Non-target site mechanism of metribuzin tolerance in induced tolerant mutants of narrow-leafed lupin (Lupinus angustifolius L.)

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Contents Vol 63(5)

doi:10.1071/CP12065

Plant Sciences, Sustainable Farming Systems & Food Quality

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Protocols in ecological and environmental plant physiology

Non-target site mechanism of metribuzin tolerance in induced tolerant mutants of narrow-leafed lupin (Lupinus angustifolius L.)

Gang Pan ^A ^B, Ping Si ^A ^D, Qin Yu ^C, Jumin Tu ^B and Stephen Powles ^C

^A Centre for Legumes in Mediterranean Agriculture (CLIMA), Faculty of Natural and Agricultural Sciences, The University of Western Australia, Crawley, WA 6009, Australia.
^B Department of Agronomy, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, 310029, China.
^C Australian Herbicide Resistance Initiative (AHRI), School of Plant Biology, The University of Western Australia, Crawley, WA 6009, Australia.
^D Corresponding author. Email: ping.si@uwa.edu.au

Crop and Pasture Science 63(5) 452-458 http://dx.doi.org/10.1071/CP12065
Submitted: 24 February 2012 Accepted: 28 June 2012 Published: 25 July 2012





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Abstract

Narrow-leafed lupin (Lupinus angustifolius L.) is an important grain legume crop in Australia. Metribuzin is an important herbicide used to control weeds in lupin crops. This study investigated metribuzin tolerance mechanism in narrow-leafed lupin by comparing two induced mutants (Tanjil-AZ-33 and Tanjil-AZ-55) of higher metribuzin tolerance with the susceptible wild type. Sequencing of the highly conserved region of the chloroplast psbA gene (target site) revealed that the sequences of the wild type and the mutants were identical and therefore metribuzin tolerance is not target site based. Photosynthetic activity was measured and the leaf photosynthesis of the two tolerant mutants was initially inhibited after metribuzin treatment, but recovered within 2.5 days whereas that of the susceptible plants remained inhibited. The photosynthetic measurements confirmed the target site chloroplast was susceptible and the tolerance mechanism is non-target site based. Investigation with known cytochrome P450 monooxygenase inhibitors (omethoate, malathion and phorate) showed that tolerance could be reversed in both mutants, indicating the tolerance mechanism in two tolerant mutants may involve cytochrome P450 enzymes. Interestingly, the inhibitor tridiphane reversed metribuzin tolerance of only one of the two tolerant mutants, indicating diversity in metribuzin tolerance mechanisms in narrow-leafed lupin. These results signify that further investigation of metribuzin metabolism in these plants is warranted. In conclusion, metribuzin tolerance mechanism in lupin mutants is non-target site based, likely involving P450-mediated metribuzin metabolism.

Additional keywords: cytochrome P450 inhibitor, Lupinus angustifolius L., metribuzin tolerance, photosynthesis, psbA gene sequence.

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