Effects of Herbicides on Growth and Extractable Phenylalanine Ammonia-Lyase Activity in Light- and Dark-Grown Soybean (Glycine max) Seedlings

Abstract

Effects of 16 herbicides representing 14 herbicide classes on growth and extractable phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) were examined in light- and dark-grown soybean [Glycine max(L.) Merr. ‘Hill’] seedlings. High purity (96 to 100%) herbicides were supplied via aqueous culture at various concentrations: 0.5 mM amitrole (3-amino-s-triazole), 0.1 mM atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine], 0.07 mM diclofop-methyl {methyl ester of 2-[4-(2,4-dichlorophenoxy)phenoxy] propanoicacid}, 0.5 mM DSMA (disodium methanearsonate), 0.2 mM fenuron (1,1-dimethyl-3-phenylurea), 0.05 mM fluridone {1-methyl-3-phenyl-[3-(trifluoromethyl)phenyl]-4(1H)-pyridinone}, 0.5 mM MH (1,2-dihydro-3,6-pyridazinedione), 0.5 mM metribuzin [4-amino-6-tert-butyl-3-(methylthio)-as-triazin-5(4H)-one], 1.8 μM nitralin [4-(methylsulfonyl)-2,6-dinitro-N,N-dipropylaniline], 0.5 mM norflurazon [4-chloro-5-(methylamino)-2-(α,α,α-trifluoro-m-tolyl)-3(2H)-pyridazinone], 0.05 mM paraquat (1,1′-dimethyl-4,4′-bipyridinium ion), 0.15 mM perfluidone {1,1,1-trifluoro-N-[2-methyl-4-(phenylsulfonyl)phenyl] methanesulfonamide}, 0.2 mM propanil (3′,4′-dichloropropionanilide), 0.1 mM propham (isopropyl carbanilate), 0.5 mM TCA (trichloroacetic acid), and 0.05 mM 2,4-D [(2,4-dichlorophenoxy)acetic acid]. Dark-grown soybean seedlings (3-day-old) were transferred to control solutions (2 mM CaSO4) or to herbicide solutions (in 2 mM CaSO4) and grown at 25 C in continuous white light (200 μE•m-2•s-1) or continuous darkness until harvested 24 or 48 h after transfer. After 48 h, growth (fresh weight, dry weight, elongation) was inhibited by most of the chemicals. Other signs of toxicity (necrosis, secondary root stunting, and root tip swelling) were noted for some treatments. Roots were most affected, although hypocotyls were generally not changed. Hypocotyl elongation was stimulated by atrazine, fluridone, and norflurazon after 48 h light. Extractable PAL activity from soybean axes was decreased by atrazine, fenuron, metribuzin, norflurazon, propanil, propham, and 2,4-D. Amitrole and paraquat were the only herbicides that increased extractable PAL activity. Other compounds tested had no effect on the enzyme. None of the herbicides significantly affected in vitro PAL activity.