The composition of external lipids from adult whiteflies,Bemisia tabaci and Trialeurodes vaporariorum

The total surface lipids, including the wax particles, of the adult whiteflies of Bemisia tabaci and Trialeurodes vaporariorum were characterized. At eclosion, there were similar amounts of long-chain hydrocarbons, aldehydes, alcohols and wax esters. Within a few hours post-eclosion, long-chain aldehydes and long-chain alcohols were the dominant surface lipid components, C₃₄ on B. tabaci and C₃₂ on T. vaporariorum. Hydrocarbons, mainly n-alkanes, were minor components of the surface lipids. The major wax esters were C₄₆ on B. tabaci and C₄₂ on T. vaporariorum. The major acid and alcohol moieties in the wax esters of B. tabaci were C₂₀ and C₂₆, respectively, and of T. vaporariorum were C₂₀ and C₂₂, respectively. Both B. tabaci and T. vaporariorum had a minor wax ester composed of the fatty acid C_18:1 esterified to the major alcohols, C₃₄ and C₃₂, respectively. Bemisia were readily distinguished from Trialeurodes based on the composition of their wax particles and/or their wax esters; however, no differentiating surface lipid components were detected between biotypes A and B of B. tabaci.     

Loss of the Transit Peptide and an Increase in Gene Expression of an Ancestral Chloroplastic Carbonic Anhydrase Were Instrumental in the Evolution of the Cytosolic C4 Carbonic Anhydrase in Flaveria

C₄ photosynthesis has evolved multiple times from ancestral C₃ species. Carbonic anhydrase (CA) catalyzes the reversible hydration of CO₂ and is involved in both C₃ and C₄ photosynthesis; however, its roles and the intercellular and intracellular locations of the majority of its activity differ between C₃ and C₄ plants. To understand the molecular changes underlying the evolution of the C₄ pathway, three cDNAs encoding distinct β-CAs (CA1, CA2, and CA3) were isolated from the leaves of the C₃ plant Flaveria pringlei. The phylogenetic relationship of the F. pringlei proteins with other embryophyte β-CAs was reconstructed. Gene expression and protein localization patterns showed that CA1 and CA3 demonstrate high expression in leaves and their products localize to the chloroplast, while CA2 expression is low in all organs examined and encodes a cytosolic enzyme. The roles of the F. pringlei enzymes were considered in light of these results, other angiosperm β-CAs, and Arabidopsis (Arabidopsis thaliana) “omics” data. All three F. pringlei CAs have orthologs in the closely related C₄ plant Flaveria bidentis, and comparisons of ortholog sequences, expression patterns, and intracellular locations of their products indicated that CA1 and CA2 have maintained their ancestral role in C₄ plants, whereas modifications to the C₃ CA3 gene led to the evolution of the CA isoform that catalyzes the first step in the C₄ photosynthetic pathway. These changes included the loss of the chloroplast transit peptide and an increase in gene expression, which resulted in the high levels of CA activity seen in the cytosol of C₄ mesophyll cells.     

Role of fatty acids in cold adaptation of Antarctic psychrophilic Flavobacterium spp.

Cold-loving microorganisms developed numerous adaptation mechanisms allowing them to survive in extremely cold habitats, such as adaptation of the cell membrane. The focus of this study was on the membrane fatty acids of Antarctic Flavobacterium spp., and their adaptation response to cold-stress. Fatty acids and cold-response of Antarctic flavobacteria was also compared to mesophilic and thermophilic members of the genus Flavobacterium. The results showed that the psychrophiles produced more types of major fatty acids than meso- and thermophilic members of this genus, namely C_15:1 iso G, C_15:0 iso, C_15:0 anteiso, C_15:1 ω6c, C_15:0 iso 3OH, C_17:1 ω6c, C_16:0 iso 3OH and C_17:0 iso 3OH, summed features 3 (C_16:1 ω7cand/or C_16:1 ω6c) and 9 (C_16:0 10-methyl and/or C_17:1 iso ω9c). It was shown that the cell membrane of psychrophiles was composed mainly of branched and unsaturated fatty acids. The results also implied that Antarctic flavobacteria mainly used two mechanisms of membrane fluidity alteration in their cold-adaptive response. The first mechanism was based on unsaturation of fatty acids, and the second mechanism on de novo synthesis of branched fatty acids. The alteration of the cell membrane was shown to be similar for all thermotypes of members of the genus Flavobacterium.     

Differences in drought sensitivities and photosynthetic limitations between co-occurring C3 and C4 (NADP-ME) Panicoid grasses

Background and Aims

The success of C₄ plants lies in their ability to attain greater efficiencies of light, water and nitrogen use under high temperature, providing an advantage in arid, hot environments. However, C₄ grasses are not necessarily less sensitive to drought than C₃ grasses and are proposed to respond with greater metabolic limitations, while the C₃ response is predominantly stomatal. The aims of this study were to compare the drought and recovery responses of co-occurring C₃ and C₄ NADP-ME grasses from the subfamily Panicoideae and to determine stomatal and metabolic contributions to the observed response.

Methods

Six species of locally co-occurring grasses, C₃ species Alloteropsis semialata subsp. eckloniana, Panicum aequinerve and Panicum ecklonii, and C₄ (NADP-ME) species Heteropogon contortus, Themeda triandra and Tristachya leucothrix, were established in pots then subjected to a controlled drought followed by re-watering. Water potentials, leaf gas exchange and the response of photosynthetic rate to internal CO₂ concentrations were determined on selected occasions during the drought and re-watering treatments and compared between species and photosynthetic types.

Key Results

Leaves of C₄ species of grasses maintained their photosynthetic advantage until water deficits became severe, but lost their water-use advantage even under conditions of mild drought. Declining C₄ photosynthesis with water deficit was mainly a consequence of metabolic limitations to CO₂ assimilation, whereas, in the C₃ species, stomatal limitations had a prevailing role in the drought-induced decrease in photosynthesis. The drought-sensitive metabolism of the C₄ plants could explain the observed slower recovery of photosynthesis on re-watering, in comparison with C₃ plants which recovered a greater proportion of photosynthesis through increased stomatal conductance.

Conclusions

Within the Panicoid grasses, C₄ (NADP-ME) species are metabolically more sensitive to drought than C₃ species and recover more slowly from drought.     

Microbial Oxidation of Hydrocarbons and Related Compounds by Whole-Cell Suspensions of the Methane-Oxidizing Bacterium H-2

Previously, a thermophilic obligate methane-oxidizing bacterium, H-2 (type I), was isolated in our laboratory. H-2 is a new type of methylotroph because of the G+C content of DNA; it uses both the ribulose monophosphate pathway and the serine pathway for carbon assimilation and possesses a new quinone. In addition, we found that resting cell suspensions of H-2 had the ability to oxidize a variety of compounds different from the other methane-oxidizing bacteria as follows. (i) C₁ to C₈n-alkanes are hydroxylated and further oxidized, yielding mixtures of the corresponding alcohols, aldehydes, acids, and ketones. Liquid alkanes are transformed through a different oxidative pathway from that of gaseous ones. (ii) Both gaseous (C₂ to C₄) and liquid (C₅, C₆) n-alkenes are oxidized to their corresponding 1,2-epoxides. (iii) Liquid monochloro and dichloro n-alkanes (C₅, C₆) are oxidized, yielding their corresponding acids or haloacids. (iv) Diethyl ether is oxidized to acetic acid; no ethanol and acetaldehyde are detected. (v) Cyclic and aromatic compounds are also oxidized. (vi) Secondary alcohols (C₃ to C₁₀) are oxidized to their corresponding methyl ketones.     

Surface wax of Andreaea and Pogonatum species

The mosses Andreaea rupestris, Pogonatum aloides and P. urnigerum contain surface waxes in amounts of 0.05–0.12% dry wt. The waxes consisted of esters (C₃₈-C₅₄), primary alcohols (C₂₀-C₃₂), free fatty acids (C₁₆-C₃₀), and alkanes (C₂₁-C₃₁). Additionally, aldehydes (C₂₂-C₃₀) were major constituents in the wax of P. urnigerum. The classes and their chain length distributions in the surface waxes of these mosses are comparable to those of epicuticular waxes of higher plants.     

C3 and C4 leaf anatomy types in Camphorosmeae (Camphorosmoideae,Chenopodiaceae)

Complementary to our previous project on the molecular phylogeny of Camphorosmeae, the leaf anatomy of ca. 35 species including all non-Australian and selected Australian species was studied by use of light microscopy. Nine anatomical leaf types were described, compared to previous classifications, and discussed with regard to their putative evolution on the background of phylogenetic trees. Particular emphasis was given to the relationships between the C₃ and C₄ leaf types: Chenolea type (C₃), Eokochia type (C₃), Neokochia type (C₃), Sedobassia type (C₃/C₄ intermediate), Bassia prostrata type (C₄), B. muricata type (C₄), B. eriantha type, B. lasiantha type (C₄), Camphorosma type (C₄). The main results and conclusions were: (1) Two unusual new C₃ leaf types: Chenolea with microfenestrate chlorenchyma, Eokochia with unique complex vascular bundles; (2) Sedobassia interpreted as anatomically C₃/C₄ intermediate by kranz-like bundle sheath cells is the first C₃/C₄ intermediate in Camphorosmeae and found in a derived position; (3) Neokochia type detected as the likely starting point for all four C₄ leaf types and for the C₃/C₄ intermediate; (4) hypodermis of C₄ types originated from outermost chlorenchyma layer of C₃ types and lost multiple times during further evolution; (5) atriplicoid Bassia. lasiantha type without water storage tissue evolved from kochioid B. muricata type; (6) two independent gains of C₄ photosynthesis, one in Bassia and one in Camphorosma; (7) depending on the lineage, leaf architecture remains comparatively stable (Australian Camphorosmeae) or shows an unexpected plasticity (Bassia scoparia group).     

Transfer of C(4) Photosynthetic Characters through Hybridization of Flaveria Species

Transfer of C₄ photosynthetic traits was studied through hybridization of Flaveria trinervia (Spreng.) Mohr (C₄) and Flaveria brownii A.M. Powell (C₄-like) with Flaveria linearis Lag. (C₃-C₄) and the C₃ species Flaveria pringlei Gandoger (C₃). Fertility was low, based on irregular chromosome pairing and low pollen stainability, except in F. brownii × F. linearis which had bivalent pairing and 76% stainable pollen. Hybrids had apparent photosynthesis values of 71 to 148% of the midparental means, while the CO₂ compensation concentration was similar to the C₄ or C₄-like parent, except in hybrids having the C₃ species F. pringlei as a parent. Inhibition of apparent photosynthesis by O₂, and phosphoenolpyruvate carboxylase and NADP-malic enzyme activities and subunit levels in the hybrids were closer to the C₃ or C₃-C₄ parent. The species F. brownii and F. trinervia were equal in their capacity to transfer reduced O₂ inhibition of AP and CO₂ compensation concentration values to hybrids with F. linearis (C₃-C₄), although hybrids with F. trinervia had higher PEPC activity. The O₂ inhibition of AP was correlated with the logarithm of activities of phosphoenolpyruvate carboxylase (r = −0.95) and NADP-malic enzyme (r = −0.87). These results confirm that C₄ traits can be transferred by hybridization of C₃-C₄ and C₄ or C₄-like species, with a higher degree of C₄ photosynthesis than exists in C₃-C₄ species, and at least in F. brownii × F. linearis, fertile progeny are obtained.     

Getting the most out of natural variation in C4 photosynthesis

C₄ photosynthesis is a complex trait that has a high degree of natural variation, involving anatomical and biochemical changes relative to the ancestral C₃ state. It has evolved at least 66 times across a variety of lineages and the evolutionary route from C₃ to C₄ is likely conserved but not necessarily genetically identical. As such, a variety of C₄ species are needed to identify what is fundamental to the C₄ evolutionary process in a global context. In order to identify the genetic components of C₄ form and function, a number of species are used as genetic models. These include Zea mays (maize), Sorghum bicolor (sorghum), Setaria viridis (Setaria), Flaveria bidentis, and Cleome gynandra. Each of these species has different benefits and challenges associated with its use as a model organism. Here, we propose that RNA profiling of a large sampling of C₄, C₃–C₄, and C₃ species, from as many lineages as possible, will allow identification of candidate genes necessary and sufficient to confer C₄ anatomy and/or biochemistry. Furthermore, C₄ model species will play a critical role in the functional characterization of these candidate genes and identification of their regulatory elements, by providing a platform for transformation and through the use of gene expression profiles in mesophyll and bundle sheath cells and along the leaf developmental gradient. Efforts should be made to sequence the genomes of F. bidentis and C. gynandra and to develop congeneric C₃ species as genetic models for comparative studies. In combination, such resources would facilitate discovery of common and unique C₄ regulatory mechanisms across genera.     

Photosynthetic and photorespiratory characteristics of flaveria species

The genus Flaveria shows evidence of evolution in the mechanism of photosynthesis as its 21 species include C₃, C₃-C₄, C₄-like, and C₄ plants. In this study, several physiological and biochemical parameters of photosynthesis and photorespiration were measured in 18 Flaveria species representing all the photosynthetic types. The 10 species classified as C₃-C₄ intermediates showed an inverse continuum in level of photorespiration and development of the C₄ syndrome. This ranges from F. sonorensis with relatively high apparent photorespiration and lacking C₄ photosynthesis to F. Among the intermediates, the photosynthetic CO₂ compensation points at 30°C and 1150 micromoles quanta per square meter per second varied from 9 to 29 microbars. The values for the three C₄-like species varied from 3 to 6 microbars, similar to those measured for the C₄ species. The activities of the photorespiratory enzymes glycolate oxidase, hydroxypyruvate reductase, and serine hydroxymethyltransferase decreased progressively from C₃ to C₃-C₄ to C₄-like and C₄ species. On the other hand, most intermediates had higher levels of phosphenolpyruvate carboxylase and NADP-malic enzyme than C₃ species, but generally lower activities compared to C₄-like and C₄ species. The levels of these C₄ enzymes are correlated with the degree of C₄ photosynthesis, based on the initial products of photosynthesis. Another indication of development of the C₄ syndrome in C₃-C₄ Flaveria species was their intermediate chlorophyll a/b ratios. The chlorophyll a/b ratios of the various Flaveria species are highly correlated with the degree of C₄ photosynthesis suggesting that the photochemical machinery is progressively altered during evolution in order to meet the specific energy requirements for operating the C₄ pathway. In the progression from C₃ to C₄ species in Flaveria, the CO₂ compensation point decreased more rapidly than did the decrease in O₂ inhibition of photosynthesis or the increase in the degree of C₄ photosynthesis. These results suggest that the reduction in photorespiration during evolution occurred initially by refixation of photorespired CO₂ and prior to substantive reduction in O₂ inhibition and development of the C₄ syndrome. However, further reduction in O₂ inhibition in some intermediates and C₄-like species is considered primarily due to the development of the C₄ syndrome. Thus, the evolution of C₃-C₄ intermediate photosynthesis likely occurred in response to environmental conditions which limit the intercellular CO₂ concentration first via refixation of photorespired CO₂, followed by development of the C₄ syndrome.     

Photosynthesis, Morphology, Leaf Anatomy, and Cytogenetics of Hybrids between C(3) and C(3)/C(4)Panicum Species

The Laxa group of the Panicum genus contains species which have CO₂ exchange and anatomical characteristics intermediate to C₃ and C₄ photosynthetic types (C₃/C₄), and also species characterized as C₃. Hybrids were made between two of the C₃/C₄ species and two C₃ species. Carbon dioxide exchange and morphological, leaf anatomical, and cytogenetic characteristics of F₁ hybrids between Panicum milioides Nees. ex Trin (C₃/C₄) and P. laxum Mez. (C₃), P. spathellosum Doell (C₃/C₄) and P. boliviense Hack. (C₃), and P. spathellosum and P. laxum were studied. There were no consistent differences in apparent photosynthesis, although two of the three hybrids had higher net CO₂ uptake than the C₃ parent. Values of inhibition of apparent photosynthesis by 21% O₂, CO₂ loss in the light, and CO₂ compensation concentration for the hybrids were between those of the parents. All three hybrids showed leaf anatomical traits, especially organelle quantities in the bundle sheath cells, between those of their respective parents. Linear regression of CO₂ compensation concentration on the percentage of mitochondria and chloroplasts in vascular bundle sheaths of the parents and hybrids gave correlation coefficients of −0.94. This suggests that the reduction in CO₂ loss in the C₃/C₄ species, and to a lesser degree in the F₁ hybrids, was due to development of organelles and perhaps a higher proportion of leaf photorespiration in bundle sheaths. The overall morphology of the hybrids was so different from the parents that they could be described as new taxonomic forms. The chromosomes in the hybrids were mainly unpaired or paired as bivalents indicating possible homology between some parental genomes.     

Cell Envelope Phospholipid Composition of Burkholderia multivorans

Burkholderia multivorans causes opportunistic pulmonary infections in cystic fibrosis and immunocompromised patients. The purpose of the present study was to determine the nature of the phospholipids and their fatty acid constituents comprising the cell envelope membranes of strains isolated from three disparate sources. A conventional method for obtaining the readily extractable lipids fraction from bacteria was employed to obtain membrane lipids for thin-layer chromatographic and gas chromatography-mass spectrophotometric analyses. Major fatty acid components of the B. multivorans readily extractable lipid fractions included C_16:0 (palmitic acid), C_16:1 (palmitoleic acid), and C_18:1 (oleic acid), while C_14:0 (myristic acid), ΔC_17:0 (methylene hexadecanoic acid), C_18:0 (stearic acid), and ΔC_19:0 (methylene octadecanoic acid) were present in lesser amounts. Fatty acid composition differed quantitatively among strains with regard to C_16:0, C_16:1, ΔC_17:0, C_18:1, and ΔC_19:0 with the unsaturated:saturated fatty acid ratios being significantly less in a cystic fibrosis type strain than either environmental or chronic granulomatous disease strains. Phospholipids identified in all B. multivorans strains included lyso-phosphatidylethanolamine, phosphatidylglycerol, phosphatidylethanolamine, and diphosphatidylglycerol in similar ratios. These data support the conclusion that the cell envelope phospholipid profiles of disparate B. multivorans strains are similar, while their respective fatty acyl substituent profiles differ quantitatively under identical cultivation conditions.     

Leaf anatomy of c(3)-c(4) species as related to evolution of c(4) photosynthesis

This study was undertaken to examine the degree of Kranz anatomy development in the species intermediate to C₃ and C₄ types (C₃-C₄) in Panicum, Neurachne, Flaveria, and Moricandia. In each genus, C₃ and/or C₄ species were used for comparison. Leaf transections from each species were examined by light and transmission electron microscopy. The percentages of leaf photosynthetic cell profiles partitioned to bundle sheaths were higher in C₄ than in C₃ species, while C₃-C₄ species tended to be in between. However, percentages for C₃-C₄ species in Moricandia and some C₃-C₄Flaveria species were not greater than C₃. When expressed on a cell profile area basis, C₃-C₄ species partitioned more photosynthetic tissue to bundle sheaths than C₃ species in Moricandia, but not in Flaveria. Neurachne minor S. T. Blake (C₃-C₄) partitioned a very small portion of cell profile area to the inner bundle sheaths (5%) compared to Neurachne munroi F. Muell (C₄) (21%). The percentage of organelles partitioned to bundle sheaths was much greater in C₃-C₄ than in C₃ species. The average C₃ percentages for mitochondria plus peroxisomes were 19, 8, and 19.5% for Neurachne, Flaveria, and Moricandia, respectively, compared to 41, 29, and 46.5% for the C₃-C₄ species. The CO₂ compensation concentration was negatively related to the partitioning of tissue to bundle sheaths and to the percentage of organelles in bundle sheaths. It is concluded that all of the C₃-C₄ species examined have developed some degree of Kranz anatomy and that this altered anatomy is involved in their reduced apparent photorespiration.     

Acetate esters of saturated and unsaturated alcohols (C12–C20) are major components in Dufour glands of Bracon cephi and Bracon lissogaster (Hymenoptera: Braconidae), parasitoids of the wheat stem sawfly,Cephus cinctus (Hymenoptera: Cephidae)

Chemistry of Dufour glands associated with the venom complex in Bracon cephi (Gahan) and Bracon lissogaster Muesebeck (Hymenoptera; Braconidae), two parasitoids of the wheat stem sawfly, Cephus cinctus Norton (Hymenoptera: Cephidae), was examined by solid phase micro-extraction (SPME) and gas chromatography/mass spectrometry. Homologous series of five chemical classes were detected in individual glands from each species. Major classes included: (1) acetate esters of saturated and unsaturated primary alcohols with parent chain lengths from C₁₂ to C₂₀. Hexadecanyl acetate, octadecanyl acetate, and octadecenyl acetate were major components in B. cephi. The composition of the acetate series in B. lissogaster was similar except that the octadecanyl acetate was only a minor component. (2) A homologous series of monoenes from C_23:1 to C_35:1 were detected in both species, with C_29:1, C_31:1 and C_33:1 being the major components. Dienes from C_31:2 to C_35:2 and trienes (C_33:3–C_35:3) were also detected in both species. (3) A homologous series of n-alkanes from C₁₉ to C₃₁ was detected in both species. n-Tricosane was the major component. Minor components in both species included homologous series of both mono- and dimethyl branched alkanes. The Dufour gland hydrocarbon components in both B. cephi and B. lissogaster have some similarities to the composition of cuticular hydrocarbons of their host C. cinctus, a species with a complex pheromonal signaling system.     

Degree of C(4) Photosynthesis in C(4) and C(3)-C(4)Flaveria Species and Their Hybrids : I. CO(2) Assimilation and Metabolism and Activities of Phosphoenolpyruvate Carboxylase and NADP-Malic Enzyme

The degree of C₄ photosynthesis was assessed in four hybrids among C₄, C₄-like, and C₃-C₄ species in the genus Flaveria using ¹⁴C labeling, CO₂ exchange, ¹³C discrimination, and C₄ enzyme activities. The hybrids incorporated from 57 to 88% of the ¹⁴C assimilated in a 10-s exposure into C₄ acids compared with 26% for the C₃-C₄ species Flaveria linearis, 91% for the C₄ species Flaveria trinervia, and 87% for the C₄-like Flaveria brownii. Those plants with high percentages of ¹⁴C initially fixed into C₄ acids also metabolized the C₄ acids quickly, and the percentage of ¹⁴C in 3-phosphoglyceric acid plus sugar phosphates increased for at least a 30-s exposure to ¹²CO₂. This indicated a high degree of coordination between the carbon accumulation and reduction phases of the C₄ and C₃ cycles. Synthesis and metabolism of C₄ acids by the species and their hybrids were highly and linearly correlated with discrimination against ¹³C. The relationship of ¹³C discrimination or ¹⁴C metabolism to O₂ inhibition of photosynthesis was curvilinear, changing more rapidly at C₄-like values of ¹⁴C metabolism and ¹³C discrimination. Incorporation of initial ¹⁴C into C₄ acids showed a biphasic increase with increased activities of phosphoenolpyruvate carboxylase and NADP-malic enzyme (steep at low activities), but turnover of C₄ acids was linearly related to NADP-malic enzyme activity. Several other traits were closely related to the in vitro activity of NADP-malic enzyme but not phosphoenolpyruvate carboxylase. The data indicate that the hybrids have variable degrees of C₄ photosynthesis but that the carbon accumulation and reduction portions of the C₄ and C₃ cycles are well coordinated.     

Kinetic properties of phosphoenolpyruvate carboxylase from c(3), c(4), and c(3)-c(4) intermediate species of flaveria (asteraceae)

Flaveria cronquistii (C₃), F. chloraefolia (C₃-C₄), F. floridana (C₃-C₄), F. pubescens (C₃-C₄), F. anomala (C₃-C₄), F. linearis (C₃-C₄), F. brownii (C₄), F. palmeri (C₄), F. trinervia (C₄) and F. australasica (C₄), comprising 10 out of the 21 known species of the genus Flaveria (Asteraceae), were included in a comparative study of the kinetic and regulatory properties of green leaf phosphoenolpyruvate (PEP) carboxylase. At least three kinetically distinct enzyme-forms were identified on the basis of their affinities for PEP and the degree of allosterism with respect to this substrate. The kinetic properties of PEP carboxylase of most of the species seemingly were modified in vivo depending on the growth conditions of the plants. K_m(PEP_free)-values of the enzyme from the five C₃-C₄ intermediate species ranged from 6 micromolar (F. chloraefolia, low light-grown) to 38 micromolar (F. pubescens, high light-grown). In contrast, the K_m for PEP of PEP carboxylase from the C₃ species F. cronquistii (13 micromolar) apparently was not influenced by growth conditions. The response of the enzyme from the C₃ and C₃-C₄ species was hyperbolic in all cases. A second isoform with a lower affinity for PEP (88-100 micromolar), but also hyperbolic kinetics was found in the C₄ species F. brownii, whereas in the three other C₄ species examined a PEP carboxylase with a still lower affinity for PEP (187-221 micromolar) and sigmoidal kinetics was present. These isozyme-related kinetic data were supported by analyses of the elution behavior of the enzyme during anion-exchange chromatography on DEAE-Trisacryl M. The results are discussed with respect to the evolution of C₄ photosynthesis in the Flaveria genus.     

Rapid Biomass Quality Determination of Corn Stover Using Near-Infrared Reflectance Spectroscopy

Near-infrared reflectance spectroscopy (NIRS) has been used extensively in the forage industry for rapid measurement of forage constituents and could be useful for determining quality of biomass feedstocks at the point of delivery. In previous work, we developed an assay that partitions feedstock carbohydrates based on their availability to be converted to fermentable sugars, including non-structural carbohydrates (C _N), biochemically available carbohydrates (C _B) with an associated first-order availability rate constant (k _B), and unavailable carbohydrates (C _U ). Additional quality parameters measured included neutral detergent lignin (NDL), total available carbohydrates (C _A), and total carbohydrates (C _T). We evaluated the variability of biomass quality parameters in a set of corn stover samples and developed calibration equations for determining parameter values using NIRS. Fifty-two corn stover samples harvested in Iowa and Wisconsin in 2005 and 2006 were analyzed using a high-throughput assay for determining feedstock quality for biochemical conversion. Non-structural carbohydrates ranged from 84 to 155?g?kg^?1 dry matter (DM); C _B ranged from 354 to 557?g?kg^?1 DM; k _B ranged from 0.199 to 0.330?h^?1; C _A ranged from 463 to 699?g?kg^?1 DM, and NDL ranged from 32 to 74?g?kg^?1 DM. Significant differences (P?<?0.0001) among samples were observed for all parameters, except k _B. Near-infrared reflectance spectroscopy calibration equations were developed for C _N, C _B, C _A, C _U , C _T, and NDL. It was not possible to generate a meaningful calibration equation for k _B. There is significant variability within the corn stover population for several key quality-related carbohydrate and lignin constituents which can be predicted reliably using NIRS.     

Cometabolism of Methyl tertiary Butyl Ether and Gaseous n-Alkanes by Pseudomonas mendocina KR-1 Grown on C5 to C8 n-Alkanes

Pseudomonas mendocina KR-1 grew well on toluene, n-alkanes (C₅ to C₈), and 1° alcohols (C₂ to C₈) but not on other aromatics, gaseous n-alkanes (C₁ to C₄), isoalkanes (C₄ to C₆), 2° alcohols (C₃ to C₈), methyl tertiary butyl ether (MTBE), or tertiary butyl alcohol (TBA). Cells grown under carbon-limited conditions on n-alkanes in the presence of MTBE (42 μmol) oxidized up to 94% of the added MTBE to TBA. Less than 3% of the added MTBE was oxidized to TBA when cells were grown on either 1° alcohols, toluene, or dextrose in the presence of MTBE. Concentrated n-pentane-grown cells oxidized MTBE to TBA without a lag phase and without generating tertiary butyl formate (TBF) as an intermediate. Neither TBF nor TBA was consumed by n-pentane-grown cells, while formaldehyde, the expected C₁ product of MTBE dealkylation, was rapidly consumed. Similar K_s values for MTBE were observed for cells grown on C₅ to C₈ n-alkanes (12.95 ± 2.04 mM), suggesting that the same enzyme oxidizes MTBE in cells grown on each n-alkane. All growth-supporting n-alkanes (C₅ to C₈) inhibited MTBE oxidation by resting n-pentane-grown cells. Propane (K_i = 53 μM) and n-butane (K_i = 16 μM) also inhibited MTBE oxidation, and both gases were also consumed by cells during growth on n-pentane. Cultures grown on C₅ to C₈ n-alkanes also exhibited up to twofold-higher levels of growth in the presence of propane or n-butane, whereas no growth stimulation was observed with methane, ethane, MTBE, TBA, or formaldehyde. The results are discussed in terms of their impacts on our understanding of MTBE biodegradation and cometabolism.     

Équilibres conformationnels de glucides au niveau de liaisons σ SP-SP: Partie V. Dérivés 2,3,-O-isopropylidène de pyranosides hybrides sp en C-4. Comparaison avec leurs analogues saturés

The conformation in solution of derivatives of methyl hexopyranosides has been studied by n.m.r. The esters of methyl 2,3-O-isopropylidene-α-D-manno- and -talopyranosides as well as their 4-deoxy-4-C-methyl analog having a manno configuration exist mainly in a flattened (^4,0F) chair conformation (⁴C₁). The presence in the talo epimer of the 4-deoxy-4-C-methyl analog of the bulky methyl group on the endo side of the bicyclic system results in a skew form (³S₁). The methyl 4-deoxy-2,3-O-isopropylidene-4-C-methylene-α-D-lyxo-hexopyranosides monosubstituted at C-4′ adopt, in solution, a conformation close to ³S₁, whichever their configuration (cis or trans) at the double bond, as indicated by their allylic coupling constants.