Detection of transgenic and endogenous plant DNA in rumen fluid, duodenal digesta, milk, blood, and feces of lactating dairy cows

The objective was to determine the presence or absence of transgenic and endogenous plant DNA in ruminal fluid, duodenal digesta, milk, blood, and feces, and if found, to determine fragment size. Six multiparous lactating Holstein cows fitted with ruminal and duodenal cannulas received a total mixed ration. There were two treatments (T). In T1, the concentrate contained genetically modified (GM) soybean meal (cp4epsps gene) and GM corn grain (cry1a[b] gene), whereas T2 contained the near isogenic non-GM counterparts. Polymerase chain reaction analysis was used to determine the presence or absence of DNA sequences. Primers were selected to amplify small fragments from single-copy genes (soy lectin and corn high-mobility protein and cp4epsps and cry1a[b] genes from the GM crops) and multicopy genes (bovine mitochondrial cytochrome b and rubisco). Single-copy genes were only detected in the solid phase of rumen and duodenal digesta. In contrast, fragments of the rubisco gene were detected in the majority of samples analyzed in both the liquid and solid phases of ruminal and duodenal digesta, milk, and feces, but rarely in blood. The size of the rubisco gene fragments detected decreased from 1176 bp in ruminal and duodenal digesta to 351 bp in fecal samples.

Binding of pentagalloyl glucose to two globular proteins occurs via multiple surface sites

The interaction between pentagalloyl glucose (PGG) and two globular proteins, bovine serum albumin (BSA) and ribulose-1,5-bisphosphate carboxylase oxygenase (rubisco), was investigated by isothermal titration calorimetry (ITC). ITC data fit to a binding model consisting of two sets of multiple binding sites, which reveal similarities in the mode of binding of PGG to BSA and rubisco. In both cases, the interaction is characterized by a high number of binding sites, which suggests that binding occurs by a surface adsorption mechanism that leads to coating of the protein surface, which promotes aggregation and precipitation of the PGG-protein complex. This model was confirmed by turbidimetry analysis of the PGG-BSA interaction. Analysis of tryptophan fluorescence quenching during the interaction of PGG with BSA suggests that binding of PGG leads to some conformational changes that are energetically closer to the unfolded state of the BSA structure, because small red shifts in the resulting emission spectra were observed.

The transcriptome of Populus in elevated CO2

The consequences of increasing atmospheric carbon dioxide for long-term adaptation of forest ecosystems remain uncertain, with virtually no studies undertaken at the genetic level. A global analysis using cDNA microarrays was conducted following 6 yr exposure of Populus × euramericana (clone I-214) to elevated [CO2] in a FACE (free-air CO2 enrichment) experiment.• Gene expression was sensitive to elevated [CO2] but the response depended on the developmental age of the leaves, and < 50 transcripts differed significantly between different CO2 environments. For young leaves most differentially expressed genes were upregulated in elevated [CO2], while in semimature leaves most were downregulated in elevated [CO2].• For transcripts related only to the small subunit of Rubisco, upregulation in LPI 3 and downregulation in LPI 6 leaves in elevated CO2 was confirmed by anova. Similar patterns of gene expression for young leaves were also confirmed independently across year 3 and year 6 microarray data, and using real-time RT–PCR.• This study provides the first clues to the long-term genetic expression changes that may occur during long-term plant response to elevated CO2.

Quantitative plant proteomics

Quantitation is an inherent requirement in comparative proteomics and there is no exception to this for plant proteomics. Quantitative proteomics has high demands on the experimental workflow, requiring a thorough design and often a complex multi-step structure. It has to include sufficient numbers of biological and technical replicates and methods that are able to facilitate a quantitative signal read-out. Quantitative plant proteomics in particular poses many additional challenges but because of the nature of plants it also offers some potential advantages. In general, analysis of plants has been less prominent in proteomics. Low protein concentration, difficulties in protein extraction, genome multiploidy, high Rubisco abundance in green tissue, and an absence of well-annotated and completed genome sequences are some of the main challenges in plant proteomics. However, the latter is now changing with several genomes emerging for model plants and crops such as potato, tomato, soybean, rice, maize and barley. This review discusses the current status in quantitative plant proteomics (MS-based and non-MS-based) and its challenges and potentials. Both relative and absolute quantitation methods in plant proteomics from DIGE to MS-based analysis after isotope labeling and label-free quantitation are described and illustrated by published studies. In particular, we describe plant-specific quantitative methods such as metabolic labeling methods that can take full advantage of plant metabolism and culture practices, and discuss other potential advantages and challenges that may arise from the unique properties of plants.

Relationship between condensed tannin structures and their ability to precipitate feed proteins in the rumen

Abstract BACKGROUND Tannins can bind to and precipitate protein by forming insoluble complexes resistant to fermentation and with a positive effect on protein utilisation by ruminants. Three protein types, Rubisco, rapeseed protein and bovine serum albumin (a single high-molecular weight protein), were used to test the effects of increasing concentrations of structurally different condensed tannins on protein solubility/precipitation. RESULTS Protein type (PT) influenced solubility after addition of condensed tannins (P < 0.001) in the order: Rubisco < rapeseed < BSA (P < 0.05). The type of condensed tannin (CT) affected protein solubility (P = 0.001) with a CT × PT interaction (P = 0.001). Mean degree of polymerisation, proportions of cis- versus trans-flavanol subunits or prodelphinidins versus procyanidins among CTs could not explain precipitation capacities. Increasing tannin concentration decreased protein solubility (P < 0.001) with a PT × CT concentration interaction. The proportion of low-molecular weight rapeseed proteins remaining in solution increased with CT concentration but not with Rubisco. CONCLUSIONS Results of this study suggest that PT and CT type are both of importance for protein precipitation but that the CT structures investigated did not allow identification of parameters that contribute most to precipitation. It is possible that the three-dimensional structures of tannins and proteins may be more important factors in tannin–protein interactions. © 2013 Society of Chemical Industry

PHYLOGENETIC AFFINITIES OF "CHANTRANSIA" STAGES IN MEMBERS OF THE BATRACHOSPERMALES AND THOREALES (RHODOPHYTA)

This study evaluated the phylogenetic relationship among samples of ""Chantransia"" stage of the Batrachospermales and Thoreales from several regions of the world based on sequences of two genes-the plastid-encoded RUBISCO LSU gene (rbcL) and the nuclear SSU ribosomal DNA gene (SSU rDNA). All sequences of ""Chantransia macrospora"" were shown to belong to Batrachospermum macrosporum based on both molecular markers, confirming evidence from previous studies. In contrast, nine species are now associated with ""Chantransia pygmaea,"" including seven species of the Batrachospermales and two of the Thoreales. Therefore, the presence of ""C. macrospora"" in a stream can be considered reliable evidence that it belongs to B. macrosporum, whereas the occurrence of ""C. pygmaea"" does not allow the recognition of any particular species, since it is associated with at least nine species. Affinities of ""Chantransia"" stages to particular taxa were congruent for 70.5% of the samples comparing the rbcL and SSU analyses, which were associated with the same or closely related species for both markers. Sequence divergences have been reported in the ""Chantransia"" stage in comparison to the respective gametophyte, and this matter deserves further attention.

Phylogenetic relationships in Kumanoa (Batrachospermales, Rhodophyta) species in Brazil with the proposal of Kumanoa amazonensis sp nov.

This Study evaluated the species-level taxonomy and phylogenetic relationship among Kumanoa species from Brazil with other regions of the world based on the plastid-encoded RUBISCO large Subunit gene (rhcL). Partial rbcL sequences were obtained for 11 Kulnanoa specimens. Eight species are recognised from Brazil on the basis of molecular and morphological data: seven previously described (K abilii, K ambignia, K. breviarticulata, K. cipoensis, K. equisetoidea, K. globospora and K procarpa) and a new species here proposed (K. amazonensis sp. nov. Necchi & Vis). The new species has reduced and dense whorls but differs from the two closest related species in lacking secondary fascicles. Previously proposed infrageneric categories were not supported by the molecular data. Species described and endemic (K. breviarticulata, K. cipoensis, K equiseloidea and K. procarpa) to Brazil are not grouped together but are variously related to other species from North America, Europe and Australasia. With the species recognised in this study using molecular and morphological data and those previously distinguished by morphology, 13 species of Kumanoa are Currently documented from Brazil.

Growth and photosynthetic down-regulation in Coffea arabica in response to restricted root volume

Coffee (Coffea arabica L.) plants were grown in small (3-L), medium (10-L) and large (24-L) pots for 115 or 165 d after transplanting (DAT), which allowed different degrees of root restriction. Effects of altered source : sink ratio were evaluated in order to explore possible stomatal and non-stomatal mechanisms of photosynthetic down-regulation. Increasing root restriction brought about large and general reductions in plant growth associated with a rising root : shoot ratio. Treatments did not affect leaf water potential or leaf nutrient status, with the exception of N content, which dropped significantly with increasing root restriction even though an adequate N supply was available. Photosynthesis was severely reduced when plants were grown in small pots; this was largely associated with non-stomatal factors, such as decreased Rubisco activity. At 165DAT contents of hexose, sucrose, and amino acids decreased in plants grown in smaller pots, while those of starch and hexose-P increased in plants grown in smaller pots. Photosynthetic rates were negatively correlated with the ratio of hexose to free amino acids, but not with hexose content. Activities of acid invertase, sucrose synthase, sucrose-P synthase, fructose-1,6- bisphosphatase, ADP-glucose pyrophosphorylase, starch phosphorylase, glyceraldehyde-3-P dehydrogenase, PPi : fructose-6-P 1-phosphotransferase and NADP : glyceraldehyde-3-P dehydrogenase all decreased with severe root restriction. Glycerate-3-P : Pi and glucose-6-P : fructose-6-P ratios decreased accordingly. Photosynthetic down-regulation was unlikely to have been associated directly with an end-product limitation, but rather with decreases in Rubisco. Such a down-regulation was largely a result of N deficiency caused by growing coffee plants in small pots.

Phylogenetic relationships in Kumanoa (Batrachospermales, Rhodophyta) species in Brazil with the proposal of Kumanoa amazonensis sp nov.

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Molecular systematics of Thorea (Rhodophyta, Thoreales) species in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Phylogenetic relationship of Sirodotia species (Batrachospermales, Rhodophyta) in North and South America

Sequence data from the RUBISCO large subunit (rbcL) plastid gene and nuclear small-subunit ribosomal DNA (SSU rDNA) were examined for five samples of Sirodotia delicatula from southeastern Brazil. Data from six North American samples previously identified as S. huillensis and S. suecica were also included in the analysis. Molecular data supported the continued recognition of these three species as separate entities, although one of the North American collections was misidentified. These results were shown to be congruent with morphology, chromosome number and geographic distribution. S. delicatula is more closely related to S. huillensis, both occurring in tropical-subtropical regions, than either to S. suecica with a temperate-boreal distribution. There was little rbcL variation within S. delicatula from Brazil and Costa Rica (the latter a collection previously identified as S. huillensis), with the six samples sequenced diverging from each other by 0-8 bp (0-0.67%). SSU rDNA data set did not provide sufficient resolution to infer phylogenetic relationships among the species of this group due to the low rates of variation (5 bp). Sirodotia was a well-supported clade (100% bootstrap or 1.00 a posteriori probability) based on rbcL sequences. Thus, the results confirm that Sirodotia is a monophyletic group within the Batrachospermales and we continue to recognize it at the generic level. The species S. delicatula, S. huillensis and S. suecica are morphologically and genetically distinct.

PHYLOGENETIC AFFINITIES of "CHANTRANSIA" STAGES IN MEMBERS of THE BATRACHOSPERMALES and THOREALES (RHODOPHYTA)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Trocas gasosas, índices fisiológicos e acúmulo iônico em plantas jovens de Annona emarginata (Schltdl.) H. Rainer em solução nutritiva

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Sistemática da seção Virescentia do gênero Batrachospermum (Rhodophyta, Batrachospermales) no Brasil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Desenvolvimento, trocas gasosas, rendimento e composição de óleo essencial de Mentha piperita L. cultivada em solução nutritiva com variação dos níveis de N, P, K e Mg

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)