Review: role of carbon sources for in vitro plant growth and development

In vitro plant cells, tissues and organ cultures are not fully autotrophic establishing a need for carbohydrates in culture media to maintain the osmotic potential, as well as to serve as energy and carbon sources for developmental processes including shoot proliferation, root induction as well as emission, embryogenesis and organogenesis, which are highly energy demanding developmental processes in plant biology. A variety of carbon sources (both reducing and non-reducing) are used in culture media depending upon genotypes and specific stages of growth. However, sucrose is most widely used as a major transport-sugar in the phloem sap of many plants. In micropropagation systems, morphogenetic potential of plant tissues can greatly be manipulated by varying type and concentration of carbon sources. The present article reviews the past and current findings on carbon sources and their sustainable utilization for in vitro plant tissue culture to achieve better growth rate and development.     

Role of Gene Duplications in the Adaptation of Salmonella Typhimurium to Growth on Limiting Carbon Sources

Duplication-containing cells are selected when growth of Salmonella typhimurium is limited by the availability of any one of several carbon and energy sources. Under conditions of extreme starvation, growth occurs almost exclusively in the duplication-containing fraction of the population. Cells with duplications of one large segment of the chromosome are repeatedly selected regardless of which of these carbon sources limits growth. The duplicated chromosomal segment encodes the transport systems for all of these carbon sources. This duplication is not selected during growth on a carbon source for which the permease is not included within the duplication segment. This suggests that the growth advantage conferred by the duplication may be due to increased transport of the limiting carbon source. Inclusion of the permease alone is not sufficient to explain the growth advantage of the duplications, since other common duplications that include the permease are not selected.     

Microbial growth on substitutable substrates: characterizing the consumer-resource relationship

In this article, we consider the growth of microorganisms on mixtures of carbon sources and characterize the consumer-resource relationship for this system. The characteristic features observed for the growth of a single microorganism on a pair of carbon sources allow a representation of this relationship based on a general paradigm for resource classification. This representation is verified using a comprehensive model for microbial growth on carbon sources. The results show that for the same pair of carbon sources the qualitative nature of the consumer-resource relationship changes with changing specific growth rate, and therefore a change in the identity of the rate-limiting substrate(s). (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 77-90, 1997.     

Kinetics of growth and substrate consumption of Escherichia coli ML 30 on two carbon sources.

When E. coli ML 30 is grown in batch culture on a mineral salt medium containing a mixed carbon source of glucose and pyruvate, there is no sequential utilization of the carbon sources. The consumption of glucose and pyruvate takes place simultaneously with reciprocal influence (inhibition) on rates of substrate uptake. The specific growth rate is greater than mupmax for pyruvate but smaller than musmax for glucose. In the paper three cases of kinetics of growth and of substrate consumption at several combinations of initial substrate concentrations are considered. A mathematical model is proposed and investigated. The model allows to describe the growth on glucose or on pyruvate not only as singular carbon sources, but also as a mixed carbon source with reciprocal inhibition on rates of substrate uptake. By data fitting parameters of growth and substrate consumption were found.     

Carbon utilization profiles of Fusarium virguliforme isolates

Fusarium virguliforme is the cause of sudden death syndrome in soybean. Physiological variability among isolates of the fungus is unknown. One way to measure physiologic variability is to analyze growth on different carbon sources. The carbon source utilization profiles of 18 F. virguliforme isolates were examined using the Biolog FF 96-well microplate, which contains 95 different carbon sources. The utilization of dextrin,D-mannitol, maltotriose,D-lactic acid methyl ester, N-acetyl-D-galactosamine, salicin, D-trehalose, and L-alanine differed significantly among isolates (P = 0.05). Carbon sources were grouped into 3 clusters based on their ability to promote growth of F. virguliforme, after calculating Euclidean distances among them. About 12% of the carbon sources promoted a high amount of mycelial growth, 39% promoted a medium amount of growth, and 49% promoted a low amount of mycelial growth; the latter was not significantly different from the water blank control. A hierarchical tree diagram was produced for the 18 isolates based on their carbon source utilization profiles using Ward's hierarchical analysis method. Two main clusters of isolates were formed. One cluster represented greater average mycelial growth on all of the carbon sources than the other cluster. In this study, variability in carbon source utilization among F. virguliforme isolates was evident, but the results were not associated with geographic origin of the isolates, year collected, or published data on aggressiveness. Additional research is needed to determine if these carbon utilization profiles are associated with other biological characteristics, like spore germination, propagule formation, and saprophytic competitiveness.     

紫云英根际微生物碳源利用多样性研究

为探究绿肥根际调控效果的原因,采用碳素利用法-BIOLOG生态版,分析紫云英根际微生物种群结构和功能多样性。结果表明:紫云英根际土壤微生物总活性比非根际高,其主要贡献者是以氨基酸、糖类、酯类和醇类四大碳源为生的微生物,而与以磷酸盐、羧酸和胺类为碳源的微生物关系不大。进一步分析发现:紫云英根际产生了以α-酮丁酸、2-羟基安息香酸、4-羟基安息香酸、D-半乳糖醛酸、α-环式糊精、D-木糖、β-甲基-D-葡萄糖、D-甘露醇和L-精氨酸作为碳源的新微生物种群,完全抑制了以γ-羟基丁酸和苯乙基胺为碳源的微生物种群活性;根泌物抑制了以D-苹果酸、衣康酸和肝糖为碳源的微生物活性,而对以D-化纤二糖、N-乙酰基-D-葡萄胺、丙酮酸甲酯、吐温40、吐温80、L-苯基丙氨酸、L-天门酰胺酸和腐胺为碳源的微生物有促进作用,但对以D-氨基葡糖酸、α-D-乳糖和I-赤藻糖醇为碳源的微生物活性无显著影响;另外,即使是以同一类物质为碳源的微生物种群,在紫云英根际的表现也不完全一样,如以D-乳酸γ-内酯和L-丝氨酸为碳源的可快速培养微生物受到抑制,可慢速培养的微生物活性则有提高,而以甘氨酰基-L-谷氨酸为碳源的微生物的表现正好与之相反。     

Mutants of yeast with altered oxidative energy metabolism: selection and genetic characterization

Isolation of a series of mutants, characterized by decreased ability to utilize nonfermentable carbon sources for growth and presence of all cytochromes, is reported. A total of 161 mutants, showing deficient growth on glycerol but able to reduce 2,3,5-triphenyltetrazolium chloride, were isolated, purified, and characterized by ability to grow on various carbon sources. Mutants showing decreased growth were examined by low-temperature spectroscopy, and the 35 strains shown to possess all cytochromes were retained for further studies. These strains were characterized by growth on various nonfermentable carbon sources, relative yield on glucose medium, and respiration (Q(O2)) of glucose and ethyl alcohol. Genetic studies revealed that at least 19 of the 35 mutants are the result of mutation in single nuclear genes. Furthermore, at least 11 complementation groups are represented among these 19 mutants. Mutants within two complementation groups were shown to be very similar in various properties. These studies demonstrate that a large number of nuclear genes control oxidative energy metabolism and that the characteristics of mutants of the general class are extremely diverse.     

Kinetics of the simultaneous utilization of sugar mixtures by Escherichia coli in continuous culture.

In natural environments heterotrophic microorganisms encounter complex mixtures of carbon sources, each of which is present at a concentration of a few micrograms per liter or even less. Under such conditions no significant growth would be expected if cells utilized only one of the available carbon compounds, as suggested by the principle of diauxic growth. Indeed, there is much evidence that microbial cells utilize many carbon compounds simultaneously. Whereas the kinetics of single-substrate and diauxic growth are well understood, little is known about how microbial growth rates depend on the concentrations of several simultaneously utilized carbon sources. In this study this question was answered for carbon-limited chemostat growth of Escherichia coli fed with mixtures of up to six sugars; the sugars used were glucose, galactose, maltose, ribose, arabinose, and fructose. Independent of the mixture composition and dilution rate tested, E. coli utilized all sugars simultaneously. Compared with growth with a single sugar at a particular growth rate, the steady-state concentrations were consistently lower during simultaneous utilization of mixtures of sugars. The steady-state concentrations of particular sugars depended approximately linearly on their contributions to the total carbon consumption rate of the culture. Our experimental data demonstrate that the simultaneous utilization of mixtures of carbon sources enables heterotrophic microbes to grow relatively fast even in the presence of low environmental substrate concentrations. We propose that the observed reductions in the steady-state concentrations of individual carbon sources during simultaneous utilization of mixtures of carbon sources by heterotrophic microorganisms reflect a general kinetic principle.     

Improved ε‐poly‐l‐lysine productivity partly resulting from rapid cell growth in cultures using a glucose–glycerol mixed carbon source

Productivity enhancements with mixed carbon sources are usually accompanied by simultaneous improvement of cell growth. However, whether the enhanced cell growth in mixed carbon sources influences the biochemical productivity of ε‐poly‐l ‐lysine (ε‐PL) still remains unclear. In this study, we investigated the effect of growth rate on the ε‐PL productivity in a glucose–glycerol mixed carbon source. Based on the typical ε‐PL fed batch fermentation, chemostat culture and relevant physiological analyses were carried out. The ε‐PL productivity was positively correlated to the growth rate ranging from 0.02 to 0.06 h^?1. The primary metabolism activity was enhanced at higher growth rate, providing sufficient precursor l ‐lysine and energy for ε‐PL production. Meanwhile, these two key elements were equally important for biomass production, which could be quickly produced when the cells were fast growing. In addition, rapid growth also strengthened the antioxidant capacity of cells to defend potential oxidative stress. The positive correlation between the growth rate and ε‐PL productivity indicated that the improvement of ε‐PL productivity in the mixed carbon source was partly attributed to the simultaneously enhanced cell growth. Information obtained may provide references for further studies on other secondary biochemicals’ production using mixed carbon sources.     

Synthesis of oxaloacetate in Bacillus subtilis mutants lacking the 2-ketoglutarate dehydrogenase enzymatic complex.

Bacillus subtilis mutants deficient in the 2-ketoglutarate dehydrogenase enzymatic complex required aspartate for growth at wild-type rates on carbon sources for which synthesis of the degradative enzymes is sensitive to catabolite repression (e.g., poor carbon sources), but did not require aspartate for growth on carbon sources which exert catabolite repression (e.g., good carbon sources). Measurement of metabolite pools in a mutant lacking the 2-ketoglutarate dehydrogenase active complex showed that the aspartate requirement for growth on poor carbon sources resulted from a deficiency in intracellular oxaloacetate pools even through pyruvate carboxylase was present at levels corresponding to those in wild-type cells. The oxaloacetate deficiency most likely resulted from the inability of the mutant to regenerate oxaloacetate from citrate due to the enzymatic block in the tricarboxylic acid cycle. Mutants in the enzymes of the dicarboxylic acid half of the citric acid cycle similarly required aspartate for wild-type growth in minimal medium. These results suggested that the complete turning of the tricarboxylic acid cycle is involved in the maintainance of oxaloacetate levels in B. subtilis. The ability of the mutants lacking the 2-ketoglutarate dehydrogenase enzymatic complex to grow at wild-type rates on media containing good carbon sources in the absence of exogenous aspartate is not understood.     

A carbon source-dependent antimetabolite sensitivity test in Bacillus megaterium B71 for isolation of ethionine resistant mutants

Alteration of carbon sources significantly altered the analogue sensitivity of Bacillus megaterium B71. DL-Ethionine (ETN) was highly inhibitory with glucose, mannitol, sucrose, citrate, glycerol and arabinose. DL-Norleucine, L-homoserine and S(2'-aminoethyl)-L-cysteine were either highly inhibitory, slightly inhibitory or non-inhibitory depending on the carbon sources used. Maltose markedly overcame the inhibitory effect of ETN in liquid culture. Uninhibited growth was poor on citrate and arabinose when compared with other carbon sources. Six carbon sources showing comparable growth were used to determine minimum inhibitory concentrations (MIC) of the analogues. The MIC of ETN was highest (450 μg/ml) with maltose and lowest (4 μg/ml) with mannitol. ETN sensitivity was inversely related to the endogenous L-methionine pool size, and was relatively low with mannitol which was used to isolate ETN resistant mutants of B. megaterium B71. The best mutant BUE-118 produced 435 μg/ml of L-methionine.     

Effect of carbon source on growth,nitrogenase and uptake hydrogenase activities of Frankia isolates from Casuarina sp.

The effect of different carbon sources on the growth of Frankia isolates for Casuarina sp. was studied. In addition, regulation of nitrogenase and uptake hydrogenase activity by carbon sources was investigated. For each of the three isolates, JCT287, KB5 and HFPCcI3, growth was greatest on the carbon sources pyruvate and propionate. In general the carbon sources which gave the greatest growth gave the highest levels of nitrogenase activity, but repressed the activity of uptake hydrogenase. The regulation of growth, uptake hydrogenase activity and nitrogenase activity is discussed.     

Increased antimetabolite sensitivity with variation of carbon source during growth.

In Serratia marcescens, analogs of leucine (norleucine), methionine (alpha-methylmethionine), histidine (3-amino-1,2,4-triazolealanine), tyrosine (p-aminophenylalanine), and tryptophan (7-methylindole) are conditional inhibitors of growth; inhibition occurs during the metabolism of some carbon sources but not with others. A further increase in sensitivity to growth inhibition by these analogs can be accomplished through the use of particular combinations of carbon sources present in the inoculum and in the subsequent analog-containing culture medium. Variable sensitivity to analog-mediated inhibition of growth observed during growth on glucose, glycerol, fructose, or citrate correlated inversely with the intracellular pool sizes of the amino acids cognate to the analogs used. The above-cited results, in conjunction with previous results obtained with Pseudomonas aeruginosa and Bacillus subtilis, involve diverse biochemical pathways and suggest that nutritional manipulation to alter the pattern of carbon flow in microorganisms is a generally useful means to accomplish increased sensitivity to growth inhibition by metabolite analogs.     

马铃薯干腐病菌硫色镰孢的生物学特性

从碳源、氮源、酸碱度及生长温度等方面对引起马铃薯干腐病的硫色镰孢Fusarium sulphureum的生物学特性进行了研究。结果表明,该病原菌在不同发育阶段对营养和环境条件的要求存在差异。在固体培养基上,菌落生长最佳碳源为葡萄糖、麦芽糖,最佳氮源为蛋白胨,pH8?为最佳;在液体培养基中,菌丝体生长以麦芽糖为最佳碳源,以硝酸钠为最佳氮源,pH6?为最佳;在分生孢子萌发阶段,在以羧甲基纤维素钠、蛋白胨和谷氨酸为碳、氮源的营养液中,分生孢子萌发率最高,最适pH 6–8。该病菌最适生长温度为25℃,分生孢子致死     

Utilization of Carbon, Nitrogen and Sulphur Compounds by Kabatiella zeae Narita et Hiratsuka

Abstract Morphogenesis of two K. zeae isolates (V7486 and ZP24986) on carbon, nitrogen and sulphur solid nutritive media after 2, 10, 18 and 28 days of incubation were studied. Highly significant differences were found between the isolates in regard to the use of compounds containing these elements. On carbon sources the fungus generally developed smaller colonies of branchy, loose, creamy-brown appearance, with abundant sporulation and abnormal conidia. Sucrose and D-glucose were the best carbon sources for K. zeae because both isolates had the maximum growth, abundant to medium abundant sporulation and mostly typical conidia. K. zeae colonies on organic and inorganic nitrogen and on sulphur sources too were leathery, creased, predominantly pink. Organic nitrogen sources lead to maximum growth and abundant sporulation, whereas ammonium and nitrate compounds caused intermediary growth and medium to very poor fungus sporulation. Addition of different sulphur sources, except sodium thiosulphate and thiocarbamide, did not affect the growth and sporulation of V7486. The growth of ZP 24986 depended on potassium and ammonium sulphate and sodium bisulphate.     

Effect of Carbon Source on Enzymes and Metabolites of Arginine Metabolism in Neurospora

The levels of enzymes and metabolites of arginine metabolism were determined in exponential cultures of Neurospora crassa grown on various carbon sources. The carbon sources decreased in effectiveness (as determined by generation times) in the following order: sucrose, acetate, glycerol, and ethanol. The basal and induced levels of the catabolic enzymes, arginase (EC 3.5.3.1) and ornithine transaminase (EC 2.6.1.13), were lower in mycelia grown on poor carbon sources. Arginase was more sensitive to variations in carbon source than was ornithine transaminase. Induction of both enzymes was sensitive to nitrogen metabolite control, but this sensitivity was reduced in mycelia grown on glycerol or ethanol. The pools of arginine and ornithine were reduced in mycelia grown in unsupplemented medium containing poor carbon sources, but the biosynthetic enzyme ornithine transcarbamylase (EC 2.1.3.3) was not derepressed. The arginine pools were similar, regardless of carbon source, in mycelia grown in arginine-supplemented medium. The ornithine pool was reduced by growth on poor carbon sources. The rate of arginine degradation was proportional to the level of arginase in both sucrose- and glycerol-grown mycelia. The distribution of arginine between cytosol and vesicles was only slightly altered by growth on glycerol instead of sucrose. The slightly smaller cytosolic arginine concentration did not appear to be sufficient to account for the alterations in basal and induced enzyme levels. The results suggest a possible carbon metabolite effect on the expression or turnover of a variety of genes for enzymes of arginine metabolism in Neurospora.     

Deciphering functional redundancy and energetics of malate oxidation in mycobacteria

Oxidation of malate to oxaloacetate, catalyzed by either malate dehydrogenase (Mdh) or malate quinone oxidoreductase (Mqo), is a critical step of the tricarboxylic acid cycle. Both Mqo and Mdh are found in most bacterial genomes, but the level of functional redundancy between these enzymes remains unclear. A bioinformatic survey revealed that Mqo was not as widespread as Mdh in bacteria but that it was highly conserved in mycobacteria. We therefore used mycobacteria as a model genera to study the functional role(s) of Mqo and its redundancy with Mdh. We deleted mqo from the environmental saprophyte Mycobacterium smegmatis, which lacks Mdh, and found that Mqo was essential for growth on nonfermentable carbon sources. On fermentable carbon sources, the Δmqo mutant exhibited delayed growth and lowered oxygen consumption and secreted malate and fumarate as terminal end products. Furthermore, heterologous expression of Mdh from the pathogenic species Mycobacterium tuberculosis shortened the delayed growth on fermentable carbon sources and restored growth on nonfermentable carbon sources at a reduced growth rate. In M. tuberculosis, CRISPR interference of either mdh or mqo expression resulted in a slower growth rate compared to controls, which was further inhibited when both genes were knocked down simultaneously. These data reveal that exergonic Mqo activity powers mycobacterial growth under nonenergy limiting conditions and that endergonic Mdh activity complements Mqo activity, but at an energetic cost for mycobacterial growth. We propose Mdh is maintained in slow-growing mycobacterial pathogens for use under conditions such as hypoxia that require reductive tricarboxylic acid cycle activity.     

Original Articles: Sources of Dissolved Organic Carbon Supporting Planktonic Bacterial Production in the Tidal Freshwater Hudson River

Planktonic bacterial production in the tidal freshwater Hudson River is a major component of secondary productivity and is uncoupled from planktonic primary productivity. There are several major sources of allochthonous dissolved organic carbon (DOC) whose potential contribution to heterotrophic bacterial growth was examined with bioassays. Supply of DOC from the upper Hudson drainage basin and a large tributary in the mid-Hudson together comprise 70 kT DOC/year, which is the bulk of the DOC load to the tidal freshwater Hudson River. Two contrasting tidal wetlands contribute DOC to the main-stem river but were only a few percent of the tributary load even during summer low-flow conditions. The quantity of DOC released from fine sediments was intermediate to the other two loadings considered. Bacterial growth in bioassays receiving water from the sources varied, but differences in thymidine incorporation between reference and DOC sources were small, usually less than 2 nmol/L/h. Similarity in thymidine incorporation suggests that all sources of DOC were capable of supporting bacterial growth at approximately equal rates. Seasonal shifts in carbon availability were clear in several cases, for example, greater growth on wetland-derived DOC at times of peak plant productivity. Seasonal differences in tributary DOC bioavailability were not large despite the well-known seasonality of tributary inputs. Activities of a suite of extracellular enzymes were used as a biologically based characterization of DOC from the various sources. Shifts in allocation among enzymes were apparent, indicating that there are biologically relevant differences in composition among the sources. Fluorescence characteristics and absorbance per unit carbon also varied among sources, providing an independent confirmation of compositional differences among sources. The absence of large differences in bacterial productivity among sources suggests that growth is supported by a wide range of DOC, and the relative importance of the sources is probably related to the quantitative differences in inputs. Efforts to classify carbon supplies to ecosystems must recognize that organism plasticity in carbon use and physical mixing processes will both act to homogenize what might initially appear to be quite distinctive carbon inputs. Received 15 April 1997; accepted 17 February 1998     

不同碳、氮营养对球孢白僵菌生物学特性和抑菌活性的影响

研究了不同碳营养和氮营养对球孢白僵菌菌丝生长、分生孢子产生量、菌丝生物量及其次生代谢产物的抑菌活性.结果表明,球孢白僵菌对单糖、双糖、多糖等碳营养及有机氮和无机氮等氮营养均能够利用,但利用程度存在一定差异.其中,球孢白僵菌的菌丝生长以白砂糖和蛋白胨最快,以甘露醇和硫酸铵最慢;分生孢子产生量以白砂糖和硝酸钾最多,以甘露醇...