Relationship between protein kinase C and derepression of different enzymes.

The PKC1 gene in the yeast Saccharomyces cerevisiae encodes for protein kinase C which is known to control a MAP kinase cascade consisting of different kinases: Bck1, Mkk1 and Mkk2, and Mpk1. This cascade affects the cell wall integrity but the phenotype of pkc1Delta mutants suggests additional targets that have not yet been identified [Heinisch et al., Mol. Microbiol. 32 (1999) 671-680]. The pkc1Delta mutant, as opposed to other mutants in the MAP kinase cascade, displays defects in the control of carbon metabolism. One of them occurs in the derepression of SUC2 gene after exhaustion of glucose from the medium, suggesting an involvement of Pkc1p in the derepression process that is not shared by the downstream MAP kinase cascade. In this work, we demonstrate that Pkc1p is required for the increase of the activity of enzymatic systems during the derepression process. We observed that Pkc1p is involved in the derepression of invertase and alcohol dehydrogenase activities. On the other hand, it seems not to be necessary for the derepression of the enzymes of the GAL system. Our results suggest that Pkc1p is acting through the main glucose repression pathway, since introduction of an additional mutation in the PKC1 gene in yeast strains already presenting mutations in the HXKII or MIG1 genes does not interfere with the typical derepressed phenotype observed in these single mutants. Moreover, our data indicate that Pkc1p participates in this process through the control of the cellular localization of the Mig1 transcriptional factor.

Evidence for involvement of Saccharomyces cerevisiae protein kinase C in glucose induction of HXT genes and derepression of SUC2.

The PKC1 gene in the yeast Saccharomyces cerevisiae encodes protein kinase C that is known to control a mitogen-activated protein (MAP) kinase cascade consisting of Bck1, Mkk1 and Mkk2, and Mpk1. This cascade affects the cell wall integrity but the phenotype of Pkc1 mutants suggests additional targets which have not yet been identified. We show that a pkc1Delta mutant, as opposed to mutants in the MAP kinase cascade, displays two major defects in the control of carbon metabolism. It shows a delay in the initiation of fermentation upon addition of glucose and a defect in derepression of SUC2 gene after exhaustion of glucose from the medium. After addition of glucose the production of both ethanol and glycerol started very slowly. The V(max) of glucose transport dropped considerably and Northern blot analysis showed that induction of the HXT1, HXT2 and HXT4 genes was strongly reduced. Growth of the pkc1Delta mutant was absent on glycerol and poor on galactose and raffinose. Oxygen uptake was barely present. Derepression of invertase activity and SUC2 transcription upon transfer of cells from glucose to raffinose was deficient in the pkc1Delta mutant as opposed to the wild-type. Our results suggest an involvement of Pkc1p in the control of carbon metabolism which is not shared by the downstream MAP kinase cascade.

First Report of Rhizoctonia solani AG-7 on Potato in Mexico.

Among isolates of Rhizoctonia solani Kühn collected as sclerotia from potato (Solanum tuberosum L.) tubers harvested from the Toluca Valley in 1995 was one isolate (MexND) of anastomosis group 7 (AG-7). Virulence of this and other isolates of R. solani representing AG-3 (four isolates) or AG-7 (four isolates), was determined on developing potato plants (cv. Russet Burbank) in growth chamber studies at 10 and 21°C by methods used previously (1). A control treatment was inoculated with sterile disks of potato dextrose agar (PDA). Seed tubers were placed on a layer of pasturized sand-soil mix near the bottom of 6- × 25-cm black plastic tubes, then covered with 2 to 3 cm of the sand-soil mix. Seed tubers used in virulence tests carried no R. solani as they had been surface disinfested by immersing for 2 min in a 1.85% solution of formaldehyde. Inoculum, in the form of one 7-mm agar disk cut from the growing edge of appropriate colonies growing on PDA, was placed 2 to 3 cm above the seed and covered with 7 to 10 cm of sand-soil mix (1). Each treatment had five replications. Plants were harvested after control plants had emerged, 29 days at 21°C and 36 days at 10°C, and damage to roots and shoots was assessed according to Carling and Leiner (1). All four isolates of AG-3, one from the U.S. and three from Mexico, caused major damage to roots and shoots at both temperatures. The two isolates of AG-7 from the U.S. caused superficial discoloration to shoots at both temperatures but the two isolates of AG-7 from Japan caused no damage to shoots. None of the four isolates of AG-7 damaged roots at either temperature. MexND caused no damage to roots or shoots at 21°C but produced lesions (< 1 mm in diameter) on roots and shoots at 10°C. Thus, MexND caused less damage to potato than AG-3, more damage than other isolates of AG-7, and appears more aggressive at lower temperatures. This is the first reported occurrence of R. solani AG-7 in Mexico and its first reported recovery from a potato tuber-borne sclerotium. Reference: (1) D. E. Carling and R. H. Leiner. Phytopathology 80:930, 1990.