Invertase: understanding changes in the photosynthetic and carbohydrate metabolism of barley leaves infected with powdery mildew



Infection of barley leaves with powdery mildew results in an increase in the activity of acid invertase, concomitant with an accumulation of glucose, fructose and sucrose in the infected leaf; this increase is confined to the mesophyll cells. The rate of photosynthesis is controlled by different factors depending upon the experimental conditions under which it is measured. In saturating light and ambient CO2, photosynthesis is determined to a large extent by Rubisco whereas, in saturating light and saturating CO2. it is mainly determined by the rate of end-product synthesis (Pi-limitation). The rate of photosynthesis was measured under these conditions to reveal which of the partial processes was most affected in mildewed leaves. Under conditions of saturating light and ambient CO2, the rate of photosynthesis declined in mildewed leaves from 3 d after inoculation, suggesting that carboxylation had been affected. However, the maximum capacity for photosynthesis, measured at saturating CO2 and irradiance, increased in mildewed leaves for the first 3 d after infection and then decreased to below control values on days 5 and 7, suggesting that Pi was not limiting photosynthesis. This hypothesis was investigated by measuring changes in photosynthetic intermediates and in the activity and amount of key enzymes of the Calvin cycle as infection progressed. There was a decline in the activity of the stromal fru-1, 6-bisPase, Rubisco and NADP-GAPDH in mildewed leaves. These decreases were consistent with changes in the ratio of metabolites. As infection progressed, there was an increase in the ratio of Rul,5P2: PGA and triose-P: Rul,5P2, the first indicating a restriction in the carboxylation of CO2 and the second a restriction in the regeneration of Rul,5P2, The PGA: triose-P ratio was similar in control and mildewed leaves until day 7 when it decreased, suggesting that the reduction of PGA to triose-P was not affected by the disease. There was little evidence of Pi-limitation in the mildewed leaves; the amount of Pi in infected leaves was similar to controls, infected leaves showed no secondary oscillations during a transition from dark to light and there was a reduction in the amount of PGA in infected leaves. We suggest that the high concentration of carbohydrates, resulting from the increase in invertase activity, causes directly or indirectly a gradual down-regulation of the Calvin cycle leading to aft inhibition of photosynthesis.