Emanation of water from underground plant parts

Summary A device was designed which used gypsum and electrical conductivity as a means of detecting and quantitizing the amount of water emanating from underground plant parts under natural conditions. Use of this device with the Common French Bean demonstrated that considerable amounts of moisture emanated from the underground part of the stem. The amount of water emanated was found to be influenced by environmental conditions, differences in soil type and degree of soil compaction. When plants were grown in an environment of approximately 100 per cent relative air humidity and at a temperature of 28 to 30°C, 115 to 125 µl of water emanated from an 11 mm length of stem in a 24-hour period, in contrast to 70 to 80 µl of water emanating from plants grown at 50 to 60 per cent relative air humidity and at a temperature of 18 to 20°C. Water emanation was more rapid in a compacted soil than loose soil and also appeared to be dependent on the amount of sand in a soil. Whereas 115 to 125 µl of water emanated in a silt loam soil, only 35 to 45 µl emanated in a loamy sand soil and less than 10 µl in washed sea sand during a 24-hour period. The sensitivity of the conductivity unit to changes in moisture and the utility of its design enables its employment under varied environmental conditions and offers a technique for plant pathologists to study the effect of both the below- and aboveground environments on moisture emanation from underground plant parts and the resultant effect on the associated microflora.Senior author was visiting Research Fellow and NATO Science Fellow, sponsored by the Netherlands Organization for the Advancement of Pure Research (ZWO), Sept. 16, 1964 to Sept. 15, 1965. Present address: Phytopathologisch Laboratorium W.C.S. Baarn, The Netherlands.     

Pseudomonas siderophores: A mechanism explaining disease-suppressive soils

The membrane-bound hydrogenase ffomPseudomonas pseudoflava GA3 was purified up to a specific activity of 172 μmol H₂ oxidized/min and mg protein and a yield of 31%. The enzyme has a molecular weight of 98,000, consists of two different subunits (65,000 and 30,000), and contains 6 atoms iron and 6 molecules of acid-labile sulfide per molecule of enzyme. The isoelectric point was determined to be 6.5. The enzyme was stable under nitrogen, oxygen, and air atmosphere and unstable under hydrogen. The purified hydrogenase was able to reduce only a few of artificial electron acceptors, i.e., pyocyanine, methylene blue, phenazinemethosulfate, benzylviologen, and dichlorophenolindophenol.     

SMA CARNI-VAL Trial Part I: Double-Blind,Randomized, Placebo-Controlled Trial of L-Carnitine and Valproic Acid in Spinal Muscular Atrophy

Background

Valproic acid (VPA) has demonstrated potential as a therapeutic candidate for spinal muscular atrophy (SMA) in vitro and in vivo.

Methods

Two cohorts of subjects were enrolled in the SMA CARNIVAL TRIAL, a non-ambulatory group of “sitters” (cohort 1) and an ambulatory group of “walkers” (cohort 2). Here, we present results for cohort 1: a multicenter phase II randomized double-blind intention-to-treat protocol in non-ambulatory SMA subjects 2–8 years of age. Sixty-one subjects were randomized 1∶1 to placebo or treatment for the first six months; all received active treatment the subsequent six months. The primary outcome was change in the modified Hammersmith Functional Motor Scale (MHFMS) score following six months of treatment. Secondary outcomes included safety and adverse event data, and change in MHFMS score for twelve versus six months of active treatment, body composition, quantitative SMN mRNA levels, maximum ulnar CMAP amplitudes, myometry and PFT measures.

Results

At 6 months, there was no difference in change from the baseline MHFMS score between treatment and placebo groups (difference = 0.643, 95% CI = −1.22–2.51). Adverse events occurred in >80% of subjects and were more common in the treatment group. Excessive weight gain was the most frequent drug-related adverse event, and increased fat mass was negatively related to change in MHFMS values (p = 0.0409). Post-hoc analysis found that children ages two to three years that received 12 months treatment, when adjusted for baseline weight, had significantly improved MHFMS scores (p = 0.03) compared to those who received placebo the first six months. A linear regression analysis limited to the influence of age demonstrates young age as a significant factor in improved MHFMS scores (p = 0.007).

Conclusions

This study demonstrated no benefit from six months treatment with VPA and L-carnitine in a young non-ambulatory cohort of subjects with SMA. Weight gain, age and treatment duration were significant confounding variables that should be considered in the design of future trials.

Trial Registry

Clinicaltrials.gov {"type":"clinical-trial","attrs":{"text":"NCT00227266","term_id":"NCT00227266"}}NCT00227266     

Structure and function of methanotrophic communities in a landfill-cover soil

In landfill-cover soils, aerobic methane-oxidizing bacteria (MOB) convert CH(4) to CO(2), mitigating emissions of the greenhouse gas CH(4) to the atmosphere. We investigated overall MOB community structure and assessed spatial differences in MOB diversity, abundance and activity in a Swiss landfill-cover soil. Molecular cloning, terminal restriction-fragment length polymorphism (T-RFLP) and quantitative PCR of pmoA genes were applied to soil collected from 16 locations at three different depths to study MOB community structure, diversity and abundance; MOB activity was measured in the field using gas push-pull tests. The MOB community was highly diverse but dominated by Type Ia MOB, with novel pmoA sequences present. Type II MOB were detected mainly in deeper soil with lower nutrient and higher CH(4) concentrations. Substantial differences in MOB community structure were observed between one high- and one low-activity location. MOB abundance was highly variable across the site [4.0 × 10(4) to 1.1 × 10(7) (g soil dry weight)(-1)]. Potential CH(4) oxidation rates were high [1.8-58.2 mmol CH(4) (L soil air)(-1) day(-1) ] but showed significant lateral variation and were positively correlated with mean CH(4) concentrations (P < 0.01), MOB abundance (P < 0.05) and MOB diversity (weak correlation, P < 0.17). Our findings indicate that Methylosarcina and closely related MOB are key players and that MOB abundance and community structure are driving factors in CH(4) oxidation at this landfill.     

Circadian methane oxidation in the root zone of rice plants

In the root zone of rice plants aerobic methanotrophic bacteria catalyze the oxidation of CH₄ to CO₂, thereby reducing CH₄ emissions from paddy soils to the atmosphere. However, methods for in situ quantification of microbial processes in paddy soils are scarce. Here we adapted the push–pull tracer-test (PPT) method to quantify CH₄ oxidation in the root zone of potted rice plants. During a PPT, a test solution containing CH₄ ± O₂ as reactant(s), Cl^? and Ar as nonreactive tracers, and BES as an inhibitor of CH₄ production was injected into the root zone at different times throughout the circadian cycle (daytime, early nighttime, late nighttime). After a 2-h incubation phase, the test solution/pore-water mixture was extracted from the same location and rates of CH₄ oxidation were calculated from the ratio of measured reactant and nonreactive tracer concentrations. In separate rice pots, O₂ concentrations in the vicinity of rice roots were measured throughout the circadian cycle using a fiber-optic sensor. Results indicated highly variable CH₄ oxidation rates following a circadian pattern. Mean rates at daytime and early nighttime varied from 62 up to 451 μmol l^?1 h^?1, whereas at late nighttime CH₄ oxidation rates were low, ranging from 13 to 37 μmol l^?1 h^?1. Similarly, daytime O₂ concentration in the vicinity of rice roots increased to up to 250% air saturation, while nighttime O₂ concentration dropped to below detection (<0.15% air saturation). Our results suggest a functional link between root-zone CH₄ oxidation and photosynthetic O₂ supply.     

Recovery of in-situ methanotrophic activity following acetylene inhibition

Methane (CH₄) is the second most important greenhouse gas after carbon dioxide (CO₂). To understand CH₄ cycling, quantitative information about microbial CH₄ oxidation in soils is essential. Field methods such as the gas push-pull test (GPPT) to quantify CH₄ oxidation are often used in combination with specific inhibitors, such as acetylene (C₂H₂). Acetylene irreversibly binds to the enzyme methane monooxygenase, but little is known about recovery of CH₄ oxidation activity after C₂H₂ inhibition in situ, which is important when performing several experiments at the same location. To assess recovery of CH₄ oxidation activity following C₂H₂ inhibition, we performed a series of GPPTs over 8 weeks at two different locations in the vadose zone above a petroleum hydrocarbon-contaminated aquifer in Studen, Switzerland. After 4 weeks a maximum recovery of 30% and 50% of the respective initial activity was reached, with a subsequent slight drop in activity at both locations. Likely, CH₄ oxidation activity and CH₄ concentrations were too low to allow for rapid recovery following C₂H₂ inhibition at the studied locations. Therefore, alternative competitive inhibitors have to be evaluated for application in conjunction with GPPTs, especially for sites with low activity.     

Improvement of DNA transfer frequency and transposon mutagenesis of Erwinia carotovora subsp. betavasculorum

The production of antibiotics and their role in microbial competition under natural conditions can be readily studied by the use of transposon mutants. Several antibiotic-producing strains of Erwinia carotovora subsp. betavasculorum were unable to accept foreign DNA. A plasmid delivery system was developed, using ethyl methanesulfonate mutagenesis, which entailed isolating E. carotovora subsp. betavasculorum mutants able to accept foreign DNA and transfer it to other strains. This enabled transposon mutagenesis of a wild-type antibiotic-producing strain of E. carotovora subsp. betavasculorum. Twelve antibiotic-negative mutants were isolated, and one of these showed a reduction in antibiotic production in vitro. Many of these mutants also showed a reduction in their ability to macerate potato tissue. The mutants were classified into four genetic groups on the basis of their genetic and phenotypic characteristics, indicating that several genes are involved in antibiotic biosynthesis by E. carotovora subsp. betavasculorum.     

Deoxyribonucleic acid relatedness of 21 strains of Xanthomonas species and pathovars

The relationship of 17 Xanthomonas campestris pathotype strains, three additional X. campestris strains, and the type strain of Xanthomonas albilineans were examined by DNA-DNA hybridization tests. The results coupled with those of a previous study (Hildebrand et al. 1990) support the hypothesis that X. campestris does not constitute a single bacterial species. There were low levels of DNA-DNA reassociation among many of the different pathovars examined. Six clusters of related pathovars were discerned. In addition, four of the pathovars were only distantly related to each other and to the six clusters. Xanthomonas albilineans was not closely related to any of the other xanthomonads tested.
Mapping and superimposing the botanical families of the host plants upon a three-dimensional genomic distance matrix of the xanthomonads confirms previous observations that pathovars that infect plants of the same botanical family do not necessarily belong to the same genomic group. Six legume-infecting pathovars cluster within one genomic group, but one pathovar, X. cam. pv. pisi is only distantly related to this group. There was also no genomic relationship between X. cam. pv. oryzicola and X. albilineans both of which infect Gramineae. Consequently, pathogenicity toward members of the same plant family is not a good indicator of the genomic relationships among xanthomonads nor is it a good taxonomic determinant.     

Response of methanotrophic activity and community structure to temperature changes in a diffusive CH4/O2 counter gradient in an unsaturated porous medium

Microbial methane oxidation is a key process in the global methane cycle. In the context of global warming, it is important to understand the responses of the methane-oxidizing microbial community to temperature changes in terms of community structure and activity. We studied microbial methane oxidation in a laboratory-column system in which a diffusive CH₄/O₂ counter gradient was maintained in an unsaturated porous medium at temperatures between 4 and 20 °C. Methane oxidation was highly efficient at all temperatures, as on average 99 ± 0.5% of methane supplied to the system was oxidized. The methanotrophic community that established in the model system after initial inoculation appeared to be able to adapt quickly to different temperatures, as methane emissions remained low even after the system was subjected to abrupt temperature changes. FISH showed that Type I as well as Type II methanotrophs were probably responsible for the observed activity in the column system, with a dominance of Type I methanotrophs. Cloning and sequencing suggested that Type I methanotrophs were represented by the genus Methylobacter while Type II were represented by Methylocystis . The results suggest that in an unsaturated system with diffusive substrate supply, direct effects of temperature on apparent methanotrophic activity and community may be of minor importance. However, this remains to be verified in the field.