Chlorophyll fluorescence protocol for quick detection of triazinone resistant Chenopodium album L

Sugar beet growers in Europe are more often confronted with an unsatisfactory control of Chenopodium album L. (fat-hen), possibly due to the presence of a triazinone resistant biotype. So far, two mutations on the psbA-gene, i.e. Ser264-Gly and Ala251-Val, are known to cause resistance in C. album to the photosystem II-inhibiting triazinones metamitron, a key herbicide in sugar beet, and metribuzin. The Ser264-Gly biotype, cross-resistant to many other photosystem II-inhibitors like the triazines atrazine and terbuthylazine, is most common. The second resistant C. album biotype, recorded in Sweden, is highly resistant to triazinones but only slightly cross-resistant to terbuthylazine. Since farmers should adapt their weed control strategy when a resistant biotype is present, a quick and cheap detection method is needed. Therefore, through trial and error, a protocol for detection with chlorophyll fluorescence measurements was developed and put to the test. First, C. album leaves were incubated in herbicide solution (i.e. 0 microM, 25 microM metribuzin, 200 microM metamitron or 25 microM terbuthylazine) during three hours under natural light. After 30 minutes of dark adaptation, photosynthesis yield was measured with Pocket PEA (Hansatech Instruments). In Leaves from sensitive C. album, herbicide treatment reduces photosynthesis yield due to inhibition of photosynthesis at photosystem II. This results in a difference of photosynthesis yield between the untreated control and herbicide treatment. Based on the relative photosynthesis yield (as a percentage of untreated), a classification rule was formulated: C. album is classified as sensitive when its relative photosynthesis yield is less than 90%, otherwise it is resistant. While metribuzin, and to a lesser extent, metamitron treatment allowed a quick detection of triazinone resistant C. album, terbuthylazine treatment was able to distinguish the Ser264-Gly from the Ala251-Val biotype. As a final test, 265 plants were classified with the protocol. Simultaneously, a CLeaved Amplified Polymorphic Sequence (CAPS)-analysis was conducted on the same plants to verify the presence of the Ser264-Gly mutation. Only one mismatch was found when results of both detection methods were compared. The test results illustrate that this protocol provides a reliable, quick and cheap alternative for DNA-analysis and bio-assays to detect the triazinone resistant C. album biotypes.