The starch-sugar hypothesis

Several workes have observed (ref. Devlin, 1969) that the starch content of guard cells is high
in the dark and low in light. It has been observed that cell sap has a pH value of 6-7 when sto-
mata are open compared to 4-5 when they are closed. From these observations it can be
concluded that stomatal opening in the light is caused by increase in pH which favours the
hydrolysis of starch to sugar in guard cells and thus lowers the osmotic potential. The pH
changes in the guard cell affected by light were thought to occur as a result of photosynthesis.
The reduction of CO2 (an acid) concentration in the guard cells and surrounding tissues that
results from its utilization in photosynthesis causes a rise in pH. On return to darkness,
photosynthesis stops, and the concentration of CO2 rises as a result of respiration. This results
in a decrease in pH (Devlin, 1969). However, there has been criticism that changes in CO2
concentrations observed are insuffucient to cause the observed change in acidity which may
be two units (Sutcliffe, 1979).
B. What is the solute and where does it come from?
1. Carbohydrates, such as sucrose
Since guard cells are the only epidermal cells with chloroplasts, plant physiologists
have long hypothesized that sucrose and related carbohydrates are osmotic regulators
of guard cells. For example, the starch content of guard cell chloroplasts decreases as
the stomata open. This idea, the "starch-sugar hypothesis", was the first postulated
mechanism for guard cell activity. It lost popularity after the role of potassium ions
was discovered, but most now agree that both sugar and potassium ions play a role in
guard cell regulation. Sucrose seems to be especially important in closing guard cells.
see graph in text/class
Where does the sucrose come from? (a) hydrolysis of starch in the GC chloroplasts.
In other words, an indirect product of photosynthesis (evidence: starch grains
disappear during opening); or (b) a direct product of carbon fixation (photosynthesis).

2. Malate
Malate is an organic acid (C4). You may already be familiar with its role in the
Kreb's cycle in the mitochondria. In plants, malate is also derived from the hydrolysis
of starches. The enzyme phosophoenolpyruvate carboxylase (PEPase) binds carbon
dioxide (actually bicarbonate ions) to phosphoenolpyruvate (PEP; 3-carbon atoms; an
intermediate in glycolysis) to produce oxaloacetate (C4) which is then reduced to
malate and stored in the vacuole.
3. Chloride ions
Chloride ions are transported into the cell from the apoplast via a Cl-/H+ symport in
which a proton gradient is used to "drag" the chloride into the cell.
4. Potassium ions
This appears to be the primary osmotic agent, especially for opening the GC in the
morning. The potassium comes comes from surrounding cells. Evidence: (a) if you
strip epidermis from a leaf, which breaks many epidermal cells but not the more
resistant GC, the GC will only open if K+ is provided in the medium; (b) potassium
concentrations increase in the guard cells upon opening (see Table 1)

Table 1: Potassium in the stomatal aperture of Commelina

communis

Open Closed
 K+ (mol) in GC 0.45 0.10

K+ (mol) in epidermal cells 0.07 0.45

Thus, we can modify our original scheme:

stoma closed (GC flaccid) → sucrose/potassium/malate/chloride ions → lower Ψs →
decrease Ψw → water uptake (osmosis) → increase pressure → stoma open (GC turgid)
To close the stoma, the reverse process occurs. However, time course studies indicate that
potassium uptake is associated with opening of the stomata in the morning, but sucrose loss is
more closely associated with closure in the afternoon. Thus, the final modification to our
scheme:
stoma closed (GC flaccid) → potassium and chloride ion uptake, malate synthesis → lower
Ψs → decrease Ψw → water uptake (osmosis) from subsidiary cells → pressure increases
→ stoma open (GC turgid) → ||||| → sucrose (potassium, chloride, malate) decreases → Ψs
increases → Ψw increases → water loss → pressure decreases → stoma closed (GC flaccid)
Stomatal Opening Based on The Starch-Sugar Hypothesis
Stomatal Closing Based on The Starch-Sugar Hypothesis

• during the day, photosynthesis occurs

• Concentration of CO2 decreases, pH increases
• Starch is converted to sugar
• sugar concentration in guard cells increases
• water potential in the guard cells decreases
• water from subsidiary cells enter the guard cells
by osmosis
• guard cells become turgid
• Stomata open

• At night, no photosynthesis
• Concentration of CO2 increases, pH
decreases
• Sugar is converted to starch
• sugar concentration in guard cells
decreases
• water potential in the guard cells increases
• water leaves the guard cells by osmosis
• guard cells become flaccid
• Stomata close