Asparagusate dehydrogenases and lipoyl dehydrogenase from asparagus mitochondria. Physical, chemical, and enzymatic properties.

Asparagusate dehydrogenases I and II and lipoyl dehydrogenase have been obtained in homogeneous state from asparagus mitochondria. They are flavin enzymes with 1 mol of FAD/mol of protein. Asparagusate dehydrogenases I and II and lipoyl dehydrogenase have s20,w of 6.22 S, 6.39 S, and 5.91 S, respectively, and molecular weights of 111,000, 110,000, and 95,000 (sedimentation equilibrium) or 112,000, 112,000, and 92,000 (gel filtration). They are slightly acidic proteins with isoelectric points of 6.75, 5.75, and 6.80. Both asparagusate dehydrogenases catalyzed the reaction Asg(SH)2 + NAD+ equilibrium AsgS2 + NADH + H+ and exhibit lipoyl dehydrogenase and diaphorase activities. Lipoyl dehydrogenase is specific for lipoate and has no asparagusate dehydrogenase activity. NADP cannot replace NAD in any case. Optimum pH for substrate reduction of the three enzymes are near 5.9. Asparagusate dehydrogenases I and II have Km values of 21.5 mM and 20.0 mM for asparagusate and 3.0 mM and 3.3 mM for lipoate, respectively. Lipoyl dehydrogenase activity of asparagusate dehydrogenases is enhanced by NAD and surfactants such as lecithin and Tween 80, but asparagusate dehydrogenase activity is not enhanced. Asparagusate dehydrogenases are strongly inhibited by mercuric ion, p-chloromercuribenzoic acid, and N-ethylmaleimide. Amino acid composition of the three enzymes is presented and discussed.     

Alicyclic acid metabolism in plants 8. Association, of two enzymes of the shikimate pathway in shoots of Phaseolus mungo seedlings

The association of two enzymes involved in the shikimate pathway,3-dehydroquinate hydro-lyase (EC 4.2.1.10 [EC] ) and shikimate: NADPoxidoreductase (EC 1.1.1.25 [EC] ), was studied with shoots of etiolated4-day-old Phaseolus mungo seedlings. The enzymes were not separableby ammonium sulfate fractionation, sucrose density gradientcentrifugation, polyacrylamide gel electrophoresis and chromatographyon Sephadex G-100 and DEAE-Sephadex A-50. The results are discussedin relation to the channelling function of metabolites in thealicyclic acid metabolism in higher plants. (Received October 28, 1975; )     

Evidence for the expression of morphological and biochemical characteristics of C3-photosynthesis in chlorohyllous callus cultures of Zea mays

A Zea mays callus culture containing chlorophyll was established and grown photomixotrophically. Cell chloroplast structure, and pigment and soluble protein contents were examined. Expression of some key enzymes of C₄ carbon metabolism was compared with that of etiolated (heterotrophic) and green photoautotrophic leaves. Chlorophyll content of the callus was 15–20% that of green leaves. Soluble protein content of callus was half that of leaf cells. Electron microscopic observations showed that green callus cells contained only typical granal chloroplasts. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.38) activities in green callus were ca 30% those of green leaves but 2–3 times higher than in etiolated leaves. Quantitative enzyme protein determination, using antibodies specific to maize leaf Rubisco showed that the chloroplastic carboxylase represented about 7% of total soluble protein in green callus, in parallel to its low chlorophyll content. The specific activity of Rubisco in callus and leaves was unchanged. Phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) activity in green callus was about 20% that of green leaves and similar to that measured in etiolated leaves. Apparent K_m (PEP) values (0.08 mM) for PEPC isolated from green callus and etiolated leaves were very different from values (0.5 mM) obtained with PEPC from green leaves. These kinetic characteristics together with the absence of inhibition by malate and activation by glucose-6-phosphate suggest that the properties of PEPC isolated from green callus and etiolated maize leaves are very similar to those of PEPPC from C₃ plants. Using PEPC antibodies specific to green maize leaf enzyme, immunotitration of PEPC preparations containing identical enzyme units allowed complete precipitation of the green leaf enzyme with increasing antibody volumes. In contrast, 60–70% of the activity of PEPC from etiolated and green callus was inhibited, suggesting low affinity for the maize green leaf PEPC antiserum (typical C₄ form). Ouchterlony double diffusion tests revealed only partial recognition of PEPC in green callus and etiolated leaves. NAD-malate dehydrogenase (NAD-MDH, EC 1.1.1.37) activity in callus was 2 and 3 times higher, respectively, than in etiolated and green leaves. NADP-malic enzyme (NADP-ME, EC 1.1.1.40) activity in callus cultures was much lower than in green leaves. All our data support the hypothesis that cultures of fully dedifferentiated chlorophyllous tissues of Zea mays possess a C₃-like metabolism.     

Polyamine oxidase activity and polyamine content in maize during seed germination

Polyamine oxidase (PAO, EC 1.5.3.3) activity and polyamine content in the cell wall and soluble fractions obtained from embryos, endosperms and shoots and roots of etiolated or green seedlings of maize ( Zea mays L. cv. WF9) during the first 7 days of germination were investigated. Polyamine content was also determined in the trichloroacetic acid-soluble (free polyamines) and trichloroacetic acid insoluble (bound polyamines) fraction obtained from the same tissues. PAO activity, determined by the radiometric method based on the recovery of the labelled reaction product 1-pyrroline, was mostly localized in the cell wall fraction. The activity was very low in embryos and endosperms and present in traces in roots. In etiolated shoots PAO activity increased sharply, while in green shoots it was low and increased slowly. No polyamines were found in the cell wall fraction and only putrescine was detected in the soluble fraction, with the exception of the embryo, where spermidine and spermine were also present. In the TCA-soluble fraction of embryos, putrescine increased during imbibition, while spermidine and spermine decreased; in the endosperm no relevant changes in polyamines occurred. In the same fraction of green and etiolated seedlings, putrescine increased, giving a peak at days 3–5, while spermidine decreased to very low levels. The amount of bound polyamines was 1–4% of the free ones. The pattern of PAO activity seems to be unrelated to endogenous free polyamine content, which is the same in shoots and roots of etiolated and green seedlings. Enzyme activity, very low in ungerminated seeds, increased continuously during the progression of germination, especially in etiolated shoots, indicating a possible involvement in cell wall formation.     

Effect of malformin on phytochrome reactions in Vigna and Avena

The rate of destruction of the far red absorbing form of phytochrome(Pfr) in green or etiolated cuttings of Vigna radiata was slowerin the presence of malformin than in its absence. Malforminhad no effect on the accumulation of total phytochrome in thedark, or on the reaccumulation of phytochrome after destructionin red light. The amount of photoconversion of the red absorbingform of phytochrome (Pr) to Pfr or Pfr to Pr by given dosesof red or far red radiation was slightly but consistently lessin malformin-treated cuttings of V. radiata than in controls.Malformin had no effect on the rate of destruction or photoconversionof phytochrome in etiolated shoots of Avena sativa. The decreasein destruction rate of Pfr by malformin in V. radiata may contributeto the inhibition of dark abscission by malformin after lighttreatment. (Received October 3, 1979; )     

Immunochemical detection with rabbit polyclonal and mouse monoclonal antibodies of different pools of phytochrome from etiolated and green Avena shoots

While two monoclonal antibodies directed to phytochrome from etiolated oat (Avena sativa L.) shoots can precipitate up to about 30% of the photoreversible phytochrome isolated from green oat shoots, most precipitate little or none at all. These results are consistent with a report by J.G. Tokuhisa and P.H. Quail (1983, Plant Physiol. 72, Suppl., 85), according to which polyclonal rabbit antibodies directed to phytochrome from etiolated oat shoots bind only a small fraction of the phytochrome obtained from green oat shoots. The immunoprecipitation data reported here indicate that essentially all phytochrome isolated from green oat shoots is distinct from that obtained from etiolated oat shoots. The data indicate further that phytochrome from green oat shoots might itself be composed of two or more immunochemically distinct populations, each of which is distinct from phytochrome from etiolated shoots. Phytochrome isolated from light-grown, but norflurazon-bleached oat shoots is like that isolated from green oat shoots. When light-grown, green oat seedlings are kept in darkness for 48 h, however, much, if not all, of the phytochrome that reaccumulates is like that from etiolated oat shoots. Neither modification during purification from green oat shoots of phytochrome like that from etiolated oat shoots, nor non-specific interference by substances in extracts of green oat shoots, can explain the inability of antibodies to recognize phytochrome isolated from green oat shoots. Immunopurified polyclonal rabbit antibodies to phytochrome from etiolated pea (Pisum sativum L.). shoots precipitate more than 95% of the photoreversible phytochrome obtained from etiolated pea shoots, while no more than 75% of the pigment is precipitated when phytochrome is isolated from green pea shoots. These data indicate in preliminary fashion that an immunochemically unique pool of phytochrome might also be present in extracts of green pea shoots.Abbreviation ELISA enzyme-linked immunosorbent assay - mU milliunit - Pfr far-red-absorbing form of phytochrome - Pr red-absorbing form of phytochrome     

Nonreversible d-Glyceraldehyde 3-Phosphate Dehydrogenase of Plant Tissues

Preparations of TPN-linked nonreversible d-glyceraldehyde 3-phosphate dehydrogenase (EC 1.2.1.9), free of TPN-linked reversible d-glyceraldehyde 3-phosphate dehydrogenase, have been obtained from green shoots, etiolated shoots, and cotyledons of pea (Pisum sativum), cotyledons of peanut (Arachis hypogea), and leaves of maize (Zea mays). The properties of the enzyme were similar from each of these sources: the Km values for d-glyceraldehyde 3-phosphate and TPN were about 20 μm and 3 μm, respectively. The enzyme activity was inhibited by l-glyceraldehyde 3-phosphate, d-erythrose 4-phosphate, and phosphohydroxypyruvate. Activity was found predominantly in photosynthetic and gluconeogenic tissues of higher plants. A light-induced, phytochrome-mediated increase of enzyme activity in a photosynthetic tissue (pea shoots) was demonstrated. Appearance of enzyme activity in a gluconeogenic tissue (endosperm of castor bean, Ricinus communis) coincided with the conversion of fat to carbohydrate during germination. In photosynthetic tissue, the enzyme is located outside the chloroplast, and at in vivo levels of triose-phosphates and pyridine nucleotides, the activity is probably greater than that of DPN-linked reversible d-glyceraldehyde 3-phosphate dehydrogenase. Several possible roles for the enzyme in plant carbohydrate metabolism are considered.     

Spectral sensitivity of the flowering response in green and etiolated Lemna paucicostata T-101

Light sensitivity in the reversal of the far-red inhibitionof flowering in etiolated Lemna paucicostata T-101 was 10- (612nm) to 33-fold (660 nm) higher than that of green plants. Theaction spectrum of this response for etiolated plants showeda peak near 660 nm, but for green plants it was near 612 nm.Thus, the effect of light on flowering is influenced greatlyby the screening effect of photosynthetic pigments. (Received July 24, 1980; )     

Spectral sensitivity of the flowering response in green and etiolated Lemna paucicostata T-101

Light sensitivity in the reversal of the far-red inhibitionof flowering in etiolated Lemna paucicostata T-101 was 10- (612nm) to 33-fold (660 nm) higher than that of green plants. Theaction spectrum of this response for etiolated plants showeda peak near 660 nm, but for green plants it was near 612 nm.Thus, the effect of light on flowering is influenced greatlyby the screening effect of photosynthetic pigments. (Received July 24, 1980; )     

Changes in Enzyme Regulation during Growth of Maize: I. Progressive Desensitization of Homoserine Dehydrogenase during Seedling Growth

The sensitivity of homoserine dehydrogenase (EC 1.1.1.3) to inhibition by the feed-back modifier, l-threonine, was examined in preparations derived from etiolated shoots, roots, and lightgrown tissues of Zea mays L. var. earliking. A progressive decrease in enzyme sensitivity was observed during seedling growth. Enzyme derived from internode tissue retained a greater sensitivity to the effector than enzyme derived from apical portions of etiolated shoots, whereas enzyme from root tips was characteristically more sensitive than that prepared from mature cells of the root. Enzyme desensitization occurred rapidly during culture of excised shoots and the activities of both homoserine dehydrogenase and aspartokinase (EC 2.7.2.4) declined during shoot culture under a variety of conditions. The initial enzyme levels and the characteristic sensitivity of homoserine dehydrogenase were preserved during culture at 5 to 7 C, but desensitization was not prevented by inclusion of cycloheximide in the culture medium.Results of control experiments provide evidence that desensitization occurs in vivo. No alteration of the enzyme properties was detected during extraction or concentration of sensitive or insensitive enzyme or during coextraction of enzyme from mixed populations of different age shoots; nor was a differential distribution of inhibitors or activators indicated during assay of mixed preparations. The change in enzyme sensitivity was apparent under a variety of assay conditions and was not accompanied by changes in the apparent affinity of the enzyme for the substrate, homoserine. It is suggested that systematic changes in the regulatory characteristics of certain enzymes could be an important level of metabolic regulation during cellular differentiation.Three forms of maize homoserine dehydrogenaase were detected after acrylamide gel electrophoresis of samples derived from 72-hr shoots. Similar analysis of samples from older shoots revealed a broad asymmetric band of enzyme activity, suggesting that changes in the relative distribution of specific forms of the enzyme could be related to the growth-dependent changes in the sensitivity of maize homoserine dehydrogenase.     

Comparative Enzymology of the Glyceraldehyde 3-Phosphate Dehydrogenases from Pisum sativum

Glyceraldehyde 3-phosphate dehydrogenases (EC 1.2.1.12 and 1.2.1.13) have been purified from the seed, root, etiolated, and green shoot of peas (Pisum sativum). These enzymes are tetramers of 140,000 daltons, with subunits of 35,000 daltons. The enzymes differ in isoelectric point. The seed enzyme has a pI of 5.1, and the root enzyme has a pI of 4.5. The cytoplasmic enzyme from etiolated shoots is slightly acidic with a pI of 5.7 to 6.1 and is found in two separable forms. The chloroplast enzyme (from green shoots) is most basic with a pI of 8.0.     

Some properties of ferredoxin-nitrite reductase from green shoots of bean and an immunological comparison with nitrite reductase from roots and etiolated shoots

Ferredoxin-nitrite reductase (EC 1.7.7.1), an enzyme which catalyzes the 6-electron reduction of nitrite to ammonia, has been isolated from green shoots of bean (Phaseolus angularis). The isolated enzyme (GR-NiR), having a molecular mass of 68 000, showed 1.4 times higher ferredoxin-dependent activity than methyl viologen-linked activity. The enzyme was homogeneous by polyacrylamide gel electrophoresis (PAGE). In the oxidized form, the enzyme had absorption maxima at 275, 393 (Soret band), 535 and 571 (α band) nm, indicating that siroheme is involved in the catalysis of nitrite reduction. The absorbance ratios, A₃₉₃ : A₂₇₅ and A₅₇₁ : A₃₉₃ were 0.26 and 0.32, respectively. Antibody against the isolated enzyme was raised in rabbits. Analysis of the antiserum by immunodiffusion and immunoelectrophoresis suggested that it was a specific antiserum against GR-NiR. Using the antiserum, immunodiffusion and immunoprecipitation procedures were employed to compare the immunological similarity of NiR from green shoots, etiolated shoots and roots of bean. These tests revealed that the three forms of assimilatory NiR have antigenic determinants in common.     

Some Biochemical Characteristics of Guard Cell and Mesophyll Cell Protoplasts from Commelina communis L.

Guard cell and mesophyll cell protoplasts of Commelina communisL., were isolated and used to investigate their various biochemicalcharacteristics. Contamination of the samples by other celltypes was very low and viability of the protoplasts, assessedby the use of neutral red, Evans blue and fluorescein diacetate,was high (89–98%). Mesophyll cell protoplasts containedmore chlorophyll (x 47), more soluble protein (x 10), more totalN (x 36) and more DNA (x 9) than guard cell protoplasts. Theabsorption spectra of protoplast extracts were similar for bothcell types except that below 400 nm there was a large increasein absorption by the guard cell protoplast extract. In guardcell protoplast extracts, high levels of activity of phosphoenolpyruvatecarboxylase (E.C. 4.1.1.31 [EC] ), NAD malate dehydrogenase (E.C.1.1,1.37), NADP malic enzyme (E.C. 1.1.1.40 [EC] ) and carbonic anhydrase(E.C. 4.2.1.1 [EC] ) were detected while only low levels of pyruvate-orthophosphatedikinase (E.C. 2.7.9.1 [EC] ) activity were detected. Glycollate oxidase(E.C. 1.1.3.1 [EC] ), ribulose-l,5-bisphosphate carboxylase (E.C 4.1.1.39 [EC] ),NADP malate dehydrogenase (E.C. 1.1.1.82 [EC] ) and NAD malic enzyme(E.C. 1.1.1.39 [EC] ) were not detected in guard cell protoplast extracts.High levels of ribulose-1, 5-bisphosphate carboxylase, glycollateoxidase, NAD malate dehydrogenase and carbonic anhydrase weredetected in mesophyll cell protoplast extracts which is typicalof C₃ plants. A pathway of carbon flow during stomatal openingand closing is proposed. Key words: Carbon metabolism, Commelina communis, guard cell protoplasts, mesophyll cell protoplasts, stomata     

Fatty Acids in Chloroplasts and Leaves

The fatty acid composition of green and white leaf tissue inAcer negundo, Zea mais, and Ilex aquifolium, of green and yellowtissue in Ligustrum ovatifolium and of etiolated and green tissuein Vicia faba has been determined. The mesophytic green leavesexamined show a general similarity in fatty acid composition,characterized by a high concentration of non-conjugated octadecatrienoicacid. Chloroplasts were isolated from Vicia and Acer and containan even higher concentration of this acid and only traces ofnon-conjugated octadecadienoic acid. Conjugated diene and trieneacids occur in traces in chioroplasts, but are also found innon-green leaf tissue. The fats of non-green leaves are in generalmore saturated than those from green tissue but vary considerablyin composition. The relationship between fat composition andplastid development is discussed.     

Asparagusic acid, a new plant growth inhibitor in etiolated young asparagus shoots

Yellow prisms of asparagusic acid, with a molecular formulaof C₄H₆O₂S₂ were isolated from etiolated asparagus tissues (Asparagusofficinalis L.). This acid inhibits growth in lettuce and otherseedlings when applied in concentrations of 6.67x10^–7Mto 6.67xl0^–7M. The extent of activity was very similarto that of abscisic acid. ¹ A well known shift reagent in the NMR spectrum (1). (Received April 12, 1972; )     

不同生态型东南景天对土壤中Cd的生长反应及吸收积累的差异性

 采用盆栽方法研究了两种生态型东南景天(Sedum alfredii)对土壤中不同含量Cd(即对照, 12.5, 25, 50, 100, 200, 300, 400 mg&;#8226;kg^－1)的生 长反应、吸收和积累Cd的差异性。结果表明,土壤添加重金属Cd后,矿山生态型东南景天生长正常,地上部和根系Cd含量随着土壤中Cd含量的 增加而增加,在400 mg&;#8226;kg^－1 Cd处理下含量分别高达2 900和500 mg&;#8226;kg^－1,其地上部显著大于根部;然而,土壤添加Cd后,非矿山生态型东 南景天的生长受到抑制,地上部和根部的生物量显著降低。当土壤Cd含量为50～100 mg&;#8226;kg^－1 时,非矿山生态型东南景天的地上部和根系Cd含 量随着土壤中Cd含量的增加而增加,而且根系Cd含量则大于地上部。当土壤Cd≤50 mg&;#8226;kg^－1时,矿山生态型东南景天根系Cd含量比非矿山生态 型高 ,但当土壤Cd≥100 mg&;#8226;kg^－1,两者之间无显著差异;然而,但在同一Cd处理水平下,矿山生态型东南景天地上部Cd含量总是高于非矿山 生态型。这些结果表明,矿山生态型东南景天有很强的忍耐和吸收土壤Cd的能力,再次证明其为一种Cd超积累植物。     

Immunoprecipitation of phytochrome from green Avena by rabbit antisera to phytochrome from etiolated Avena

Thirty-nine antiserum preparations from eight rabbits were screened for their ability to precipitate the immunochemically distinct phytochrome that is obtained from green oat (Avena sativa L.) shoots. The antisera were obtained from rabbits immunized with either proteolytically degraded, but still photoreversible, 60-kDa (kilodalton) phytochrome, or approx. 120-kDa phytochrome, both of which were purified from etiolated oat shoots. The ability of these antisera to precipitate phytochrome from green oats was independent of the size of phytochrome used for immunization. While crude antisera immunoprecipitated as much as 80% of the phytochrome isolated from green oat shoots, antibodies immunopurified from these sera with a column of highly purified, approx. 120-kDa phytochrome from etiolated oats precipitated no more than about 5–10%.Abbreviations kDa kilodalton - mU milliunit     

Respiration of Malate and Glutamate in Symbiotic Frankia Prepared from Alnus incana

Frankia vesicle clusters were prepared from root nodules ofAlnus incana (L.) Moench inoculated either with a local sourceof Frankia or with Frankia Cpll. The capacity of vesicle clustersto respire was investigated by respirometric and enzymologicalstudies. Simultaneous addition of malate, glutamate, and NAD⁺supported respiration in both types of Frankia, though at asmaller rate compared to the substrates NADH or 6-phosphogluconate.The saturating concentrations of malate and glutamate were alsomuch higher than with the other substrates. No respiration wassupported by succinate. Activity of the enzymes malate dehydrogenase(EC 1.1.1.37 [EC] ) and glutamate oxaloacetate transaminase (EC 2.6.1.1 [EC] )was demonstrated in crude extracts from both types of symbioticFrankia. Their maximum rates were high enough to account forthe respiration of malate and glutamate. This respiration wasinhibited by mersalylic acid, an inhibitor of the dicarboxylateshuttle in mitochondria, but it was shown that inhibition ofrespiration could be due to a direct effect on the enzymes.We conclude that respiration of malate and glutamate is mostlikely mediated by malate dehydrogenase and glutamate oxaloacetatetransaminase, but no explicit evidence for or against the presenceof a dicarboxylate carrier was found. The utilization of respiratorysubstrates was largely similar in the two types of Frankia,except for some differences in maximum rates and cofactor dependency. Key words: Actinorhizal symbioses, Alnus, dicarboxylate shuttle, Frankia, reducing power, respiration     

Effect of shade treatment on biosynthesis of catechins in tea plants

When tea plants were shaded with black lawn cloth for severaldays in the field, the accumulations of (—)-epicatechin,(—)-epicatechin-3-gallate, (—)-epigallocatechinand (—)-epigallocatechin-3-gallate decreased in newlydeveloping tea shoots. Radioactive tracer studies showed thatthe conversions of glucose-U-¹⁴C, shikimic acid-G-¹⁴C and phenylalanine-U-¹⁴Cinto (—)-epicatechin and (—)-epigallocatechin moietieswere depressed by the shade treatment for tea plants but theincorporation of trans-cinnamic acid-3-¹⁴C was not affected.The treatment was found to have no significant effect on theactivities of phospho-2-keto-3-deoxy-heptonate. aldolase (EC.4.1.2.15 [EC] ), 3-dehydroquinate synthase (EC. 4.6.1.3 [EC] ), 3-dehydroquinatedehydratase (EC. 4.2.1.10 [EC] ), shikimate dehydrogenase (EC. 1.1.1.25 [EC] )and trans-cinnamate 4-monooxygenase (EC. 1.14.13.11 [EC] ) in theshoots, whereas the activity of phenylalanine ammonia-lyase(EC. 4.3.1.5 [EC] ) clearly decreased. (Received March 17, 1980; )     

Activities of enzymes involved in lignification during the postharvest storage of etiolated asparagus spears

The content of lignin and the activities of 5 enzymes involved in lignification were monitored along the length of etiolated spears of asparagus ( Asparagus officinalis L., INRA Fl male hybrid n°156) stored for 22 h with their base in air (control), water or water containing the ethylene antagonist, silver thiosulfate (STS). At the time of harvest the lignin content increased basipetally, as did the activity of all the enzymes studied, viz., phenylalanine ammonia lyase (PAL; EC 4.3.1.5), hydroxycinnamate: CoA ligase (HCoAL; EC 6.2.1.12), cinnamoyl-CoA reductase (CCR: EC 1.2.1.44), cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) and syringaldazine oxidase (SyrOx. a peroxidase [POD; EC 1.11.1.7] with syringaldazine as substrate). Neither the lignin content nor the activity of any enzyme changed in the spear apex during storage, regardless of treatment. In the spear base. all enzyme activities decrased during the first 2 to 4 h in every storage treatment. Subsequently. PAL and HCoAL activities remained constant. whereas the activities of CAD and SyrOx gradually increased. Lignification in the spear base was not affected by storage in air. However, storage in water increased lignin formation and SyrOx activity, whereas treatment with STS prevented both of these increases. The results indicate that postharvest lignification in etiolated asparagus spears is caused primarily by enhanced SyrOx activity, and that ethylene is involved in the control of this activity.