GROWTH AND DEVELOPMENT OF REPRODUCTIVE AND VEGETATIVE TISSUES OF BOUGAINVILLEA CULTURED IN VITRO AS A FUNCTION OF CARBOHYDRATE

Inflorescence meristems and vegetative tissues, excised from noninduced Bougainvillea ‘San Diego Red’ plants, were cultured in vitro in media containing either 3% fructose, glucose or sucrose as carbon sources. Growth and development of young leaves were equivalent whether sucrose or fructose was used whereas floret initiation on inflorescence meristems was much greater when fructose or glucose was the carbon sources. Brief (1-3 days) exposure of inflorescence meristems to fructose at the beginning of culture and subsequent transfer to sucrose did not increase development over continuous culture in sucrose. Longer exposures (4-7 days) to fructose with subsequent transfer to sucrose did, however, increase the percentage of meristems developing florets, but such treatment did not increase development to the same level as those exposed to fructose for the entire period in vitro. During the first 18 days of culture, growth of meristems in sucrose was linear while that in fructose was exponential. There was no difference in carbohydrate requirements for floret initiation on meristems excised from short-day induced or noninduced plants, suggesting that induction does not enhance the ability of meristems to utilize sucrose.     

β-Fructofuranosidase and sucrose phosphorylase of rumen bacterium <Emphasis Type="Italic">Pseudobutyrivibrio ruminis</Emphasis> strain 3

The subject of this study was the fructan and sucrose degrading enzymes of bacterium Pseudobutyrivibrio ruminis strain 3. It was stated that cell extract from bacteria growing on inulin contained β-fructofuranosidase (EC 3.2.1.80 and/or EC 3.2.1.26) and sucrose phosphorylase (EC 2.4.1.7), while the bacteria maintained on sucrose showed only phosphorylase. Partially purified β-fructofuranosidase digested inulooligosaccharides and sucrose to fructose or fructose and glucose, respectively, but was unable to degrade the long chain polymers of commercial inulin and Timothy grass fructan. Digestion rate of inulooligosaccharides fit Michaelis–Menten kinetics with V_max 5.64 μM/mg/min and K_m 1.274%, respectively, while that of sucrose was linear. Partially purified sucrose phosphorylase digested only sucrose. The digestion products were fructose, glucose-1P and free glucose. The reaction was in agreement with Michaelis–Menten kinetics. The V_max were 0.599 and 0.584 μM/mg/min, while K_m were 0.190 and 0.202% for fructose release and glucose-1P formation, respectively, when bacteria grew on inulin. The V_max were, however, 1.37 and 1.023 μM/mg/min, while K_m were 0.264 and 0.156%, if bacteria were grown on sucrose. The free glucose was hardly detectable for the enzyme originated from inulin grown bacteria, but glucose levels ranged from 0.05 to 0.25 μM/mg/min, when cell extract from bacteria grown on sucrose was used. Release of free glucose was observed when no inorganic phosphate was present in reaction mixture.     

Influence of glucose,fructose and sucrose as carbon sources on kinetics and stoichiometry of lysine production by <Emphasis Type="Italic">Corynebacterium glutamicum</Emphasis>

Batch cultivations of l-lysine-producing Corynebacterium glutamicum ATCC 21253 were carried out on the different carbon sources, glucose, sucrose and fructose. The time profiles of substrate and product concentrations were evaluated to compare kinetics and stoichiometry of lysine production. The lysine yield (mol C/mol C) on glucose was 8% higher than on sucrose and 30% higher than on fructose. The highest final biomass concentration of 5.0 g/l was obtained on glucose, whereas fructose and sucrose yielded 20% less biomass. Compared to glucose, fructose resulted in significantly higher respiration rates, a higher substrate uptake rate but a lower lysine production rate during the cultivation process. This was probably due to a higher tricarboxylic cycle activity combined with a lower activity of the pentose phosphate pathway. On sucrose, specific rates and yields differed significantly from those on fructose and glucose. Transport and metabolism of sucrose, therefore, are not a simple superposition of its building blocks, glucose and fructose. Journal of Industrial Microbiology & Biotechnology (2002) 28, 338–343 DOI: 10.1038/sj/jim/7000252 Received 28 November 2001/ Accepted in revised form 06 March 2002     

Morphometric variation in a hybrid zone of two subspecies of Gerris costae (Heteroptera: Gerridae) in the Maritime Alps

Two morphologically distinct subspecies of Gerris costae form a contact zone extending from the Maritime to the Western Alps. Within this area canonical trend surface analysis revealed a geographic pattern of morphometric variation consistent with topography. From North to South, and from high to low elevation there was a transition from G. c. costae-like phenotypes to phenotypes resembling pure G. c. fieberi. The same pattern was confirmed with canonical variate analysis not taking geographic location into account; it is therefore not an artifact of trend surface analysis. Comparisons of the pattern of morphometric variation of laboratory-reared offspring with the pattern of their parents sampled from natural populations show that geographic variation is mostly determined genetically. Intermediate individuals from field populations probably are natural hybrids between the two subspecies, because laboratory-reared hybrids were intermediate between the offspring of pure strains. We did not find increased morphometric variation within the contact zone. This suggests unimpeded introgression and is in contrast with an increase in size variability that is predicted to be associated with a transition between uni- and bivoltine forms.     

Sorbitol production by Zymomonas mobilis

Summary High resolution ¹³C Nuclear Magnetic Resonance (NMR) spectroscopy has been employed to determine the chemical composition of the unknown major products in a sucrose or fructose plus glucose fermentation to ethanol by the bacterium Zymmonas mobilis. When grown on these sugars Z.mobilis was found to produce significant amounts of sorbitol, up to 43 g·l^-1 for strain ZM31 when grown on 250 g·l^-1 sucrose.The production of sorbitol and decrease of glucose, fructose, or sucrose was followed throughout batch fermentations by NMR and HPLC. Sorbitol was shown to be derived only from fructose by [¹⁴C]-feeding experiments. Additionally ³¹P NMR spectroscopy was utilized to determine the concentrations of both glucose 6-phosphate and fructose 6-phosphate relative to their respective concentrations in Z.mobilis cells fermenting glucose or fructose alone.It is suggested that free glucose inside the cell inhibits fructokinase. Free intracellular fructose may then be reduced to sorbitol via a dehydrogenase type enzyme. Attempts to grow Z.mobilis on sorbitol were unsuccessful, as were experiments to induce growth via mutagenesis.This work was supported in part by the National Energy Research, Development and Demonstration Council of Australia     

Variability for Free Sugars and Organic Acids in Capsicum chinense

Variability in the concentrations of the chemical constituents that contribute to fruit flavor in Capsicum chinense is poorly documented in the scientific literature. We surveyed fruit of 216 landraces and cultivated varieties of Capsicum chinense acquired from North, Central, and South America, and analyzed these for concentrations of the simple sugars sucrose, glucose, and fructose, and citric, malic, succinic, fumaric, and ascorbic acids. Concentrations (mg/100 g Fresh Weight (FW) of whole fruit) of sucrose, glucose, and fructose in fruit of this species ranged from 0.0 to 150, 68 to 701, and 101 to 823, respectively. The total of these sugars (sucrose+glucose+fructose) ranged from 198 to 1543 mg/100 g FW. Concentrations of organic acids ranged from 0.0 (not detected) to 818, 430, 340, and 232 mg/100 g FW for citric, malic, fumaric, and succinic acids, respectively. However, the relative ranking in the concentrations of the individual acids was genotype‐dependent. Total ascorbic‐acid values ranged from 30 to 1466 mg/100 g FW. These data serve to document the range in the concentrations of individual sugars and acids present in mature C. chinense fruit and suggest that this variability may lend itself to studies involving the synthesis and/or metabolism of compounds associated with fruit flavor.     

Sugar utilization by yeast during fermentation

Summary When glucose and fructose are fermented separately, the uptake profiles indicate that both sugars are utilized at similar rates. However, when fermentations are conducted in media containing an equal concentration of glucose and fructose, glucose is utilized at approximately twice the rate of fructose. The preferential uptake of glucose also occurred when sucrose, which was first rapidly hydrolyzed into glucose and fructose by the action of the enzyme invertase, was employed as a substrate. Similar results were observed in the fermentation of brewer's wort and wort containing 30% sucrose and 30% glucose as adjuncts. In addition, the high levels of glucose in the wort exerted severe catabolite repression on maltose utilization in theSaccharmyces uvarum (carlsbergensis) brewing strain. Kinetic analysis of glucose and fructose uptake inSaccharomyces cerevisiae revealed aK _m of 1.6 mM for glucose and 20 mM for fructose. Thus, the yeast strain has a higher affinity for glucose than fructose. Growth on glucose or fructose had no repressible effect on the uptake of either sugar. In addition, glucose inhibited fructose uptake by 60% and likewise fructose inhibited, glucose uptake by 40%. These results indicate that glucose and fructose share the same membrane transport components.     

Phylogeographic differentiation in the mitochondrial control region in the koala, Phascolarctos cinereus (Goldfuss 1817)

The koala, Phascolarctos cinereus, is a geographically widespread species endemic to Australia, with three currently recognized subspecies: P.c. adustus, P.c. cinereus, and P.c. victor. Intraspecific variation in the mitochondrial DNA (mtDNA) control region was examined in over 200 animals from 16 representative populations throughout the species’ range. Eighteen different haplotypes were defined in the ≈ 860 bp mtDNA control region, as determined by heteroduplex analysis/temperature gradient gel electrophoresis (HDA/TGGE). Any single population typically possessed only one or two haplotypes yielding an average within-population haplotypic diversity of 0.180 ± 0.003, and nucleotide diversity of 0.16%. Overall, mtDNA control region sequence diversity between populations averaged 0.67%, and ranged from 0% to 1.56%. Nucleotide divergence between populations averaged 0.51%, and ranged from 0% to 1.53%. Neighbour-joining methods revealed limited phylogenetic distinction between geographically distant populations of koalas, and tentative support for a single evolutionarily significant unit (ESU). This is consistent with previous suggestions that the morphological differences formalized by subspecific taxonomy may be interpreted as clinal variation. Significant differentiation in mtDNA-haplotype frequencies between localities suggested that little gene flow currently exists among populations. When combined with microsatellite analysis, which has revealed substantial differentiation among koala populations, we conclude that the appropriate short-term management unit (MU) for koalas is the local population.     

DYNAMICS OF A HYBRID ZONE IN PHLOX: AN EXPERIMENTAL DEMOGRAPHIC INVESTIGATION

The dynamics of a narrow hybrid zone between Phlox drummondii ssp. drummondii and ssp. mcallisteri was investigated using an experimental demographic approach. Three populations of each subspecies as well as those from the hybrid zone were involved. Experimental plots containing seeds from each of the nine populations were established in each of the eight sites. Germination, survivorship to reproduction, and fecundity were monitored at 4–6 wk intervals. Finite rates of increase were estimated from these data. There were significant site effects for each demographic variable and rate of increase as well as significant population effects for germination and finite rate of increase. Although there were population effects, no significant differences were found between plant types in sites of the subspecies or the hybrid zone. We reject the hypothesis that the narrow width of the hybrid zone is the result of hybrid disadvantage. We propose that the narrow width is a result of restricted gene flow and discuss the supporting evidence.     

Gene flow across secondary contact zones of the Emys orbicularis complex in the Western Mediterranean and evidence for extinction and re‐introduction of pond turtles on Corsica and Sardinia (Testudines: Emydidae)

European pond turtles represent a phylogeographically deeply structured complex of distinct taxa. Here, we use mitochondrial DNA sequences (cytochrome b gene) and eight polymorphic microsatellite loci to investigate genetic differentiation and gene flow of Sicilian, Corsican and Sardinian pond turtles and of subspecies involved in two secondary contact zones in the Pyrenean region and Southern Italy. Mitochondrial and microsatellite differentiation is largely concordant in populations from the core regions of the distribution ranges of the studied taxa. Both marker systems provide no evidence for gene flow between Sicilian pond turtles (Emys trinacris) and Southern Italian subspecies of E. orbicularis. By contrast, in the contact zones limited gene flow occurs between distinct subspecies of E. orbicularis. Although the Southern Italian contact zone is significantly older than the Pyrenean contact zone of Holocene age, patterns of asymmetric introgression are similar. Introgressive hybridization leads to the exchange of mitochondria, but microsatellite data indicate only a few individuals with mixed ancestry. This suggests that incipient isolating mechanisms maintain largely discrete nuclear genomic gene pools. Furthermore, this implies that Southern Italy acted as a hotspot rather than as a melting pot of genetic diversity during the last glacial. Pond turtles from Corsica and Sardinia are not differentiated from continental populations of the subspecies E. o. galloitalica, neither in the mitochondrial nor in the quickly evolving microsatellite markers. As the fossil record argues for a continuous presence of pond turtles on both islands since the Middle Pleistocene, this suggests that the native island populations became extinct and the extant turtles were later introduced by prehistoric settlers. The lack of genetic differentiation of pond turtles from Corsica and Sardinia supports the view that the subspecies described from these islands are not valid.     

Improvement of the catharanthine productivity in hairy root cultures ofCatharanthus roseus by using monosaccharides as a carbon source

Summary Sucrose, glucose, and fructose as carbon sources in culture medium were assessed in hairy root cultures ofCatharanthus roseus. The cultures preferentially consumed sucrose, resulting in about 40% (dry wt) higher growth rate. However, fructose enhanced the cathranthine yield about two-fold. The elevated yield was not seemingly ascribed to the higher osmolarity per unit weight of fructose than sucrose. A two stage culture using sucrose (1st) and fructose (2nd) improved volumetric yields of catharanthine about two-fold, i.e. 41 mg/l.     

Genetic relationships of hellbenders in the Ozark highlands of Missouri and conservation implications for the Ozark subspecies (<Emphasis Type="Italic">Cryptobranchus alleganiensis bishopi</Emphasis>)

The hellbender (Cryptobranchus alleganiensis) is an obligately aquatic salamander that is in decline due to habitat loss and disease. Two subspecies of hellbender have been described based on morphological characteristics: C. a. alleganiensis (eastern subspecies) and C. a. bishopi (Ozark hellbender). Current conservation strategies include captive propagation for restorative releases even though information regarding the current levels of genetic variability and structure within populations is not sufficient to effectively plan for conservation of the genetic diversity of the species. To investigate patterns of population structure in the hellbender, we genotyped 276 hellbenders from eight Missouri River drainages, representing both subspecies. Our results showed low levels of within-drainage diversity but strong population structure among rivers, and three distinct genetic clusters. F _ST values ranged from 0.00 to 0.61 and averaged 0.40. Our results confirmed previous reports that C. a. bishopi and C. a. alleganiensis are genetically distinct, but also revealed an equidistant relationship between two groups within C. a. bishopi and all populations of C. a. alleganiensis. Current subspecies delineations do not accurately incorporate genetic structure, and for conservation purposes, these three groups should be considered evolutionarily significant units.     

THE SUGARS OF SOME FLORIDA NECTARS

The quantitative composition of major sugars in the nectars of 35 plant species was compared with that of 12 fruit juices. The use of specific quantitative assays for glucose, fructose, and sucrose made it possible to analyze large samples without quantitative chromatographic separations. Sucrose ranged from 0–99%. In most nectars, glucose and fructose were present in approximately equal amounts. No evidence was found for the presence of maltose, raffinose, and other oligosaccharides.     

Fructose production from sucrose and high fructose syrup: A mycelial fungal system

Summary The use of Mucor sp. M105 and Fusarium sp. F5 in the production of fructose from sugarcane sucrose and high fructose syrup (HFS) was investigated. Although Mucor sp. could not utilize sucrose as the sole carbon and energy source for cell growth, Mucor sp. preferentially utilized glucose in a glucose:fructose (1:1) mixture during fermentation to ethanol. In contrast, Fusarium sp. utilized sucrose as sole carbon source by secretion of extracellular hydrolytic enzymes that degraded the disaccharide. In Fusarium sp., glucose formation in the medium was faster than fructose. Due to the low consumption rate of fructose, this substrate remained in the fermentation broth. The application of these biological systems for the production of fructose from either sucrose or HFS is discussed.     

Carbon catabolite repression of invertase during batch cultivations of Saccharomyces cerevisiae: the role of glucose, fructose, and mannose

When Saccharomyces cerevisiae are grown on a mixture of glucose and another fermentable sugar such as sucrose, maltose or galactose, the metabolism is diauxic, i.e. glucose is metabolized first, whereas the other sugars are metabolized when glucose is exhausted. This phenomenon is a consequence of glucose repression, or more generally, catabolite repression. Besides glucose, the hexoses fructose and mannose are generally also believed to trigger catabolite repression. In this study, batch fermentations of S. cerevisiae in mixtures of sucrose and either glucose, fructose or mannose were performed. It was found that the utilization of sucrose is inhibited by concentrations of either glucose or fructose higher than 5 g/l, and thus that glucose and fructose are equally capable of exerting catabolite repression. However, sucrose was found to be hydrolyzed to glucose and fructose, even when the mannose concentration was as high as 17 g/l, indicating, that mannose is not a repressing sugar. It is suggested that the capability to trigger catabolite repression is connected to hexokinase PII, which is involved in the in vivo phosphorylation of glucose and fructose. Received: 5 May 1998 / Received revision: 3 August 1998 / Accepted: 8 August 1998     

Clinal genetic variation and the 'rare allele phenomenon' in random mating populations of <Emphasis Type="Italic">Urophora cardui</Emphasis> (Diptera: Tephritidae)

In the present study we investigate a contact zone between two population groups of the tephritid fly Urophora cardui. We investigate scenarios that may have produced the genetic differentiation of the two groups, and we describe the 'rare allele phenomenon' from the contact zone. The rare allele phenomenon refers to alleles that are found at high frequency in contact zones but are rare or lacking outside the contact zone. The phenomenon is often observed in hybrid zones between subspecies of limited reproductive compatibility, but seldom in populations with random mating. Clinal genetic variation was observed at three loci in the contact zone. Three alleles at the locus Aat showed steep clines, between 20--70 km wide. A rare Aat-A allele occurred at high frequency in the centre of the contact zone. Two further loci, Hk and Pgd, showed less steep clinal genetic variation, the transition being in and slightly south of the centre of the Aat cline. Populations showed Hardy--Weinberg proportions and there was no evidence for linkage dis-equlibrium. These findings suggest random mating and gradual introgression between the population systems, which may originate from at least two range expansions. Aat's steep clines and rare allele may indicate selection on Aat alleles, although we presently can not quantify any agents. Because U. cardui experiences random mating in the contact zone with no apparent 'hybrid' incompatibility, mating experiments offer the possibility for future enquiries about the genetic basis of the rare allele phenomenon.     

Seasonal Dynamic of Nonstructural Saccharides in a Rhizomatous Grass Calamagrostis Epigeios

Seasonal dynamic of total nonstructural saccharides (TNS) and individual saccharides (starch, sucrose, glucose, fructose, fructans) was followed in rhizomes and stem bases of Calamagrostis epigeios (L.) Roth at two types of meadows communities in the South Moravia (Czech Republic): cnidion and molinion alliances, which differ in their water regime. The TNS were formed mainly by fructans and starch, while glucose, sucrose and fructose were low. The amount of TNS in rhizomes and stem bases of plants from wet cnidion site was higher than in plants from drier molinion site. The seasonal trends of all saccharides were similar in the both sites. During growing season (June to October) the main storage sugar was fructan (18 – 21 % of dry biomass). At the beginning of September the content of fructan decreased to 10 – 12 % and simultaneously the content of sucrose increased from 1 to 3 %. This may increase frost resistance. The content of TNS in the stem bases was lower than in the rhizomes. During winter time the stem bases contained 2 to 2.5 % of sucrose. Plant height and aboveground biomass were also higher in molinion site.     

A hybrid zone between two subspecies of Chorthippus parallelus. X-chromosome variation through a contact zone

Two grasshopper subspecies, Chorthippus parallelus parallelus (Cpp) and Chorthippus parallelus erythropus (Cpe), meet along the Pyrenees where they hybridize and produce a hybrid zone. A contact zone located in the Col de Portalet has been analyzed for the distribution of chromosome markers on the sex (X) chromosome in pure and hybrid populations. C-banding allowed us to distinguish both pure subspecific forms and recombinant forms, and to demonstrate their different frequencies through the contact zone. Interestingly, the distal C-band (P) on the X chromosome that characterizes Cpp individuals occurs at very high frequency through the zone and then drops sharply, close to where pure Cpe populations are found. A novel interstitial C-band (H) has been found, probably derived from that characterizing Cpe individuals (E). This marker band is only present in hybrid populations. These data are discussed in the light of the dynamics of the hybrid zone.     

Subspecific relationships and genetic structure in the spotted owl

Hierarchical genetic structure was examined in the three geographically-defined subspecies of spotted owl (Strix occidentalis) to define relationships among subspecies and quantify variation within and among regional and local populations. Sequences (522 bp) from domains I and II of the mitochondrial control region were analyzed for 213 individuals from 30 local breeding areas. Results confirmed significant differences between northern spotted owls and the other traditional geographically defined subspecies but did not provide support for subspecific level differences between California and Mexican spotted owls. Divergence times among subspecies estimated with a 936 bp portion of the cytochrome b gene dated Northern and California/Mexican spotted owl divergence time to 115,000–125,000 years ago, whereas California/Mexican spotted owl divergence was estimated at 15,000 years ago. Nested clade analyses indicated an association between California spotted owl and Mexican spotted owl haplotypes, implying historical contact between the two groups. Results also identified a number of individuals geographically classified as northern spotted owls (S. o. caurina) that contained haplotypes identified as California spotted owls (S. o. caurina). Among all northern spotted owls sampled (n=131), 12.9% contained California spotted owl haplotypes. In the Klamath region, which is the contact zone between the two subspecies, 20.3% (n=59) of owls were classified as California spotted owls. The Klamath region is a zone of hybridization and speciation for many other taxa as well. Analyses of population structure indicated gene flow among regions within geographically defined subspecies although there was significant differentiation among northern and southern regions of Mexican spotted owls. Among all areas examined, genetic diversity was not significantly reduced except in California spotted owls where the southern region consists of one haplotype. Our results indicate a stable contact zone between northern and California spotted owls, maintaining distinct subspecific haplotypes within their traditional ranges. This supports recovery efforts based on the traditional subspecies designation for the northern spotted owl. Further, although little variation was found between California and Mexican spotted owls, we suggest they should be managed separately because of current isolation between groups.     

Floral traits and mating systems in sister species of <Emphasis Type="Italic">Nicotiana</Emphasis>: interpopulational variability and sympatry effects

Mating systems of Angiosperms are important determinants of population genetic structure and evolutionary potential. Nicotiana longiflora and N. plumbaginifolia are self-compatible, sister species, with contrasting floral morphology and can be found in allopatry and sympatry in North Argentina. In two sympatric and 10 nearby allopatric populations we studied their natural interpopulational variability and sympatry effects on corolla length and anther–stigma distance. We also estimated seed set by selfing and via pollinators using pollination treatments. Both corolla length and anther–stigma distance varied significantly among N. longiflora, but not among N. plumbaginifolia populations. We did not detect an effect of sympatry in either species for any of the floral traits studied. Pollination treatments suggest that N. longiflora is mainly an outcrosser, although selfing occurs at some extent. Seed set attributed to pollinators was significantly higher in N. longiflora whereas most seeds in N. plumbaginifolia were sired through self-pollination. In N. plumbaginifolia, selfing seems to assure reproduction in sympatric populations, where floral visitors have a strong preference for N. longiflora. Corolla length was significantly negatively correlated with an increase in the percentage of self-seeds estimated by pollination treatments. Within N. longiflora, anther–stigma distance showed a positive correlation with selfing.