Distribution, frequency and molecular basis of clethodim and quizalofop resistance in brome grass (Bromus diandrus).

BACKGROUND: Brome grass (Bromus diandrus Roth) is prevalent in the southern and western cropping regions of Australia, where it causes significant economic damage. A targeted herbicide resistance survey was conducted in 2020 by collecting brome grass populations from 40 farms in Western Australia and subjecting these samples to comprehensive herbicide screening. One sample (population 172-20), from a field that had received 12 applications of clethodim over 20 years of continuous cropping, was found to be highly resistant to the acetyl-CoA carboxylase (ACCase)-inhibiting herbicides clethodim and quizalofop, and so the molecular basis of resistance was investigated. RESULTS: All 31 individuals examined from population 172-20 carried the same resistance-endowing point mutation causing an aspartate-to-glycine substitution at position 2078 in the translated ACCase protein sequence. A wild-type susceptible population and the resistant population had similar expression levels of plastidic ACCase genes. The level of resistance to quizalofop, either standalone or in mixture with clethodim, in population 172-20 was lower under cooler growing conditions. CONCLUSION: Target-site resistance to ACCase-inhibiting herbicides, conferred by one ACCase mutation, was selected in all tested brome plants infesting a field with a history of repeated clethodim use. This mutation appears to have been fixed in the infesting population. Notably, clethodim resistance in this population was not detected by the farmer, and a high future incidence of quizalofop resistance is anticipated. Herbicide resistance testing is essential for the detection of evolving weed resistance issues and to inform effective management strategies. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Effect of Cobalt Chloride on Fermentation of Alfalfa and Smooth Bromegrass Hays by Horse Cecal Microorganisms.

Effects of Co on fiber digestibility in horses are largely unknown. Our objective was to evaluate effects of Co chloride on in vitro gas production, VFA production, and dry matter (DM) disappearance (IVDMD) using cecal fluid from 4 cannulated Quarter Horses. Five grams DM of alfalfa or smooth bromegrass hay were provided as substrate with Co added at 0.0, 0.5, 5.0, 25.0, or 50.0 mg/kg substrate DM. Ten milliliters cecal fluid and 140 mL McDougall's buffer were incubated in duplicate, and pH, VFA concentrations, and IVDMD were measured after 48 hours. Gas production, used as an indicator of fermentative activity, was recorded every 15 minutes. Terminal pH was not affected by Co or forage type (P > .19). There was greater IVDMD in cultures containing alfalfa (29.2%) compared with smooth bromegrass hay (19.4%; P < .01). There was a forage × Co interaction (P < .05) in which gas production was greater in cultures containing alfalfa, and 5 mg Co/kg substrate DM led to greater gas production than 50 mg Co/kg substrate DM in cultures containing brome (P < .05). Gas production was affected by forage, Co, and time (P < .01). Production of individual and total VFA, as well as acetate:propionate (A:P) ratio were increased in cultures containing alfalfa (P < .05); however, there were no effects of Co on VFA concentrations (P > .05). While gas production was influenced by Co in cultures containing brome, no effects of Co were observed for pH, VFA, or IVDMD.

Biochemical and genetic polymorphism of Bromopsis inermis populations under chronic radiation exposure.

MAIN CONCLUSION: For the subsequent assessment of the genetic mechanisms responsible for the resistance of plants to chronic irradiation, the analysis of RAPD-cDNA with the subsequent isolation, cloning, and sequencing of expressed polymorphic sequences is a promising technique. A study was conducted on Bromopsis inermis populations that have been growing for a long time in the EURT area. Using RAPD primers, we studied the genetic spectra of plants. In analysing the UPGMA algorithm, we identified two well-distinguishable clusters with a high level of bootstrap support (> 85%): background samples hit the first, and impact samples hit the second. Our data indicate a decrease in diversity in the most polluted population, as well as the appearance of new alleles in chronically irradiated samples of the B. inermis. Smooth brome seedlings were characterised by the content of anthocyanins, comparable with other types of cereals. In the gradient of chronic irradiation, the relative content of anthocyanins was not significantly changed. For the first time, the partial nucleotide sequences of the key genes of anthocyanin biosynthesis (Chi and F3h) in the brome were determined, these sequences were found to be 191 and 356 bp in length, respectively, and were cloned and sequenced. Three copies of the Chi gene were identified in the B. inermis genome. One copy (BiChi-1) clustered with the sequences of the Aegilops tauschii gene (D genome), and the other two copies (BiChi-2 and BiChi-3) formed a separate cluster in the Pooideae subfamily adjacent to Hordeum vulgare. In the copy of BiChi-1, a complete deletion of intron 1 was detected. For the F3h gene, one copy of the B. inermis gene was obtained, which forms a separate branch in the subfamily Pooideae.

Posttranslational mutagenesis: A chemical strategy for exploring protein side-chain diversity.

Posttranslational modification of proteins expands their structural and functional capabilities beyond those directly specified by the genetic code. However, the vast diversity of chemically plausible (including unnatural but functionally relevant) side chains is not readily accessible. We describe C (sp3)-C (sp3) bond-forming reactions on proteins under biocompatible conditions, which exploit unusual carbon free-radical chemistry, and use them to form Cß-Cγ bonds with altered side chains. We demonstrate how these transformations enable a wide diversity of natural, unnatural, posttranslationally modified (methylated, glycosylated, phosphorylated, hydroxylated), and labeled (fluorinated, isotopically labeled) side chains to be added to a common, readily accessible dehydroalanine precursor in a range of representative protein types and scaffolds. This approach, outside of the rigid constraints of the ribosome and enzymatic processing, may be modified more generally for access to diverse proteins.

Polycyclic aromatic hydrocarbons in biomass-burning emissions and their contribution to light absorption and aerosol toxicity.

In recent years, brown carbon (BrC) has been shown to be an important contributor to light absorption by biomass-burning atmospheric aerosols in the blue and near-ultraviolet (UV) part of the solar spectrum. Emission factors and optical properties of 113 polycyclic aromatic hydrocarbons (PAHs) were determined for combustion of five globally important fuels: Alaskan, Siberian, and Florida swamp peat, cheatgrass (Bromus tectorum), and ponderosa pine (Pinus ponderosa) needles. The emission factors of total analyzed PAHs were between 1.9±0.43.0±0.6 and 9.6±1.2-42.2±5.4mgPAHkg(-1)fuel for particle- and gas phase, respectively. Spectrophotometric analysis of the identified PAHs showed that perinaphthenone, methylpyrenes, and pyrene contributed the most to the total PAH light absorption with 17.2%, 3.3 to 10.5%, and 7.6% of the total particle-phase PAH absorptivity averaged over analyzed emissions from the fuels. In the gas phase, the top three PAH contributors to BrC were acenaphthylene (32.6%), anthracene (8.2%), and 2,4,5-trimethylnaphthalene (8.0%). Overall, the identified PAHs were responsible for 0.087-0.16% (0.13% on average) and 0.033-0.15% (0.11% on average) of the total light absorption by dichloromethane-acetone extracts of particle and gas emissions, respectively. Toxic equivalency factor (TEF) analysis of 16 PAHs prioritized by the United States Environmental Protection Agency (EPA) showed that benzo(a)pyrene contributed the most to the PAH carcinogenic potency of particle phase emissions (61.8-67.4% to the total carcinogenic potency of Σ16EPA PAHs), while naphthalene played the major role in carcinogenicity of the gas phase PAHs in the biomass-burning emission analyzed here (35.4-46.0% to the total carcinogenic potency of Σ16EPA PAHs). The 16 EPA-prioritized PAHs contributed only 22.1±6.2% to total particle and 23.4±11% to total gas phase PAH mass, thus toxic properties of biomass-burning PAH emissions are most likely underestimated.

Versatile (bio)functionalization of bromo-terminated phosphonate-modified porous aluminum oxide.

Porous aluminum oxide (PAO) is a nanoporous material used for various (bio)technological applications, and tailoring its surface properties via covalent modification is a way to expand and refine its application. Specific and complex chemical modification of the PAO surface requires a stepwise approach in which a secondary reaction on a stable initial modification is necessary to achieve the desired terminal molecular architecture and reactivity. We here show that the straightforward initial modification of the bare PAO surface with bromo-terminated phosphonic acid allows for the subsequent preparation of PAO with a wide scope of terminal reactive groups, making it suitable for (bio)functionalization. Starting from the initial bromo-terminated PAO, we prepared PAO surfaces presenting various terminal functional groups, such as azide, alkyne, alkene, thiol, isothiocyanate, and N-hydroxysuccinimide (NHS). We also show that this wide scope of easily accessible tailored reactive PAO surfaces can be used for subsequent modification with (bio)molecules, including carbohydrate derivatives and fluorescently labeled proteins.

From lows to highs: using low-resolution models to phase X-ray data.

The study of virus structures has contributed to methodological advances in structural biology that are generally applicable (molecular replacement and noncrystallographic symmetry are just two of the best known examples). Moreover, structural virology has been instrumental in forging the more general concept of exploiting phase information derived from multiple structural techniques. This hybridization of structural methods, primarily electron microscopy (EM) and X-ray crystallography, but also small-angle X-ray scattering (SAXS) and nuclear magnetic resonance (NMR) spectroscopy, is central to integrative structural biology. Here, the interplay of X-ray crystallography and EM is illustrated through the example of the structural determination of the marine lipid-containing bacteriophage PM2. Molecular replacement starting from an ~13 Å cryo-EM reconstruction, followed by cycling density averaging, phase extension and solvent flattening, gave the X-ray structure of the intact virus at 7 Å resolution This in turn served as a bridge to phase, to 2.5 Å resolution, data from twinned crystals of the major coat protein (P2), ultimately yielding a quasi-atomic model of the particle, which provided significant insights into virus evolution and viral membrane biogenesis.

Dietary sulfur concentration affects rumen hydrogen sulfide concentrations in feedlot steers during transition and finishing.

Angus steers (n = 96; 321 ± 29 kg BW) were used to determine how previous exposure to increased dietary S would affect ruminal hydrogen sulfide concentrations ([H(2)S]) in the feedlot, to investigate the effects of dietary S on ruminal [H(2)S] during transition and finishing, and to determine if dietary S affects the glutathione status of finishing cattle. Steers were strip-grazed on smooth bromegrass (Bromus inermis L.) over a 35 d period and received a dry distillers grains plus solubles (DDGS) supplement at 1% of BW (DM basis) that contained either 0.50% S (LS; n = 4 plots) or the DDGS supplement with an additional 0.30% S from sodium sulfate (0.80% S in supplement; HS; n = 4 plots). On d 36 steers were moved from the pastures to feedlot pens with one-half of the steers on each treatment in the pasture period remaining on the same treatment during the feedlot period and half being switched to the other treatment (n = 6 pens). For the first 10 d in the feedlot, steers were fed hay ad libitum and 1% BW of the DDGS supplement representing their new treatment, followed by transition to finishing diets. Dietary S of transition and finishing diets were 0.2% to 0.3% S for LS and 0.5% to 0.6% S for HS. No interaction between pasture and feedlot treatment was observed (P ≥ 0.50), so data for the feedlot period were pooled by feedlot treatment (n = 12 pens). Rumen [H(2)S] were measured on d 35 of the pasture period and on d 46 while receiving ad libitum hay and supplement at 6 h after the feeding of the supplement and after 7 d on each of the 3 transition diets (d 53, 60, and 67) and on d 93, 126, and 155 of the study after receiving the finishing diet for 26, 59, and 88 d at 6 h after feeding. Ruminal [H(2)S] did not differ between treatment while steers were fed the supplement on forage-based diets. However, ruminal [H(2)S] of HS-fed steers was greater (P < 0.05) than LS-fed steers when transition diets and the finishing diets were fed. Relative to S intake, ruminal [H(2)S] increased disproportionally after 26 d on the finishing diet. This was followed by a decrease in [H(2)S] on d 59 of finishing, although S intake was increased (P < 0.05) compared with d 26 of finishing. It appears that factors other than S intake alone contribute to ruminal [H(2)S]. The amount of glutathione in the liver of steers did not differ (P = 0.47) because of dietary S, but the concentration of oxidized glutathione increased (P = 0.03) in HS-fed compared with LS-fed steers, suggesting that the potential for oxidative stress in cattle fed high-S diets may warrant further investigation.

Functional characterization of a sucrose:fructan 6-fructosyltransferase of the cold-resistant grass Bromus pictus by heterelogous expression in Pichia pastoris and Nicotiana tabacum and its involvement in freezing tolerance.

We have previously reported the molecular characterization of a putative sucrose:fructan 6-fructosyltransferase (6-SFT) of Bromus pictus, a graminean species from Patagonia, tolerant to cold and drought. Here, this enzyme was functionally characterized by heterologous expression in Pichia pastoris and Nicotiana tabacum. Recombinant P. pastoris Bp6-SFT showed comparable characteristics to barley 6-SFT and an evident fructosyltransferase activity synthesizing bifurcose from sucrose and 1-kestotriose. Transgenic tobacco plants expressing Bp6-SFT, showed fructosyltransferase activity and fructan accumulation in leaves. Bp6-SFT plants exposed to freezing conditions showed a significantly lower electrolyte leakage in leaves compared to control plants, indicating less membrane damage. Concomitantly these transgenic plants resumed growth more rapidly than control ones. These results indicate that Bp6-SFT transgenic tobacco plants that accumulate fructan showed enhanced freezing tolerance compared to control plants.

Karyotype and nuclear DNA content of hexa-, octo-, and duodecaploid lines of Bromus subgen. Ceratochloa

The subgenus Ceratochloa of the genus Bromus includes a number of closely related allopolyploid forms or species that present a difficult taxonomic problem. The present work combines data concerning chromosome length, heterochromatin distribution and nuclear genome size of different 6x, 8x and 12x accessions in this subgenus. Special attention is paid to the karyotype structure and genomic constitution of duodecaploid plants recently found in South America. Hexaploid lineages possess six almost indistinguishable genomes and a nuclear DNA content between 12.72 pg and 15.10 pg (mean 1Cx value = 2.32 pg), whereas octoploid lineages contain the same six genomes (AABBCC) plus two that are characterized by longer chromosomes and a greater DNA content (1Cx = 4.47 pg). Two duodecaploid accessions found in South America resemble each other and apparently differ from the North American duodecaploid B. arizonicus as regards chromosome size and nuclear DNA content (40.00 and 40.50 pg vs. 27.59 pg). These observations suggest that the South American duodecaploids represent a separate evolutionary lineage of the B. subgenus Ceratochloa, unrecognized heretofore.

Effects of dried distillers grains and equivalent undegradable intake protein or ether extract on performance and forage intake of heifers grazing smooth bromegrass pastures.

Crossbred heifers (n = 120; BW = 368 kg, SD = 39 kg) were used to determine effects of dried distillers grains (DDG) and relative contributions of undegradable intake protein (UIP) and fat (ether extract, EE) in DDG on ADG and forage intake (FI). Heifers rotationally grazed six 3.5-ha, smooth bromegrass paddocks (IVDMD = 65.7%, CP = 20.8%, UIP = 2.17%, DM basis). Heifers were blocked by previous ADG and allotted to treatments in a 3 x 3 + 1 factorial design. Factors were source and level of supplementation. Supplements were as follows: 1) DDG (UIP = 15.8%, EE = 9.67%), 2) corn gluten meal (CGM; UIP = 31.6%, EE = 0.83%), or 3) corn oil (OIL; UIP = 0.74%, EE = 19.3%). Amounts of DDG were 750, 1,500, or 2,250 g/d, whereas amounts of CGM and OIL were 375, 750, or 1,125 g/ d. Supplements containing CGM and OIL were fed in amounts that provided UIP and EE, respectively, equivalent to those of the DDG. Contrasts of interest were DDG vs. CGM and DDG vs. OIL. Control heifers were fed 250 g/d of a supplement containing corn bran and molasses (UIP = 0.92%, EE = 1.13%). Heifers were supplemented individually. Treatments were separated by regressing the response variables on grams of nutrient (DM, UIP, or EE) intake per kilogram of BW, because not all heifers consumed their allotment of supplement. Supplemental DDG resulted in a linear increase in ADG (P < 0.01), whereas CGM tended to increase ADG (P = 0.14) but at a rate that was 39% of that for DDG, representing a response to MP. Supplementation of OIL did not affect ADG (P = 0.25) and tended to result in ADG less than that of DDG (P = 0.09). Supplementation with DDG had no effect (P = 0.63) on FI when predicted by the use of chromic oxide but tended (P = 0.07) to decrease FI when it was predicted from ADG using NE equations. Despite the differences between methods in the significance of the effect of DDG, the rates of substitution agreed (-0.50 and -0.45 for chromic oxide and NE equations, respectively), suggesting that the chromic oxide method was less sensitive in assessing FI. Supplementation with CGM decreased FI (P < 0.01), but FI for CGM did not differ from that of DDG when the chromic oxide method was used (P = 0.19). Corn oil had no effect on FI (P = 0.42). Increased ADG and decreased FI observed from DDG supplementation is not independently explained by UIP or EE contained in DDG.

Xenopus Xp54 and human RCK/p54 helicases functionally replace yeast Dhh1p in brome mosaic virus RNA replication.

By using a Brome mosaic virus (BMV)-Saccharomyces cerevisiae system, we previously showed that the cellular Lsm1p-7p/Pat1p/Dhh1p decapping-activator complex functions in BMV RNA translation and replication. As a first approach in investigating whether the corresponding human homologues play a similar role, we expressed human Lsm1p (hLsm1p) and RCK/p54 in yeast. Expression of RCK/p54 but not hLsm1p restored the defect in BMV RNA translation and replication observed in the dhh1Delta and lsm1Delta strains, respectively. This functional conservation, together with the common replication strategies of positive-stranded RNA viruses, suggests that RCK/p54 may also play a role in the replication of positive-stranded RNA viruses that infect humans.

Identification and immunogold localization of a novel bromegrass (Bromus inermis Leyss) peroxisome channel protein induced by ABA, cold and drought stresses, and late embryogenesis.

A cDNA (BG-15) was isolated through differential screening of a cDNA library made from an ABA-treated bromegrass (Bromus inermis Leyss) suspension cell culture. The 819 bp pair cDNA encoded a 174 amino acid polypeptide with a calculated molecular mass of 18.08 kD and isolectric point of 7.50. The deduced amino acid sequences for the cDNA were 29.5% and 32.6% homologous to the known amino acid-selective channel proteins of the chloroplastic outer membrane in pea and barley, but were highly homologous (55.6% to 83.2%) to the putative membrane channel proteins from rice and Arabidopsis. Immunogold localization demonstrated that the channel protein encoded by this cDNA was present on the peroxisome membrane. High stringency southern analysis revealed that 1 to 2 copies of the peroxisomal channel protein (PCP) genes were present in the bromegrass genome. Northern and Western blots revealed that the PCP gene was responsive to both cold and drought stresses, and was rapidly induced by ABA (75 microM). The transcript of the PCP gene also accumulated during late embryogenesis, but declined rapidly during germination. Data taken together, responsiveness of the PCP to cold and drought stresses, and accumulation during late embryogenesis suggest this novel peroxisomal channel protein is associated with sugar and fatty acid metabolism through fatty acid import or succinate export from peroxisome during desiccation tolerance and energy metabolism.

Synthesis and analytical characteristics of new progesterone skeleton haptens.

This article describes the synthesis of new conjugated progesterone-bovine serum albumin substances, used to obtain specific antisera toward this hormone and for progesterone assay kits. In general, these kits are designed to be used on the farm or at the local veterinary clinics.

Quantitative assessment of the effects of agricultural practices designed to reduce 137Cs and 90Sr soil-plant transfer in meadows.

Agricultural practices (ploughing and reseeding, addition of lime and fertiliser) were tested as a feasible remediation strategy to reduce 137Cs (RCs) and 90Sr (RSr) soil-plant transfer in natural meadows in areas affected by the Chernobyl fallout. Field experiments were carried out for 2 years at six sites, covering dry and wet meadows. Observed results at field scale showed that ploughing plus reseeding provoked the main reduction in RSr transfer, with no further reduction after liming, while ploughing + reseeding + K fertiliser led to the maximum decrease in RCs transfer at most sites. The direct effects of agricultural practices on the exchange complex and soil solution composition were quantified by subsequent soil analyses. At the doses applied, lime did not affect the Ca + Mg concentrations in the exchange complex and soil solution of the ploughed soils, thus suggesting that the decrease in RSr transfer on treated plots was mainly due to the changes in the plant species after reseeding. With respect to RCs, changes in the K+NH4+ concentrations in the exchange complex and soil solution were consistent with changes in soil-plant transfer. Finally, RSr and RCs soil-plant transfer in ploughed plots was well predicted from soil properties, such as the solid-liquid distribution coefficient, the ionic composition of the soil solution and the exchangeable cations, with Pearson correlation coefficients of 0.98 and 0.86, respectively, between calculated and experimental log transfer factors.

A lipid transfer protein gene BG-14 is differentially regulated by abiotic stress, ABA, anisomycin, and sphingosine in bromegrass (Bromus inermis).

The objective was to investigate the expression of a lipid transfer protein gene (LTP) both in bromegrass (Bromus inermis) cells and seedlings after exposure to abiotic stresses, abscisic acid (ABA), anisomycin, and sphingosine. A full-length cDNA clone BG-14 isolated from bromegrass suspension cell culture encodes a polypeptide of 124 amino acids with typical LTP characteristics, such as a conserved arrangement of cysteine residues. During active stages of cold acclimation LTP expression was up-regulated, whereas at the final stage of cold acclimation LTP transcript level declined to pre-acclimation level. A severe drought stress induced the LTP gene; yet, LTP expression doubled 3 d after re-hydration. Both temperature and heat shock duration influence LTP induction; however temperature is the primary factor. Treatment with NaCl stimulated accumulation of LTP mRNA within 15 min and the transcripts remained at elevated levels for the duration of the salinity stress. Most interestingly, Northern blots showed LTP was rapidly induced not only by ABA, but also by anisomycin and sphingosine in suspension cell cultures. Of the three chemicals, ABA induced the most rapid and highest response in LTP expression as well as highest freezing tolerance, whereas sphingosine was the least active for both LTP expression and freezing tolerance.

Bromus fasciculatus presl--a third diploid progenitor of Bromus section Genea allopolyploids (Poaceae).

Allozyme variation of ten heterozymes of seven enzymes among five accessions of a rare diploid Bromus fasciculatus was analysed with the use of PAGE and compared with that for six other species of section Genea of the genus Bromus. Allozymes charasteristic for diploids B. tectorum and B. fasciculatus are combined in fixed heterozygous phenotypes of tetraploid B. rubens. Fixed heterozygous phenotypes of tetraploid B. madritensis combine one allozyme of B. fasciculatus with another of diploid B. sterilis at each of the loci studied. Of the three diploids studied, only B. sterilis fits well for a role of a genome donor for the polyploid B. diandrus-rigidus complex. Bromus fasciculatus thus appears to be a diploid ancestor for the two tetraploids of section Genea, B. rubens and B. madritensis.