Inhibition of Tyrosine Aminotransferase by β-N-Oxalyl-l-α,β-Diaminopropionic Acid, the Lathyrus sativus Neurotoxin

Abstract: Species differences in susceptibility are a unique feature associated with the neurotoxicity of β-N-oxalyl-l -α,β-diaminopropionic acid (l -ODAP), the Lathyrus sativus neurotoxin, and the excitotoxic mechanism proposed for its mechanism of toxicity does not account for this feature. The present study examines whether neurotoxicity of l -ODAP is the result of an interference in the metabolism of any amino acid and if it could form the basis to explain the species differences in susceptibility. Thus, Wistar rats and BALB/c (white) mice, which are normally resistant to l -ODAP, became susceptible to it following pretreatment with tyrosine (or phenylalanine), exhibiting typical neurotoxic symptoms. C57BL/6J (black) mice were, however, normally susceptible to l -ODAP without any pretreatment with tyrosine. Among the various enzymes associated with tyrosine metabolism examined, the activity of only tyrosine aminotransferase (TAT) was inhibited specifically by l -ODAP. The inhibition was noncompetitive with respect to tyrosine (K_i = 2.0 ± 0.1 mM) and uncompetitive with respect to α-ketoglutarate (K_i = 8.4 ± 1.5 mM). The inhibition of TAT was also reflected in a marked decrease in the rate of oxidation of tyrosine by liver slices, an increase in tyrosine levels of liver, and also a twofold increase in the dopa and dopamine contents of brain in l -ODAP-injected black mice. The dopa and dopamine contents in the brain of only l -ODAP-injected white mice did not show any change, whereas levels of these compounds were much higher in tyrosine-pretreated animals. Also, the radioactivity associated with tyrosine, dopa, and dopamine arising from [¹⁴C]tyrosine was twofold higher in both liver and brain of l -ODAP-treated black mice. Thus, a transient increase in tyrosine levels following the inhibition of hepatic TAT by l -ODAP and its increased availability for the enhanced synthesis of dopa and dopamine and other likely metabolites (toxic?) resulting therefrom could be the mechanism of neurotoxicity and may even underlie the species differences in susceptibility to this neurotoxin.     

Effects of Application Strategies of Fumigant and Nonfumigant Nematicides on Cantaloupe Grown in Deep Sand Soils in Florida

A 2-year study was conducted in which three treatment tactics of oxamyl (at planting application, application every 2 weeks, and rescue applications, as determined by crop symptoms) were compared to fumigant treatments with methyl bromide, 1,3-dichloropropene (1,3-D), and 1,3-D plus chloropicrin for management of Meloidogyne spp. In 2002, treatments that included 1,3-D produced higher yields as determined both by number and weight of marketable fruit. All treatment tactics relying solely on oxamyl, at planting, scheduled treatments, and rescue, were not different from untreated controls for both marketable yield and number of fruit. Gall ratings in 2002 were lowest for 1,3-D at the 112-liters/ha rate, followed by 1,3-D at 84 liters/ha with and without oxamyl. All treatments of oxamyl, except when combined with 1,3-D, had gall ratings not different from untreated plots. In 2004, treatments of methyl bromide and 1,3-D plus chloropicrin had the highest total number of both marketable fruit and highest marketable yields. All treatment strategies relying solely on oxamyl had yields equivalent to the untreated controls. Mean root-gall ratings were lowest for methyl bromide plus chloropicrin and 1,3-D plus chloropicrin treatments. Root-gall ratings for all treatment tactics relying solely on oxamyl were not different from untreated controls.     

Sieve element occlusion provides resistance against Aphis gossypii in TGR-1551 melons

Feeding behavior and plant response to feeding were studied for the aphid Aphis gossypii Glover on susceptible and resistant melons(cv.Iroquois and TGR-1551,respectively).Average phloem phase bout duration on TGR-1551 was<7% of the duration on Iroquois.Sixty-seven percent of aphids on TGR-1551 never produced a phloem phase that attained ingestion(EPG waveform E2)in contrast to only 7% of aphids on Iroquois.Average bout duration of waveform E2(scored as zero if phloem phase did not attain E2)on TGR-1551 was<3% of the duration on Iroquois.Conversely,average bout duration of EPG waveform El(sieve element salivation)was 2.8 times greater on TGR-1551 than on Iroquois.In a second experiment,liquid nitrogen was used to rapidly cryofix leaves and aphids within a few minutes after the aphids penetrated a sieve element.Phloem near the penetration site was then examined by confocal laser scanning microscopy.Ninety-six percent of penetrated sieve elements were occluded by protein in TGR-1551 in contrast to only 28% in Iroquois.Usually in TGR-1551,occlusion was also observed in nearby nonpenetrated sieve elements.Next,a calcium channel blocker,trivalent lanthanum,was used to prevent phloem occlusion in TGR-1551,and A.gossypii feeding behavior and the plants phloem response were compared between lanthanum-treated and control TGR-1551.Lanthanum treatment eliminated the sieve element protein occlusion response and the aphids readily ingested phloem sap from treated plants.This study provides strong evidence that phloem occlusion is a mechanism for resistance against A.gossypii in TGR-1551.     

Phosphorus uptake and translocation in salt-stressed melon plants

Melon seedlings (Cucumis melo L. cv.Galia) were grown hydroponically to study the effect of salinity (80 mmol/LNaCl) on phosphate (Pi) uptake and translocation at two levels of Pi (25 μmol/L and 1 mmol/L). Net uptake rates of Pi were determined by depletionof the medium and by plant content. Salinity decreased Pi uptake at low Pi (high affinity uptake mechanism), 25 μmol/L, although no specific competitive inhibition of Pi uptake by Cl⁻ was observed. When plants were grown with high Pi (1 mmol/L), the uptake of Pi through the low affinity system was increased by 80 mmol/L NaCl. Salinity also reduced the phosphorus flux, as Pi, through the xylem. It is hypothesised that high levels of NaCl decrease the mobility of Pi stored in vacuoles, and as a result, inhibit export from this storage compartment to other parts of the plant.     

Effect of temperature on the growth of individual cucumber fruits

In order to study the effect of temperature on the growth of individual fruits in cucumber (cucumis sativus L. cv. Corona), fruits were grown at 17. 5. 20,25 and 30°C continuously or the fruit temperature was changed from 17. 5 to 27.5°C or vice versa. Fruit development appeared to be closely related to the temperature sum. When the growth of a fruit was not constrained by assimilate supply, a decrease in growing period with increasing temperature was more than compensated for by a strong increase in growth rate, resulting in an increase in final fruit weight. However, when the growth of a fruit was constrained by assimilate supply, the increase in growth rate with increasing temperature was small and did not compensate for the decrease in growing period, resulting in a decrease in final fruit weight. Determinations of cell number and size showed that the effect of temperature on fruit growth was due to effects on cell expansion rather than on cell division. When growth was not constrained by assimilate supply. However, when assimilate supply did constrain fruit growth the number of cells per fruit decreased with increasing temperature, while the effect on cell size was negligible. In all stages of fruit development, the growth rate of a cucumber fruit responded within one day to a change in temperature. It was not irreversibly impaired by a low temperature (17. 5°C) during the early development of a fruit. A high temperature treatment (27. 5°C), however, had a great effect on the growth rate of a fruit after the temperature treatment had terminated. At all stages of fruit development (even before anthesis) a period of four days at 27. 5°C resulted in a pronounced stimulation of the growth rate afterwards at 17. 5°C.     

Photoinhibition of radish (Raphanus sativus L) seed germination: control of growth potential by cell-wall yielding in the embryo

The biophysical mechanism underlying photoinhibition of radish (Raphanus sativus L.) seed germination was investigated using three cultivars differing in sensitivity to continuous irradiation with far-red light (high-irradiance reaction of phytochrome). Sensitivity of germination to the inhibitory action of light was assessed by probing germination under osmotic stress (incubation in media of low water potentials adjusted with polyethylene glycol 6000) and expressed in terms of ‘germination potential’ (positive value of the water potential at which germination is inhibited by 50%). Far-red light decreases the germination potential to various degrees in the different cultivars, reflecting the light-sensitivity of germination in water. Removal of the seed coat increases the germination potential by a constant amount in darkness and light. It is concluded that germination depends on the expansive force of the embryo which can be drastically diminished by far-red light. Seed-coat constraint and expansive force of the embryo interact additively on the level of the germination potential. Photoinhibition of germination was accompanied by an inhibition of water uptake into the seed. Analysis of seed water relations showed that osmotic pressure and turgor assumed higher levels in photoinhibited seeds, compared to seeds germinating in darkness, while the water potential was close to zero under both conditions. Far-red light produced a shift (to less negative values) in the curve relating water-uptake rate to external water potential, i.e. a reduction in the driving force for water uptake. It is concluded that photoinhibition of germination results from the maintenance of a high threshold of cell-wall extensibility in the embryo.     

Melon resistance to the aphid Aphis gossypii: behavioural analysis and chemical correlations with nitrogenous compounds

In the melon, the Vat (monogenic, dominant) resistance gene governs both an antixenotic reaction to the melon aphid Aphis gossypii Glover (Homoptera, Aphididae) and a resistance to non-persistent virus transmission, restricted to this vector species. We investigated the behavioural features and tissue localisation of the antixenosis resistance by the electrical penetration graph technique (EPG, DC system). We also compared the chemical composition in amino compounds and proteins of the phloem sap collected from two isogenic lines of melon (Cucumis melo L.), carrying the Vat gene or not. All behavioural and chemical data indicated that this resistance is constitutive. EPG analysis clearly showed that access to phloem, although delayed by alterations in pathway activities, was not impaired in terms of frequency of access or initiation of feeding. The most striking feature was, however, a very reduced duration of ingestion from phloem of resistant plants, making this compartment one of the tissues where the effects of the Vat gene are unambiguously expressed. This was confirmed by clear differential activity of phloem extracts in artificial no-choice bioassays. Chemical analyses have shown that phloem saps from the two isogenic lines were extremely similar in profiles of ninhydrin positive compounds, and contained a low total amount of free amino acids (less than 10 mM). Out of more than 40 distinguishable peaks in the chromatograms (protein and non-protein amino acids, as well as small peptides), only five differentiated the two genotypes. Two of them were increased in the resistant genotype: glutamic acid and a major unknown peak, probably a non-protein amino acid (different from pyrazolyl-alanine, a Cucumis-specific amino acid). The three others were depressed in resistant plants, and included the sulphur amino acid cystine and a peptide peak partly composed of the cysteine-containing peptide glutathione (reduced form). Sap collection also showed that phloem exudation rates, as well as total protein and glutathione levels, were depressed in phloem sap from resistant plants. Such data are all indicative of a modified phloem-sealing physiology, linked to sulfhydryl oxidation processes, in plants carrying the Vat gene. The originality of the mechanism of Vat resistance to aphids is discussed.     

细菌性角斑病菌诱导黄瓜叶片水杨酸的积累

对黄瓜(Cucumis sativa L.)幼苗第一片真叶接种细菌性角斑病菌(Pseudomonas syringae pv.lachy-mans)后,利用TLC和HPLC方法分离测定了不同时间内游离态和结合态水杨酸(SA)的含量。结果表明,接种显著提高了第一片真叶游离态SA含量,增加了30％～197％,结合态SA的含量变化与游离态相似,提高了79％～240％,但是接种后第三天游离态SA和结合态SA含量都下降。接种还诱导了非接种第二片真叶中SA的积累:游离态SA含量在接种初期增加了29％～46％,接种后第三天达到峰值,第五天回复到正常水平,结合态SA含量在接种初期没有变化,接种后第四至六天也显著增加,提高了62％～107％。