Biochemical and biophysical characterization of herbicide-resistant mutants of the unicellular cyanobacterium,Anacystis nidulans R2

Two herbicide-resistant mutants of the unicellular cyanobacterium, Anacystis nidulans R2, were obtained by mutagenesis with N-methyl-N′-nitro-N-nitrosoguanidine. These mutants, A. nidulans R2D1 and R2D2, were selected by growth of mutagenized cells in the presence of 10^?6 M and 10^?5 M 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU), respectively. Both were found to be cross-resistant to 2-chloro-4-ethylamino-6-isopropylamino-s-triazine (atrazine) and 2-n-heptyl-4-hydroxyquinoline-n-oxide (HQNO) by measurement of Photosystem II activity in the presence of the inhibitors. The DCMU-resistance trait from each mutant was transferred to a wild-type genetic background by DNA-mediated transformation of A. nidulans cells. The two resulting transformants, A. nidulans R2D1-X1 and R2D2-X1, were similar to the original mutants with respect to DCMU- and HQNO-resistance. However, both exhibited increased sensitivity to atrazine relative to the mutants from which they were derived. Polyacrylamide gel electrophoretic analysis revealed that the mutants and transformants were deficient in a 34 kDa, surface-exposed polypeptide which was present in the wild-type strain; the transformants exhibited a new polypeptide of 35.5 kDa which was also highly surface-exposed.