ACCase mutations in Avena sterilis populations and their impact on plant fitness.

Avena sterilis (sterile oat) populations originating from wheat-growing regions of Greece, developed resistance to fenoxaprop, clodinafop and other herbicides. The partial ACCase gene sequence revealed six point mutations (Ile-1781-Leu, Trp-1999-Cys, Trp-2027-Cys, Ile-2041-Asn, Asp-2078-Gly, and Cys-2088-Arg) in 24 out of the 26 resistant (R) populations, confirming the molecular mechanism of resistance to ACCase-inhibiting herbicides. However, DNA sequence of two R populations did not reveal any known ACCase mutations, suggesting possible presence of unknown mutation or metabolism-based mechanism of resistance. The Cys-2088-Arg mutation is the first record for ACCase mutant conferring target-site resistance in A. sterilis worldwide. The evaluation of 12 R and 6 susceptible (S) populations under non-competitive field conditions did not indicate consistent mean growth rate differences, whereas the pot evaluation of the same (12 R and 6 S) populations grown in competition with wheat or in pure stands showed significant growth (fresh weight and panicle number) differences between six S populations and between six R populations containing the same ACCase mutation (Ile-2041-Asn). Finally, one S and five R (Trp-1999-Cys, Trp-2027-Cys, Ile-2041-Asn, Asp-2078-Gly, and Cys-2088-Arg) populations grown under field competitive conditions indicated fresh weight and panicle number differences in competition with other populations as compared with pure stands. These findings suggest clearly that the inconsistent fitness differences between R and S A. sterilis populations are not related with the ACCase resistance trait but they may result from other non-resistance fitness traits selected in their different geographical locations.

In the wild hybridization of annual Datura species as unveiled by morphological and molecular comparisons.

BACKGROUND: The present work aimed to verify whether intermediate variants were natural crosses between Datura species (D. stramonium forms and D. ferox). Their existence has been long ago insinuated but has not been studied using morphological features and molecular tools. The variants differed in stem coloring, upper bearing forks, and fruit characters. RESULTS: Principal Components Analysis of 11 morphological characteristics showed that D. ferox and D. stramonium (forms stramonium and tatula) were quite different and the putative hybrids were intermittent. The D. ferox × D. stramonium f. tatula was closer to the latter of its parents. Sequencing analysis revealed identical amplified trnL intron in all variants and a 100% homology with D. stramonium accession number EU580984.1 suggested that this plastid cannot discern Datura variants. However, genomic analysis with URP markers indicated that the hybrids had >60% genetic makeup similarity with both parents suggesting that the intermediate variants were putative inter-specific hybrids. Moreover, the dendrogram stemmed from cluster analysis of the fingerprint profile of variants placed D. stramonium and D. ferox in different branches indicating their genetic differentiation from each other as well as from their hybrids. CONCLUSIONS: The findings suggest that the natural hybridization of annual Datura species occurs. Extrapolating, this hybridization could be the first step for speciation. More possibly, it can alter population composition, its weediness and adaptability to local conditions.

Target-site mutation associated with cross-resistance to ALS-inhibiting herbicides in late watergrass (Echinochloa oryzicola Vasing.).

BACKGROUND: Studies were carried out to elucidate the mechanism of resistance to ALS-inhibiting herbicides in 29 Echinochloa accessions from water-seeded rice fields of northern Greece and to discriminate the Echinochloa species. RESULTS: Two E. oryzicola accessions were found to be cross-resistant to penoxsulam, bispyribac-sodium, imazamox, foramsulfuron, nicosulfuron and rimsulfuron, whereas all accessions were susceptible (S) to profoxydim. Sequencing of the ALS gene revealed that resistant (R) accessions had a Trp574Leu mutation, which was also confirmed by TspRI endonuclease digestion. Use of cpDNA sequence comparison analysis of Echinochloa species discriminated successfully E. crus-galli and E. oryzicola accessions. CONCLUSION: This is the first report of Echinochloa oryzicola cross-resistance to ALS-inhibiting herbicides as a result of Trp574Leu mutation. The cpDNA sequence comparison analysis is a reliable tool for discrimination of conventionally classified E. crus-galli and E. oryzicola accessions.

Corn poppy (Papaver rhoeas) cross-resistance to ALS-inhibiting herbicides.

BACKGROUND: Papaver rhoeas (L.) has evolved resistance to tribenuron in winter wheat fields in northern Greece owing to multiple Pro(197) substitutions. Therefore, the cross-resistance pattern to other sulfonylurea and non-sulfonylurea ALS-inhibiting herbicides of the tribenuron resistant (R) and susceptible (S) corn poppy populations was studied by using whole-plant trials and in vitro ALS catalytic activity assays. RESULTS: The whole-plant trials revealed that tribenuron R populations were also cross-resistant to sulfonylureas mesosulfuron + iodosulfuron, chlorsulfuron and triasulfuron. The whole-plant resistance factors (RFs) calculated for pyrithiobac, imazamox and florasulam ranged from 12.4 to > 88, from 1.5 to 28.3 and from 5.6 to 25.4, respectively, and were lower than the respective tribenuron RF values (137 to > 2400). The ALS activity assay showed higher resistance of the ALS enzyme to sulfonylurea herbicides (tribenuron > chlorsulfuron) and lower resistance to non-sulfonylurea ALS-inhibiting herbicides (pyrithiobac > florasulam ≈ imazamox). CONCLUSION: These findings indicate that Pro(197) substitution by Ala, Ser, Arg or Thr in corn poppy results in a less sensitive ALS enzyme to sulfonylurea herbicides than to other ALS-inhibiting herbicides. The continued use of sulfonylurea herbicides led to cross-resistance to all ALS-inhibiting herbicides, making their use impossible in corn poppy resistance management programmes.