Sucrose metabolism during the growth of Camellia japonica pollen

The free sugar in the mature pollen grains of Camellia japonica is almost all sucrose and the sucrose content decreases rapidly during pollen growth. The activity of soluble invertase increases during culturing and a high constant activity was found at the later stages of pollen tube growth. By contrast, the level of sucrose synthetase activity remains constant during pollen growth and that of wall-bound invertase activity is very low. Cycloheximide has little effect on the activity of these enzymes. Exogenous sucrose or glucose was simultaneously incorporated into the pollen grains when they absorbed water and swelled. The free sugar levels in growing pollen depend on the nature of the exogenous sugar. The sugar metabolism in the pollen at the stage of germination differs from that during tube growth, the latter being particularly influenced by exogenous sugar.     

Enzymes of starch metabolism in Nicotiana tabacum callus

Activities of enzymes of starch metabolism were determined in tobacco callus grown in light or darkness and in the presence or absence of gibberellic acid. There was a higher rate of starch turnover in light-grown cultures, as judged by the activities of synthetic and degradative enzymes. Gibberellic acid-treated tissues contained a lower level of starch than the corresponding control tissue. This decrease could be correlated with the activity of phosphorylase. Cultured tissue and seedling material were found to have comparable levels of activity for the starch metabolizing enzymes.     

Carbohydrate oxidation during Nicotiana tabacum callus growth

Tobacco callus was cultured in light or dark, with or without giberellic acid, and with various carbon sources in the medium, and the growth rate and activities of some enzymes of the Embden-Meyerhof-Parnas and pentose phosphate pathway were determined. No changes in the specific activities of enzymes of either pathway could be correlated with growth but there was a light-dependent stimulation of the pentose phosphate pathway enzymes examined.     

Distribution in excised Lycopersicum esculentum roots of the principal enzymes involved in sucrose metabolism

Sucrose synthetase and sucrose phosphate synthetase could not be detected in 7-day-old excised tomato roots grown in sucrose. These roots, however, possessed a highly active acid invertase and a neutral invertase of low activity. The distribution of the cell wall-located acid invertase along the root axis appeared to be related to growth. This was not the case for the soluble enzyme. The possible functions of these two enzymes are discussed.     

Sucrose absorption and synthesis by excised Lycopersicon esculentum roots

Asymmetrically-labelled sucrose was absorbed intact by excised roots of tomato, grown in sucrose. Glucose-grown roots possessed sucrose synthetase and sucrose phosphate synthetase activity.     

Kinetin induced changes in cell wall composition of tobacco callus

Culture of tobacco callus on high or low kinetin in light or darkness leads to changed tissue texture and associated changes in cell wall composition. In particular, friable callus (low kinetin, darkness) cell walls have a greater extensin content and an altered composition of arabinose and xylose containing hemicelluloses compared with cell walls of compact callus (high kinetin, darkness). The possible importance of these differences in determining callus friability is discussed.     

Isolation and characterization of two isoperoxidases from tobacco tissue cultures

Two isoperoxidases (A_f and C_n) from the medium of tobacco tissue suspension culture WR-132 grown in darkness have been purified to apparent homogeneity and partially characterized. C_n and A_f have MWs of ca 30 000 and 54 000, respectively. A_f has ca 5.1% carbohydrate, but none could be detected in C_n. Both isoperoxidases appear to follow simple Michaelis-Menten kinetics with respect to guaiacol as the substrate. The K_ms for guaiacol are 4 and 13.3 mM for Af and C_n, respectively, while both isoperoxidases have a pH optimum at 6.5. C_n, is dissimilar to other isoperoxidases from tobacco tissue cultures, but A_f is very similar to isoperoxidase A₃ from W-38 tobacco tissue culture.     

Selection of high nicotine-producing cell lines of tobacco callus by single-cell cloning

Single-cell clones of Nicotiana tabacum callus showed wide variation in the production of nicotine. An efficient screening of numerous clones was made possible by the adoption of the ‘cell squash method’ devised for estimating the approximate alkaloid content of a small piece of callus. From these clones, several cell lines with higher yield of nicotine (1.0–3.4% of dry wt) have been isolated by repeated clonal selection.     

Sucrose metabolism during endosperm development in wheat (Triticum aestivum)

This work reports changes in sucrose synthase and invertase activities throughout endosperm development in wheat, together with the associated substrates and metabolites, sucrose, UDP, glucose, fructose and UDP-glucose. Throughout endosperm development, sucrose synthase had consistently higher activity than invertase and indeed invertase activity did not change appreciably. The observed variation in pattern and amounts of glucose and fructose present during the mid- and late stages of endosperm development confirmed the suggestion that invertase was not the preferred pathway of sucrose catabolism. Kinetic parameters for sucrose synthase were determined in crude extracts. Estimates of UDP and sucrose concentrations suggest that sucrose synthase is unlikely to achieve its potential maximum velocity. This limitation may however be overcome in part by the apparent excess catalytic activity measured during endosperm development.     

Sucrose synthetase from triploid quaking aspen callus

Sucrose synthetase and UDPG pyrophosphorylase were found in crude extracts of dark-grown triploid quaking aspen callus. The sucrose synthetase was like that from other plants, but it appeared to be membrane-bound. The activity also occurred in extracts prepared from callus of other tree species.     

Sucrose accumulation and enzyme activities in callus culture of sugarcane

The activities of sucrose phosphate synthase (SPS), sucrose synthase (SUSY), neutral invertase (NI) and soluble acid invertase (SAI) were measured in callus cultures of four Mexican sugarcane cultivars (Saccharum spp.) with a different capacity to accumulate sucrose in stem parenchyma cells. The results indicated that sucrose accumulation in callus was positively correlated to the activity of SPS and SUSY and negatively to the activity of SAI and NI while SPS explained most of the variation found for sucrose accumulation and NI least.The research was funded by the department of Biotechnology and Bioengineering CINVESTAV Mexico City, and F. G.-M. received grant-aided support from CONACyT, Mexico.     

Comparison of isoperoxide patterns in tobacco cell culture and in the intact plant

Analysis by ion-exchange chromatography of the enzymes from cultured tobacco cells and root or leaf tissues of the tobacco plant revealed that the cultured cells contain exclusively cationic peroxidases and the leaf tissues mainly anionic and neutral peroxidases.     

Significance of caffeic acid-O-methyltransferase in lignification of cultured tobacco cells

Caffeic acid-O-methyltransferase, activity was assayed in the callus and suspension cultured cells of tobacco. Lignification of cells was observed only in a kinetin (10^?5 M) culture and not in an auxin (10^?6 M 2,4-D or 10^?5 M IBA) culture. Enzyme activity in the kinetin cultured cells was much higher than in the stock culture and the rise in enzyme activity coincided with the onset of lignification.     

甘肃黄花烟草愈伤组织在生长与衰老期间呼吸途径的动态变化

实验结果表明：烟划愈伤组织在生长和衰老期间,总呼吸速率（Ｖｔ）分别在１１ｄ和１９ｄ出现２次呼吸跃升;细胞色素途径的运行（ρ＇Ｖｃｙｔ）与Ｖｔ的变化几乎一致,表明细胞色素途径仍组织主要的电子传递途径;交替途径容量（Ｖａｌｔ）及其与Ｖｔ的比值（Ｖａｌｔ／Ｖｔ）：在１５ｄ前不断上升,而在１５－１９ｄ之间处于稳定水平后下降。交替途径运行（ρＶａｌｔ）及其对Ｖｔ的贡献（ρＶａｌｔ／Ｖｔ）却与Ｖａｌｔ变化趋势     

Phosphodiesterases of tobacco cell cultures and the intact plant

Tobacco cells grown in suspension culture contained a higher specific activity of acid phosphodiesterase than tissues of the intact plant. The activity of the cells increased with age. The presence of alkaline phosphodiesterase both in the cultured cells and the intact plant was demonstrated, and the properties were partially characterized.     

Properties of peroxidases from tobacco cell suspension culture

An enzyme preparation from suspension cultured tobacco cells oxidized IAA only in the presence of added cofactors, Mn²⁺ and 2,4-dichlorophenol, and showed two pH optima for the oxidation at pH 4·5 and 5·5. Effects of various phenolic compounds and metal ions on IAA oxidase activity were examined. The properties of seven peroxidase fractions separated by column chromatography on DEAE-cellulose and CM-Sephadex, were compared. The peroxidases were different in relative activity toward o-dianisidine and guaiacol. All the peroxidases catalysed IAA oxidation in the presence of added cofactors. The pH optima for guaiacol peroxidation were very similar among the seven isozymes, but the optima for IAA oxidation were different. The anionic and neutral fractions showed pH optima near pH 5·5, but the cationic isozymes showed optima near pH 4·5. With guaiacol as hydrogen donor, an anionic peroxidase (A-1) and a cationic peroxidase (C-4) were very different in H₂O₂ concentration requirements for their activity. Peroxidase A-1 was active at a wide range of H₂O₂ concentrations, while peroxidase C-4 showed a more restricted H₂O₂ requirement. Gel filtration and polyacrylamide gel studies indicated that the three cationic peroxidases have the same molecular weight.     

Changes in isoperoxidases during shoot formation in tobacco callus

Summary Shoot formation in tobacco (Nicotiana tabacum L.) callus is accompanied by an increase in peroxidase activity which takes a form similar to a sigmoid curve. The “stationary” phase coincide with the period of organ formation. Characteristic changes in isoperoxidase pattern are found in the shoot-forming part of the callus. These changes are different from those in the nonshoot-forming part or in gibberellin-treated tissue, which does not form shoots.     

Comparison of tryptic peptide maps of eight isoperoxidases from tobacco tissue cultures

Eight isoperoxidases from tobacco suspension culture WR-132 (termed C_n, C₃, C₄, A_c, and A_f) and tobacco callus culture W-38 (termed A₁, A₂, and A₃) have been subjected to trypsin digestion followed by peptide mapping. The peptide maps of isoperoxidases A_f and A₃ are identical. All other isoperoxidases do not appear to be dramatically dissimilar in certain portions of their sequence, since many matching peptides have been found when various isoperoxidases are cross-compared. However, only two, and possibly three, highly homologous peptides are present in all of the isoperoxidases.     

Stress compounds in tobacco callus infiltrated by Pseudomonas solanacearum

Two sesquiterpenoids, phytuberin and phytuberol, have been identified in tobacco callus infiltrated by Pseudomonas solanacearum.     

Coniferyl alcohol, sinapyl alcohol and scopoletin in tobacco callus tissue during growth of a subculture

Coniferyl and sinapyl alcohols were isolated, identified and quantitatively determined as unbound (or weakly bound) phenylpropanoids in neutral hot-water extracts of Nicotiana tabacum L. callus tissue. This is the first identification of these alcohols in cultured tobacco callus. Scopoletin was also detected in these extracts, and it was the most abundant of these three phenylpropanoids with concentrations that ranged from 50–119 μg/g dry wt. Coniferyl alcohol (17–34 μg/g dry wt.) and sinapyl alcohol (23–35 μg/g dry wt.) were present in nearly equimolar concentration ratios and at levels which were about half those determined for unbound (or weakly bound) scopoletin. The amount of scopoletin extracted increased about 10 times when 1 M HCl-50% methanol - 0.3% ascorbic acid was used as the extractant. This indicated that most scopoletin moieties were strongly bound, perhaps by acid-hydrolyzable linkage. Coniferyl alcohol and sinapyl alcohol were not found in the acid extracts, presumably because they were acid-labile. In general, the concentration of each endogenous unbound (or weakly bound) phenylpropanoid appeared to remain relatively constant throughout the growth phase of the subculture. The only exceptions to this were the relatively higher concentrations of scopoletin and coniferyl alcohol present during the initial 0–2 weeks of subculture.