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Plant Cell, Vol. 12, 1703-1718, September 2000, Copyright © 2000, American Society of Plant Physiologists

MAP Kinase and Protein Kinase A–Dependent Mobilization of Triacylglycerol and Glycogen during Appressorium Turgor Generation by Magnaporthe grisea

Eckhard Thinesa, Roland W. S. Webera, and Nicholas J. Talbota
a School of Biological Sciences, University of Exeter, Washington Singer Laboratories, Exeter EX4 4QG, United Kingdom

Correspondence to: Nicholas J. Talbot, N.J.Talbot{at}exeter.ac.uk (E-mail), 1392-264668 (fax)

Magnaporthe grisea produces an infection structure called an appressorium, which is used to breach the plant cuticle by mechanical force. Appressoria generate hydrostatic turgor by accumulating molar concentrations of glycerol. To investigate the genetic control and biochemical mechanism for turgor generation, we assayed glycerol biosynthetic enzymes during appressorium development, and the movement of storage reserves was monitored in developmental mutants. Enzymatic activities for glycerol generation from carbohydrate sources were present in appressoria but did not increase during development. In contrast, triacylglycerol lipase activity increased during appressorium maturation. Rapid glycogen degradation occurred during conidial germination, followed by accumulation in incipient appressoria and dissolution before turgor generation. Lipid droplets also moved to the incipient appressorium and coalesced into a central vacuole before degrading at the onset of turgor generation. Glycogen and lipid mobilization did not occur in a {Delta}pmk1 mutant, which lacked the mitogen-activated protein kinase (MAPK) required for appressorium differentiation, and was retarded markedly in a {Delta}cpkA mutant, which lacks the catalytic subunit of cAMP-dependent protein kinase A (PKA). Glycogen and lipid degradation were very rapid in a {Delta}mac1 sum1-99 mutant, which carries a mutation in the regulatory subunit of PKA, occurring before appressorium morphogenesis was complete. Mass transfer of storage carbohydrate and lipid reserves to the appressorium therefore occurs under control of the PMK1 MAPK pathway. Turgor generation then proceeds by compartmentalization and rapid degradation of lipid and glycogen reserves under control of the CPKA/SUM1-encoded PKA holoenzyme.




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