Capturing Plant Metabolome with Direct-Immersion in Vivo Solid Phase Microextraction of Plant Tissues

This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://pubs.acs.org/doi/abs/10.1021/acs.analchem.5b03684 .For the first time, an in vivo sampling mode of direct immersion–solid phase microextraction (DI-SPME) was employed to capture the metabolome of living plant specimens, using apple (Malus × domestica Borkh.) as a model system. Metabolites were extracted from apple tissues and introduced by thermal desorption into a comprehensive two-dimensional gas chromatography–time-of-flight mass spectrometry instrument. The feasibility of this sampling approach, based on exploitation of microextraction principles, including negligible depletion of free analyte concentrations, solventless sampling and sample preparation, and on-site compatibility, was determined in global metabolite analysis. Rather than adopting an approach of traditional sample preparation, requiring metabolism quenching and laborious sample preparation, the objective of the study was to capture the metabolome in vivo, evaluate the feasibility of the approach to provide unbiased extraction coverage, and compare analytical precision when different SPME sampling modes are employed. The potential of in vivo DI-SPME in quantitative plant metabolomics was assessed by evaluating changes in metabolic fingerprints in response to fruit maturation. The in vivo SPME sampling approach has been demonstrated as capable of sampling living systems with high reproducibility, considering that nearly 50% of hundreds of evaluated compounds included in the determination of analytical performance met the 15% RSD FDA criterion. Esters were extracted with high repeatability (% RSD for hexyl butanoate and butyl butanoate of 16.5 and 5.9, respectively, from 9 determinations in 3 apples) and found to be upregulated in response to apple fruit maturation.Natural Sciences and Engineering Research Council of Canada (NSERC

Combining Metabolic Engineering and Electrocatalysis: Application to the Production of Polyamides from Sugar

Biorefineries aim to convert biomass to a spectrum of products ranging from biofuels to specialty chemicals. To achieve economically sustainable conversion it is crucial to streamline the catalytic and downstream processing steps. Here we report a route that integrates bio- and chemical catalysis to convert glucose into bio-based unsaturated nylon 6,6. An engineered strain of Saccharomyces cerevisiae, with the highest reported muconic acid titer of 559.5 mg L-1 in yeast, was used as the initial biocatalyst to convert glucose into muconic acid. Without any separation, muconic acid was further electrocatalytically hydrogenated to 3-hexenedioic acid with 94% yield, despite the presence of all the biogenic impurities. Bio-based unsaturated nylon 6,6 (unsaturated polyamide 6,6) was finally obtained by polymerization of 3-hexenedioic acid with hexamethylenediamine, demonstrating the integrated design of bio-based polyamides from glucose

Kepler-4B: A Hot Neptune-Like Planet of A G0 Star Near Main-Sequence Turnoff

Early time-series photometry from NASA's Kepler spacecraft has revealed a planet transiting the star we term Kepler-4, at R.A. = 19(h)02(m)27.(s)68, delta = +50 degrees 08'08 '' 7. The planet has an orbital period of 3.213 days and shows transits with a relative depth of 0.87 x 10(-3) and a duration of about 3.95 hr. Radial velocity (RV) measurements from the Keck High Resolution Echelle Spectrometer show a reflex Doppler signal of 9.3(-1.9)(+1.1) m s(-1), consistent with a low-eccentricity orbit with the phase expected from the transits. Various tests show no evidence for any companion star near enough to affect the light curve or the RVs for this system. From a transit-based estimate of the host star's mean density, combined with analysis of high-resolution spectra, we infer that the host star is near turnoff from the main sequence, with estimated mass and radius of 1.223(-0.091)(+0.053) M(circle dot) and 1.487(-0.084)(+0.071) R(circle dot).We estimate the planet mass and radius to be {M(P), R(P)} = {24.5 +/- 3.8 M(circle plus), 3.99 +/- 0.21 R(circle plus)}. The planet's density is near 1.9 g cm(-3); it is thus slightly denser and more massive than Neptune, but about the same size.W. M. Keck FoundationNASA's Science Mission DirectorateAstronom

Transit Timing Observations from Kepler: VII. Confirmation of 27 planets in 13 multiplanet systems via Transit Timing Variations and orbital stability

We confirm 27 planets in 13 planetary systems by showing the existence of statistically significant anti-correlated transit timing variations (TTVs), which demonstrates that the planet candidates are in the same system, and long-term dynamical stability, which places limits on the masses of the candidates---showing that they are planetary. %This overall method of planet confirmation was first applied to \kepler systems 23 through 32. All of these newly confirmed planetary systems have orbital periods that place them near first-order mean motion resonances (MMRs), including 6 systems near the 2:1 MMR, 5 near 3:2, and one each near 4:3, 5:4, and 6:5. In addition, several unconfirmed planet candidates exist in some systems (that cannot be confirmed with this method at this time). A few of these candidates would also be near first order MMRs with either the confirmed planets or with other candidates. One system of particular interest, Kepler-56 (KOI-1241), is a pair of planets orbiting a 12th magnitude, giant star with radius over three times that of the Sun and effective temperature of 4900 K---among the largest stars known to host a transiting exoplanetary system.Comment: 12 pages, 13 figures, 5 tables. Submitted to MNRA

Kepler Observations of Transiting Hot Compact Objects

Kepler photometry has revealed two unusual transiting companions orbiting an early A-star and a late B-star. In both cases the occultation of the companion is deeper than the transit. The occultation and transit with follow-up optical spectroscopy reveal a 9400 K early A-star, KOI-74 (KIC 6889235), with a companion in a 5.2 day orbit with a radius of 0.08 Rsun and a 10000 K late B-star KOI-81 (KIC 8823868) that has a companion in a 24 day orbit with a radius of 0.2 Rsun. We infer a temperature of 12250 K for KOI-74b and 13500 K for KOI-81b. We present 43 days of high duty cycle, 30 minute cadence photometry, with models demonstrating the intriguing properties of these object, and speculate on their nature.Comment: 12 pages, 3 figures, submitted to ApJL (updated to correct KOI74 lightcurve

Persistent starspot signals on M dwarfs: multi-wavelength Doppler observations with the Habitable-zone Planet Finder and Keck/HIRES

Young, rapidly-rotating M dwarfs exhibit prominent starspots, which create quasiperiodic signals in their photometric and Doppler spectroscopic measurements. The periodic Doppler signals can mimic radial velocity (RV) changes expected from orbiting exoplanets. Exoplanets can be distinguished from activity-induced false positives by the chromaticity and long-term incoherence of starspot signals, but these qualities are poorly constrained for fully-convective M stars. Coherent photometric starspot signals on M dwarfs may persist for hundreds of rotations, and the wavelength dependence of starspot RV signals may not be consistent between stars due to differences in their magnetic fields and active regions. We obtained precise multi-wavelength RVs of four rapidly-rotating M dwarfs (AD Leo, G 227-22, GJ 1245B, GJ 3959) using the near-infrared (NIR) Habitable-zone Planet Finder, and the optical Keck/HIRES spectrometer. Our RVs are complemented by photometry from Kepler, TESS, and the Las Cumbres Observatory (LCO) network of telescopes. We found that all four stars exhibit large spot-induced Doppler signals at their rotation periods, and investigated the longevity and optical-to-NIR chromaticity for these signals. The phase curves remain coherent much longer than is typical for Sunlike stars. Their chromaticity varies, and one star (GJ 3959) exhibits optical and NIR RV modulation consistent in both phase and amplitude. In general, though, we find that the NIR amplitudes are lower than their optical counterparts. We conclude that starspot modulation for rapidly-rotating M stars frequently remains coherent for hundreds of stellar rotations, and gives rise to Doppler signals that, due to this coherence, may be mistaken for exoplanets.Comment: Accepted for publication in the Astrophysical Journa