The essential oil of Kochia scoparia (L.) Schrad. as a potential repellent against stored-product insects.

Chemical composition of the essential oil from Kochia scoparia (L.) Schrad. (syn. Bassia scoparia (L.) A. J. Scott) was analyzed in quality and quantity by GC-MS and GC-FID. Repellent activities of the essential oil from K. scoparia (KSEO) were evaluated against two common species of stored-product insects Tribolium castaneum Herbst and Liposcelis bostrychophila Badonnel. Results indicated that KSEO mainly consisted of eugenol, ß-caryophyllene, and α-humulene, accounting for 75.6%, 8.2%, and 1.4% of the total oil, respectively. KSEO and the three major components were repellent to T. castaneum and L. bostrychophila adults. Notably, KSEO exerted significant effects, comparable to the positive control DEET at 2 and 4 h post-exposure. Eugenol at 63.17-2.53 nL/cm2 exhibited high percentage repellency ranging from 96 to 70% against L. bostrychophila during 4-h exposure. To gain further insights into the repellent activity, molecular docking simulation was performed with eugenol as the ligand and an odorant binding protein TcOBPC12 (gene: TcOBP10B) from the model insect T. castaneum as the receptor. Docking calculation results revealed that TcOBPC12 had binding affinity to eugenol (△G = - 4.52 kcal/mol) along with a hydrogen bond of 0.18 nm (1.8 Å) long forming between them, which could be an important target protein associated with identifying volatile repellent molecules. This work highlights the promising potential of KSEO as a botanical repellent for controlling stored-product insects.

Mitigation of salinity stress effects on kochia (Bassia scoparia L.) biomass productivity using biochar application.

Salinity continues to be a key factor limiting food security and agricultural sustainability in arid and semi-arid countries. Biochar has been promoted to reduce the risk of saline irrigation. In a controlled study, physiological and morphological growth factors of kochia plants that were irrigated with tap water (S1) and saline water (S2) were assessed to identify the ameliorative effects of biochar amendment to the soil at different levels (B1: 0%, B2: 2%, B3: 5%, and B4: 10%. According to our findings, salinity stress negatively affected morphological and physiological growth parameters of kochia plants by decreasing the fresh and dry weight (25% and 28%, respectively), plant height (30%), circumference (46%), total chlorophyll (51%), and relative water content (29%) when compared to the controls. Furthermore, electrolyte leakage increased considerably (19%) due to salt stress. Significant morphological and physiological growth enhancements were seen at all biochar levels in comparison to the control treatment, with the highest level increasing plant height by 55%, circumference by 76%, total chlorophyll concentrations by 121%, and relative water content by 28%. Furthermore, it resulted in a 36% reduction in the stressed plants' electrolyte leakage. The findings demonstrated biochar's benefits in reducing salinity's negative effects on kochia plants.

• This study provides new data about the specifying the impact of using biochar on salinity concentration and the growth parameters of kochia plants. This investigation demonstrated a significant results in terms of that the salinity stress relative to using biochar.• Biochar is crucial for stimulating and activating biochemical defensive mechanisms against salt stress; yet, research in this area is lacking.• Biochar has shown that it is crucial to stimulate biochemical defense mechanisms against salinity stress• It was found that using biochar as a soil supplement improved morphological, physiological, and biochemical characteristics of the kochia plant by increasing fresh and dried weight per plant, plant height, plant circumference, chlorophyll concentrations, and relative water content while lowering electrolyte leakage in stressed kochia plants. This research will aid in increasing kochia's early development and stand establishment in saline circumstances.

Kochia scoparia L., a newfound candidate halophyte, for phytoremediation of cadmium-contaminated saline soils.

In recent years, heavy metal pollution in saline soil is increasingly severe due to the rapid development of industry and agriculture. Halophytes can survive at higher concentrations of salt and heavy metal, which make them suitable candidates for the phytoremediation of heavy metals in saline soils. In the present study, the halophyte plant Kochia scoparia (L.) Schrad. seedlings were exposed to different doses of Cd (0, 5, 10, 30 mg/kg) and NaCl (0, 200, 400, 800 mM) to explore its tolerance and phytoremediation ability for Cd. There was no significant toxic effect of Cd on the K. scoparia seedlings. NaCl reduced the biomass of K. scoparia compared with the control, but did not show any visible toxic symptom. Furthermore, Cd accumulation in K. scoparia is mainly distributed in the shoot; especially when exposed to low-Cd (5 mg/kg) treatment, the accumulation of Cd in the shoots was up to 5.42-22.25 mg/kg, which was 3.18-53.4 times of that in the roots. Moreover, the contents of glutathione and oxalate in plants increased gradually with the increase of NaCl concentration. Under the treatment of 800 mM NaCl without Cd, the content of glutathione reached the highest 51.21 µg/g, and the proportion of oxalate reached the highest 28.76% under the treatment of 30 mg/kg Cd with 400 Mm NaCl. Finally, we also found the significant alterations of cadmium chemical forms in rhizosphere soil with the addition of NaCl. Overall, K. scoparia could be an efficient and valuable candidate for the phytoextraction of low-Cd (5 mg/kg)-contaminated saline soil.

Genomic-based epidemiology reveals independent origins and gene flow of glyphosate resistance in Bassia scoparia populations across North America.

Genomic-based epidemiology can provide insight into the origins and spread of herbicide resistance mechanisms in weeds. We used kochia (Bassia scoparia) populations resistant to the herbicide glyphosate from across western North America to test the alternative hypotheses that (i) a single EPSPS gene duplication event occurred initially in the Central Great Plains and then subsequently spread to all other geographical areas now exhibiting glyphosate-resistant kochia populations or that (ii) gene duplication occurred multiple times in independent events in a case of parallel evolution. We used qPCR markers previously developed for measuring the structure of the EPSPS tandem duplication to investigate whether all glyphosate-resistant individuals had the same EPSPS repeat structure. We also investigated population structure using simple sequence repeat markers to determine the relatedness of kochia populations from across the Central Great Plains, Northern Plains and the Pacific Northwest. We found that the original EPSPS duplication genotype was predominant in the Central Great Plains where glyphosate resistance was first reported. We identified two additional EPSPS duplication genotypes, one having geographical associations with the Northern Plains and the other with the Pacific Northwest. The EPSPS duplication genotype from the Pacific Northwest seems likely to represent a second, independent evolutionary origin of a resistance allele. We found evidence of gene flow across populations and a general lack of population structure. The results support at least two independent evolutionary origins of glyphosate resistance in kochia, followed by substantial and mostly geographically localized gene flow to spread the resistance alleles into diverse genetic backgrounds.

Corn starch and methylcellulose edible films incorporated with fireweed (Chamaenerion angustifolium L.) extract: Comparison of physicochemical and antioxidant properties.

The properties of edible films derived from corn starch (CS) and methylcellulose (MC) supplemented with fireweed extract (FE; 0.0125-0.05% w/w) were analyzed. Due to their more crystalline structure, the MC films were significantly stronger (~26 MPa) than the CS films (~4 MPa). In turn, CS produced films with lower water vapor permeability (WVP, 50.12-51.74 vs. 56.52-59.10 g mm m-2 d-1 kPa-1). The hydrothermally-disrupted starch granules contributed to high roughness and opacity of the CS films. The FE-supplemented films exhibited an intensive yellow color and improved the UV-absorbing effect. FE delayed starch retrogradation, as indicated by the reduced crystallinity and slightly improved transparency of the CS films. Incorporation of FE significantly enhanced the released radical scavenging activity (RSA) of the films, while did not affect the WVP and mechanical properties. Due to better FE-trapping capacity, the CS-based films exhibited lower antioxidant activity (RSA60min = 2.21-19.75%) as compared to the MC counterparts (RSA60min = 4.87-38.31%).

Effect of EPSPS gene copy number and glyphosate selection on fitness of glyphosate-resistant Bassia scoparia in the field.

The widespread evolution of glyphosate-resistant (GR) Bassia scoparia in the U.S. Great Plains poses a serious threat to the long-term sustainability of GR sugar beet. Glyphosate resistance in B. scoparia is due to an increase in the EPSPS (5-enolpyruvyl-shikimate-3-phosphate) gene copy number. The variation in EPSPS gene copies among individuals from within a single GR B. scoparia population indicated a differential response to glyphosate selection. With the continued use of glyphosate in GR sugar beet, the effect of increasing glyphosate rates (applied as single or sequential applications) on the fitness of GR B. scoparia individuals with variable EPSPS gene copies was tested under field conditions. The variation in EPSPS gene copy number and total glyphosate rate (single or sequential applications) did not influence any of the reproductive traits of GR B. scoparia, except seed production. Sequential applications of glyphosate with a total rate of 2214 g ae ha-1 or higher prevented seed production in B. scoparia plants with 2-4 (low levels of resistance) and 5-6 (moderate levels of resistance) EPSPS gene copies. Timely sequential applications of glyphosate (full recommended rates) can potentially slow down the evolution of GR B. scoparia with low to moderate levels of resistance (2-6 EPSPS gene copies), but any survivors (highly-resistant individuals with ≥ 8 EPSPS gene copies) need to be mechanically removed before flowering from GR sugar beet fields. This research warrants the need to adopt ecologically based, multi-tactic strategies to reduce exposure of B. scoparia to glyphosate in GR sugar beet.

Momordin Ic induces G0/1 phase arrest and apoptosis in colon cancer cells by suppressing SENP1/c-MYC signaling pathway.

Momordin Ic (MI) is a natural pentacyclic triterpenoid enriched in various Chinese natural medicines such as the fruit of Kochia scoparia (L.) Schrad. Studies have shown that MI presents antitumor properties in liver and prostate cancers. However, the activity and potential mechanisms of MI against colorectal cancer remain elusive. Here, we showed that MI inhibited cell proliferation with G0/1 phase cell cycle arrest in colon cancer cells. Moreover, it was observed that MI increased apoptosis compared to untreated cells. Further investigation showed that the SUMOylation of c-Myc was enhanced by MI and led to the down-regulated protein level of c-Myc, which is involved in regulating cell proliferation and apoptosis. SENP1 has been demonstrated to be critical for the SUMOylation of c-Myc. Meanwhile, knockdown of SENP1 by siRNA abolished the effects of MI on c-Myc level and cell viability in colon cancer cells. Together, these results revealed that MI exerted an anti-tumor activity in colon cancer cells via SENP1/c-Myc signaling pathway. These finding provide an insight into the potential of MI for colon cancer therapy.

Molecular Identification and Phylogenetic Analysis of the Traditional Chinese Medicinal Plant Kochia scoparia Using ITS2 Barcoding.

Kochia scoparia has high medicinal and economic value. However, with similar morphological features, adulterants and some closely related species of K. scoparia are increasingly sold in the medicinal markets, leading to potential safety risks. In this study, 128 internal transcribed spacer 2 (ITS2) sequences were collected to distinguish K. scoparia from its closely related species and adulterants. Then, sequence alignment, sequence characteristics analysis, and genetic distance calculations were performed using MEGA 6.06 software, and the phylogenetic trees were reconstructed using both MEGA 6.06 and IQ-Tree software. Finally, the secondary structure of ITS2 was modeled using the prediction tool in the ITS2 database. The results showed that ITS2 sequences of K. scoparia ranged in length from 226 to 227 bp, with a mean GC content of 55.3%. The maximum intraspecific distance was zero, while the minimum interspecific distance from closely related species and adulterants was 0.009 and 0.242, respectively. Kochia scoparia formed an independent clade in the phylogenetic trees, and its secondary structure exhibited enough variation to be separated from that of other species. In summary, ITS2 can be used as a mini-barcode for distinguishing K. scoparia from closely related species and adulterants. Its phylogenetic trees could illustrate the evolutionary process of K. scoparia in the Camphorosmeae. The phylogenetic results using ITS2 barcode further supported the internationally recognized revised classifications of Kochia and Bassia genera as a combined Bassia genus, together with the establishment of new genera Grubovia and Sedobassia, which we suggest is accepted by the Flora of China. Graphical abstract .

In Vitro Human Skin Penetration, Antioxidant and Antimicrobial Activity of Ethanol-Water Extract of Fireweed (Epilobium angustifolium L.).

Epilobium angustifolium L. is applied as an antiseptic agent in the treatment of skin diseases. However, there is a lack of information on human skin penetration of active ingredients with antioxidative potential. It seems crucial because bacterial infections of skin and subcutaneous tissue are common and partly depend on oxidative stress. Therefore, we evaluated in vitro human skin penetration of fireweed ethanol-water extracts (FEEs) by determining antioxidant activity of these extracts before and after penetration study using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and Folin-Ciocalteu methods. Microbiological tests of extracts were done. The qualitative and quantitative evaluation was performed using gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC-UV) methods. The in vitro human skin penetration using the Franz diffusion chamber was assessed. The high antioxidant activity of FEEs was found. Gallic acid (GA), chlorogenic acid (ChA), 3,4-dihydroxybenzoic acid (3,4-DHB), 4-hydroxybenzoic acid (4-HB), and caffeic acid (CA) were identified in the extracts. The antibacterial activities were found against Serratia lutea, S. marcescens, Bacillus subtilis, B. pseudomycoides, and B. thuringiensis and next Enterococcus faecalis, E. faecium, Streptococcus pneumoniae, Pseudomonas aeruginosa, and P. fluorescens strains. In vitro penetration studies showed the penetration of some phenolic acids and their accumulation in the skin. Our results confirm the importance of skin penetration studies to guarantee the efficacy of formulations containing E. angustifolium extracts.

A dicamba resistance-endowing IAA16 mutation leads to significant vegetative growth defects and impaired competitiveness in kochia (Bassia scoparia)†.

BACKGROUND: Precise quantification of the fitness cost of synthetic auxin resistance has been impeded by lack of knowledge about the genetic basis of resistance in weeds. Recent elucidation of a resistance-endowing IAA16 mutation (G73N) in the key weed species kochia (Bassia scoparia), allows detailed characterization of the contribution of resistance alleles to weed fitness, both in the presence and absence of herbicides. Different G73N genotypes from a segregating resistant parental line (9425) were characterized for cross-resistance to dicamba, 2,4-d and fluroxypyr, and changes on stem/leaf morphology and plant architecture. Plant competitiveness and dominance of the fitness effects was quantified through measuring biomass and seed production of three F2 lines in two runs of glasshouse replacement series studies. RESULTS: G73N confers robust resistance to dicamba but only moderate to weak resistance to 2,4-D and fluroxypyr. G73N mutant plants displayed significant vegetative growth defects: (i) they were 30-50% shorter, with a more tumbling style plant architecture, and (ii) they had thicker and more ovate (versus lanceolate and linear) leaf blades with lower photosynthesis efficiency, and 40-60% smaller stems with less-developed vascular bundle systems. F2 mutant plants had impaired plant competitiveness, which can lead to 80-90% less biomass and seed production in the replacement series study. The pleiotropic effects of G73N were mostly semidominant (0.5) and fluctuated with the environments and traits measured. CONCLUSION: G73N is associated with significant vegetative growth defects and reduced competitiveness in synthetic auxin-resistant kochia. Management practices should target resistant kochia's high vulnerability to competition in order to effectively contain the spread of resistance.

Investigating the presence of compensatory evolution in dicamba resistant IAA16 mutated kochia (Bassia scoparia)†.

BACKGROUND: Lack of fitness costs has been reported for multiple herbicide resistance traits, but the underlying evolutionary mechanisms are not well understood. Compensatory evolution that ameliorates resistance costs, has been documented in bacteria and insects but rarely studied in weeds. Dicamba resistant IAA16 (G73N) mutated kochia was previously found to have high fecundity in the absence of competition, regardless of significant vegetative growth defects. To understand if costs of dicamba resistance can be compensated through traits promoting reproductive success in kochia, we thoroughly characterized the reproductive growth and development of different G73N kochia biotypes. Flowering phenology, seed production and reproductive allocation were quantified through greenhouse studies, floral (stigma-anthers distance) and seed morphology, as well as resulting mating and seed dispersal systems were studied through time-course microcopy images. RESULTS: G73N covaried with multiple phenological, morphological and ecological traits that improve reproductive fitness: (i) 16-60% higher reproductive allocation; (ii) longer reproduction phase through early flowering (2-7 days); (iii) smaller stigma-anthers separation (up to 60% reduction of herkogamy and dichogamy) that can potentially promote selfing and reproductive assurance; (iv) 'winged' seeds with 30-70% longer sepals that facilitate long-distance seed dispersal. CONCLUSION: The current study demonstrates that costs of herbicide resistance can be ameliorated through coevolution of other fitness penalty alleviating traits. As illustrated in a hypothetical model, the evolution of herbicide resistance is an ongoing fitness maximization process, which poses challenges to contain the spread of resistance. © 2020 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Health awareness and the transition towards clean cooking fuels: Evidence from Rajasthan.

Ensuring affordable, reliable, sustainable and modern energy for all by 2030 is part of the internationally agreed Sustainable Development Goals (SDG7). With roughly 3 billion people still lacking access to clean cooking solutions in 2017, this remains an ambitious task. The use of solid biomass such as wood and cow dung for cooking causes household air pollution resulting in severe health hazards. In this context, the Indian government has set up a large program promoting the use of liquefied petroleum gas (LPG) in rural areas. While this has led millions of households to adopt LPG, a major fraction of them continues to rely heavily on solid biomass for their daily cooking. In this paper, we evaluate the effect of simple health messaging on the propensity of these households to use LPG more regularly. Our results from rural Rajasthan are encouraging. They show that health messaging increases the reported willingness to pay for LPG, and substantially increases actual consumption. We measure this based on a voucher, which can only be used if LPG consumption is doubled until a certain deadline. Households exposed to health messaging use the voucher about 30% more often than households exposed to a placebo treatment. We further show that the impact of our very brief, but concrete health messaging is close to the effect of a 10% price reduction for a new LPG cylinder. Finally, our study raises some interesting questions about gender-related effects that would be worth consideration in future research.

[Effects of Kochia scoparia-Brassica rapa rotation on Cd uptake by Brassica rapa].

In this study, pot and field experiments were conducted to study the enrichment of soil cadmium by Kochia scoparia. Further, rotations in pot experiments were carried out with four varieties of Brassica rapa to verify the remediation effect of Kochia scoparia on cadmium contamination in soil. The enrichment capacity of Kochia scoparia was leaf > root > stem with bioconcentration factors (BCFCd) of 15.07, 5.44 and 2.96, respectively. The total cadmium in soil decreased by 6.02% to 13.60% after planting Kochia scoparia, and the activities of soil urease and acid phosphatase also increased. The results of pot cultivation shows that the above-ground cadmium content of Brassica rapa in Kochia scoparia-Brassica rapa rotation system decreased by 17.21% on average compared with the control group without rotation, whereas the biomass increased slightly, and the above-ground translocation factors (TFCd) did not change significantly. These results suggest that the rotation of Brassica rapa with Kochia scoparia could increase the yield of Brassica rapa, and effectively reduce the cadmium content in edible parts of Brassica rapa, toward the purpose of realizing the green agricultural concept of "harnessing while producing".

Kochia scoparia seed extract suppresses VEGF-induced angiogenesis via modulating VEGF receptor 2 and PI3K/AKT/mTOR pathways.

Context: Kochia scoparia (L.) Schrad (Amaranthaceae), known as a traditional medicine in China, Japan and Korea, is reported to have various biological activities. However, K. scoparia seed extract (KSE) functional roles on angiogenesis and prostate cancer inhibition have not been elucidated. Objective: This study elucidates the effects of KSE on vascular endothelial growth factor (VEGF)-induced angiogenesis in human umbilical vein endothelial cells (HUVECs) and inhibition of proliferation in prostate cancer cells. Materials and methods: HUVECs were treated with 10-20 µg/mL of KSE and 20-50 ng/mL of VEGF for 12-72 h. Anti-angiogenesis properties of KSE were determined by wound healing, trans-well, tube formation, rat aortic ring assay and western blotting. Prostate cancer and normal cells were incubated with 10-250 µg/mL of KSE for 24 h, and cell viability was measured by SRB assay. Phenolic compounds in KSE were analyzed using a HPLC-PDA system. Results: IC50 for cell viability of HUVECs, LNCaP, PC-3, RC-58T and RWPE-1 by KSE were 30.64, 89.25, 123.41, 141.62 and >250 µg/mL, respectively. Treatment with KSE (20 µg/mL) significantly suppressed VEGF-induced migration, invasion and capillary-like structure formation of HUVECs and microvessel sprouting from rat aortic rings. In addition, KSE down-regulated PI3K/AKT/mTOR levels and phosphorylation of VEGF receptor 2 in HUVECs. 3-OH-tyrosol (1.63 mg/g) and morin hydrate (0.17 mg/g) were identified in KSE. Conclusions: KSE inhibits angiogenesis in HUVECs as well as proliferation in human prostate cancer cells, suggesting KSE may be useful herbal medicine for preventing progression of prostate cancer and angiogenesis.

The Draft Genome of Kochia scoparia and the Mechanism of Glyphosate Resistance via Transposon-Mediated EPSPS Tandem Gene Duplication.

Increased copy number of the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene confers resistance to glyphosate, the world's most-used herbicide. There are typically three to eight EPSPS copies arranged in tandem in glyphosate-resistant populations of the weed kochia (Kochia scoparia). Here, we report a draft genome assembly from a glyphosate-susceptible kochia individual. Additionally, we assembled the EPSPS locus from a glyphosate-resistant kochia plant by sequencing select bacterial artificial chromosomes from a kochia bacterial artificial chromosome library. Comparing the resistant and susceptible EPSPS locus allowed us to reconstruct the history of duplication in the structurally complex EPSPS locus and uncover the genes that are coduplicated with EPSPS, several of which have a corresponding change in transcription. The comparison between the susceptible and resistant assemblies revealed two dominant repeat types. Additionally, we discovered a mobile genetic element with a FHY3/FAR1-like gene predicted in its sequence that is associated with the duplicated EPSPS gene copies in the resistant line. We present a hypothetical model based on unequal crossing over that implicates this mobile element as responsible for the origin of the EPSPS gene duplication event and the evolution of herbicide resistance in this system. These findings add to our understanding of stress resistance evolution and provide an example of rapid resistance evolution to high levels of environmental stress.

[Screening of pyrene degradation bacteria strain from the Kochia scoparia roots and its potential for promoting growth]. / ç„¦åŒ–åŽ‚åœ°è‚¤æ ¹å† è§£èŠ˜ç»†èŒçš„ç­›é€‰åŠä¿ƒç”Ÿæ½œåŠ›.

The endophytic bacteria were isolated from the roots of polycyclic aromatic hydrocarbon (PAHs)-tolerant plant. We investigated their ability of PAHs degradation and plant growth promo-ting, with the aim to provide theoretical support for bacterial-plant cooperative soil remediation. Kochia scoparia living in coking plant area were selected for strains isolation. Eight endophytic bacteria strains, which used pyrene and 1-aminocyclopropane-1-carboxylic acid (ACC) as sole carbon and nitrogen source, were isolated from the roots of K. scoparia. Three endophytic bacteria, KSE4, KSE7 and KSE8, displayed high degradation efficiency in pyrene degradation experiment. They were identified as Bacillus sp., Pseudomonas sp., and Sphingobacterium sp., respectively. The abili-ties of those three strains to produce ACC deaminase and their effects on seed germination of K. scoparia were examined under pyrene stress through liquid culture tests. The results showed that the activity of ACC deaminase decreased with increasing pyrene concentration (0-15 mg·L-1). KSE7 had the strongest promotion effect. When pyrene concentration reached to 15 mg·L-1, the germination rate and bud length of K. scoparia increased by 44.8% and 61.1%, respectively. Our results indicated that KSE7 is a promising bacterial strain for soil remediation in coking plant area.

Water-soluble exudates from seeds of Kochia scoparia exhibit antifungal activity against Colletotrichum graminicola.

Plant seed exudates are composed of complex mixtures of chemicals with potential for bioactive compounds with antimicrobial properties. This study focused on kochia (Kochia scoparia), one of many weedy plant species considered invasive in many agricultural systems. Extraction of compounds in water yielded an exudate mass equivalent to 7% of the original seed mass used. Water-soluble exudates were tested against 16 known plant pathogens in disk diffusion assays and kochia exudates were found to inhibit Colletotrichum graminicola, the fungal causative agent of anthracnose and stalk rot in maize. The narrow range of fungi found as targets suggested the mechanism of inhibition may be specific rather than broadly antifungal. A decline in viability of cells over four orders of magnitude occurred within six hours of exposure to exudate. The minimum inhibitory concentration was 3.125 mg L-1. Hyphae formation in C. graminicola appeared inhibited following exposure to the exudate. Small molecular weight compounds as determined by GC/MS analysis showed high relative amounts of the sugars fructose, galactopyranose, glucose, and sorbitol, along with moderate proportions of organic acids and amino acids. Protein content averaged 0.7% in the standard concentration (100 mg mL-1) used for inhibition assays. Size fractionation of the exudate and subsequent disk diffusion assays revealed bioactive fractions with compounds in the MW range <5 kDa. To the best of our knowledge, this study is the first to show promising bioactivity against C. graminicola that was associated with water-extractable compounds from a common weed species. The results suggest that seeds of persistent plant species with long-lived seed banks like kochia may have potential for use in the discovery of compounds active in inhibiting fungal pathogens.

In silico identification of genetic mutations conferring resistance to acetohydroxyacid synthase inhibitors: A case study of Kochia scoparia.

Mutations that confer herbicide resistance are a primary concern for herbicide-based chemical control of invasive plants and are often under-characterized structurally and functionally. As the outcome of selection pressure, resistance mutations usually result from repeated long-term applications of herbicides with the same mode of action and are discovered through extensive field trials. Here we used acetohydroxyacid synthase (AHAS) of Kochia scoparia (KsAHAS) as an example to demonstrate that, given the sequence of a target protein, the impact of genetic mutations on ligand binding could be evaluated and resistance mutations could be identified using a biophysics-based computational approach. Briefly, the 3D structures of wild-type (WT) and mutated KsAHAS-herbicide complexes were constructed by homology modeling, docking and molecular dynamics simulation. The resistance profile of two AHAS-inhibiting herbicides, tribenuron methyl and thifensulfuron methyl, was obtained by estimating their binding affinity with 29 KsAHAS (1 WT and 28 mutated) using 6 molecular mechanical (MM) and 18 hybrid quantum mechanical/molecular mechanical (QM/MM) methods in combination with three structure sampling strategies. By comparing predicted resistance with experimentally determined resistance in the 29 biotypes of K. scoparia field populations, we identified the best method (i.e., MM-PBSA with single structure) out of all tested methods for the herbicide-KsAHAS system, which exhibited the highest accuracy (up to 100%) in discerning mutations conferring resistance or susceptibility to the two AHAS inhibitors. Our results suggest that the in silico approach has the potential to be widely adopted for assessing mutation-endowed herbicide resistance on a case-by-case basis.

Reactive oxygen species trigger the fast action of glufosinate.

MAIN CONCLUSION: Glufosinate is primarily toxic to plants due to a massive light-dependent generation of reactive oxygen species rather than ammonia accumulation or carbon assimilation inhibition. Glutamine synthetase (GS) plays a key role in plant nitrogen metabolism and photorespiration. Glufosinate (C5H12NO4P) targets GS and causes catastrophic consequences leading to rapid plant cell death, and the causes for phytoxicity have been attributed to ammonia accumulation and carbon assimilation restriction. This study aimed to examine the biochemical and physiological consequences of GS inhibition to identify the actual cause for rapid phytotoxicity. Monocotyledonous and dicotyledonous species with different forms of carbon assimilation (C3 versus C4) were selected as model plants. Glufosinate sensitivity was proportional to the uptake of herbicide between species. Herbicide uptake also correlated with the level of GS inhibition and ammonia accumulation in planta even with all species having the same levels of enzyme sensitivity in vitro. Depletion of both glutamine and glutamate occurred in glufosinate-treated leaves; however, amino acid starvation would be expected to cause a slow plant response. Ammonia accumulation in response to GS inhibition, often reported as the driver of glufosinate phytotoxicity, occurred in all species, but did not correlate with either reductions in carbon assimilation or cell death. This is supported by the fact that plants can accumulate high levels of ammonia but show low inhibition of carbon assimilation and absence of phytotoxicity. Glufosinate-treated plants showed a massive light-dependent generation of reactive oxygen species, followed by malondialdehyde accumulation. Consequently, we propose that glufosinate is toxic to plants not because of ammonia accumulation nor carbon assimilation inhibition, but the production of reactive oxygen species driving the catastrophic lipid peroxidation of the cell membranes and rapid cell death.

Cross-resistance to dicamba, 2,4-D, and fluroxypyr in Kochia scoparia is endowed by a mutation in an AUX/IAA gene.

The understanding and mitigation of the appearance of herbicide-resistant weeds have come to the forefront of study in the past decade, as the number of weed species that are resistant to one or more herbicide modes of action is on the increase. Historically, weed resistance to auxin herbicides has been rare, but examples, such as Kochia scoparia L. Schrad (kochia), have appeared, posing a challenge to conventional agricultural practices. Reports of dicamba-resistant kochia populations began in the early 1990s in areas where auxin herbicides were heavily utilized for weed control in corn and wheat cropping systems, and some biotypes are resistant to other auxin herbicides as well. We have further characterized the auxin responses of one previously reported dicamba-resistant biotype isolated from western Nebraska and found that it is additionally cross-resistant to other auxin herbicides, including 2,4-dichlorophenoxyacetic acid (2,4-D) and fluroxypyr. We have utilized transcriptome sequencing and comparison to identify a 2-nt base change in this biotype, which results in a glycine to asparagine amino acid change within a highly conserved region of an AUX/indole-3-acetic acid (IAA) protein, KsIAA16. Through yeast two-hybrid analysis, characterization of F2 segregation, and heterologous expression and characterization of the gene in Arabidopsis thaliana, we show that that the single dominant KsIAA16R resistance allele is the causal basis for dicamba resistance in this population. Furthermore, we report the development of a molecular marker to identify this allele in populations and facilitate inheritance studies. We also report that the resistance allele confers a fitness penalty in greenhouse studies.