Changes in L-phenylalanine ammonia-lyase activity and isoflavone phytoalexins accumulation in soybean seedlings infected with Sclerotinia sclerotiorum

2013 ◽  
Vol 8 (9) ◽  
pp. 921-929
Author(s):  
Djordje Malenčić ◽  
Jelena Cvejić ◽  
Vesna Tepavčević ◽  
Mira Bursać ◽  
Biljana Kiprovski ◽  
...  

AbstractSoybean [Glycine max (L.) Merr.] cultivars (Meli, Alisa, Sava and 1511/99) were grown up to V1 phase (first trifoliate and one node above unifoliate) and then inoculated with Sclerotinia sclerotiorum (Lib.) de Bary under controlled conditions. Changes in L-phenylalanine ammonia-lyase (PAL) activity and isoflavone phytoalexins were recorded 12, 24, 48 and 72 h after the inoculation. Results showed an increase in PAL activity in all four examined soybean cultivars 48 h after the inoculation, being the highest in Alisa (2-fold higher). Different contents of total daidzein, genistein, glycitein and coumestrol were detected in all samples. Alisa and Sava increased their total isoflavone content (33.9% and 6.2% higher than control, respectively) as well as 1511/99, although 48 h after the inoculation its content decreased significantly. Meli exhibited the highest rate of coumestrol biosynthesis (72 h after the inoculation) and PAL activity (48 h after the inoculation). All investigated cultivars are invariably susceptible to this pathogen. Recorded changes could point to possible differences in mechanisms of tolerance among them.

Weed Science ◽  
1981 ◽  
Vol 29 (4) ◽  
pp. 433-439 ◽  
Author(s):  
Robert E. Hoagland ◽  
Stephen O. Duke

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.


Weed Science ◽  
1983 ◽  
Vol 31 (6) ◽  
pp. 845-852 ◽  
Author(s):  
Robert E. Hoagland ◽  
Stephen O. Duke

The effects of 16 herbicides from 14 different chemical classes on levels of soluble protein, hydroxyphenolics, anthocyanin, and chlorophyll were determined in light- and dark-grown soybean [Glycine max(L.) Merr. ‘Hill’] seedlings. Growth-reducing concentrations of the herbicides were supplied to 3-day-old dark-grown soybean seedlings in liquid culture. Soluble protein (per axis) was reduced by fluridone {1-methyl-3-phenyl-5-[3-(trifluoromethyl)phenyl]-4(1H)-pyridinone}, paraquat (1,1′-dimethyl-4,4′-bipyridinium ion), perfluidone {1,1,1-trifluoro-N-[2-methyl-4-(phenylsulfonyl)phenyl] methanesulfonamide}, and propanil (3′,4′-dichloropropionanilide) 24 or 48 h after treatment. In light-grown plants, soluble hydroxyphenolic compound levels were decreased on a per axis basis after 48 h by all chemical treatments except by atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine], the methyl ester of diclofop {2-[4-(2,4-dichlorophenoxy)phenoxy] propanoate}, DSMA (disodium methanearsonate), fluridone, MH (1,2-dihydro-3,6-pyridazinedione), nitralin [4-(methylsulfonyl)-2,6-dinitro-N,N-dipropylaniline], TCA (trichloroacetic acid), and 2,4-D [(2,4-dichlorophenoxy)acetic acid]. Total chlorophyll content in hypocotyls of these seedlings was decreased by fluridone, metribuzin [4-amino-6-tert-butyl-3-(methylthio)-as-triazin-5 (4H)-one], norflurazon [4-chloro-5-(methylamino)-2-(α,α,α-trifluoro-m-tolyl)-3 (2H)-pyridazinone], paraquat, and 2,4-D, but others had no significant effect. Anthocyanin accumulation in hypocotyls of 48-h light-grown seedlings was decreased by atrazine, fenuron (1,1-dimethyl-3-phenylurea), metribuzin, norflurazon, paraquat, propanil, and propham (isopropyl carbanilate). Phenylalanine ammonia-lyase (PAL) activity (previously reported work) was positively correlated with anthocyanin levels in tissues after treatment with these 16 herbicides, but not with glyphosate [N-(phosphonomethyl)glycine]. There was, however, no correlation between extractable PAL activity and chlorophyll, soluble hydroxyphenolic compounds, or soluble protein content. These results indicated that extracted PAL activities usually reflect relative in vivo activities and that PAL activity is limiting to phenylpropanoid synthesis in vivo.


1982 ◽  
Vol 60 (6) ◽  
pp. 741-745 ◽  
Author(s):  
Jeffrey C. Suttle ◽  
Donald R. Schreiner

The effects of the herbicide DPX-4189 (2-chloro-N-((4-methoxy-6-methyl-1,3,5-triazin-2-yl)aminocarbonyl)benzenesulfonamide) on anthocyanin accumulation, phenylalanine ammonia lyase (PAL) activity, and ethylene production in seedlings of soybean (Glycine max L.) were investigated. Application of 1 μg DPX-4189 per plant led to an increase in anthocyanin content in soybean hypocotyls. The increase in anthocyanin content became evident 4 days after application of the herbicide. Accompanying the increase in anthocyanin content was an eightfold increase in extractable PAL activity. An increase in endogenous ethylene evolution also accompanied the increase in anthocyanin content. Application of silver nitrate (an inhibitor of ethylene action) to herbicide-treated seedlings did not prevent the increase in anthocyanin content. Application of 2-chloroethylphosphonic acid (an ethylene-releasing compound) to soybean seedlings stimulated PAL activity but had no effect on anthocyanin content. These results indicated that ethylene did not play a role in DPX-4189 mediated anthocyanin accumulation.


HortScience ◽  
1993 ◽  
Vol 28 (9) ◽  
pp. 937-938 ◽  
Author(s):  
Phillip N. Miklas ◽  
Kenneth F. Grafton ◽  
Phillip E. McClean

We investigated the partial physiological resistance (PPR) of common beans (Phaseolus vulgaris L.) to white mold disease caused by Sclerotinia sclerotiorum (Lib.) deBary. The activity of phenylalanine ammonia-lyase (PAL) was measured in detached stems inoculated with a growing mycelium of the pathogen. Noninoculated detached stems and whole plants were included as controls. Five bean cultivars-Upland, Bunsi, Sierra, UI-114, and Montcalm-and one breeding line-NY 5394-were tested; all varied in PPR to white mold disease. Greater PAL activity in the resistant NY 5394 than in the susceptible `Upland' suggests that PAL activity may be involved in the PPR of common beans to S. sclerotiorum.


Weed Science ◽  
1989 ◽  
Vol 37 (6) ◽  
pp. 743-747 ◽  
Author(s):  
Robert E. Hoagland

Three-day-old soybean seedlings were treated with acifluorfen via liquid culture [50 μM in dark, 1 μM in light (200 μE·m–2·s–1)]. Root elongation in the dark was inhibited only slightly (6%) after 96 h. In the light, acifluorfen inhibited root elongation after 48 h; after 96 h, inhibition was 32%. Hypocotyl length was not affected in either the light or dark. Soluble hydroxyphenolic content per axis was unaffected in the dark but was reduced by about 13 to 25% at 24 to 96 h in the light. Extractable phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) activity per axis was unaltered by herbicide in the dark. A brief transient increase in PAL activity (4 to 24 h) in axes of light-grown seedlings in the presence of acifluorfen was followed by reduced enzyme levels compared to light-grown controls at 72 and 96 h. Although light is required for maximal herbicide action, there was no effect on anthocyanin or chlorophyll accumulation or on the chlorophyll a/b ratio in hypocotyls or light-grown seedlings.


HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1127c-1127
Author(s):  
M.A. Ritenour ◽  
M.E. Saltveit ◽  
M.J. Ahrens

Russet spotting (RS) is an important postharvest disorder in Iceberg lettuce (Lactuca sativa L.). Previous studies showed that RS is induced by exposure to ∼5 ppm (ul/l) ethylene at ∼5C for 3 days and is characterized by the appearance of 1 to 2 mm diam. oval, brown sunken spots along the midrib. Increases in phenylalanine ammonia-lyase (PAL) activity and phenolic content are highly correlated with RS development. Ethylene-induced PAL activity is much less at higher (12C) or lower (0C) temperatures. In this study isolated whole leaves were exposed to a log series of ethylene concentrations from 0.1 to 10 ppm at temperatures from 0.0C to 20C for up to 8 days. Tissue was transferred among these various treatments to investigate the kinetics of PAL induction, activity and deactivation, phenolic accumulation, and RS development. A subjective evaluation was then made of RS development using a 1 to 9 scoring system in which 1 was no RS, and then PAL activity and phenolic content were measured. Preliminary results indicate that ethylene-induced PAL activity was decreased more rapidly upon transfer to temperatures above 10C than to 0.0C. Accumulation of phenolic compounds and development of RS paralleled each other, and were positively related to PAL activity. Practical implications of these results will be discussed.


2014 ◽  
Vol 57 (2) ◽  
pp. 279-286
Author(s):  
Khalequz Zaman

Comparison of SB<sub>1</sub> and SB<sub>3</sub> cell lines in both suspension and callus cultures showed that the latter, which produce much more dark brown pigments, had higher levels of condensed tannins. They also had higher total phenolics and flavonoid contents and L-phenylalanine ammonia lyase activity.


2004 ◽  
Vol 51 (3) ◽  
pp. 625-634 ◽  
Author(s):  
Mihai Nita-Lazar ◽  
Alain Heyraud ◽  
Claude Gey ◽  
Isabelle Braccini ◽  
Yvette Lienart

Activation of the phenolic pathway is known to be part of a defense response against cell wall-derived elicitors from pathogens. Many examples of a defense response by increasing the synthesis of phenolic compound against the elicitor were demonstrated in the past, but the elicitor structure has so far been poorly characterized. Our results indicate that a disaccharide fraction containing the following structure: alpha-D-mannopyranosyl (1-->2)alpha/beta-D-glucopyranosyl and alpha-D-mannopyranosyl (1-->x) inositol, isolated from Fusarium oxysporum L., promotes rapid and transient phenylalanine ammonia lyase activity in Rubus fructicosus cells at nanomolar concentration. The disaccharides were isolated by size-exclusion chromatography directly from extracts obtained by alkaline treatment of F. oxysporum mycelium. Their structure was determined by 500-MHz-1H-NMR spectroscopy combined with methylation analysis and fast atom bombardment mass spectrometry.


2014 ◽  
Vol 57 (1) ◽  
pp. 177-186 ◽  
Author(s):  
Khalequz Zaman

Through this preliminary experiment examining different color expressions in two soybean cell lines, SB<sub>1</sub>, and SB<sub>3</sub> several important features were observed. SB<sub>3</sub> cells, which produce much more dark brown pigments in both suspension and callus cultures, had higher total phenolics and flavonoid contents as well as higher L-phenylalanine ammonia lyase activity.


2011 ◽  
Vol 80 (2) ◽  
pp. 99-104 ◽  
Author(s):  
Marcin Horbowicz ◽  
Ryszard Kosson ◽  
Danuta Koczkodaj ◽  
Lesław B. Lahuta

The present paper discusses the effects of α-aminooxyacetic acid (AOA) on contents of polyamines, anthocyanins, photosynthetic pigments and phenylalanine ammonia-lyase activity in seedlings of common buckwheat (<em>Fagopyrum esculentum</em> Moench). AOA clearly decreased light-induced formation of anthocyanins and inhibited PAL activity in buckwheat hypocotyls, although a slight stimulatory effect on anthocyanins content in buckwheat cotyledons was observed. AOA declined the contents of chlorophylls <em>a</em> and <em>b</em> and total carotenoids in buckwheat cotyledons. The results show that AOA inhibits phenylpropanoids biosynthesis in buckwheat hypocotyls, and suppress photosynthesis in cotyledons. Moreover, the experiments show that AOA enhances the level of free putrescine in hypocotyls and the level of spermidine in buckwheat cotyledons. AOA also diminished the content of putrescine in cotyledons, but did not affect its level in buckwheat hypocotyls. AOA also substantially declined the level of cadaverine in buckwheat cotyledons, and did not affect its content in hypocotyls. Differences in effect of AOA on anthocyanins and polyamines accumulation indicate various physiological roles of the compounds in buckwheat hypocotyls and cotyledons.


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