Periodic carbon flushing to roots of Quercus rubra saplings affects soil respiration and rhizosphere microbial biomass

Patterns of root/shoot carbon allocation within plants have been studied at length. The extent, however, to which patterns of carbon allocation from shoots to roots affect the timing and quantity of organic carbon release from roots to soil is not known. We employed a novel approach to study how natural short-term variation in the allocation of carbon to roots may affect rhizosphere soil biology. Taking advantage of the semi-determinate phenology of young northern red oak (Quercus rubra L.), we examined how pulsed delivery of carbon from shoots to roots affected dynamics of soil respiration as well as microbial biomass and net nitrogen mineralization in the rhizosphere. Young Q. rubra exhibit (1) clear switches in the amount of carbon allocated below-ground that are non-destructively detected simply by observing pulsed shoot growth above-ground, and (2) multiple switches in internal carbon allocation during a single growing season, ensuring our ability to detect short-term effects of plant carbon allocation on rhizosphere biology separate from longer-term seasonal effects. In both potted oaks and oaks rooted in soil, soil respiration varied inversely with shoot flush stage through several oak shoot flushes. In addition, upon destructive harvest of potted oaks, microbial biomass in the rhizosphere of saplings with actively flushing shoots was lower than microbial biomass in the rhizosphere of saplings with shoots that were not flushing. Given that plants have evolved with their roots in contact with soil microbes, known species-specific carbon allocation patterns within plants may provide insight into interactions among roots, symbionts, and free-living microbes in the dynamic soil arena.     

Contribution of Glucosinolate Transport to Arabidopsis Defense Responses

Accumulation of glucosinolates, a class of defense-related secondary metabolites found almost exclusively in the Capparales, is induced in response to a variety of biological stresses. It is often assumed that elevated glucosinolate levels result from de novo biosynthesis, but glucosinolate transport from other parts of the plant to the site of herbivory or pathogen infection can also contribute to the defense response. Several studies with Arabidopsis and other crucifers have demonstrated that glucosinolates from vegetative tissue are transported to developing seeds. Here we discuss evidence that long-chain aliphatic glucosinolates are transported to the site of herbivory in response to Myzus persicae (green peach aphid) feeding on Arabidopsis.Key Words: glucosinolate, transport, graft, Arabidopsis, Myzus persicae, aphid     

The role of nitric oxide in the regulation of the systemic and pulmonary vasculature of the rattlesnake, <Emphasis Type="Italic">Crotalus durissus terrificus</Emphasis>

The functional role of nitric oxide (NO) was investigated in the systemic and pulmonary circulations of the South American rattlesnake, Crotalus durissus terrificus. Bolus, intra-arterial injections of the NO donor, sodium nitroprusside (SNP) caused a significant systemic vasodilatation resulting in a reduction in systemic resistance (Rsys). This response was accompanied by a significant decrease in systemic pressure and a rise in systemic blood flow. Pulmonary resistance (Rpul) remained constant while pulmonary pressure (Ppul) and pulmonary blood flow (Qpul) decreased. Injection of L-Arginine (L-Arg) produced a similar response to SNP in the systemic circulation, inducing an immediate systemic vasodilatation, while Rpul was unaffected. Blockade of NO synthesis via the nitric oxide synthase inhibitor, L-NAME, did not affect haemodynamic variables in the systemic circulation, indicating a small contribution of NO to the basal regulation of systemic vascular resistance. Similarly, Rpul and Qpul remained unchanged, although there was a significant rise in Ppul. Via injection of SNP, this study clearly demonstrates that NO causes a systemic vasodilatation in the rattlesnake, indicating that NO may contribute in the regulation of systemic vascular resistance. In contrast, the pulmonary vasculature seems far less responsive to NO.     

Delivery of antibodies to the cytosol: Debunking the myths

The use of antibodies to target their antigens in living cells is a powerful analytical tool for cell biology research. Not only can molecules be localized and visualized in living cells, but interference with cellular processes by antibodies may allow functional analysis down to the level of individual post-translational modifications and splice variants, which is not possible with genetic or RNA-based methods. To utilize the vast resource of available antibodies, an efficient system to deliver them into the cytosol from the outside is needed. Numerous strategies have been proposed, but the most robust and widely applicable procedure still remains to be identified, since a quantitative ranking of the efficiencies has not yet been done. To achieve this, we developed a novel efficiency evaluation method for antibody delivery based on a fusion protein consisting of a human IgG₁ Fc and the recombination enzyme Cre (Fc-Cre). Applied to suitable GFP reporter cells, it allows the important distinction between proteins trapped in endosomes and those delivered to the cytosol. Further, it ensures viability of positive cells and is unsusceptible to fixation artifacts and misinterpretation of cellular localization in microscopy and flow cytometry. Very low cytoplasmic delivery efficiencies were found for various profection reagents and membrane penetrating peptides, leaving electroporation as the only practically useful delivery method for antibodies. This was further verified by the successful application of this method to bind antibodies to cytosolic components in living cells.     

The composition of amino acid-peptide mixtures obtained during hydrolysis of proteins

The composition of the amino acid and peptide fraction of the protein hydrolyzate and its dependence upon the spray-drying process at elevated temperatures were investigated by GLC and GLC-MS. Spray-drying leads to decrease in the Glu and Met contents, to transformation of Cys into cystine and to formation of lactames (pyroglutamic acid and pyrrolidone) and diketopiperazines. Some dipeptides, acetylproline, paraffines, fatty acid pyridinecarboxylic and lactic acids, ethylene glycol were identified in the fraction. The incidental occurrence of these compounds should be taken into account when investigating the biological activity of protein hydrolyzates.     

Helminths of Cnemidophorus ruthveni (Squamata: Teiidae) from Bonaire, Netherlands Antilles, with description of a new species of Alaeuris (Nematoda: Pharyngodonidae)

Alaeuris rinconensis n. sp. (Oxyuroidea, Pharyngodonidae) from the large intestine of the Bonaire whiptail lizard, Cnemidophorus ruthveni, is described and illustrated. Alaeuris rinconensis n. sp. represents the 13th Neotropical species assigned to the genus. It is most similar to Alaeuris iguanae in that only these 2 Neotropical species have postbulbar excretory pores; in A. iguanae, a tail filament is absent; in A. rinconensis, a tail filament is present. Two additional helminth species were found, i.e., the cestode Oochoristica iguanae and a nematode, Ozolaimus megatyphlon. Cnemidoporus ruthveni represents a new host record for the latter 2 species.     

Effects of the number of markers per haplotype and clustering of haplotypes on the accuracy of QTL mapping and prediction of genomic breeding values

The aim of this paper was to compare the effect of haplotype definition on the precision of QTL-mapping and on the accuracy of predicted genomic breeding values. In a multiple QTL model using identity-by-descent (IBD) probabilities between haplotypes, various haplotype definitions were tested i.e. including 2, 6, 12 or 20 marker alleles and clustering base haplotypes related with an IBD probability of > 0.55, 0.75 or 0.95. Simulated data contained 1100 animals with known genotypes and phenotypes and 1000 animals with known genotypes and unknown phenotypes. Genomes comprising 3 Morgan were simulated and contained 74 polymorphic QTL and 383 polymorphic SNP markers with an average r² value of 0.14 between adjacent markers. The total number of haplotypes decreased up to 50% when the window size was increased from two to 20 markers and decreased by at least 50% when haplotypes related with an IBD probability of > 0.55 instead of > 0.95 were clustered. An intermediate window size led to more precise QTL mapping. Window size and clustering had a limited effect on the accuracy of predicted total breeding values, ranging from 0.79 to 0.81. Our conclusion is that different optimal window sizes should be used in QTL-mapping versus genome-wide breeding value prediction.     

Decomposition and Peat Accumulation in Rich Fens of Boreal Alberta, Canada

Fens are important components of Canada’s western boreal forests, occupying about 63% of the total peatland area and storing about 65% of the peatland carbon. Rich fens, dominated by true moss-dominated ground layers, make up more than half of the fens in the region. We studied organic matter accumulation in three rich fens that represent the diversity in structural types. We used in situ decomposition socks, a new method that examines actual decomposition throughout the upper peat profile over an extended period of time. We coupled our carbon loss data with macrofossil analyses and dated peat profiles using ²¹⁰Pb. Across the three rich fens and in the top 39 cm of the peat column, dry mass increases on average 3.1 times. From our dry mass loss measurements, we calculate that annual mass loss from the top 39 cm varies from 0.52 to 1.08 kg m². Vertical accumulation during the past 50 years has varied from 16 to 32 cm and during these 50 years, organic matter accumulation has averaged 174 g m⁻² y⁻¹ compared to 527 g m² y⁻¹ dry mass loss, with additional mass losses of 306 g m² y⁻¹ from peat between 50 and 150 years of age. Organic matter accumulation from our rich fens compares well with literature values from boreal bogs, whereas peat bulk densities increase about three times within the uppermost 40 cm, much more than in bogs. Hence, rich fens accumulate peat not because the plant material is especially hard to decompose, is acidic, or has the catotelm especially close to the surface, but because dense, rapidly produced inputs outweigh the relatively rapid decomposition process of the upper peat column. Author Contributions: DHV conceived study; KS, KW, SF, & DHV performed research; DHV, KW analyzed data; DHV, KW contributed new methods; DHV, KW wrote the paper.