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Physiological and Molecular Characterization of Mutation-Derived Imidazolinone Resistance in Spring Wheat

^a Crop Development Centre, Dep. of Plant Sciences, Univ. of Saskatchewan, 51 Campus Drive, Saskatoon, SK, Canada, S7N 5A8
^b BASF Plant Sciences, Research Triangle Park, Raleigh, NC 27709 USA
^c DNA LandMarks Inc., 84 Richelieu Street, Saint-Jean-sur-Richelieu, QC, Canada, J3B 6X3

View larger version (13K): [in a new window]   Fig. 1. In vitro inhibition of AHAS activity in resistant lines TealIMI 10A (), TealIMI 11A (), TealIMI 15A (), BW755 () and susceptible line CDC Teal () by imazethapyr (µM). Lines represent the fitted line of data from four independent experiments with two replications in each.

View larger version (13K): [in a new window]   Fig. 2. In vitro inhibition of AHAS activity in resistant lines TealIMI 10A (), TealIMI 11A (), TealIMI 15A (), BW755 () and susceptible line CDC Teal () by chlorsulfuron (nM). Lines represent the fitted line of data from four independent experiments with two replications in each. Data points represent the means at each concentration of chlorsulfuron.

View larger version (12K): [in a new window]   Fig. 3. In vitro inhibition of AHAS activity in resistant lines TealIMI 10A (), TealIMI 11A (), TealIMI 15A (), BW755 () and susceptible line CDC Teal () by valine + leucine (µM). Lines represent the fitted line of data from four independent experiments with two replications in each. Data points represent the means of each treatment.

View larger version (45K): [in a new window]   Fig. 4. MspI digestion of a 617-bp product amplified from plasmid DNA extracted from recombinant clones containing the imi1, imi2, and imi3 genes cloned from CDC Teal. The three genes could be distinguished on the basis of polymorphic restriction patterns following digestion of the 617-bp amplicon. The 617-bp sequence of imi1 lacks an MspI digestion site (Lanes 2, 9, 10), whereas imi2 possesses four restriction sites, resulting in fragments 63, 75, 171, and 223 bp in length (Lanes 3, 4, 8). The 617-bp sequence of imi3 possesses a single restriction site, resulting in fragments 446 and 171 bp in length (Lanes 1, 5, 6, 7). Identical restriction patterns where noted in all resistant lines (Data not shown).

View larger version (30K): [in a new window]   Fig. 5. DNA sequence alignment of partial catalytic subunit genes amplified from genomic DNA of CDC Teal, TealIMI 10A, TealIMI 11A, TealIMI 15A, and BW755. Only relevant sequence is presented with complete sequences available on GenBank (accession numbers presented on figure). Imi1 and Imi2 and wild-type sequences (1816 bp) were PCR cloned by means of primers AHAS17FWD and AHAS30REV. Nucleotide sequences of Imi3 and imi3 (617 bp) were PCR cloned by means of primers AHAS21FWD and AHAS26REV. The ruler represents the amino acid residue no. of AHAS from O. sativa. Single nucleotide polymorphisms between the three partial wheat catalytic subunit genes are indicated by a box.

View larger version (44K): [in a new window]   Fig. 6. MspI digestion of a 617-bp product amplified from genomic DNA of wild-type Chinese Spring (Lane 1) and aneuploid stocks (Lanes 2–10) by means of primers AHAS21FWD and AHAS26REV: Lane 2: Nullisomic 6A; Terasomic 6B; Lane 3: Nullisomic 6B; Tetrasomic 6D; Lane 4: Nullisomic 6D; Tetrasomic 6A; Lane 5: Ditelosomic 6AS; Lane 6: Ditelosomic 6AL; Lane 7: Ditelosomic 6BS; Lane 8: Ditelosomic BL; Lane 9: Ditelosomic 6DS; Lane 10: Ditelosomic 6DL.