Nitrogen fixation and poly-β-hydroxybutyric acid content in bacteroids ofRhizobium lupini andRhizobium leguminosarum

Summary Experiments carried out in a sand culture have demonstrated that during the growth ofVicia faba andLupinus luteus inoculated with effective strains of Rhizobium, and when the behaviour of bacteroids isolated from nodules ofLupinus luteus, Pisum sativum andVicia faba which had been inoculated with effective and ineffective strains and when comparisons were made between bacteriods isolated from effective nodules ofVicia faba andLupinus luteus either at midday or at midnight there is a reverse correlation between the intensity of nitrogen fixation and respiration, on the one hand, and the content of poly--hydroxybutyric acid (PHB), on the other. This evidence suggests an important role played by PHB in the supply of symbiotic fixation with energy and carbon substrates.Glucose and -hydroxybutyrate were the best substrates for PHB synthesis in the suspension of bacteroids of an effective strain ofR. lupini at all stages of plant growth. At the stage of active nitrogen fixation (flowering) PHB was actively synthesized in the presence of succinate. In the absence of exogenous substrates the polymer degraded, the process being enhanced in the presence of ammonium ions. When ammonium was added together with glucose, PHB synthesis did not occur and at the flowering stage the polymer broke down particularly rapidly. re]19760505     

Physiology of nitrogen fixation in Azospirillum lipoferum Br 17 (ATCC 29 709)

Changes of cellular activities during batch cultures with Azospirillum lipoferum strain Br 17 (ATCC 29 709) were observed within the growth cycle, at optimal pO₂ (0.002–0.003 atm). The relative growth rate for cells growing with N₂ as sole nitrogen source during log phase was =0.13 h^-1 and the doubling time was 5.3 h. Nitrogenase activity was not accompanied by hydrogen evolution at any growth stage, and a very active uptake hydrogenase was demonstrated. The hydrogenase activity increased towards the end of the growth period when glucose became limiting and N₂ fixation reached its maximal specific activity. Oxygen consumption and oxygen tolerance at the various growth stages, increased simultaneously with the uptake hydrogenase activity indicating a possible role of this enzyme in an oxygen protection mechanism of A. lipoferum nitrogenase. The efficiency of nitrogen fixation expressed as mg total nitrogen fixed in cells and supernatant per g glucose consumed, was 20 at the early log phase and increased to 48 at the late log phase. About 25% of the total fixed nitrogen was recovered in the culture supernatant.Abbreviations DOT Dissolved oxygen tension - PHB Poly--hydroxybutyric acid - O.D. Optical density (560 nm) - A.T.C.C. American type culture collection - NTA Nitrilotriacetic acid Graduate student of the Universidade Federal Rural do Rio de Janeiro, Brazil     

A rapid gas chromatographic method for the determination of poly-β-hydroxybutyric acid in microbial biomass

Summary The gas chromatographic method for the determination of poly--hydroxybutyric acid (PHB) consists of a mild acid or alkaline methanolysis of poly--hydroxybutyric acid directly without previous extraction of PHB from the cells; this is followed by gas chromatography of the 3-hydroxybutyric acid methylester. The method is characterized by high accuracy and excellent reproducibility, permitting determinations as low as 10^–5 g/l. Only 4 h is required from sampling from the fermenter till completion of the PHB determination.     

Kinetics and effect of nitrogen source feeding on production of poly-β-hydroxybutyric acid by fed-batch culture

Summary A kinetic study of the production of poly--hydroxybutyric acid (PHB) by a fed-batch culture of Protomonas extorquens showed that a nitrogen source was necessary even in the PHB production phase. The effect of ammonia feeding on PHB production was consequently investigated. The nitrogen source (ammonia water) was supplied at a low constant feeding rate after the growth phase in which cell mass concentration reached 60 g/l. Feeding with a small quantity of ammonia resulted in a more rapid increase in intracellular PHB content than was the case without ammonia feeding. Excessive feeding of ammonia, however, caused not only degradation of accumulated PHB but also reduction of microbial PHB synthetic activity.     

Differentiation of nodules of Glycine max

Plants of Glycine max var. Caloria, infected as 14 d old seedlings with a defined titre of Rhizobium japonicum 3Il b85 in a 10 min inoculation test, develop a sharp maximum of nitrogenase activity between 17 and 25 d after infection. This maximum (14±3 nmol C₂H₄ h^-1 mg nodule fresh weight^-1), expressed as per mg nodule or per plant is followed by a 15 d period of reduced nitrogen fixation (20–30% of peak activity). 11 d after infection the first bacteroids develop as single cells inside infection vacuoles in the plant cells, close to the cell wall and infection threads. As a cytological marker for peak multiplication of bacteroids and for peak N₂-fixation a few days later the association of a special type of nodule mitochondria with amyloplasts is described. 20 d after inoculation, more than 80% of the volume of infected plant cells is occupied by infection vacuoles, mostly containing only one bacteroid. The storage of poly--hydroxybutyrate starts to accumulate at both ends of the bacteroids. Non infected plant cells are squeezed between infected cells (25d), with infection vacuoles containing now more than two (up to five) bacteroids per section. Bacteroid development including a membrane envelope is also observed in the intercellular space between plant cells. 35 d after infection, more than 50% of the bacteroid volume is occupied by poly--hydroxybutyrate. The ultrastructural differentiation is discussed in relation to some enzymatic data in bacteroids and plant cell cytoplasm during nodule development.     

Nitrogen fixation by the hydrogen-oxidizing bacterium Alcaligenes latus

The aerobic hydrogen-oxidizing bacterium Alcaligenes latus represented by three strains was found to be able to grow with dinitrogen as the sole nitrogen source: The doubling time of total (Kjeldahl) nitrogen during growth on glucose at 30°C under an atmosphere containing 2% (v/v) oxygen in dinitrogen amounted to 39 h, while that in the presence of ammonium was 3 h. Nitrogen fixation did apparently not occur under air. During diazotrophic growth the cells accumulated up to 75% (w/dry weight) poly--hydroxybutyric acid. The efficiency of nitrogen fixation varied between 10 and 15 mg N per g glucose utilized. The specific nitrogenase activity measured in the acetylene reduction assay amounted to 5–17 nmol C₂H₄ formed per min and mg protein.     

Nutritional effects of non-steady state soil salinity on a salt-tolerant wheat cultivar

Summary Biological nitrogen fixation is considered an important trait of cowpeas (Vigna unguiculata (L.) Walp. var. California Blackeye No. 5) for economical production yet the process does not alone provide the quantity of nitrogen required by the plant for maximum productivity. Two experiments were undertaken to determine the potential of an increase in nodule mass and number of bacteroids resulting in increased nitrogen fixation. Cowpeas were grown in a glasshouse for 7 weeks under conditions forcing near total dependence on biological nitrogen fixation for growth. Nodule mass on the roots was varied by inoculating seeds with various ratios of effective and ineffective rhizobia that could be identified serologically and by the color of nodule formed. The results of both experiments demonstrated a linear relationship between total nodule mass formed by the effective rhizobia and quantity of nitrogen fixed. The regression coefficients were high in both experiments (r=0.99^** and 0.91^**). The relationship between total nitrogen fixed and total number of bacteroids of the effective strain was not consistent. In one experiment the regression coefficient was 0.93^** but in the other experiment it was 0.65^**. From these results it appears that there is good potential for increasing nitrogen fixation in cowpeas by increasing nodule mass. An increase in nodule mass would also result in an increase in the number of bacteroids.     

Isolation of pea nodule bacteroids utilizing silica sol (Percoll) density gradient centrifugation

Isolation of bacteroids from effective (Fix⁺) and ineffective (Fix^–) pea nodules, inoculated withRhizobium leguminosarum K, were performed by a density gradient centrifugation method using silica sol (Percoll). Only one zone (=1.064–1.072; n-zone) was recognized in the Fix⁺ nodule which contained typical Y-shaped bacteroids while two zones (n-zone and =1.125–1.145; n'-zone) were obtained from the Fix^– nodule. The cells in the n'-zone, which are long rods differed morphologically from free-living cells at any growth phase (=1.108–1.125; f-zone and =1.074–1.078; f'-zone), and differed from Y-shaped bacteroids by cell density. The esterase isozyme pattern of bacteroids in the n-and n'-zones also showed clear differences from that of f-and f'-zone of free-living cells.     

Expression of the Alcaligenes eutrophus poly(β-hydroxybutyric acid)-synthetic pathway in Pseudomonas sp.

The 12.5-kb EcoRI restriction fragment PP1 of Alcaligenes eutrophus strain H16, which encodes for -ketothiolase, NADP-dependent acetoacetyl-CoA reductase and poly(-hydroxybutyric acid)-synthase was mobilized to six different species of the genus Pseudomonas belonging to the rRNA homology group I. Pseudomonas aeruginosa, P. fluorescens, P. putida, P. oleovorans, P. stutzeri and P. syringae, which are unable to synthesize and accumulate poly(-hydroxybutyric acid), PHB, were employed as recipients. Whereas the A. eutrophus PHB-synthetic enzymes were only marginally expressed in P. stutzeri, they were readily expressed in the other species. For example, the specific activity of PHB-synthase was 1.8 U/g protein in transconjugants of P. stutzeri but was between 21 and 77 U/mg protein in transconjugants of the other species. All recombinant strains harboring plasmid pVK101::PP1 except those of P. stutzeri accumulated PHB; the PHB content of the cells grown on gluconate under nitrogen limitation varied between 8 and 24.3% of the cellular dry mass.Abbreviations PHB poly(-hydroxybutyric acid) - PHA poly(hydroxyalkanoic acid)     

Optimized production of L-β-hydroxybutyric acid by a mutant of Yarrowia lipolytica

A mutant strain of Yarrowia lipolytica was developed which produced 8.0 g l--hydroxybutyric acid l^–1 from butyric acid in a batch culture. The optimum culture conditions in the fermenter for maintenance of a high cell activity, determined by chemostat analyses, were a specific growth rate of 0.06 h^–1, a glucose concentration of 2.0 g l^–1, and a butyric acid concentration of 8.1 g l^–1. A fed-batch fermentation was performed under these conditions resulting in an l--hydroxybutyric acid yield of 31 g l^–1.     

Development of nodules of Glycine max infected with an ineffective strain of Rhizobium japonicum

Bacteroids in ineffective (nitrogenase negative) nodules of Glycine max, infected with Rhizobium japonicum 61-A-24, as compared to those in effective nodules are characterized by reduced specific activities of alanine dehydrogenase to 15%, of 3-hydroxybutyrate dehydrogenase to 50%, and an increase of glutamine synthetase to 400%. In the plant cytoplasm of ineffective nodules, glutamine synthetase activity is reduced to 10–30%, glutamate dehydrogenase to 50–70%, and the aspartate aminotransferase and alanine aminotransferase are enhanced to 120–200%, depending on the age of the nodules. The total pool of soluble amino acids is reduced to 52 mol per g nodule fresh weight, as compared to 186 mol in effective nodules, with a replacement of asparagine (42 mol% of the amino acids) by an unknown amino compound. This compound is absent in nitrogenase, repressed and derepressed, free-living Rhizobium japonicum cells and in the uninfected root tissue. In nitrogenase derepressed, as compared to the repressed free-living cells of Rhizobium japonicum 61-A-101, arginine shows the most obvious change with a reduction to less than one tenth. The ultrastructure of the ineffective nodule is different from the effective organ even in the early stages. The membrane envelopes of the infection vacuoles are decomposing in heavily infected cells within 18 to 20 d after infection. In lightly infected cells very large vacuoles develop with only a few bacteroids inside. No close associations of cristae-rich mitochondria with amyloplasts are observed as in effective nodules. The uninfected cells keep their large starch granules even 40 d after infection. Some poly--hydroxybutyrate accumulation in the bacteroids is observed but only in the early stages, and it is almost absent in old nodules (40 d). At this age the infected cells are obviously compressed by uninfected cells, whereas in effective nodules with nitrogenase activity and leghaemoglobin formation, the infected cells have a much higher osmotic pressure than the neighbouring uninfected cells.Abbreviations PHBA poly--hydroxybutyric acid Prof. Dr. A. Pirson on the occasion of his 70th birthday     

Nitrogen fixation by Nodularia spumigena Mertens (Cyanobacteriaceae). 2: Laboratory studies

The effects of changes in diurnal light patterns, salinity, and phosphorus on nitrogen fixation (as measured by acetylene reduction) by Nodularia spumigena Mertens were examined. As well, the effects of added inorganic nitrogen on growth, nitrogen fixation and heterocyt frequencies, and changes in nitrogen fixation and heterocyst frequencies during the growth cycle of Nodularia in cultures were determined.The diurnal pattern of nitrogenase activity in Nodularia was primarily light-induced, though dark activity did occur. Nitrogenase activity following a period of darkness exceeded the normal light rate (> 90 compared to 50 nmol · C₂H₂ reduced · ml^–1 · h^–1). Nitrogen fixation was reduced by high and very low salinities (5 to 10 was the optimum range), and added phosphorus stimulated nitrogenase in P-starved cells. Added nitrogen (ammonium or nitrate) had no effect on the growth of Nodularia, but in short term studies, ammonium completely inhibited nitrogenase activity. Heterocyst frequencies were greatest in the log phase of growth (to 40 per mm). During stationary phase, nitrogenase activity was negligable.     

Production of poly-β-hydroxybutyrate from methanol: characterization of a new isolate of Methylobacterium extorquens

Summary Poly--hydroxybutyric acid (PHB) and similar bacterial polyesters are promising candidates for the development of environment-friendly, totally biodegradable plastics. The use of methanol, one of the cheapest noble substrates available, may help to reduce the cost of producing such bioplastics. As a first step, a culture collection of 118 putative methylotrophic microorganisms was obtained from various soil samples without any laboratory enrichment step to favour culture diversity. The most promising culture was selected based on rapidity of growth and PHB accumulation and later identified as Methylobacterium extorquens. This isolate was obtained from soml contaminated regularly with used oil products for some 40 years. Concentrations of methanol greater than 8 g/l affected growth significantly and the methanol concentration was optimal at 1.7 g/l. PHB concentrations averaged 25–30% (w/v) of dry weight under non-optimized conditions. Controlling methanol concentration, using an open-loop configuration, led to biomass levels of 9–10 g/l containing 30–33% PHB while preventing methanol accumulation. The new isolate was also able to produce the co-polymer PHB/poly--hydroxyvalerate (PHV) using the mixture methanol + valerate. The PHV-to-PHB ratio was about 0.2 at the end of the fermentation. An average molecular mass varying between 2 and 3 × 10⁵ Da was obtained for three PHB samples using two different measurement methods.Publication number NRCC No. 33672 Offprint requests to: D. Groleau     

Some characteristics of Methylococcus mobilis sp. nov.

A methane-assimilating coccus was isolated from a continuous culture. Under certain conditions, in pure culture the cells grew in sarcina-like, refractile clusters.That these clusters resembled multiple-bodied cysts was confirmed by electron microscopy. In addition, tube-like structures, not previously reported in methane-oxidizing bacteria, were found. Motile cells, with one or, rarely, two flagella were occasionally observed. Methane and methanol were exclusively assimilated as the sole source of carbon and energy. Formaldehyde, formate, lower alkanes, alcohols and aldehydes, with the exception of acetaldehyde, were oxidized by cell suspensions. The presence of a number of key enzymes in cell-free extracts suggested that the ribulose-monophosphate pathway of carbon assimilation is active. This agreed with the internal membrane structure found (type I organism).Nitrate, ammonia, peptone, yeast extract, asparagine, aspartic acid and glutamic acid were assimilated as sources of nitrogen. No nitrogen fixation could be demonstrated.The cells contained 9.1–9.7% nitrogen and 7–10% lipid, dependent on the growth conditions. Poly--hydroxybutyric acid could not be detected. The guanine+cytosine (G+C) content of the DNA was found to be 56.3%. As a name for the organism Methylococcus mobilis sp. nov. is proposed.Abbreviations used G+C Guanine+Cytosine     

Nitrogen nutrition and the development of biochemical functions associated with nitrogen fixation and ammonia assimilation of nodules on cowpea seedlings

During early development (up to 18 d after sowing) of nodules of an effective cowpea symbiosis (Vigna unguiculata (L.) Walp cv. Vita 3: Rhizobium strain CB756), rapidly increasing nitrogenase (EC 1.7.99.2) activity and leghaemoglobin content were accompanied by rapid increases in activities of glutamine synthetase (EC 6.3.1.2), glutamate synthase (EC 2.6.1.53), enzymes of denovo purine synthesis (forming inosine monophosphate) xanthine oxidoreductase (EC 1.2.3.2), urate oxidase (EC 1.7.3.3), phosphoenolpyruvate carboxylase (EC 4.1.1.31) and led to increased export of ureides (allantoin and allantoic acid) to the shoot of the host plant in the xylem. Culturing plants with the nodulated root systems maintained in the absence of N₂ (in 80 Ar: 20 O₂, v/v) had little effect on the rates of induction and increase in nitrogenase activity and leghaemoglobin content but, in the absence of N₂ fixation and consequent ammonia production by bacteroids, there was no stimulation of activity of enzymes of ammonia assimilation or of the synthesis of purines or ureides. Addition of NO ₃ ^- (0.1–0.2 mM) relieved host-plant nitrogen deficiency caused by the Ar: O₂ treatment but failed to increase levels of enzymes of N metabolism in either the bacteroid or the plant-cell fractions of the nodule. Premature senescence in Ar: O₂-grown nodules occurred at 18–20 d after sowing, and resulted in reduced levels of nitrogenase activity and leghaemoglobin but increased the activity of hydroxybutyrate oxidoreductase (EC 1.1.1.30).     

Effects of constitutive expression of nitrate reductase in transgenic Nicotiana plumbaginifolia L. in response to varying nitrogen supply

Transformed Nicotiana plumbaginifolia plants with constitutive expression of nitrate reductase (NR) activity were grown at different levels of nitrogen nutrition. The gradients in foliar NO ₃ ^– content and maximum extractable NR activity observed with leaf order on the shoot, from base to apex, were much decreased as a result of N-deficiency in both the transformed plants and wild type controls grown under identical conditions. Constitutive expression of NR did not influence the foliar protein and chlorophyll contents under any circumstances. A reciprocal relationship between the observed maximal extractable NR activity of the leaves and their NO ₃ ^– content was observed in plants grown in nitrogen replete conditions at low irradiance (170 mol photons·m^–2 ·s^–1). This relationship disappeared at higher irradiance (450 mol photons·m^–2·S^–1) because the maximal extractable NR activity in the leaves of the wild type plants in these conditions increased to a level that was similar to, or greater than that found in constitutive NR-expressors. Much more NO ₃ ^– accumulated in the leaves of plants grown at 450 mol photons·m^–2·s^–1 than in those grown at 170 mol photons·m^–2·s^–1 in N-replete conditions. The foliar NO ₃ ^– level and maximal NR activity decreased with the imposition of N-deficiency in all plant types such that after prolonged exposure to nitrogen depletion very little NO ₃ ^– was found in the leaves and NR activity had decreased to almost zero. The activity of NR decreased under conditions of nitrogen deficiency. This regulation is multifactoral since there is no regulation of NR gene expression by NO ₃ ^– in the constitutive NR-expressors. We conclude that the NR protein is specifically targetted for destruction under nitrogen deficiency. Consequently, constitutive expression of NR activity does not benefit the plant in terms of increased biomass production in conditions of limiting nitrogen.Abbreviations Chl chlorophyll - N nitrogen - NR NADH-nitrate reductase - WT wild type     

Nitrogen fixation and metabolism by groundwater-dependent perennial plants in a hyperarid desert

The Central Asian Taklamakan desert is characterized by a hyperarid climate with less than 50 mm annual precipitation but a permanent shallow groundwater table. The perched groundwater (2–16 m) could present a reliable and constant source of nitrogen throughout the growing season and help overcome temporal nitrogen limitations that are common in arid environments. We investigated the importance of groundwater and nitrogen fixation in the nitrogen metabolism of desert plants by assessing the possible forms and availability of soil N and atmospheric N and the seasonal variation in concentration as well as isotopic composition of plant N. Water availability was experimentally modified in the desert foreland through simulated flooding to estimate the contribution of surface water and temporally increased soil moisture for nutrient uptake and plant–water relations. The natural vegetation of the Taklamakan desert is dominated by plants with high foliar nitrogen concentrations (2–3% DM) and leaf nitrate reductase activity (NRA) (0.2–1 mol NO₂^– g^–1 FW h^–1). There is little evidence that nitrogen is a limiting resource as all perennial plants exhibited fast rates of growth. The extremely dry soil conditions preclude all but minor contributions of soil N to total plant N so that groundwater is suggested as the dominant source of N with concentrations of 100 M NO₃^–. Flood irrigation had little beneficial effect on nitrogen metabolism and growth, further confirming the dependence on groundwater. Nitrogen fixation was determined by the ¹⁵N natural abundance method and was a significant component of the N-requirement of the legume Alhagi, the average contribution of biologically fixed nitrogen in Alhagi was 54.8%. But nitrogen fixing plants had little ecological advantage owing to the more or less constant supply of N available from groundwater. From our data we conclude that the perennial species investigated have adapted to the environmental conditions through development of root systems that access groundwater to satisfy demands for both water and nutrients. This is an ecologically favourable strategy since only groundwater is a predictable and stable resource.     

Relationship between bacteroid poly-beta-hydroxybutyrate accumulation and nodule functioning in the Galega orientalis-Rhizobium galegae symbiosis under diamine treatment

Exposure of Galega orientalis plants to diamines putrescine (Put) and cadaverine (Cad) at concentrations from 0.01 to 2.0 m M significantly altered carbon and nitrogen metabolism in their root nodules. Correlative studies of bacteroid poly- β -hydroxybutyrate (PHB) content and acetylene-reduction capacity of the nodules revealed a negative relationship between these parameters. Utilisation of PHB deposits by bacteroids and high acetylene reduction activity was observed when applying low diamine concentrations. The increase in PHB accumulation in response to high diamine levels was accompanied by a considerable decline in nodule nitrogenase activity. Supplying isolated Galega bacteroids with various diamine concentrations significantly modified bacteroid oxygen consumption, which might be associated with alterations in carbon flux to the bacteroids. Finally, modulation of the bacteroid content upon Put and Cad treatment was examined. The results are discussed in terms of possible causes of the diamine-induced changes in nodule metabolism.     

Nitrogen fixation in perennial forage legumes in the field

Nitrogen acquisition is one of the most important factors for plant production, and N contribution from biological N₂ fixation can reduce the need for industrial N fertilizers. Perennial forages are widespread in temperate and boreal areas, where much of the agriculture is based on livestock production. Due to the symbiosis with N₂-fixing rhizobia, perennial forage legumes have great potential to increase sustainability in such grassland farming systems. The present work is a summary of a large number of studies investigating N₂ fixation in three perennial forage legumes primarily relating to ungrazed northern temperate/boreal areas. Reported rates of N₂ fixation in above-ground plant tissues were in the range of up to 373 kg N ha^–1 year^–1 in red clover (Trifolium pratense L.), 545 kg N ha^–1 year^–1 in white clover (T. repens L.) and 350 kg N ha^–1 year^–1 in alfalfa (Medicago sativa L.). When grown in mixtures with grasses, these species took a large fraction of their nitrogen from N₂ fixation (average around 80%), regardless of management, dry matter yield and location. There was a large variation in N₂ fixation data and part of this variation was ascribed to differences in plant production between years. Studies with experiments at more than one site showed that also geographic location was an important source of variation. On the other hand, when all data were plotted against latitude, there was no simple correlation. Climatic conditions seem therefore to give as high N₂ fixation per ha and year in northern areas (around 60°N) as in areas with a milder climate (around 40°N). Analyzing whole plants or just above-ground plant parts influenced the estimate of N₂ fixation, and most reported values were underestimated since roots were not included. Despite large differences in environmental conditions, such as N fertilization and geographic location, N₂ fixation (Nfix; kg N per ha and year) was significantly (P<0.001) correlated to legume dry matter yield (DM; kg per ha and year). Very rough, but nevertheless valuable estimations of Nfix in legume/grass mixtures (roots not considered) are given by Nfix = 0.026DM + 7 for T. pratense, Nfix = 0.031DM + 24 for T. repens, and Nfix = 0.021DM + 17 for M. sativa.     

Nature of light requirement for the flowering of Chenopodium rubrum L. (Ecotype 60° 47′N)

It has been shown that induction of flowering in Chenopodium rubrum L. (ecotype 60° 47 N) seedlings in BCJ light conditions is intensity dependent (Cumming, 1969, Canad. J. Bot. 47, 1247–1250) and that, this intensity dependence is not based on photosynthesis (Sawhney and Cumming, 1971, Can. J. Bot. 49, 2133–2137). Since BCJ light emits a high proportion of energy in the far-red, and the High Energy Reaction (HER) has its action maxima in the far-red and blue regions of the spectrum, we tested the involvement of HER in the light induction of flowering in C. rubrum. Our results show that optimum intensities of blue light are effective in inducing flowering in C. rubrum. Red light exposure does not lead to flower induction. We suggest the HER may be involved in the flower induction of C. rubrum in light. However, when high energy in blue and/or farred is provided in presence of energy between 500–700 nm wavebands, there is no flowering in C. rubrum. We suggest that flower inducing activity of HER may be counteracted by flower inhibitory action of red wavebands.This paper constitutes a part of a Ph. D. thesis submitted to the University of Western Ontario, London, Ontario