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The involvement of the relA and rsh genes in themorphological and physiological differentiation of Streptomycesclavuligerus was evaluated with the relA and rsh genesmutants. The morphological differentiation of S. clavuligeruswas greatly affected by the disruption of the relA gene, but notvery much by the disruption of the rsh gene. The alteredmorphological characteristics were completely restored bythe complementation of the corresponding disrupted genes.Thus, it was apparent that the mycelial morphology andclavulanic acid production were severely affected by thedisruption of the relA gene. Production of clavulanic acid inthe submerged batch culture and glycerol-limited chemostatshowed that production was inversely related to the specificgrowth rate in the wild-type strain. However, the productionof clavulanic acid in the .relA and .rsh null mutants wascompletely abolished. Therefore, it seems plausible that thestringent response of S. clavuligerus to starvation for aminoacids is governed mainly by RelA, rather than Rsh, and thatthe (p)ppGpp synthesized immediately after the depletion ofamino acids triggers the initiation of pathways for bothmorphological and physiological differentiation in this species.

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