Explore the vast range of titles available.

In response to several kinds of abiotic stress, plants greatly increase their accumulation of free amino acids. Although stress-induced proline increases have been studied the most extensively, the fold-increase of other amino acids, in particular branched-chain amino acids (BCAAs; leucine, isoleucine, and valine), is often higher than that of proline. In Arabidopsis thaliana (Arabidopsis), BCAAs accumulate in response to drought, salt, mannitol, polyethylene glycol, herbicide treatment, and nitrogen starvation. Plants that are deficient in abscisic acid signaling accumulate lower amounts of BCAAs, but not proline and most other amino acids. Previous bioinformatic studies had suggested that amino acid synthesis, rather than protein degradation, is responsible for the observed BCAA increase in osmotically stressed Arabidopsis. However, whereas treatment with the protease inhibitor MG132 decreased drought-induced BCAA accumulation, inhibition of BCAA biosynthesis with the acetolactate synthase inhibitors chlorsulfuron and imazapyr did not. Additionally, overexpression of BRAN-CHED-CHAIN AMINO ACID TRANSFERASE2 (BCAT2), which is upregulated in response to osmotic stress and functions in BCAA degradation, decreased drought-induced BCAA accumulation. Together, these results demonstrate that BCAA accumulation in osmotically stressed Arabidopsis is primarily the result of protein degradation. After relief of the osmotic stress, BCAA homeostasis is restored over time by amino acid degradation involving BCAT2. Thus, drought-induced BCAA accumulation is different from that of proline, which is accumulated due to de novo synthesis in an abscisic acid-independent manner and remains elevated for a more prolonged period of time after removal of the osmotic stress.

We previously reported Agrobacterium-mediated transformation methods for the liverwort Marchantia polymorpha using the hygromycin phosphotransferase gene as a marker for selection with hygromycin. In this study, we developed three additional markers for M. polymorpha transformation: the gentamicin 3'-acetyltransferase gene for selection with gentamicin; a mutated acetolactate synthase gene for selection with chlorsulfuron; and the neomycin phosphotransferase II gene for selection with G418. Based on these four marker genes, we have constructed a series of Gateway binary vectors designed for transgenic experiments on M. polymorpha. The 35S promoter from cauliflower mosaic virus and endogenous promoters for constitutive and heat-inducible expression were used to create these vectors. The reporters and tags used were Citrine, 3×Citrine, Citrine-NLS, TagRFP, tdTomato, tdTomato-NLS, GR, SRDX, SRDX-GR, GUS, ELuc(PEST), and 3×FLAG. These vectors, designated as the pMpGWB series, will facilitate molecular genetic analyses of the emerging model plant M. polymorpha.

A field survey was conducted in 2014 to determine the response of Palmer amaranth (Amaranthus palmeri S. Wats.) populations from 16 counties in southcentral Kansas to commonly used herbicides. The main objective was to evaluate the sensitivity of 28 randomly collected populations to field‐use rates of glyphosate, chlorsulfuron, 2,4‐D, dicamba, atrazine, and mesotrione herbicides; and further characterize the sensitivity levels in two populations. The response of all tested populations was categorized as less sensitive (≤59% injury), moderately sensitive (60–89% injury), and susceptible (90–100% injury) based on visible injury at 21 days after treatment (DAT). Results from herbicide screening experiments indicated that 47, 36, 7, 75, and 32% of tested populations were less sensitive (putative resistant) to glyphosate, chlorsulfuron, 2,4‐D, atrazine, and mesotrione herbicide, respectively. None of the tested populations showed less sensitivity (≤59% injury) to dicamba herbicide. Whole plant dose‐response assays on two populations collected from Kiowa (KW2) and Pratt (PR8) counties indicated that the KW2 population had 3.0‐, 2.3‐, 5.8‐, and 8.4‐fold less sensitivity to 2,4‐D, chlorsulfuron, atrazine, and mesotrione, respectively, compared to a known susceptible (MHS) population. In contrast, the PR8 population was 2.0‐, 8.6‐, 10.6‐, 3.7‐, and 2.8‐fold less sensitive to 2,4‐D, glyphosate, chlorsulfuron, atrazine, and mesotrione, respectively, compared to MHS population. Overall, these results suggest that reduced sensitivity to glyphosate, chlorsulfuron, atrazine, and/or mesotrione is highly prevalent in southcentral Kansas. Growers should adopt diversified weed control methods to manage these difficult‐to‐control populations on their production fields.

To estimate the prevalence of herbicide-resistant weeds, 87 wheat and barley farms were randomly surveyed in the Canterbury region of New Zealand. Over 600 weed seed samples from up to 10 mother plants per taxon depending on abundance, were collected immediately prior to harvest (two fields per farm). Some samples provided by agronomists were tested on an ad-hoc basis. Over 40,000 seedlings were grown to the 2–4 leaf stage in glasshouse conditions and sprayed with high priority herbicides for grasses from the three modes-of-action acetyl-CoA carboxylase (ACCase)-inhibitors haloxyfop, fenoxaprop, clodinafop, pinoxaden, clethodim, acetolactate synthase (ALS)-inhibitors iodosulfuron, pyroxsulam, nicosulfuron, and the 5-enolpyruvyl shikimate 3-phosphate synthase (EPSPS)-inhibitor glyphosate. The highest manufacturer recommended label rates were applied for the products registered for use in New Zealand, often higher than the discriminatory rates used in studies elsewhere. Published studies of resistance were rare in New Zealand but we found weeds survived herbicide applications on 42 of the 87 (48%) randomly surveyed farms, while susceptible reference populations died. Resistance was found for ALS-inhibitors on 35 farms (40%) and to ACCase-inhibitors on 20 (23%) farms. The number of farms with resistant weeds (denominator is 87 farms) are reported for ACCase-inhibitors, ALS-inhibitors, and glyphosate respectively as: Avena fatua (9%, 1%, 0% of farms), Bromus catharticus (0%, 2%, 0%), Lolium spp. (17%, 28%, 0%), Phalaris minor (1%, 6%, 0%), and Vulpia bromoides (0%, not tested, 0%). Not all farms had the weeds present, five had no obvious weeds prior to harvest. This survey revealed New Zealand’s first documented cases of resistance in P . minor (fenoxaprop, clodinafop, iodosulfuron) and B . catharticus (pyroxsulam). Twelve of the 87 randomly sampled farms (14%) had ALS-inhibitor chlorsulfuron-resistant sow thistles, mostly Sonchus asper but also S . oleraceus . Resistance was confirmed in industry-supplied samples of the grasses Digitaria sanguinalis (nicosulfuron, two maize farms), P . minor (iodosulfuron, one farm), and Lolium spp. (cases included glyphosate, haloxyfop, pinoxaden, iodosulfuron, and pyroxsulam, 9 farms). Industry also supplied Stellaria media samples that were resistant to chlorsulfuron and flumetsulam (ALS-inhibitors) sourced from clover and ryegrass fields from the North and South Island.

Multiple herbicide–resistant (MHR) kochia [Bassia scoparia (L.) A.J. Scott] is a concern for farmers in the Great Plains. A total of 82 B. scoparia populations were collected from western Kansas (KS), western Oklahoma (OK), and the High Plains of Texas (TX) during fall of 2018 and 2019 (from the various locations), and their herbicide resistance status was evaluated. The main objectives were to (1) determine the distribution and frequency of resistance to atrazine, chlorsulfuron, dicamba, fluroxypyr, and glyphosate; and (2) characterize the resistance levels to glyphosate, dicamba, and/or fluroxypyr in selected B. scoparia populations. Results indicated that 33%, 100%, 48%, 30%, and 70% of the tested B. scoparia populations were potentially resistant (≥20% survival frequency) to atrazine, chlorsulfuron, dicamba, fluroxypyr, and glyphosate, respectively. A three-way premixture of dichlorprop/dicamba/2,4-D provided 100% control of all the tested populations. Dose–response studies further revealed that KS-9 and KS-14 B. scoparia populations were 5- to 10-fold resistant to dicamba, 3- to 6-fold resistant to fluroxypyr, and 4- to 5-fold resistant to glyphosate as compared with the susceptible (KS-SUS) population. Similarly, OK-10 and OK-11 populations were 10- to 13-fold resistant to dicamba and 3- to 4-fold resistant to fluroxypyr and glyphosate compared with the OK-SUS population. TX-1 and TX-13 B. scoparia populations were 2- to 4-fold resistant to dicamba, and TX-1 was 5-fold resistant to glyphosate compared with the TX-SUS population. These results confirm the first report of dicamba- and fluroxypyr-resistant B. scoparia from Oklahoma and glyphosate- and dicamba-resistant B. scoparia from Texas. These results imply that adopting effective integrated weed management strategies (chemical and nonchemical) is required to mitigate the further spread of MHR B. scoparia in the region.

Field and pot experiments were conducted in Greece to study the occurrence of resistance in silky windgrass to acetolactate synthase (ALS)- and acetyl-CoA synthase (ACCase)-inhibiting herbicides. Twenty-four populations of silky windgrass were examined in whole-plant response experiments. High levels of field-evolved resistance to chlorsulfuron (0% to 28% control in terms of fresh weight reduction) with the recommended field rates were confirmed in most silky windgrass populations. However, other ALS inhibitors, such as pyroxsulam and a premix of mesosulfuron-methyl and iodosulfuron, provided adequate control (76% to 100% in terms of fresh weight reduction) of most populations, except eight silky windgrass populations that were found to be cross-resistant to all ALS-inhibiting herbicides tested (i.e., chlorsulfuron, commercial mixture of mesosulfuron-methyl plus iodosulfuron, and pyroxsulam). Conversely, most silky windgrass populations were controlled effectively (90% to 100% in terms of fresh weight reduction) with the recommended field rates of ACCase inhibitors cycloxydim, clethodim, and pinoxaden, but five populations were also found to be resistant to clodinafop-propargyl (10% to 68% control in terms of fresh weight reduction). The ALS gene sequencing of the eight silky windgrass populations, with cross-resistance to ALS inhibitors, revealed a point mutation at the Pro-197 position, causing amino acid substitution by Ser or Thr in the ALS enzyme. Overall, chlorsulfuron and clodinafop-propargyl were selecting agents of field-evolved multiple resistance to ALS- and ACCase-inhibiting herbicides in five silky windgrass populations. As the available postemergence-applied chemistries/modes of action registered for grass weed control in cereals are rather limited, adopting integrated management practices and implementing proactive and reactive measures to delay the evolution of resistant populations is essential. Nomenclature: chlorsulfuron; pyroxsulam; mesosulfuron-methyl; iodosulfuron; cycloxydim; clethodim; pinoxaden; clodinafop-propargyl; Silky windgrass, Apera spica-venti (L.) P. Beauv.; wheat, Triticum aestivum L.

Evolution of multiple herbicide resistance in Palmer amaranth across the United States is a serious challenge for its management. Recently, a Palmer amaranth population (KCTR; Kansas Conservation Tillage Resistant) from a long-term conservation tillage research project in Kansas, United States, was found uncontrolled by several commonly used herbicides. Importantly, this field did not have a history of repeated use of some of the herbicides for which the KCTR Palmer amaranth population showed lack of control. The objectives of this study were to confirm the evolution of multiple resistances and determine possible mechanism(s) of resistance in KCTR Palmer amaranth plants. In response to post-emergence application, 28–100% of KCTR Palmer amaranth survived field recommended rates of 2,4-D, ALS-, PS II-, EPSPS-, PPO-, HPPD-inhibitor herbicides, or tank- or pre-mixture of PS II- and HPPD-inhibitor herbicides, confirming evolution of six-way resistance in this Palmer amaranth population. However, this population was found susceptible to the PS I- and glutamine synthetase inhibitor herbicides. Chlorsulfuron-, imazethapyr-, and atrazine-resistant plants did not show any previously reported mutation in ALS and psbA genes, the target sites of these herbicides, respectively. However, the survivors of glyphosate treatment showed amplification of EPSPS gene (up to 88 copies). The KCTR plants pretreated with cytochrome P450 or GST inhibitors along with atrazine, 2,4-D, lactofen, or mesotrione had significantly less biomass accumulation than those treated with herbicides alone. Plants treated with P450 inhibitor followed by imazethapyr showed moderate reduction of biomass in KCTR which was statistically similar to a susceptible Palmer amaranth population treated with imazethapyr. These results suggest predominance of metabolic resistance possibly mediated by cytochrome P450 and GST enzyme activity that may have predisposed the KCTR Palmer amaranth population to evolve resistance to multiple herbicides. This is the first report of evolution of six-way resistance in a single Palmer amaranth population. Appropriate management strategies, including integration of cultural, and mechanical, and herbicide mixtures, are warranted to control such Palmer amaranth populations.

Citrus canker disease affects citrus production. This disease is caused by Xanthomonas citri subsp. citri (Xcc). Previous studies confirmed that during Xcc infection, PthA4, a transcriptional activator like effector (TALE), is translocated from the pathogen to host plant cells. PthA4 binds to the effector binding elements (EBEs) in the promoter region of canker susceptibility gene LOB1 (EBE PthA4 -LOBP) to activate its expression and subsequently cause canker symptoms. Previously, the Cas12a/CBE co-editing method was employed to disrupt EBE PthA4 -LOBP of pummelo, which is highly homozygous. However, most commercial citrus cultivars are heterozygous hybrids and more difficult to generate homozygous/biallelic mutants. Here, we employed Cas12a/CBE co-editing method to edit EBE PthA4 -LOBP of Hamlin ( Citrus sinensis ), a commercial heterozygous hybrid citrus cultivar grown worldwide. Binary vector GFP-p1380N-ttLbCas12a:LOBP1-mPBE:ALS2:ALS1 was constructed and shown to be functional via Xcc-facilitated agroinfiltration in Hamlin leaves. This construct allows the selection of transgene-free regenerants via GFP, edits ALS to generate chlorsulfuron-resistant regenerants as a selection marker for genome editing resulting from transient expression of the T-DNA via nCas9-mPBE:ALS2:ALS1, and edits gene(s) of interest (i.e., EBE PthA4 -LOBP in this study) through ttLbCas12a, thus creating transgene-free citrus. Totally, 77 plantlets were produced. Among them, 8 plantlets were transgenic plants (#Ham GFP 1 - #Ham GFP 8), 4 plantlets were transgene-free (#Ham NoGFP 1 - #Ham NoGFP 4), and the rest were wild type. Among 4 transgene-free plantlets, three lines (#Ham NoGFP 1, #Ham NoGFP 2 and #Ham NoGFP 3) contained biallelic mutations in EBE pthA4 , and one line (#Ham NoGFP 4) had homozygous mutations in EBE pthA4 . We achieved 5.2% transgene-free homozygous/biallelic mutation efficiency for EBE PthA4 –LOBP in C. sinensis cv. Hamlin, compared to 1.9% mutation efficiency for pummelo in a previous study. Importantly, the four transgene-free plantlets and 3 transgenic plantlets that survived were resistant against citrus canker. Taken together, Cas12a/CBE co-editing method has been successfully used to generate transgene-free canker‐resistant C. sinensis cv. Hamlin in the T0 generation via biallelic/homozygous editing of EBE pthA4 of the canker susceptibility gene LOB1.

Transgene-free plant genome editing in the T0 generation is highly desirable but challenging1,2. Here we achieved such a goal using a co-editing strategy via Agrobacterium-mediated transient expression of cytosine base editor to edit ALS encoding acetolactate synthase to confer herbicide chlorsulfuron resistance as a selection marker, Cas12a/CRISPR RNA for editing gene(s) of interest, and green fluorescent protein for selecting transgene-free transformants. The biallelic/homozygous transgene-free mutation rates for target genes among herbicide-resistant transformants ranged from 1.9% to 42.1% in tomato, tobacco, potato and citrus. This co-editing strategy is particularly useful for transgene-free genome editing of vegetatively propagated and perennial plant species in the T0 generation.Huang et al. develop a potent genome editing toolkit to generate transgene-free genome-edited plants in the T0 generation by co-editing of ALS gene (without obvious fitness costs) and gene(s) of interest via Agrobacterium-mediated transient expression.

Herbicide residues in agricultural products can have adverse effects on the environment and human health, therefore, there is an urgent need to establish a sensitive, rapid, and wide-ranging detection method. In this study, haptens of phenylurea herbicides (PUs) and sulfonylurea herbicides (SUs) were analyzed and designed based on computational simulation techniques, and two high-performance broad-spectrum monoclonal antibodies against PUs and SUs were prepared. On this basis, a multi-colloidal gold immunochromatography assay (multi-CGIA) was developed to simultaneously detect 13 herbicides in wheat. The visual limit of detection (vLOD) for PUs including diuron, chlortoluron, neburon, chlorbromuron, and linuron was 1–2 µg/kg. The vLOD for SUs including metsulfuron methyl, ethametsulfuron-methyl, sulfometuron-methyl, tribenuron methyl, cinosulfuron, triasulfuron, chlorimuron-ethyl, and chlorsulfuron was 2–10 µg/kg. The results of real sample determination indicated that the multi-CGIA is accurate, stable, and reliable, and adaptable to on-site preliminary screening of actual samples.