Coordinated action of β‐galactosidases in the cell wall of embryonic axes during chickpea germination and seedling growth

The plant cell wall is a dynamic structure whose constant modification is necessary for plant cells to grow and divide. In the cell walls of chickpea (Cicer arietinum) there are at least four β‐galactosidases, whose presence and location in embryonic axes during the first 48 h of seed imbibition are discussed in this paper. We examined their roles as cell wall‐modifying enzymes in germinative and/or post‐germinative events. At the start of germination, only βV‐Gal, and to a lesser extent βIV‐Gal, appear in the axes before rupture of the testa, suggesting they are related to germination sensu stricto. Once the testa has broken, the four β‐galactosidases are involved in growth and differentiation of the axes. Immunolocation of the different proteins in axes, which in part confirms previous results in seedlings and plants, allows assignment of post‐germinative roles to βI‐Gal and βIII‐Gal as cell wall modifiers in vascular tissue elements. βIV‐Gal and βV‐Gal participate in the initial events of germination in which cell walls are involved: βV‐Gal in cell proliferation, detachment of root cap cells and initial vascular tissue differentiation; both of them in xylem maturation; and βIV‐Gal in thickening of the primary cell wall. Together with other cell wall‐modifying enzymes, such as expansins and XTH, chickpea galactosidases might function in a sequential order in turnover of the primary cell wall, allowing the elongation of embryonic axes during seed germination.     

Hydrogen peroxide metabolism in soybean embryonic axes at the onset of germination

Hydrogen peroxide steady state levels of 5 micromolar were determined in soybean (Glycine max) embryonic axes incubated for 2 hours and in axes pretreated with aminotriazole or cyanide, where these levels were 50 and 1 micromolar, respectively. The activities of catalase (105 picomoles H₂O₂ per minute per axis), peroxidase (10-44 picomoles H₂O₂ per minute per axis), glutathione peroxidase (3 picomoles H₂O₂ per minute per axis) and superoxide dismutase (3.5 units per axis), were also determined. Catalase seems to be the most important H₂O₂ consuming enzyme at the physiological concentration of H₂O₂. A short treatment with aminotriazole, while substantially increasing H₂O₂ level, did not affect the growth of the axes. The production of superoxide anion by the mitochondria isolated from soybean axes was measured from the superoxide dismutase-sensitive rate of adrenochrome formation in the presence of NADH or succinate as substrate and amounted to 1.3 and 0.8 nanomole O₂⁻ per minute per milligram protein, respectively. According to the stoichiometry of O₂⁻ and H₂O₂ dismutation reactions, it is apparent that about 0.9 to 1.5% of the total oxygen uptake proceeds through the formation of the free intermediates of the partial reduction of oxygen.     

Effect of short-chain fatty acids on the ethylene pathway in embryonic axes of Cicer arietinum during germination

The effect of short-chain saturated fatty acids (C₅–C₁₀) on the biosynthesis of ethylene in embryonic axes of chick-pea ( Cicer arietinum L.) seeds was investigated. The emergence of radicle and fresh weight of embryonic axes diminished with increasing number of carbons. The inhibition of germination caused by lower concentrations (1 m M ) of fatty acids (C₅–C₁₀) was partially reversed by exogenous 1-aminocyclopropane-1-carboxylic acid (ACC), whereas exogenous ethylene was able to overcome the inhibitory effect provoked by all concentrations (1–5 m M ) of applied fatty acids (C₅–C₁₀). Ethylene production rates, and enzyme activities of ACC synthase and ACC oxidase decreased concomitantly with the molecular mass and increasing concentration of fatty acids. The inhibitory effect of these acids on ethylene production seems to result not only from a decreased ACC synthesis, but also from an enhancement of 1-malonylamino)cyclopropane-1-carboxylic acid (MACC) synthesis.     

Early growth of wheat embryonic axes and the synthesis of RNA and DNA

The requirement for the synthesis of RNA and DNA in early germination of wheat (Triticum aestivum var Newana) embryonic axes has been studied by incubating embryos in the presence of appropriate inhibitors and monitoring both embryo growth and the rates of specific metabolic processes. Experiments with 5-fluorouridine showed that both rRNA and DNA synthesis could be curtailed by 60 to 70% without affecting embryo growth to 24 hours. Similarly, the presence of mitomycin C and methotrexate inhibited DNA synthesis 70%, with only a small effect on growth. Experiments with a range of concentrations of cordycepin and α-amanitin indicated that mRNA synthesis could be curtailed by 30 to 40% within the first 8 hours of germination with only a small effect on embryo growth. Thus, at least the initial phases of seed embryo germination are not closely linked to the synthesis of mRNA, rRNA, or DNA. Maximal sensitivity of embryo growth was obtained with cycloheximide and 2-(4-methyl-2,6-dinitroanilino)-N-methyl propionamide, supporting the idea that protein synthesis is the macromolecular process most closely linked to early germination.     

Gibberellin-like activity in cotyledons and embryonic axes during pea seed germination

Endogenous gibberellin-like activity was determined in dry pea seeds (Pisum sativum cv. Bördi), in cotyledons and axes of germinating pea seeds and also in excised cotyledons and axes. During the first two days of pea seed germination, neither the embryonic axes nor the cotyledons show a mutual influence on gibberellin activity, but this appears after 72–96 h of germination. The gibberellin-like activity m cotyledons and axes of germinating seeds increased during the same period, but it decreased in isolated axes and excised cotyledons.     

Gibberellins in the embryonic axes of tall and dwarf beans and their changes with initial growth

Ten gibberellin-like activities were detected in the dry embryonicaxes of tall (cv. Kentucky Wonder) and dwarf (cv. Masterpiece)beans (Phaseolus vulgaris L.) using the lettuce hypocotyl assayof thin-layer chromatograms; 2 in the non-acidic ethyl acetatefraction (NEI, NEII), 3 in the acidic ethyl acetate fraction(AEI, AEII, AEIII), 2 in the non-acidic n-butanol fraction (NBI,NBII) and 3 in the acidic n-butanol fraction (ABI, ABII, ABIII).There was no qualitative difference in these gibberellins betweenthe tall and dwarf axes, but all, particularly AEIII, NBII andABIII as the main gibberellins in the axes, were contained muchmore abundantly in the tall axes. In both axes the gibberellinactivities of most fractions decreased during germination.Theamounts of some gibberellins in tall axes without cotyledonswere greater than those in axes with cotyledons at 48–72hr of germination. Neither AMO-1618 nor CCC caused significantreduction in the levels of the gibberellins. Axis growth inthe early germinating period depended on the gibberellins storedin the axis, itself. (Received November 26, 1974; )     

Germination of soybean embryonic axes: nucleotide sugar metabolism and initiation of growth

UDP-Sugars comprise the dominant class of nucleotide sugars in isolated soybean axes during early germination. While "dry" axes contain 1 nanomole per axis of UDP-sugars, further synthesis is initiated upon imbibition such that the concentration of total UDP-sugars reaches 8 nanomoles per axis or roughly 1 millimolar after 10 hours, when the axes begin to elongate. The GDP-sugars are essentially absent before imbibition, accumulate rapidly for 90 min to 173 picomoles per axis, then decrease somewhat, reattaining the earlier peak level shortly before growth begins. Meanwhile, the level of ADP-sugars is unchanged. These data indicate that the 10-hour lag period preceding axis growth does not result from a diminished ability to synthesize a major category of nucleotide sugars.Relative rates of synthesis of individual UDP- and GDP-sugars were determined by incorporation of [(3)H]uridine or [(3)H]guanosine. The distribution of label in the different classes of UDP-sugars and in the single class of GDP-sugar was quantitatively similar when analyzed before, at the onset, or during early growth. It therefore seems unlikely that synthesis of a key nucleotide sugar controls the initiation of growth.The possible relevance of nucleotide sugars to growth is discussed and new methods for enzymic analysis of picomole levels of nucleotide sugars are described.     

Superoxide anion and hydrogen peroxide metabolism in soybean embryonic axes during germination.

The total rate of mitochondrial O2- production in the presence of NADH as substrate increased from 200 to 1340 pmol/min per axis between 2 and 30 h of imbibition. The activities of the enzymes involved in hydroperoxide metabolism, e.g., superoxide dismutase, catalase, peroxidase and glutathione and ascorbate peroxidases, markedly changed during the germination of soybean embryonic axes. Superoxide dismutase was the enzymatic activity affected the most during the initial stages of germination. Intracellular O2- steady-state concentration, calculated from the rate of O2- production and superoxide dismutase activity, showed a 2-fold increase from 2 x 10(-8) M to 4 x 10(-8) M in germination phase I, declined in phase II to 2 x 10(-8) M and remained constant over the rest of the incubation period. The reaction of H2O2 and luminol catalyzed by Co2+ was utilized to measure H2O2 diffused out of the soybean axes after 5 to 10 min of incubation. The catalase-sensitive luminol emission of diffusates prepared from axes previously imbibed from 2 to 30 h corresponded to a H2O2 intracellular steady-state concentration in the range of 0.3 to 0.9 microM. The activity of metal-containing antioxidant enzymes was determined in the extracellular fluid. Cell wall peroxidase activity increased from 10 to 300 mumol/min per mg protein and appears as a potentially important pathway for H2O2 utilization. Hydrogen peroxide metabolism in soybean embryonic axes during early inhibition appears to have the following main features: (a) mitochondrial membranes are the most important source of cytosolic O2- and H2O2; (b) H2O2 is regulated at a steady-state concentration of 0.3-0.9 microM; (c) catalase is the main enzyme in terms of H2O2 utilization; (d) H2O2 exo-diffusion is quantitatively important destiny of intracellular H2O2; and (e) extracellular peroxidase located at the cell wall affords an enzymatic system able to use diffused H2O2.     

The influence of plant growth regulators on the growth of the embryonic axes of red- and far-red-treated lettuce seeds

The influence of several plant growth regulators on the growth of the embryonic axes from red- and far-red-(R- and FR-)treated lettuce (Lactuca sativa L., cv. Grand Rapids) seeds was examined; as shown previously, the water potential of the axes from R-treated seeds has been lowered by 3.5–5.6 bars compared to that in axes from FR-treated ones. Kinetin and abscisic acid (ABA), when included in the incubation medium, reduced the elongation of the axes whereas fusicoccin stimulated it; however, these effects were the same in axes of both R- and FR-treated seeds. In contrast, elongation of axes from FR-treated seeds was stimulated by gibberellic acid (GA₃, but elongation of axes from R-treated ones was not affected by this hormone. This latter result indicates that gibberellins may be involved in the phytochrome-mediated growth responses in lettuce axes.When the root caps of the embryos were removed prior to light treatment, R was still able to induce a water-potential decrease in the embryonic axes, indicating that at least a portion of the active Pfr resides in the axis and not the root cap.Abbreviations ABA abscisic acid - FR far red light - GA₃ gibberellic acid - PEG polyethylene glycol - Pfr far-red-absorbing form of phytochrome - R red light     

不同生育期转双价(Bt+CpTI)基因抗虫棉根际土壤酶活性和养分含量变化

田间试验条件下,以转双价(Bt+CpTI)基因棉SGK321及其非转基因亲本常规棉石远321为研究对象,比较分析不同生育期(30、60、90、120d)转双价基因棉和亲本棉根际土壤酶活性(脲酶、碱性磷酸酶、过氧化氢酶)及速效养分(硝态氮、铵态氮和速效磷)的变化。结果表明,转双价基因棉根际土壤硝态氮、铵态氮和速效磷含量变化趋势与其亲本常规棉基本一致,但各养分的具体变化幅度因生育期不同而异。播种后30、60、120d转双价基因棉和亲本常规棉根际土壤碱性磷酸酶活性无显著差异;土壤脲酶和过氧化氢酶活性随棉花生育进程其变化趋势虽各不相同,但同一生育期转双价基因棉与亲本常规棉根际土壤脲酶和过氧化氢酶活性均无显著差异。表明,土壤酶活性和速效养分含量受转双价基因棉的影响较小,其变化主要受生育期的影响。     

A novel effect of 6-azauridine on growth and protein synthesis in wheat embryonic axes

Growth and the rate of protein synthesis in germinating wheat embryonic axes are inhibited by the analog 6-azauridine via a mechanism which is independent of the usual effect of this compound as an inhibitor of de novo synthesis of UTP. The effects on growth and protein synthesis can be separated from that on UTP biosynthesis by analyses of the kinetics by which each effect is maximized following a 1.5-h pulse with 6-azauridine, and by saturation of the responses at different doses of the analog. The inhibitions of growth and protein synthesis are apparently not mediated through the rate of poly A(+) RNA synthesis (reduced as little as 8%), but rather by an effect on translation. Since cordycepin reduces the azauridine inhibitions of growth and protein synthesis, it is suggested that these latter effects of 6-azauridine may depend upon the synthesis of an inhibitory azauridyl-RNA.     

On the variation in enzyme activities during the growth of burkitts lymphoma cells in suspension culture

• 1.1. The growth characteristics of Burkitt's lymphoma cells in suspension culture have been studied, and a mean population doubling time of 20–21 hr established for this cell line under a range of nutritional and physical conditions; data which have provided a basis for the assessment of the reproducibility of the culture techniques and conditions which were employed in the subsequent studies.
• 2.2. Activities of lactate dehydrogenase (LDH), aldolase and esterase, as well as the cellular content of total soluble protein, and the isoenzyme pattern of LDH, were monitored in randomly growing Raji cells for the duration of a complete growth cycle.
• 3.3. In this period, the temporal pattern of variation in the levels of total soluble protein were seen to reflect alterations in LDH activity during a single growth cycle.
• 4.4. The fluctuations observed in LDH activity were greater than those observed for either aldolase or esterase activity, and, from the data considered, the maximum degree of variation appeared to be confined to the initial stages of growth.
• 5.5. Extracellular levels of LDH activity remained relatively constant throughout the growth cycle. so that the large fluctuations in intracellular LDH activity could not be attributed to either secretion or leakage of the enzyme into the culture medium.
• 6.6. No gross changes in the pattern of LDH isoenzymes in these Raji cells were detected during the course of a single growth cycle.