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Studies on Claviceps parasitic on Panicum species in India.

Panicum repens and P. antidotale were found to be infected with Claviceps sp. This is the first report of ergot on P. repens. The pyrenomycete produced abundant sclerotia on the host plants. The sclerotia contained 0.71 and 0.68% alkaloids, respectively, which predominantly consisted of chanoclavine, festuclavine and agroclavine. The infected grasses were possibly mycotoxic. Submerged cultures of Claviceps strain isolated from Panicum spp. produced significant amount of chanoclavine, festuclavine and agroclavine. No pharmaceutically important alkaloid was found in sclerotia or in submerged culture.

Claviceps↗

Photosynthetic carbon metabolism in Panicum milioides, a C3-C4 intermediate species: evidence for a limited C4 dicarboxylic acid pathway of photosynthesis.

Panicum milioides, a naturally occurring species with C4-like Kranz leaf anatomy, is intermediate between C3 and C4 plants with respect to photo-respiration and the associated oxygen inhibition of photosynthesis. This paper presents direct evidence for a limited degree of C4 photosynthesis in this C3-C4 intermediate species based on: (a) the appearance of 24% of the total 14C fixed following 4 s photosynthesis in 14CO2-air by excised leaves in malate and aspartate and the complete transfer of label from the C4 acids to Calvin cycle intermediates within a 15 s chase in 12CO2-air; (b) pyruvate- or alanine-enhanced light-dependent CO2 fixation and pyruvate stimulation ote- or alanine-enhanced light-dependent CO2 fixation and pyruvate stimulation of oxaloacetate- or 3-phosphoglycerate-dependent O2 evolution by illuminated mesophyll protoplasts, but not bundle sheath strands; and (c) NAD-malic enzyme-dependent decarboxylation of C4 acids at the C-4 carboxyl position, C4 acid-dependent O2 evolution, and 14CO2 donation from (4-14C)C4 acids to Calvin cycle intermediates during photosynthesis by bundle sheath strands, but not mesophyll protoplasts. However, P. milloides differs from C4 plants in that the activity of the C4 cycle enzymes is only 15 to 30% of a C4 Panicum species and the Calvin cycle and phosphoenolpyruvate carboxylase are present in both cell types. From these and related studies (Rathnam, C.K.M. and Chollet, R. (1979) Arch. Biochem. Biophys. 193, 346-354; (1978) Biochem. Biophys. Res. Commun. 85, 801-808) we conclude that reduced photorespiration in P. milioides is due to a limited degree of NAD-malic enzyme-type C4 photosynthesis permitting an increase in pCO2 at the site of bundle sheath, but not mesophyll, ribulose-bisphosphate carboxylase-oxygenase.

Aspartic Acid↗

Characterization of Phosphoenolpyruvate Carboxykinase from Panicum maximum.

Phosphoenolpyruvate carboxykinase, EC 4.1.1.32 (PEPCK), was purified 43-fold from the grass Panicum maximum. Michaelis constants (Km) were determined for the exchange reaction, the carboxylation reaction, and the decarboxylation reaction. The Km values for oxaloacetate and ATP in the decarboxylation reaction were found to be lower than the Km values for the substrates used in the exchange reaction and in the carboxylation reaction. Phosphoenolpyruvate carboxylase was not detectable in the purified PEPCK preparation.Studies on the nucleotide specificity of the oxaloacetate decarboxylation reaction indicate that ATP serves as the best nucleotide for this reaction and that ADP is about 60% as effective as ATP. The pH optimum for decarboxylase activity is near 6.8. The decarboxylation reaction has a divalent cation requirement with both Mn(2+) and Mg(2+) needed for full activity.Temperature curves of the three PEPCK reactions indicate optimum activities between 38 and 45 C. There is a pronounced drop in the decarboxylation and carboxylation activities as the temperature is decreased from these optima. Below 30 C the energy of activation was 8.2 kcal/mol for the decarboxylation reaction.These studies are consistent with the proposal that under physiological conditions PEPCK catalyzes the decarboxylation of oxaloacetate in the bundle sheath cells of Panicum maximum leaves during C(4) dicarboxylic acid photosynthesis.

Journal Article↗

Photosynthesis in Grass Species Differing in Carbon Dioxide Fixation Pathways: III. OXYGEN RESPONSE AND ENZYME ACTIVITIES OF SPECIES IN THE LAXA GROUP OF PANICUM.

Measurements of CO(2) exchange at varying O(2) concentrations in seven grass species of the Laxa group of Panicum and activities of five photosynthetic enzymes were compared to values obtained for these characters in a cool season C(3) grass, tall fescue (Festuca arundinacea Schreb.) and a C(4) grass, P. maximum Jacq. Plants were divided into three groups on the basis of the inhibition of apparent photosynthesis by 21% O(2.) Rates of apparent photosynthesis in P. prionitis Griseb. and P. maximum were virtually unaffected by changes in O(2) concentration. In another group consisting of P. hylaeicum Mez., P. rivulare Trin., P. laxum Sw., and tall fescue apparent photosynthesis was inhibited by 28.2 to 36.0% at 21% O(2.) An intermediate inhibition of 20.6 to 23.3% at 21% O(2) was exhibited by P. milioides Nees ex Trin., P. schenckii Hack., and P. decipiens Nees ex Trin. The CO(2) compensation concentration for P. prionitis and P. maximum was low (</=6 microliters per liter CO(2) at 21% O(2)) and affected little by O(2), whereas values for P. hylaeicum, P. rivulare, P. laxum, and tall fescue were much greater, and increased almost linearly from 2 to 48% O(2.) Values for P. milioides, P. schenckii, and P. decipiens were intermediate to the other two groups. The effect of O(2) on total leaf conductance to CO(2) was similar to the C(3) grasses and the intermediate Panicums. However, estimates of photorespiration in the intermediate species were low and changed little with O(2) in comparison to estimates for the C(3) species which were higher and increased greatly with increased O(2.)Activities of phosphoenolpyruvate carboxylase were greatest in P. maximum and P. prionitis and one-fourth or less in the remaining species. Activity of ribulose bisphosphate carboxylase was 548 micromoles per mg chlorophyll per hour in tall fescue; activity in the remaining species was approximately one-fourth or less of that in tall fescue, with the exception of P. rivulare, in which it was 440 micromoles per milligram chlorphyll per hour. High activities of two C(4) decarboxylating enzymes, phosphoenolpyruvate carboxykinase and NADP-malic enzyme, were observed in P. maximum (1,988 micromoles per milligram chlorophyll per hour) and P. prionitis (125 micromoles per milligram chlorophyll per hour), respectively. Only minimal activities of decarboxylating enzymes were detected in the remaining species.

Journal Article↗

Reduced Apparent Photorespiration by the C(3)-C(4) Intermediate Species, Moricandia arvensis and Panicum milioides.

The CO(2)/O(2) specificity factor of sucrose gradient purified ribulose 1,5-bisphosphate carboxylase/oxygenase from the C(3)-C(4) intermediate plants Moricandia arvensis (79 +/- 1) and Panicum milioides (89 +/- 2) was similar to the respective values of the enzyme from the closely related C(3) species, Moricandia foetida (80 +/- 5) and Panicum laxum (86 +/- 2). Thus, the kinetic properties of this bifunctional enzyme do not explain the reduced rates of photorespiration exhibited by either of these intermediate species.Dark/light ratios for aminoacetonitrile-sensitive (14)CO(2) evolution during decarboxylation of exogenous [1-(14)C]glycine by leaf discs had values of 9.0 with M. arvensis and 11.8 with P. milioides. Equivalent ratios with M. foetida and P. laxum were 2.5 and 3.2, respectively. Similar results were obtained using [1-(14)C]glycolate as the exogenous photorespiratory substrate, with dark/light (14)CO(2) evolution ratios for the C(3)-C(4) and C(3) leaf discs averaging 6.6 and 2.0, respectively. Stimulating photosynthetic CO(2) fixation by progressively increasing photon flux density from 0 to 1900 micromoles per square meter per second caused a concomitant reduction in (14)CO(2) evolution from leaf discs of M. arvensis and P. milioides supplied with [1-(14)C]glycine. Conversely, inhibition of photosynthesis by DCMU or the Calvin cycle inhibitor dl-glyceraldehyde increased (14)CO(2) evolution in the light to rates comparable to those in the dark. The data suggest that P. milioides and M. arvensis are capable of a more efficient internal recycling of photorespiratory CO(2) via ribulose bisphosphate carboxylase/oxygenase than closely related C(3) plants, and that this may partially account for the reduced rates of apparent photorespiration by these intermediate species.

Journal Article↗

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 CO(2) exchange and anatomical characteristics intermediate to C(3) and C(4) photosynthetic types (C(3)/C(4)), and also species characterized as C(3). Hybrids were made between two of the C(3)/C(4) species and two C(3) species. Carbon dioxide exchange and morphological, leaf anatomical, and cytogenetic characteristics of F(1) hybrids between Panicum milioides Nees. ex Trin (C(3)/C(4)) and P. laxum Mez. (C(3)), P. spathellosum Doell (C(3)/C(4)) and P. boliviense Hack. (C(3)), 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 CO(2) uptake than the C(3) parent. Values of inhibition of apparent photosynthesis by 21% O(2), CO(2) loss in the light, and CO(2) 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 CO(2) 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 CO(2) loss in the C(3)/C(4) species, and to a lesser degree in the F(1) 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.

Journal Article↗

Compensation point and isotopic characteristics of c(3)/c(4) intermediates and hybrids in panicum.

Leaf CO(2) compensation points and stable hydrogen, oxygen and carbon isotope ratios were determined for Panicum species including C(3)/C(4) intermediate photosynthesis plants, hybrids between C(3)/C(4) intermediates and C(3) plants, C(3) and C(4) plants in the Panicum genus as well as several other C(3) and C(4) plants. C(3) plants had the highest compensation points, followed by hybrids, C(3)/C(4) intermediates, and C(4) plants. delta(13)C values of cellulose nitrate and saponifiable lipids from C(4) plants were about 10 per thousand higher than those observed for cellulose nitrate and saponifiable lipids of C(3)/C(4) intermediates, hybrids, and C(3) plants. Oxygen isotope ratios of cellulose as well as those of leaf water were similar for all plants. There was substantial variability in the deltaD values of cellulose nitrate among the plants studied. In contrast, such variability was not observed in deltaD values of water distilled from the leaves, nor in the deltaD values of the saponifiable lipids. Variability in deltaD values of cellulose nitrate from C(3)/C(4) intermediates, hybrids, C(3), and C(4) plants is due to fractionations occurring during biochemical reactions specific to leaf carbohydrate metabolism.

Journal Article↗

A gene cluster encoded by panicum mosaic virus is associated with virus movement.

A subgenomic RNA (sgRNA) of about 1500 nucleotides has been detected in millet plants and protoplasts infected with panicum mosaic virus (PMV). This sgRNA expressed p8, p6.6, p15, and the 26-kDa capsid protein (CP) genes during in vitro translation assays, as determined by using mutants inactivated for expression of each open reading frame. Abolishing expression of p8 and p6.6, the two 5'-proximal genes on the sgRNA, did not affect the replication of PMV in millet protoplasts, but obstructed spread in plants. As predicted for a typical cell-to-cell movement protein, p8 localized to the cell wall fraction of PMV-infected millet plants. The introduction of premature stop codons downstream of the PMV p15 start codon (p15*) abolished infectivity in planta, but did not impair replication in protoplasts. However, a delayed systemic infection in millet plants was supported by the p15aug(-) start codon mutant, which may reflect very low levels of expression from a suboptimal start codon context and/or leaky scanning to a second inframe AUG codon to express the C-terminal portion of the 15-kDa protein. PMV CP mutants had little effect on sgRNA accumulation, but were correlated with a reduction of the gRNA and the decreased expression of the 8-kDa protein in protoplasts as well as abolishment of cell-to-cell spread in plants. These results imply that the successful establishment of a PMV systemic infection in millet host plants appears to be dependent on the concerted expression of the p8, p6.6, p15, and CP genes.

Capsid↗

Isolation, characterization and expression of cDNA clones encoding a mitochondrial malate translocator from Panicum miliaceum L.

Three cDNA clones that hybridize to a partial rice cDNA that show similarity to bovine mitochondrial 2-oxoglutarate/malate translocator were isolated from leaves of Panicum miliaceum L. (proso millet), an NAD-malic enzyme-type C4 plant. The nucleotide sequences of the clones resemble each other, and some of the isolated cDNAs contained extra sequences that seemed to be introns. The predicted proteins encoded by the cDNAs have 302 amino acids and molecular weights of 32211 and 32150. The hydrophobic profile of the amino acid sequence predicted the existence of six transmembrane alpha-helices that is a common property of members in the mitochondrial transporter family. The predicted amino acid sequence showed the highest similarity with that of the 2-oxoglutarate/malate translocator from mammalian mitochondria. An expression plasmid containing the coding region of the cDNAs was used to over-express recombinant protein with a C-terminal histidine tag Escherichia coli, which was affinity purified. The antibody against the recombinant protein cross-reacted with proteins of 31-32 kDa in the membrane fraction from P. miliaceum mitochondria, but not with the chloroplast fraction. The recombinant protein reconstituted in liposomes efficiently transported malate, citrate, and 2-oxoglutarate.

Amino Acid Sequence↗

Molecular cloning of an alanine aminotransferase from NAD-malic enzyme type C4 plant Panicum miliaceum.

We have determined the nucleotide sequence of a cDNA encoding AlaAT-2, which is believed to function in the C4-pathway of Panicum miliaceum. An open reading frame (1446 bp) encodes a protein of 482 amino acid residues. The deduced amino acid sequence of AlaAT-2 shows 44.2 and 44.8% homology with the amino acid sequences of AlaATs from rat and human livers, respectively. Northern blot analysis showed that the gene encoding AlaAT-2 in mesophyll and bundle sheath cells was the same and transcribed similarly in the cells. The level of translatable mRNA for AlaAT-2 increased dramatically during greening.

Alanine Transaminase↗

Purification and characterization of alanine aminotransferase from Panicum miliaceum leaves.

Three alanine aminotransferases, two minor (AlaAT-1, AlaAT-3) and one major (AlaAT-2), were detected by native gel electrophoresis of leaf extracts from Panicum miliaceum L. AlaAT-2 was purified to homogeneity and a specific polyclonal antibody was raised against it which did not react with the other two forms of the enzyme. The enzyme, with an apparent molecular size of 102 kDa, appeared to be a dimer of a single 50-kDa polypeptide. The enzyme has a relatively broad pH optima with similar curves for the forward and reverse directions, ranging between 6.5 and 7.5. The Km values of this enzyme were 6.67, 0.15, 5.00, and 0.33 mM for alanine, 2-oxoglutarate, glutamate, and pyruvate, respectively. The activity of AlaAT-2 was found to increase markedly during leaf greening in parallel with the increase of immunochemically titrated protein, and it is suggested to function in the C4 photosynthetic cycle.

Alanine Transaminase↗

Effect of salinity and temperature on germination, growth and ion relations of Panicum turgidum Forssk.

Seed germination of Panicum turgidum was significantly affected by salinity levels, temperature and their interaction. Maximum germination was noted in the lowest saline media (25-50 mM) and distilled water at the temperature of 15-25 degrees C and 20-30 degrees C. Seeds germination was substantially delayed and reduced with an increase in NaCl to levels above 50mM. This trend was much pronounced under high levels of NaCl and incubation temperature. Low levels of NaCl (25-50 mM) stimulated shoot and root dry weights of P. turgidum seedlings. However, the highest NaCl levels (>100 mM) resulted in a significant decrease in shoot, root and total dry weights of seedlings. Intermediate degrees of temperature, 15-25 and 20-30 degrees C, resulted in a significant increase in biomass accumulation. The Na+ concentration in shoots and roots significantly increased as NaCl concentration increased. The K+ concentration in roots and K/Na ratio in shoots and roots was significantly reduced as salinity concentration increased. The K/Na ratio was greatly affected by higher NaCl concentration and incubation temperatures.

Germination↗

Internode structure and cell wall composition in maturing tillers of switchgrass (Panicum virgatum. L).

This work examined cell composition gradients in maturing tillers of switchgrass (Panicum virgatum L.) with the aim of developing baseline information on this important forage and biomass crop. Flowering tillers were collected from plants raised from seeds in a greenhouse and field, harvested at soil level and separated into internodes beginning with the node subtending the peduncle. Internodes were analyzed using microscopy, by fiber digestion, high-performance liquid chromatography and by gas chromatography-mass spectrometry to obtain anatomical and compositional data. Microscopy demonstrated the development and maturation of cortical fibers which eventually became confluent with the fiber sheath surrounding vascular bundles in the lower internodes. Detergent fiber analysis indicated increasing cellulose and lignin contents and decreases in cell solubles and hemicelluloses with increasing distance of the internodes from the top of the plant. Soluble phenolics were greatest in amounts and complexity in top internodes. The lower internodes contained greater levels of wall-bound phenolic acids, principally as 4-coumarate and ferulate.

Biomass↗

Effects of fluoride emissions on two tropical grasses: Chloris gayana and Panicum maximum cv. Colonião.

In order to detect early effects of plant contamination by fluoride emission on two tropical grasses, Chloris gayana and Panicum maximum, previously cultivated under greenhouse conditions, were exposed to a single source of fluoride emission at a station at 1.1 km from an aluminum smelter in Ouro Preto, MG, Brazil. Controls were placed at a reference station 78km from the fluoride source. During an 8-day period of exposure leaf injury, ionic permeability, photosynthetic rates, stomatal conductance, transpiration, chlorophyll a fluorescence and chlorophyll, soluble carbohydrates and fluoride contents were evaluated. Plants at the Ouro Preto station showed an increase in fluoride content, leaf injury and ionic permeability. Symptoms of injury by fluoride exposure were visible after 3-4 days in both species. High electrolyte leakage and correlation coefficients between the total ionic permeability and the fluoride content in leaves indicate a fluoride effect on the structural and/or functional integrity of the cellular membranes. Leaf fluoride injuries were quite different in the two species. In C. gayana necroses were limited to the leaf tips, while in P. maximum damages were observed in the whole leaf, suggesting a higher susceptibility of this latter species to fluoride. Nonetheless, neither grass showed statistical differences with respect to photosynthetic rates, stomatal conductance, transpiration, chlorophyll a fluorescence and chlorophyll content in leaves without any apparent fluoride injury. Plants at the Ouro Preto station showed a significant decrease in reducing sugar content between 3 and 5 days of exposure to fluoride, but thereafter reducing sugar content increased reaching the content of control plants. Fluoride exposed plants also showed a remarkable starch content reduction, remaining always much lower than those at the reference station.

Air Pollutants↗

Developmental expression of ASG- 1 during gametogenesis in apomictic guinea grass (Panicum maximum).

We have used Western blue-visualized in situ-hybridization (ISH) to monitor the expression of apomixis-specific gene-1 (ASG-1, GenBank accession number AB000809) during gametogenesis in obligate-sexual and facultative-apomictic (aposporic) genotypes of guinea grass (Panicum maximum). The in situ-analysis revealed that ASG-1 is not expressed in the ovule during early floral development in both, the facultative apomicts (A1 stage) and the obligate sexuals (S1 stage). With the appearance of the aposporous initial cell(s) in the ovule of the apomictic type (A2-1 stage), ASG-1 expression is strong and specific to this apomixis-specific cell. ASG-1 expression continued through different stages of aposporous embryo sac development (A2-2 stage), indicating that the gene may play a role in this developmental process. Regular embryo sacs in sexual types did not show hybridization signals (S2 stage). However, strong ASG-1 expression was detected in immature pollen grains and young embryos in both reproductive types, suggesting that ASG-1 may be an allele derived from the obligate-sexual wild type. Expression in pollen grains faded with maturation. In a heterologous system, using Paspalum notatum, a facultative-aposporic tropical grass (bahia grass), identical results were obtained. The results are discussed in view of the fact, that ASG-1 shows some homology to genes known to be seed- or embryo-specific or involved in processes related to cell growth.

Gametogenesis↗

Pyrene degradation in the rhizosphere of tall fescue (Festuca arundinacea) and switchgrass (Panicum virgatum L.).

A growth chamber study was conducted to investigate the fate of pyrene in the rhizosphere of tall fescue (Festuca arundinacea) and switchgrass (Panicum virgatum L.). For this study, 14C-labeled pyrene was used, and distribution of 14C activity was assessed after plant establishment. After 190 days of incubation, 37.7 and 30.4% of 14C-pyrene was mineralized in the soil planted with tall fescue and switchgrass, respectively, while 4.3% mineralization was observed for the unplanted control. Only 7.6 and 8.7% of pyrene was recovered from the soil in the two planted treatments, while 31.5% of pyrene remained in the unplanted control. Significant amounts of 14C were observed for all treatments and controls in the humic/fulvic fraction of soil at the end of the experiment. This research indicates the potential for pyrene mineralization in planted systems, although the ultimate fate of degradation byproducts is uncertain.

Biodegradation, Environmental↗

Merremia tridentata as a supplementary feed to the grass Panicum maximum for young West African Dwarf sheep.

The provision ad libitum of the browse plant Merremia tridentata as a free-choice supplement to the grass Panicum maximum led to a significant increase in the total food intake of groups of five West African Dwarf sheep, although the increase in weight gain was not significant. The protein content of M. tridentata was higher than that of the grass and its tannin content was not sufficient to render it unpalatable.

Animal Feed↗

Nitrogen dynamics in the intact grasses Poa trivialis and Panicum maximum receiving contrasting supplies of nitrogen.

The C(3) grass Poa trivialis and the C(4) grass Panicum maximum were grown in sand culture and received a complete nutrient solution with nitrogen supplied as 1.5 mol m(-3) NH(4)NO(3). (15)N tracer techniques were used to quantify the relative use of root uptake and mobilization in supplying nitrogen to growing leaves in intact plants which either continued to receive nitrogen or which received the complete nutrient solution without nitrogen. The allocation of both (15)N-labelled nitrogen uptake and unlabelled mobilized nitrogen indicated that, under their conditions of growth, the sink strength of growing leaves was relatively greater in P. maximum than P. trivialis. The supply of nitrogen by mobilization to side tillers of P. trivialis was completely stopped as the external nitrogen supply was reduced, whilst in P. maximum some allocation of mobilized nitrogen to side tillers, roots and growing leaves was maintained. In both plant species receiving an uninterrupted supply of nitrogen the allocation pattern of mobilized nitrogen differed from that of nitrogen derived from root uptake. Differences exist in the degree to which P. trivialis and P. maximum utilized uptake and mobilization to supply nitrogen to the growing leaves. In P. trivialis roots were always a net sink of mobilized nitrogen, irrespective of the external nitrogen supply. In P. maximum, roots were a net sink of mobilized nitrogen when external nitrogen was withdrawn, but exhibited both source and sink behaviour when nitrogen supply was continued.

Biological Transport↗