Control of Ribulose-1,5-diphosphate Carboxylase Activity During Expansion of Leaves of Capsicum frutescens L.

The activity of ribulose-1,5-diphosphate carboxylase per unitlaminar area increases rapidly during early stages of leaf expansionin Capsicum frutescens L. cv. California Wonder. This is followedby a decrease to a level that is constant until expansion stops. A previous suggestion that the expansion of younger leaves inthe same phyllotactic sector controlled the decrease in enzymeactivity in older expanding leaves has not been verified byexperiments involving the selective excision of leaves and cotyledons.Decrease in enzyme activity was accompanied by a fall in FractionI protein content but another chloroplastic enzyme, -aminolevulinicacid dehydrase, did not exhibit a decrease in activity. Intra-chloroplasticmechanisms, rather than the influence of other plant organs,are suggested as controlling ribulose-1,5-diphosphate carboxylaseactivity during later stages of leaf expansion.     

Production of Microconidia by Cercospora henningsii Allesch, Cause of Brown Leaf Spot of Cassava (Manihot esculenta Crantz) and Tree Cassava (Manihot glaziovii Muell.-Arg.)

Sporulation ofCercospora henningsii Allesch has been examinedunder various relative humidities, and in the presence of freewater in lesions on leaves of cassava and tree cassava. Matureconidia of the fungus on both cassava and tree cassava do notgerminate in lesions but accumulate, and under the optimum conditionsof 25–32 °C and in the presence of free water, theybud and fragment into numerous microconidia. Microconidia arecylindrical, mostly one-celled, and measure 7.5–17.5x3.7–7.5µm. Production of microconidia significantly decreasesas relative humidity decreases. Microconidia readily germinateby means of a germ tube at 100% relative humidity on both surfacesof host leaf and on glass slides. Some germ tubes form appressoriaand symptoms appear on cassava leaves inoculated with microconidia.The results are discussed in relation to possible modes of dispersalof the spores and control of the disease. Free water; sporulation; budding; microconidia     

Rhodanese in higher plants

Rhodanese activity was detected in crude leaf extracts of 12 randomly selected plant species consisting of 9 non-cyanophoric and 3 cyanophoric species. In each case, the enzyme exhibited high activity at pH 10·4 and 55°. There appeared to be no correlation between rhodanese activity and the cyanophoric nature of the plant.     

Nitrogen and Nitrate Accumulation in Species Having Different Relationships Between Nitrate Uptake and Reduction

Species differ in the relationship of nitrate reductase activityto nitrate uptake. In Capsicum annuum different diurnal patternsof leaf nitrate reductase activity and nitrate uptake have beenreported. As a consequence, the relationship of free nitratein the plant to nitrate supplied has a higher level of significancethan has reduced nitrogen to nitrate supplied. In Zea mays ithas been reported that leaf nitrate reductase activity respondsdirectly to nitrate translocation to the leaf and in this speciesthe relationship of greatest significance is reduced nitrogencontent to nitrate supplied. In both species, and also in Cucumis melo, the proportion oftotal plant free nitrate and reduced nitrogen in the roots decreases,and in the stem increases, with increasing nitrate supplied. The accumulation of free nitrate in leaves is accompanied bya quantitatively different relationship between reduced nitrogenand dry weight compared to leaves not accumulating nitrate. Capsicum annuum. L., Cucumis melo L., melon, Zea mays L., maize, sweet corn, nitrate reductase, nitrate uptake     

Purification, characterization, and localization of linamarase in cassava

We have purified cassava (Manihot esculenta) linamarase to apparent homogeneity using a simplified extraction procedure using low pH phosphate buffer. Three isozymes of cassava linamarase were identified in leaves based on differences in isoelectric point. The enzyme is capable of hydrolyzing a number of β-glycosides in addition to linamarin. The enzyme is unusually stable and has a temperature optimum of 55°C. Immunogold labeling studies indicate that linamarase is localized in the cell walls of cassava leaf tissue. Since linamarin must cross the cell wall following synthesis in the leaf for transport to the root, it is likely that linamarin must cross the cell wall in a nonhydrolyzable form, possibly as the diglucoside, linustatin. In addition, we have quantified the levels of linamarin and linamarase activity in leaves of cassava varieties which differ in the linamarin content of their roots. We observed no substantial differences in the steady state linamarin content or linamarase activity of leaves from high or low (root) cyanogenic varieties. These results indicate that the steady state levels of linamarin and linamarase in leaves of high and low cyanogenic varieties are not correlated with the varietal differences in the steady state levels of linamarin in roots.     

Studies on chlorophyllase in tea leaves III. Properties of soluble and insoluble chlorophyllases

In contrast to previous knowledge of chlorophyllase activityin higher plants, significant enzyme activity was isolated fromtea leaves in a soluble state. Soluble chlorophyllase was partially purified by proceduresincluding ammonium sulfate fractionation (Preparation I). Theinsoluble fraction was extracted, by solubilizing it with SDC,from the methanol-acetone powder of sediments of the leaf homogenate,from which the water-soluble enzyme had been completely removedby repeated extraction. This initially insoluble enzyme wasalso partially purified (Preparation II). Specific activities(mg chlorophyll a hydrolyzed per hr per mg protein, 7.2 forPreparation I, and 12.4 for Preparation II), were much higherthan those reported for other plant material. The soluble enzyme was more resistant to PCMB, lipase and heattreatment. The two enzymes differed in optimum temperature andoptimum acetone concentration needed for the reaction, but showedthe same optimum pH, and same Km value. The Km value was thesame (7 µM) for reactions with 30% and 50% acetone. These results suggest that, in spite of differences in locationand extractability, activities of the soluble and insoluble(solubilized) chlorophyllase in tea leaves are attributableto the same enzyme. (Received March 8, 1972; )     

Variation in Nitrate Reductase Activity in Lolium

Nitrate reductase activity was studied in the leaves and rootsof Lolium perenne. Growth temperatures of 8, 15, or 20 °Cdid not affect activity, but the same temperatures during assayhad differential effects on the nitroso couple used to measureenzyme activity. Activity increased with increasing light intensity,reaching a high plateau value at around 40 W m^–2. Nitratecontent of leaves, also measured in this experiment, did notvary significantly with different light intensities. Increasingnitrate in the nutrient solution up to 0.5 mM N also increasedactivity. Adding ammonium chloride at similar levels to thenitrate caused no marked repression of activity. Removal ofnitrate from the nutrient solution decreased enzyme activitywithin 24 h. Marked diurnal fluctuations occurred in activity,apparently in response to light intensity, since the nitratelevel in the plant varied little. The enzyme activity of rootswas much less than that of leaves. In the parents and progeny from a half diallel cross, the parentalgenotypes differed significantly in activity, but the numberof families involved was too small for the regression of progenyon parents (b = 1.74) and the correlation coefficient (r = 0.44NS) to achieve significance. In this experiment a significantpositive regression was obtained between nitrate reductase activityand dry matter yield.     

Ferredoxin activation by rhodanese

Rhodanese was extracted from Brassica oleracea leaves and purified 150-fold. The enzyme was shown to have optimum activity at pH 8-8.5 and a temperature range of 50-55°; a K_m of 0.4 mM at 30° for thiosulphate and cyanide. and mol. wt around 32000. The electrophoretically pure enzyme is able to produce the biological and spectral properties of ferredoxin when added to apoferredoxin in the presence of thiosulphate.     

Nitrate Reductase in the Citrus Plant: Properties, Assay Conditions and Distribution within the Plant

A nitrate reductase enzyme preparation has been obtained from citrus plant tissue extract and the optimal assay conditions have been established. The enzyme has been found to be similar or even identical to those of many other annual plants, i.e., cysteine is required for its extraction in the active state, it is DPNH-specific, and in some cases FAD-dependent. Comparisons were made between the activity measured in fragments with that of the extracted preparation of the same plant tissues. The occurrence of the enzymes was demonstrated in germinating seeds, cotyledons, rootlets, shoots, leaves and fruit of citrus plants.     

Some Factors Concerning the Centripetal Disposition of Bundle Sheath Chloroplasts during the Leaf Development of Eleusine coracana

Factors concerning the chloroplast disposition in bundle sheathcells were investigated in finger millet (Eleusine coracanaGaertn.), and NAD malic enzyme type C₄ plant with the centripetalarrangement of bundle sheath chloroplasts. Segments were cutfrom immature regions of emerging leaves in which the centripetalarrangement of bundle sheath chloroplasts had not yet been established.The leaf segments were floated on solutions with or withoutreagents. Sections were made of the segments at time intervalsand the distribution of bundle sheath chloroplasts was observedby light microscopy. The bundle sheath chloroplasts migratedto the vascular bundle and established a centripetal arrangementby 12-16 h in control solutions. Auxins, cycloheximide and cytochalasinB inhibited the disposition of bundle sheath chloroplasts whilechloramphenicol and colchicine had no effect. The inhibitoryeffect of auxins appeared only at early stages of chloroplastmigration while cycloheximide and cytochalasin B were effectiveeven at later stages. Cessation of elongation growth, cytoplasmicprotein synthesis and microfilaments seemed to be associatedwith the centripetal disposition of bundle sheath chloroplasts.Copyright1993, 1999 Academic Press Bundle sheath chloroplast, C₄ plant, chloroplast orientation, Eleusine coracana, finger millet     

Structural and Kinetic Properties of NADP-Malic Enzyme from the Inducible Crassulacean Acid Metabolism Plant Mesembryanthemum crystallinum L.

NADP-malic enzyme (EC 1.1.1.40 [EC] ), which is involved in Crassulaceanacid metabolism (CAM), was purified to electrophoretic homogeneityfrom the leaves of the inducible CAM plant Mesembryanthemumcrystallinum. The NADP-malic enzyme, which was purified 1,146-fold,has a specific activity of 68.8 µmol (mg protein)^–1min^–1. The molecular weight of the subunits of the enzymewas 64 kDa. The native molecular weight of the enzyme was determinedby gel-filtration to be 390 kDa, indicating that the purifiedNADP-malic enzyme is a hexamer of identical subunits. The optimalpH for activity of the enzyme was around 7.2. Double-reciprocalplots of the enzymatic activity as a function of the concentrationof L-malate yielded straight lines both at pH 7.2 and at pH7.8 and did not reveal any evidence for cooperativity of bindingof L-malate. The K_m value for L-malate was 0.35 mM. Hill plotsof the activity as a function of the concentration of NADP⁺indicated positive cooperativity in the binding of NADP⁺ tothe enzyme with a Hill coefficient (nH) of 2.0. An S_0.5 value(the concentration giving half-maximal activity) of 9.9 µMfor NADP⁺ was obtained. Oxaloacetate inhibited the activityof the NADP-malic enzyme. Effects of succinate and NaHCO₃ onthe activity of NADP-malic enzyme were small. (Received October 30, 1991; Accepted May 1, 1992)     

Seasonal changes in activity of the enzyme system producing cis-3-hexenal and n-hexanal from linolenic and linoleic acids in tea leaves

The enzyme system producing cis-3-hexenal, a precursor of cis-3-hexenol(leaf alcohol) and trans-2-hexenal (leaf aldehyde), from linolenicacid showed high activity in summer and no activity in winterin tea (Thea sinensis) leaves and isolated chloroplasts. Theenzyme system producing n-hexanal from linoleic acid also showedsimilar seasonal changes in activity. These changes were closelyrelated to temperature and solar radiation. Enzyme activitycould not be induced after the leaves had been cut and was notaccompanied by de novo protein synthesis. (Received July 9, 1976; )     

NADP-malic enzyme: immunolocalization in different tissues34 plant maize and the C3 plant wheat

In situimmunolocalization and Western blot analysis of separatedcellular and subcellular fractions, were used to determine thelocalization of different isoforms of NADP-malic enzyme in bothwheat (C₃) and maize (C₄) plants. In both techniques, an affinitypurified anti-(maize 62 kDa NADP-ME) lgG from the maize greenleaf isoform also reacted with a 72 kDa protein in tissues ofC4 plants as well as C3 plants. The light- inducible 62 kDaisofomi is located in bundle sheath chioroplasts of maize leaves.In etiolated leaves and in roots of maize there is evidencefor the occurrence of a 72 kDa isoform which co-migrates on2-D (SDS and isoelectric focusing) PAGE. The 72 kDa isoformis also present in low levels in green leaves. This form mayoccur in multiple intracellular compartments; but in situ immunolocalizationexperiments and Western blot and activity assays on fractionatedprotoplasts indicate that a significant amount of this isoformoccurs in plastids. With regards to C³ plants such as wheat,a 72 kDa isoform in leaves is largely confined to the chloroplastsbased on in situ immunolocalization and Western blots and enzymeactivity assays with fractionated protoplasts. In maize, itappears that the constitutive expression pattern of a possibleC₃ ancestral gene for NADP-malic enzyme has been maintained,and a high level expression of a light-inducible isoform locatedin bundle sheath chloroplasts (62 kDa) has been acquired duringits evolution. Key words: NADP-malic enzyme, Triticum aestivum, Zea mays     

Light-induced Increase in Sucrose Phosphate Synthetase Activity in Leaves of Lolium temulentum

Mature leaves of Lolium temulentum L. were assayed for sucrosephosphate synthetase activity at different times during thephotoperiod. There was a rapid increase in activity at the onsetof illumination which was not observed in leaves maintainedin darkness. The activity prior to illumination was insufficientto catalyse the rates of sucrose synthesis observed in illuminateddetached leaves; after 15 min illumination the two processeswere of similar magnitude. Lolium temulentum L., darnel, sucrose phosphate synthetase, enzyme activity, light, sucrose, starch     

A Topographic Method for Studying Uptake, Translocation and Distribution of Inorganic Ions using Two Radiotracers Simultaneously

A novel topographic method is described for the fully automatedquantitative resolution of activity of two simultaneously appliedbeta tracers in plant tissues. The advantages of this methodusing specially developed silicon detectors are demonstratedin studying the effect of direct current on the uptake and distributionof Ca²⁺ and P₁ ions in leaves and stems of Chenopodium rubrum.This treatment has recently been shown to inhibit flower inductionin this plant. Key words: Radiotracer multi-labelling, computer-controlled resolution, quantitative topographic evaluation     

In vitro somatic embryogenesis and plant regeneration of cassava

An efficient and reproducible plant regeneration system, initiated in somatic tissues, has been devised for cassava (Manihot esculenta Crantz). Somatic embryogenesis has been induced from shoot tips and immature leaves of in vitro shoot cultures of 15 cassava genotypes. Somatic embryos developed directly on the explants when cultured on a medium containing 4–16 mg/l 2,4-D. Differences were observed with respect to the embryogenic capacity of the explants of different varieties. Secondary embryogenesis has been induced by subculture on solid or liquid induction medium. Long term cultures were established and maintained for up to 18 months by repeated subculture of the proliferating somatic embryos. Plantlets developed from primary and secondary embryos in the presence of 0.1 mg/l BAP, 1mg/l GA₃, and 0.01 mg/l 2,4-D. Regenerated plants were transferred to the field, and were grown to maturity.     

Nitrate Reduction by Cassava

Nitrate reductase (NR) was assayed in vivo in cassava (Manihotesculenta Crantz). Activity in the leaves ranged from 0 to 2.51µmole of NO₃^– reduced g^–1 h^–1, withno activity in the younger leaves (leaf 1 on top). NR activitywas localized in the sides and toward the tip of the lobes ofthe leaf. (Received December 10, 1985; Accepted April 8, 1986)     

Infochemical-Mediated Niche Use by the Predatory Mites Typhlodromalus manihoti and T. aripo (Acari: Phytoseiidae)

In Africa, Typhlodromalus manihoti and T. aripo, two introduced predators of the cassava green mite Mononychellus tanajoa, occupy different parts of cassava foliage. In the present study, niche use by these two predators, as mediated by prey-induced infochemicals, was investigated. In response to prey feeding damage, cassava plant parts emit volatile blends, that attract phytoseiidae predators. When given a choice between old cassava leaves infested with M. tanajoa and either apices or young cassava leaves infested with M. tanajoa, T. aripo displayed a marked preference for odors emitted from either infested apices or infested young leaves over infested old leaves but showed no preference for odors from apices versus young leaves, all infested with M. tanajoa. Typhlodromalus manihoti did not discriminate between volatiles from the three infested cassava plant parts. Our data show that T. aripo uses differences in volatile blends released by infested cassava plant parts and restricts its fundamental niche to a realized niche, which enables coexistence with its competitor T. manihoti.