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Rodriguez, Jr., Miguel

Publications and source records attributed to Rodriguez, Jr., Miguel.

6 records
All sourcesNASA NTRSDOE OSTI
DOE OSTI2022

Biomass formation and sugar release efficiency of Populus modified by altered expression of a NAC transcription factor

Woody biomass is an important feedstock for biofuel production. Manipulation of wood properties that enable efficient conversion of biomass to biofuel reduces cost of biofuel production. Wood cell wall composition is regulated at several levels that involve expression of transcription factors such as wood-/secondary cell wall-associated NAC domains (WND or SND). In Arabidopsis thaliana, SND1 regulates cell wall composition through activation of its down-stream targets such as MYBs. The functional aspects of SND1 homologs in the woody Populus have been studied through transgenic manipulation. In this study, we investigated the role of PdWND1B, Populus SND1 sequence ortholog, in wood formation using transgenic manipulation through over-expression or silencing under the control of a vascular-specific 4-coumarate-CoA ligase (4CL) promoter. As compared with control plants, PdWND1B-RNAi plants were shorter in height, with significantly reduced stem diameter and dry biomass, whereas there were no significant differences in growth and productivity of PdWND1B over-expression plants. Conversely, PdWND1B over-expression lines showed a significant reduction in cellulose and increase in lignin content, whereas there was no significant impact on lignin content of downregulated lines. Stem carbohydrate composition analysis revealed a decrease in glucose, mannose, arabinose, and galactose, but an increase in xylose in the over-expression lines. Transcriptome analysis revealed upregulation of several downstream transcription factors and secondary cell wall related structural genes in the PdWND1B over-expression lines, partly explaining the observed phenotypic changes in cell wall chemistry. Relative to the control, glucose release efficiency and ethanol production from stem biomass was significantly reduced in over-expression lines. Our results show that PdWND1B is an important factor determining biomass productivity, cell wall chemistry and its conversion to biofuels in Populus.

09 BIOMASS FUELS↗
DOE OSTI2022

Effects of Cone Penetrometer Testing on Shallow Hydrogeology at a Contaminated Site

Penetration testing is a popular and instantaneous technique for subsurface mapping, contaminant tracking, and the determination of soil characteristics. While the small footprint and reproducibility of cone penetrometer testing makes it an ideal method for in-situ subsurface investigations at contaminated sites, the effects to local shallow groundwater wells and measurable influence on monitoring networks common at contaminated sites is unknown. Physical and geochemical parameters associated with cone penetrometer testing were measured from a transect of shallow groundwater monitoring wells adjacent to penetrometer testing. For wells screened above the depth of cone refusal, the physical advancement and retraction of the cone had a significant effect ( p < 0.01) on water level for several pushes within 10 meters of a monitoring well, and a measured increase in specific conductivity. No effect on geochemistry or water level was observed in continuous monitoring data from wells screened below the depth of cone refusal, but variability in specific conductivity from these wells during penetration testing was only a fraction of the natural variation measured during precipitation events. Continuous measurements of specific conductivity and water level demonstrated that the effects of penetration testing have limited spatial and temporal distributions with a null effect post-testing.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2020

Performance and community structure dynamics of microbial electrolysis cells operated on multiple complex feedstocks

Microbial electrolysis is a promising technology for converting aqueous wastes into hydrogen. However, substrate adaptability is an important feature, seldom documented in microbial electrolysis cells (MECs). In addition, the correlation between substrate composition and community structure has not been well established. This study used an MEC capable of producing over 10 L/L-day of hydrogen from a switchgrass-derived bio-oil aqueous phase and investigated four additional substrates, tested in sequence on a mature biofilm. The additional substrates included a red oak-derived bio-oil aqueous phase, a corn stover fermentation product, a mixture of phenol and acetate, and acetate alone. The MECs fed with the corn stover fermentation product resulted in the highest performance among the complex feedstocks, producing an average current density of 7.3 ± 0.51 A/m 2 , although the acetate fed MECs outperformed complex substrates, producing 12.3 ± 0.01 A/m 2 . 16S rRNA gene sequencing showed that community structure and community diversity were not predictive of performance, and replicate community structures diverged despite identical inoculum and enrichment procedure. The trends in each replicate, however, were indicative of the influence of the substrates. Geobacter was the most dominant genus across most of the samples tested, but its abundance did not correlate strongly to current density. High-performance liquid chromatography (HPLC) showed that acetic acid accumulated during open circuit conditions when MECs were fed with complex feedstocks and was quickly degraded once closed circuit conditions were applied. The largest net acetic acid removal rate occurred when MECs were fed with red oak bio-oil aqueous phase, consuming 2.93 ± 0.00 g/L-day. Principal component analysis found that MEC performance metrics such as current density, hydrogen productivity, and chemical oxygen demand removal were closely correlated. Net acetic acid removal was also found to correlate with performance. However, no bacterial genus appeared to correlated to these performance metrics strongly, and the analysis suggested that less than 70% of the variance was accounted for by the two components. This study demonstrates the robustness of microbial communities to adapt to a range of feedstocks and conditions without relying on specific species, delivering high hydrogen productivities despite differences in community structure. The results indicate that functional adaptation may play a larger role in performance than community composition. Further investigation of the roles each microbe plays in these communities will help MECs to become integral in the 21st-century bioeconomy to produce zero-emission fuels.

09 BIOMASS FUELS↗
DOE OSTI2020

Microbial electrolysis cell recovery after inducing operational failure conditions

Microbial Electrolysis Cells (MECs) are often documented for their ability to produce hydrogen through new sources and new configurations. However, few studies attempt to document the recovery of the bioanode after being exposed to operational failure conditions. This study attempted to compare the behavior and recovery of MECs after being exposed to four operational failure conditions: acidification, aeration, osmotic shock, and voltage reversal. For each failure condition, three time points were tested: before failure, immediately after failure, and after recovery efforts. Of the modes tested, acidification caused the largest loss in performance and operational efficiency, while osmotic shock caused the second largest loss in performance. Aeration and voltage reversal caused negligible losses in performance immediately after failure conditions and after the recovery period. Aeration and voltage reversal had a minimal effect on the removal of individual compounds. However, acetate accumulated after acidification, and propionic acid accumulated after osmotic shock. Furthermore these findings could be useful for determining if failure will require more significant repair efforts in commercially deployed devices. While acidification created permanent losses to performance, if device failures like aeration, osmotic shock, and voltage reversal are caught early (less than 15 min), full recovery without bioanode replacement is likely.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2020

Gene targets for engineering osmotolerance in Caldicellulosiruptor bescii

Abstract Background Caldicellulosiruptor bescii , a promising biocatalyst being developed for use in consolidated bioprocessing of lignocellulosic materials to ethanol, grows poorly and has reduced conversion at elevated medium osmolarities. Increasing tolerance to elevated fermentation osmolarities is desired to enable performance necessary of a consolidated bioprocessing (CBP) biocatalyst. Results Two strains of C. bescii showing growth phenotypes in elevated osmolarity conditions were identified. The first strain, ORCB001, carried a deletion of the FapR fatty acid biosynthesis and malonyl-CoA metabolism repressor and had a severe growth defect when grown in high-osmolarity conditions—introduced as the addition of either ethanol, NaCl, glycerol, or glucose to growth media. The second strain, ORCB002, displayed a growth rate over three times higher than its genetic parent when grown in high-osmolarity medium. Unexpectedly, a genetic complement ORCB002 exhibited improved growth, failing to revert the observed phenotype, and suggesting that mutations other than the deleted transcription factor (the fruR/cra gene) are responsible for the growth phenotype observed in ORCB002. Genome resequencing identified several other genomic alterations (three deleted regions, three substitution mutations, one silent mutation, and one frameshift mutation), which may be responsible for the observed increase in osmolarity tolerance in the fruR / cra -deficient strain, including a substitution mutation in dnaK , a gene previously implicated in osmoresistance in bacteria. Differential expression analysis and transcription factor binding site inference indicates that FapR negatively regulates malonyl-CoA and fatty acid biosynthesis, as it does in many other bacteria. FruR/Cra regulates neighboring fructose metabolism genes, as well as other genes in global manner. Conclusions Two systems able to effect tolerance to elevated osmolarities in C. bescii are identified. The first is fatty acid biosynthesis. The other is likely the result of one or more unintended, secondary mutations present in another transcription factor deletion strain. Though the locus/loci and mechanism(s) responsible remain unknown, candidate mutations are identified, including a mutation in the dnaK chaperone coding sequence. These results illustrate both the promise of targeted regulatory manipulation for osmotolerance (in the case of fapR ) and the challenges (in the case of fruR/cra ).

59 BASIC BIOLOGICAL SCIENCES↗