Copper deficiency in Helianthus annuus L.: developmental and metabolic consequences, stomatal function and senescence.

The effects of copper deficiency on developmental morphology and on age related metabolic activity, water utilization, stomatal response to light and leaf senescence in the sunflower, Helianthus annuus L., were monitored by sampling single leaves representing each leaf pair at designated harvest times during much of the vegetative lifespan. Plants grown in nutrient solutions lacking copper exhibited no apparent abnormalities during early vegetative growth and leaf morphology differed little from controls. There were no significant differences in rates of respiratory oxygen uptake of basal leaves between minus copper and controls. The effects of copper deficiency on rates of photosynthetic oxygen evolution were related to leaf age: rates were lower than controls in young basal leaves but higher as these leaves approached senescence. Both respiration and photosynthesis were more susceptible to KCN inhibition in minus copper leaves than in controls. There were no differences in cumulative water use per plant during the first five weeks after transfer of seedlings to nutrient solutions and only minor differences in stomatal aperture and guard cell potassium levels in ambient greenhouse light. Both minus copper and control leaves exhibited a sequential loss of stomatal function. Decay in the turgor driven opening response to light preceded loss of light dependent uptake of potassium into guard cells. Both changes in stomatal response to light occurred before major breakdown of chlorophyll. Basal leaves of sunflowers grown in minus copper nutrient solution tended to be retained longer and exhibited delayed loss of chlorophyll.

Evidence for Two Photoreactions and Possible Involvement of Phytochrome in Light-dependent Stomatal Opening.

Leaves of the xantha mutant of Helianthus annuus have a higher rate of transpiration and a lower diffusive resistance in the light than in the dark. Stomates of this nonphotosynthetic mutant open in the light and close in the dark.Comparative studies of tobacco, xantha mutant, and wild-type sunflower stomatal opening over a range of light intensities in isolated portions of the spectrum reveal two patterns of response: (a) a low intensity opening in the green and far red characterized by partial opening, absence of a threshold, and saturation of the response at low light intensities; (b) a high intensity response in the blue characterized by a threshold (intensities greater than 100 microwatts per square centimeter needed for opening) and a linear opening response at higher incident light intensities. In xantha mutant stomates only the low intensity system appears to be operational, while both low and high intensity systems are present in the wild-type sunflower and tobacco.Red light has an inhibiting effect on stomatal opening in both mutant and wild-type sunflowers. They require prior exposure to far red for opening to occur in red light. This redfar red antagonism suggests the involvement of phytochrome.

Reversal of copper inhibition in chloroplast reactions by manganese.

In the Mehler reaction, a Hill reaction utilizing molecular oxygen as the electron acceptor, rates of net oxygen uptake are stimulated by added manganous ions. Both whole cell photosynthesis and the Mehler reaction are inhibited by copper. Copper inhibition of the Mehler reaction can be reversed by manganese salts. Glutathione. which alone has no effect on Mehler reaction rates, enhances the effect of manganese in reversing copper inhibition. The effects of added Cu(2+), Cu(2+) and Mn(2+), or Cu(2+), Mn(2+), and glutathione exhibit no induction phenomena when measured manometrically. Furthermore, the order of addition of these factors is unimportant: final rates are dependent only on the composition of reaction mixtures. Compared to the Mehler reaction, conventional Hill reactions are less sensitive to copper poisoning, while certain chloroplast mediated photoxidations (e.g. the photoxidation of diketogulonic acid) are far more sensitive. In all of the chloroplast mediated photoreactions tested, manganese is effective in reducing the sensitivity to copper poisoning.

Allagochrome II. Effects of Light and Substrate on Allagochrome and Chlorogenic Acid Levels of Incubated Sunflower Leaf Discs.

Sunflower leaf discs incubated in the light on carbohydrate substrates exhibit several-fold increases in amounts of extractable allagochrome and chlorogenic acid. These changes are linear with time, and oxygen is required. The light effect saturates at approximately 600 muW/cm(2) "white" light, roughly the compensation point for photosynthesis. Red light is as effective as white light. Incubation in the dark, or in far red light, produces negligible changes in allagochrome and chlorogenic acid content.Sucrose (0.2 m) has been used as the standard substrate. At this concentration, glucose and fructose are slightly more effective. The optimum temperature range for incubation is 20 to 30 degrees . Allagochrome and chlorogenic acid values of both light and dark incubated samples decrease between 30 and 50 degrees , approaching zero at 50 degrees . Net light effects decrease to zero between 40 and 50 degrees .Of the inhibitors tested, 2,4-dinitrophenol, hydroxylamine and salicylaldoxime have no effect on light enhanced allagochrome and chlorogenic acid values except at high concentrations (which are generally deleterious to leaf tissues and cause decreased values for both dark and light incubated samples). Net light effects are completely inhibited, without changes in values of dark incubated discs, by azide (0.1 to 1 mm) and dichlorophenyldimethylurea (1 to 10 mum).Leaf tissues from the xantha mutant of Helianthus annuus do not exhibit a light effect. The absence of light effects in nonphotosynthetic leaf tissue and the inhibiting effects of photosynthetic poisons suggest that the photosynthetic apparatus is somehow involved.An hypothesis for light regulated metabolism via phenolic synthesis is discussed.