phospholipase A (PLA 2). This
enzyme hydrolytically excises the fatty acid from the C2 position of a phospholipid.
 This enzyme is implicated in the signal transduction chain and is evidenced by the activation
of PLA2 can be blocked by anti-ABP antibodies and the products of PLA 2, fatty acids and
lysophospholipids, stimulate proton secretion and cell elongation.
 Vanadate which inhibits the plasma membrane ATPase also inhibits these effects. The data
suggest that PLA2 follows ABP1 in the chain and the lipids further along.
 Protein kinase inhibitors can block the effect of both auxin and lysophospholipids on proton
secretion and cell elongation. This suggests that lipids activate the ATPase with the
involvement of a protein kinase cascade.

4. Mobilization of food reserves in developing cereal seed by gibberellins

 Germinating cereal grains secrete a range of hydrolytic enzymes including a-amylase and
proteases, which are involved in the hydrolysis of carbohydrate and protein stored in the
endosperm.
 Starch is the food reserves and needs to be mobilized into glucose.
 When the seed begins to germinate, water enters and the gibberellins are released.
 Gibberellins then move to the aleurone layer where there are involved in the secretion of a
protease. The protease then cleaves β-amylase that is sitting in the endosperm. This β-amylase
is active after cleavage and will now target starch.
 The release of gibberellins also results in the secretion of α-amylase from the aleurone layer
to the endosperm.
 The presence of starch, β-amylase and α-amylase in the endosome can now be mobilized to
glucose.
 This glucose then goes to the embryo and allow the germination process to go forward.
5. The regulation of apical dominance by auxin and cytokinin.

 The relationship between auxin and apical dominance is early evidenced via the removal of
the shoot apex and the application of auxin to the cut stump. While lateral bud development
remained suppressed in the presence of auxin.
 This occurs because of the interplay between auxin, cytokinin, isopentyl transferase(IPT) and
cytokinin oxidase(CKX).
 In an intact plant, auxin is produced in the apical meristem and tend to move down the cell.
 The transport of auxin is polar. It does this using V-type ATPases to move it through the cell
and proteins to efflux it out of the cell.
 It can get into the cell in two ways: by naturally diffusing into the cell via V-type ATPases or
by active transport via Lax proteins. Auxin can only leave the cell via PIN proteins which are
only found at the bottom of the cell away from the meristem end. Short term treatment of
auxin can lead to the accumulation of PIN protein in the plasma membrane, while long term
exposure can lead to the internalization of PIN proteins.
 When the auxin moves down the plant from the apical meristem, the PIN proteins are
induced, auxin flows down, CKX is induced and inhibits cytokinin and IPT is repressed
which means there is no cytokinin synthesis and the PINS are repressed in the lateral buds.
 When the apical bud is removed, there is no source of auxin flowing down the plant.
Cytokinin is synthesized in the stem and is transported into the dormant lateral buds and
initiates their growth. After the outgrowth of the lateral bud, synthesis is auxin occurs which
leads to the repression of IPT and the induction of CKX and PINS.

6. Evidence that cytokinins act as inhibitors of senescence:

 When cytokinins are applied to detached leaves exogenously, the early onset of senescence is
delayed.
 The next line of evidence is the relationship between endogenous cytokinin content and
senescence. When auxin is applied to detached leaves to induce root formation at the base of
the petiole, will remain healthy for weeks. This is a result of the growing root which acts as a
synthesis site for cytokinin and the hormone is to the leaf blade via the xylem. It is also
observed that there is a sharp decrease in the levels of cytokinin exportation from the root
when a mature plant enters senescence.
 The TMR gene (encodes for isopentyl transferase) was linked to a heat shock promoter. This
allowed for the cytokinin synthesis to be turned on by a brief heat shock. The plants treated
this way showed delayed senescence compared to the non-heat shocked plants.

7. The regulation of the rate of regulation of transpiration in a drought-affected plant by ABA:

 In a drought-affected plant, ABA binds to the surface of the guards' cells plasma membrane.
 The receptors activate several interconnecting pathways which come together to produce
- a rise in pH in the cytosol
- transfer of Ca2+ from the vacuole to the cytosol
 The increased levels of Ca2+ in the cytosol blocks the uptake of K+ into the guard cell while
the increase in pH stimulates the loss of Cl − and organic ions (e.g., malate2−) from the cell.
 The loss of these solutes in the cytosol reduces the osmotic pressure of the cell and thus
turgor. The stomata close and water is conserved.