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Plant Cell, Vol. 10, 297-308, Copyright © 1998, American Society of Plant Physiologists

a-Tubulin Missense Mutations Correlate with Antimicrotubule Drug Resistance in Eleusine indica

Etsuo Yamamotoa, Linghe Zenga, and W. Vance Bairda
a Department of Horticulture, Clemson University, Clemson, South Carolina 29634

Correspondence to: W. Vance Baird, vbaird{at}clemson.edu (E-mail), 864-656-4960 (fax).

Dinitroaniline herbicides are antimicrotubule drugs that bind to tubulins and inhibit polymerization. As a result of repeated application of dinitroaniline herbicides, highly resistant and intermediately resistant biotypes of goosegrass (Eleusine indica) developed in previously wild-type populations. Three {alpha}-tubulin cDNA classes (designated TUA1, TUA2, and TUA3) were isolated from each biotype. Nucleotide differences between the susceptible and the resistant (R) {alpha}-tubulins were identified in TUA1 and TUA2. The most significant differences were missense mutations that occurred in TUA1 of the R and intermediately resistant (I) biotypes. Such mutations convert Thr-239 to Ile in the R biotype and Met-268 to Thr in the I biotype. These amino acid substitutions alter hydrophobicity; therefore, they may alter the dinitroaniline binding property of the protein. These mutations were correlated with the dinitroaniline response phenotypes (Drp). Plants homozygous for susceptibility possessed the wild-type TUA1 allele; plants homozygous for resistance possessed the mutant tua1 allele; and plants heterozygous for susceptibility possessed both wild-type and mutant alleles. Thus, we conclude that TUA1 is at the Drp locus. Using polymerase chain reaction primer-introduced restriction analysis, we demonstrated that goosegrass genomic DNA can be diagnosed for Drp alleles. Although not direct proof, these results suggest that a mutation in an {alpha}-tubulin gene confers resistance to dinitroanilines in goosegrass.




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