Root cell wall polysaccharides and endodermal barriers restrict long-distance Cd translocation in the roots of Kentucky bluegrass.

Soil Cd pollution is a significant environmental issue faced by contemporary society. Kentucky bluegrass is considered a potential phytoremediation species, as some varieties have excellent cadmium (Cd) tolerance. However, the mechanisms of Cd accumulation and transportation in Kentucky bluegrass are still not fully understood. The Cd-tolerant Kentucky bluegrass cultivar 'Midnight' (M) exhibits lower Cd translocation efficiency and a higher leaf Cd concentration compared to the Cd-sensitive cultivar 'Rugby II' (R). We hypothesized that Cd translocation from roots to shoots in cultivar M is hindered by the endodermal barriers and cell wall polysaccharides; hence, we conducted Cd distribution, cytological observation, cell wall component, and transcriptomic analyses under Cd stress conditions using the M and R cultivars. Cd stress resulted in the thickening of the endodermis and increased synthesis of cell wall polysaccharides in both the M and R cultivars. Endodermis development restricted the radical transport of Cd from the root cortex to the stele, while the accumulation of cell wall polysaccharides promoted the binding of Cd to the cell wall. These changes further inhibited the long-distance translocation of Cd from the roots to the aerial parts. Furthermore, the M cultivar exhibited limited long-distance Cd translocation efficiency compared to the R cultivar, which was attributed to the enhanced development of endodermal barriers and increased Cd binding by cell wall polysaccharides. This study provides valuable insights for screening high Cd transport efficiency in Kentucky bluegrass based on anatomical structure and genetic modification.

Integrated proteomics, transcriptomics, and metabolomics offer novel insights into Cd resistance and accumulation in Poa pratensis.

Kentucky bluegrass (Poa pratensis L., KB) demonstrates superior performance in both cadmium (Cd) accumulation and tolerance; however, the regulatory mechanisms and detoxification pathways in this species remain unclear. Therefore, phenotype, root ultrastructure, cell wall components, proteomics, transcriptomics, and metabolomics were analyzed under the hydroponic system to investigate the Cd tolerance and accumulation mechanisms in the Cd-tolerant KB variety 'Midnight (M)' and the Cd-sensitive variety 'Rugby II (R)' under Cd stress. The M variety exhibited higher levels of hydroxyl and carboxyl groups as revealed by Fourier transform infrared spectroscopy spectral analysis. Additionally, a reduced abundance of polysaccharide degradation proteins was observed in the M variety. The higher abundance of glutathione S-transferase and content of L-cysteine-glutathione disulfide and oxidized glutathione in the M variety may contribute to better performance of the M variety under Cd stress. Additionally, the R variety had an enhanced content of carboxylic acids and derivatives, increasing the Cd translocation capacity. Collectively, the down-regulation of cell wall polysaccharide degradation genes coupled with the up-regulation of glutathione metabolism genes enhances the tolerance to Cd stress in KB. Additionally, lignification of the endodermis and the increase in carboxylic acids and derivatives play crucial roles in the redistribution of Cd in KB.

Chemical Composition and Phytotoxic Activity of Artemisia selengensis Turcz. Volatiles.

The chemical profile and allelopathic action of the volatiles produced by Artemisia selengensis were studied. Artemisia selengensis was found to release volatile chemicals to the environment to influence other plants' growth, which suppressed the root length of Amaranthus retroflexus and Poa annua by 50.46 % and 87.83 % under 80 g/1.5 L treatment, respectively. GC/MS analysis led to the identification of 41 compounds (by hydrodistillation, HD) and 48 compounds (by headspace solid-phase microextraction, HS-SPME), with eucalyptol (15.45 % by HD and 28.09 % by HS-SPME) being detected as the most abundant constituent. The essential oil (EO) of A. selengensis completely inhibited the seed germination of A. retroflexus and P. annua at 1 mg/mL and 0.5 mg/mL, respectively. However, eucalyptol displayed much weaker activity compared with the EO, indicating that other less abundant constituents might contribute significantly to the EO's activity. Our study is the first report on the phytotoxicity of A. selengensis EO, suggesting that A. selengensis might release allelopathic volatile agents into the environment that negatively affect other plants' development so as to facilitate its own dominance; the potential value of utilizing A. selengensis EO as an environmentally friendly herbicide is also discussed.

Choline-Mediated Lipid Reprogramming as a Dominant Salt Tolerance Mechanism in Grass Species Lacking Glycine Betaine.

Choline, as a precursor of glycine betaine (GB) and phospholipids, is known to play roles in plant tolerance to salt stress, but the downstream metabolic pathways regulated by choline conferring salt tolerance are still unclear for non-GB-accumulating species. The objectives were to examine how choline affects salt tolerance in a non-GB-accumulating grass species and to determine major metabolic pathways of choline regulating salt tolerance involving GB or lipid metabolism. Kentucky bluegrass (Poa pratensis) plants were subjected to salt stress (100 mM NaCl) with or without foliar application of choline chloride (1 mM) in a growth chamber. Choline or GB alone and the combined application increased leaf photochemical efficiency, relative water content and osmotic adjustment and reduced leaf electrolyte leakage. Choline application had no effects on the endogenous GB content and GB synthesis genes did not show responses to choline under nonstress and salt stress conditions. GB was not detected in Kentucky bluegrass leaves. Lipidomic analysis revealed an increase in the content of monogalactosyl diacylglycerol, phosphatidylcholine and phosphatidylethanolamine and a decrease in the phosphatidic acid content by choline application in plants exposed to salt stress. Choline-mediated lipid reprogramming could function as a dominant salt tolerance mechanism in non-GB-accumulating grass species.

Molecular characteristics of the first case of haloxyfop-resistant Poa annua.

Haloxyfop is one of two acetyl-coenzyme A carboxylase (ACCase) inhibitors that is recommended for controlling Poa annua. We have characterised a population of P. annua that had developed resistance to haloxyfop. This resistant population was found to be almost 20 times less sensitive to haloxyfop than a susceptible population based on percentage survival of individuals in two dose-response experiments. However, the haloxyfop-resistant population was still susceptible to clethodim. Pre-treatment of resistant individuals with a cytochrome P450 inhibitor, malathion, did not change the sensitivity level of the resistant plants to haloxyfop, suggesting that a non-target site mechanism of resistance involving enhanced metabolism, was not responsible for this resistance in P. annua. Gene sequencing showed that a target site mutation at position 2041, which replaced isoleucine with threonine in the carboxyltransferase (CT) domain of the ACCase enzyme, was associated with resistance to haloxyfop in the resistant population. An evaluation of the 3-D structure of the CT domain suggested that, unlike Asn-2041, which is the most common mutation at this position reported to date, Thr-2041 does not change the conformational structure of the CT domain. This is the first study investigating the molecular mechanism involved with haloxyfop resistance in P. annua.

Chemical Composition, Phytotoxic, Antimicrobial and Insecticidal Activity of the Essential Oils of Dracocephalum integrifolium.

The present investigation studied the chemical composition of the essential oils extracted from Dracocephalum integrifolium Bunge growing in three different localities in northwest China and evaluated the phytotoxic, antimicrobial and insecticidal activities of the essential oils as well as their major constituents, i.e., sabinene and eucalyptol. GC/MS analysis revealed the presence of 21-24 compounds in the essential oils, representing 94.17-97.71% of the entire oils. Monoterpenes were the most abundant substances, accounting for 85.30-93.61% of the oils; among them, sabinene (7.35-14.0%) and eucalyptol (53.56-76.11%) were dominant in all three oils, which occupied 67.56-83.46% of the total oils. In general, phytotoxic bioassays indicated that the IC50 values of the oils and their major constituents were below 2 µL/mL (1.739-1.886 mg/mL) against Amaranthus retroflexus and Poa annua. Disc diffusion method demonstrated that the oils and their major constituents possessed antimicrobial activity against Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, Saccharomyces cerevisiae, and Candida albicans, with MIC values ranging from 5-40 µL/mL (4.347-37.712 mg/mL). The oils, sabinene and eucalyptol also exhibited significant pesticidal activity, with the mortality rates of Aphis pomi reaching 100% after exposing to 10 µL oil/petri dish (8.694-9.428 mg/petri dish) for 24 h. To the best of our knowledge, this is the first report on the chemical composition, phytotoxic, antimicrobial and insecticidal activity of the essential oils extracted from D. integrifolium; it is noteworthy to mention that this is also the first report on the phytotoxicity of one of the major constituents, sabinene. Our results imply that D. integrifolium oils and sabinene have the potential value of being further exploited as natural pesticides.

Drought-induced injury is associated with hormonal alteration in Kentucky bluegrass.

Plant hormones play an important role in plant adaptation to abiotic stress, but hormonal responses of cool-season turfgrass species to drought stress are not well documented. This study was to investigate responses of hormones and photosynthesis to drought stress and examine if drought stress - induced hormone alteration is associated with stress tolerance in kentucky bluegrass (KBG, Poa pratensis L.). The grass was grown in a growth chamber for 6 weeks and then subjected to drought stress [40%-50% evapotranspiration (ET) replacement)] for 28 d. Drought stress caused cell membrane damage, resulting in decline in photosynthetic rate (Pn), chlorophyll content, and visual quality in KBG. Drought stressed grass had higher leaf abscisic acid (ABA), lower leaf trans-zeatin riboside (ZR), isopentenyl adenosine (iPA), and indole-3-acetic acid (IAA), but similar level of leaf gibberellin A4 (GA4) when compared to the control (well-watered). On average, drought stress treatment reduced leaf ZR by 59.1%, iPA by 50.4%, IAA by 26.7%, while increased ABA by 108.5% when compared to the control at the end of drought stress (28 d). The turf quality and photosynthetic rate was positively correlated with cytokinins and IAA, but negatively correlated with ABA and ABA/cytokinins (CK) ratio under drought stress. The results of this study suggest drought stress-induced injury to Kentucky bluegrass may be associated with hormonal alteration, and the plants with higher cytokinins and IAA and less ABA under drought stress may have better photosynthetic function and performance.

Chemical Composition and Allelopathic, Phytotoxic and Pesticidal Activities of Atriplex cana Ledeb. (Amaranthaceae) Essential Oil.

The chemical composition and allelopathic, phytotoxic and pesticidal activities of Atriplex cana Ledeb. (Amaranthaceae) essential oil were investigated. Nineteen compounds were identified via GC/MS, representing 82.3 % of the total oil, and the most abundant constituents were dibutyl phthalate (21.79 %), eucalyptol (20.14 %) and myrtenyl acetate (15.56 %). The results showed that volatile organic compounds (VOCs) released by A. cana significantly inhibited seedling growth of Amaranthus retroflexus L. and Poa annua L., and 80 g of fresh stems and leaves of A. cana in a 1.5 L airtight container almost completely suppressed the seed germination of both plants. Meanwhile, 5 µg/mL essential oil completely inhibited the seed germination of A. retroflexus, Medicago sativa L., P. annua and Echinochloa crusgalli L. Pesticidal testing revealed that the essential oil had strong behavioral avoidance and lethal effects on Aphis pomi DeGeer. Five microliters of essential oil/Petri dish treatment resulted in an 84.5 % mortality rate after 12 h, and the mortality rate reached nearly 100 % after 48 h. This report is the first one on the chemical composition as well as the biological activity of the essential oil of A. cana, and our results indicate that the oil is valuable in terms of being further exploited as a bioherbicide/insecticide.

Water transport properties of root cells contribute to salt tolerance in halophytic grasses Poa juncifolia and Puccinellia nuttalliana.

Plant water uptake and aquaporin-mediated root water transport are among the most salt-sensitive processes in most plants, but even relatively high salt concentrations do not appear to impair water transport processes in halophytes. To develop better understanding of these processes in halophytic plants, we compared the responses to NaCl of the two halophytic grasses varying in salt tolerance, Puccinellia nuttalliana and Poa juncifolia, with the glycophytic grass Poa pratensis. The plants were hydroponically grown and subjected to different NaCl concentrations for up to 10 days. At the lower NaCl concentrations, shoot and root dry weights were drastically reduced in Poa pratensis, but increased in Puccinellia nuttalliana and Poa juncifolia. The examined treatment concentrations of up to 300 mM NaCl had either no effect (Puccinellia nuttalliana) or little effect (Poa juncifolia) on the net photosynthesis and transpiration rates in plants, but severely decreased the gas exchange parameters in Poa pratensis. Similarly, to growth and gas exchange, leaf water content in Puccinellia nuttalliana was not affected even by the highest, 300 mM NaCl concentration, while Poa pratensis showed decreased shoot water content in all examined NaCl treatments and Poa juncifolia in 150 and 300 mM NaCl. Cell hydraulic conductivity in roots of Poa pratensis also showed high sensitivity to NaCl and was drastically reduced in all examined NaCl concentrations. Cell hydraulic conductivity in Poa juncifolia roots was less affected by NaCl compared with Poa pratensis and in Puccinellia nuttalliana, cell hydraulic conductivity increased in response to NaCl treatments. Both Puccinellia nuttalliana and Poa juncifolia accumulated less Na in their shoot tissues compared with Poa pratensis. The concentrations of K in the roots of Poa pratensis sharply decreased with increasing NaCl treatment concentrations while in Puccinellia nuttalliana, K root concentrations remained high in all NaCl treatments and in Poa juncifoila, root K decreased only in the 300 mM NaCl treatment. Since K efflux from the cytoplasm can contribute to the acidification of the cytoplasm, this process could potentially lead to the inhibition of aquaporin function and reduction of root hydraulic conductivity. The, significance of stable K root concentrations in the roots of halophytes should be further investigated as a possible salt tolerance mechanism that could contribute to the maintenance of aquaporin function and root water transport under salt stress conditions.

Chemical Composition and Phytotoxic Activity of Seriphidium terrae-albae (Krasch.) Poljakov (Compositae) Essential Oil.

To evaluate the potential value of Seriphidium terrae-albae (Krasch.) Poljakov essential oil as bioherbicide, its chemical composition as well as phytotoxic activity was investigated. Seventeen compounds were identified via GC/MS, representing 98.1 % of the total oil, and the most abundant constituents were α-thujone (43.18 %), ß-thujone (16.92 %), eucalyptol (17.55 %), and camphor (13.88 %). Phytotoxic assay revealed that the essential oil as well as its major constituents exhibited inhibitory activity on root and shoot growth of receiver plants in a dose-dependent manner. When the concentration reached 20 µg/mL, root length of Amaranthus retroflexus was reduced to 31.3 %, 70.6 %, 36.9 %, and 66.6 % of the control, respectively, when treated with α-thujone, eucalyptol, camphor, and the mixture of these compounds; meanwhile, root length of Poa annua was 3.0 %, 24.2 %, 0 %, and 4.4 % of the control when the same chemicals were applied. On the other hand, the essential oil showed a much stronger activity. At 1.5 µL/mL, root and shoot length of A. retroflexus and P. annua were reduced to 0.65 %, 0.5 %, and 1.53 %, 1.51 % of the control, respectively, and seed germination of A. retroflexus and P. annua was completely inhibited when the oil concentration reached 3 µg/mL and 5 µg/mL, respectively. This is the first report on the chemical composition of the essential oil of S. terrae-albae, and our results indicated that it has the potential to be further exploited as a bioherbicide.

Transcriptomics analysis of the flowering regulatory genes involved in the herbicide resistance of Asia minor bluegrass (Polypogon fugax).

BACKGROUND: Asia minor bluegrass (Polypogon fugax, P. fugax), a weed that is both distributed across China and associated with winter crops, has evolved resistance to acetyl-CoA carboxylase (ACCase) herbicides, but the resistance mechanism remains unclear. The goal of this study was to analyze the transcriptome between resistant and sensitive populations of P. fugax at the flowering stage. RESULTS: Populations resistant and susceptible to clodinafop-propargyl showed distinct transcriptome profiles. A total of 206,041 unigenes were identified; 165,901 unique sequences were annotated using BLASTX alignment databases. Among them, 5904 unigenes were classified into 58 transcription factor families. Nine families were related to the regulation of plant growth and development and to stress responses. Twelve unigenes were differentially expressed between the clodinafop-propargyl-sensitive and clodinafop-propargyl-resistant populations at the early flowering stage; among those unigenes, three belonged to the ABI3VP1, BHLH, and GRAS families, while the remaining nine belonged to the MADS family. Compared with the clodinafop-propargyl-sensitive plants, the resistant plants exhibited different expression pattern of these 12 unigenes. CONCLUSION: This study identified differentially expressed unigenes related to ACCase-resistant P. fugax and thus provides a genomic resource for understanding the molecular basis of early flowering.

Differential effects of citric acid on cadmium uptake and accumulation between tall fescue and Kentucky bluegrass.

Organic acids play an important role in cadmium availability, uptake, translocation, and detoxification. A sand culture experiment was designed to investigate the effects of citric acid on Cd uptake, translocation, and accumulation in tall fescue and Kentucky bluegrass. The results showed that two grass species presented different Cd chemical forms, organic acid components and amount in roots. The dormant Cd accumulated in roots of tall fescue was the pectate- and protein- integrated form, which contributed by 84.85%. However, in Kentucky bluegrass, the pectate- and protein- integrated Cd was only contributed by 35.78%, and the higher proportion of Cd form was the water soluble Cd-organic acid complexes. In tall fescue, citric acid dramatically enhanced 2.8 fold of Cd uptake, 3 fold of root Cd accumulation, and 2.3 fold of shoot Cd accumulation. In Kentucky bluegrass, citric acid promoted Cd accumulation in roots, but significantly decreased Cd accumulation in shoots. These results suggested that the enhancements of citric acid on Cd uptake, translocation, and accumulation in tall fescue was associated with its promotion of organic acids and the water soluble Cd-organic acid complexes in roots.

5-Aminolevulinic acid modulates antioxidant defense systems and mitigates drought-induced damage in Kentucky bluegrass seedlings.

Drought stress occurs frequently and severely as a result of global climate change, and it exerts serious effects on plants. 5-Aminolevulinic acid (5-ALA) plays a crucial role in conferring abiotic stress tolerance in plants. To enhance the drought tolerance of turfgrass and investigate the effects of 5-ALA on antioxidant metabolism and gene expression under drought stress conditions, exogenous 5-ALA was applied by foliar spraying before Kentucky bluegrass (Poa pratensis L.) seedlings were exposed to drought [induced by 10% polyethylene glycol (PEG)] stress for 20 days. 5-ALA pretreatment increased turf quality (TQ) and leaf relative water content (RWC) while reducing reactive oxygen species (ROS) production including H2O2 content and O2•- generation rate, lipoxygenase (LOX) activity, and malondialdehyde (MDA) content under drought stress. 5-ALA pretreatment maintained ascorbate (AsA) and glutathione (GSH) contents and the ASA/DHA and GSH/GSSG ratios at high levels, and it enhanced the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione peroxidase (GPX), dehydroascorbate reductase (DHAR), and glutathione reductase (GR), which are crucial for scavenging drought-induced ROS. In addition, 5-ALA upregulated the relative expression levels of Cu/ZnSOD, APX, GPX, and DHAR but downregulated those of CAT and GR under drought stress. These results indicated that the application of 5-ALA might improve turfgrass quality and promote drought tolerance in Kentucky bluegrass through reducing oxidative damage and increasing non-enzyme antioxidant levels and antioxidant enzyme activity at transcriptional and posttranscriptional levels.

Silicate application increases the photosynthesis and its associated metabolic activities in Kentucky bluegrass under drought stress and post-drought recovery.

Drought stress is the most pervasive threat to plant growth, which disrupts the photosynthesis and its associated metabolic activities, while silicate (Si) application may have the potential to alleviate the damaging effects of drought on plant growth. In present study, the role of Si in regulating the photosynthesis and its associated metabolic events in Kentucky bluegrass (cv. Arcadia) were investigated under drought stress. Drought stress and four levels (0, 200, 400, 800 mg L(-1)) of Si (Na2SiO3.9H2O) were imposed on 1-year-old plants removed from field and cultured under glasshouse conditions. After 20 days of drought stress, the plants were re-watered to reach soil field capacity for the examination of recovery on the second and the seventh day. The experiment was arranged in completely randomized design replicated four times. Drought stress severely decreased the photosynthesis, water use efficiency, stomatal conductance, cholorophyll contents, Rubisco activity, and Rubisco activation state in Kentucky bluegrass. Nevertheless, application of Si had a positive influence on all these attributes, particularly under stress conditions. As compared to control, Si application at 400 mg L(-1) recorded 78, 64, and 48 % increase in photosynthesis, Rubisco initial activity, and Rubisco total activity, respectively, at 20 days of drought. Higher photosynthesis and higher Rubisco activity in Si-applied treatments suggest that Si may have possible (direct or indirect) role in maintenance of more active Rubisco enzyme and Rubisco activase and more stable proteins for carbon assimilation under stress conditions, which needs to be elucidated in further studies.

A new amino acid substitution (Ala-205-Phe) in acetolactate synthase (ALS) confers broad spectrum resistance to ALS-inhibiting herbicides.

MAIN CONCLUSION: This is a first report of an Ala-205-Phe substitution in acetolactate synthase conferring resistance to imidazolinone, sulfonylurea, triazolopyrimidines, sulfonylamino-carbonyl-triazolinones, and pyrimidinyl (thio) benzoate herbicides. Resistance to acetolactate synthase (ALS) and photosystem II inhibiting herbicides was confirmed in a population of allotetraploid annual bluegrass (Poa annua L.; POAAN-R3) selected from golf course turf in Tennessee. Genetic sequencing revealed that seven of eight POAAN-R3 plants had a point mutation in the psbA gene resulting in a known Ser-264-Gly substitution on the D1 protein. Whole plant testing confirmed that this substitution conferred resistance to simazine in POAAN-R3. Two homeologous forms of the ALS gene (ALSa and ALSb) were detected and expressed in all POAAN-R3 plants sequenced. The seven plants possessing the Ser-264-Gly mutation conferring resistance to simazine also had a homozygous Ala-205-Phe substitution on ALSb, caused by two nucleic acid substitutions in one codon. In vitro ALS activity assays with recombinant protein and whole plant testing confirmed that this Ala-205-Phe substitution conferred resistance to imidazolinone, sulfonylurea, triazolopyrimidines, sulfonylamino-carbonyl- triazolinones, and pyrimidinyl (thio) benzoate herbicides. This is the first report of Ala-205-Phe mutation conferring wide spectrum resistance to ALS inhibiting herbicides.

Protective effect of spermidine on salt stress induced oxidative damage in two Kentucky bluegrass (Poa pratensis L.) cultivars.

To improve the salinity tolerance of turfgrass and investigate the effect of spermidine (Spd) on antioxidant metabolism and gene expression under salinity stress condition, exogenous Spd was applied before two kentucky bluegrass (Poa pratensis L.) cultivars ('Kenblue' and 'Midnight') were exposed to 200 mM sodium chloride (NaCl) stress for 28 d. Salinity stress decreased the turfgrass quality, increased the content of malonyldialdehyde (MDA), superoxide anion (O2(·-)) and hydrogen peroxide (H2O2), and enhanced activities of superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (POD) and ascorbate peroxidase (APX) and isozymes intensity in both cultivars. In addition, the expression level of Cu/ZnSOD was down-regulated in 'Kenblue' but up-regulated in 'Midnight' after salt treatment. Salinity stress also enhanced the expression of APX but inhibited the expression of CAT and POD in both cultivars. Exogenous Spd treatment alleviated the salinity-induced oxidative stress through decreasing MDA, H2O2 and O2(·-) contents in both cultivars. Besides, exogenous Spd further enhanced the activities of SOD, CAT, POD and APX accompanied with the increased intensity of specific isozymes of SOD, CAT and APX in both cultivars and POD in 'Kenblue'. Moreover, Spd further up-regulated expression levels of Cu/ZnSOD and APX, but down-regulated those of CAT and POD in both cultivars. These results indicated that exogenous Spd might improve turfgrass quality and promote the salinity tolerance in the two cultivars of kentucky bluegrass through reducing oxidative damages and increasing enzyme activity both at protein and transcriptional levels.

Potential ecological roles of flavonoids from Stellera chamaejasme.

Stellera chamaejasme L. (Thymelaeaceae), a perennial weed, distributes widely in the grasslands of Russia, Mongolia and China. The plant synthesizes various secondary metabolites including a group of flavonoids. To our knowledge, flavonoids play important roles in the interactions between plants and the environment. So, what are the benefits to S. chamaejasme from producing these flavonoids? Here, we discuss the potential ecological role of flavonoids from S. chamaejasme in protecting the plant from insects and other herbivores, as well as pathogens and competing plant species, and new data are provided on the phytotoxicity of flavonoids from S. chamaejasme toward Poa annua L.

Utilizing next-generation sequencing to study homeologous polymorphisms and herbicide-resistance-endowing mutations in Poa annua acetolactate synthase genes.

BACKGROUND: Detection of single nucleotide polymorphisms (SNPs) related to herbicide resistance in non-model polyploid weed species is fraught with difficulty owing to the gene duplication and lack of reference sequences. Our research seeks to overcome these obstacles by Illumina HiSeq read mapping, SNP calling and allele frequency determinations. Our focus is on the acetolactate synthase (ALS) gene, the target site of ALS-inhibiting herbicides, in Poa annua, an allotetraploid weed species originating from two diploid parents, P. supina and P. infirma. RESULTS: ALS contigs with complete coding regions of P. supina, P. infirma and P. annua were assembled and compared with ALS genes from other plant species. The ALS infirma-homeolog of P. annua showed higher levels of nucleotide sequence variability than the supina-homeolog. Comparisons of read mappings of P. annua and a simulated P. supina × P. infirma hybrid showed high resemblance. Two homeolog-specific primer pairs were designed and used to amplify a 1860 bp region covering all resistance-conferring codons in the ALS gene. Four P. annua populations, GN, RB, GW and LG, showed high resistance to two ALS inhibitors, bispyribac-sodium and foramsulfuron, and two populations, HD and RS, showed lower resistance in the rate-response trial. Mutations conferring Trp-574-Leu substitution were observed in the infirma-homeolog of GN and RB and in the supina-homeolog of GW and LG, but no resistance-conferring mutation was observed in the two populations of lower resistance, HD and RS. CONCLUSION: In this study we have demonstrated the use of NGS data to study homeologous polymorphisms, parentage and herbicide resistance in an allotetraploid weed species, P. annua. Complete coding sequences of the ALS gene were assembled for P. infirma, P. supina, infirma-homeolog and supina-homeolog in P. annua. A pipeline consisting of read mapping, SNP calling and allele frequency calculation was developed to study the parentage of P. annua, which provided a new perspective on this topic besides the views of morphology, karyotype and phylogeny. Our two homeolog-specific primer pairs can be utilized in future research to separate the homeologs of the ALS gene in P. annua and cover all the codons that have been reported to confer herbicide resistance.

Physiological effects of temperature on turfgrass tolerance to amicarbazone.

BACKGROUND: Amicarbazone effectively controls annual bluegrass (Poa annua L.) in bermudagrass [Cynodon dactylon (L.) Pers. × C. transvaalensis Burtt-Davy] and tall fescue (Festuca arundinacea Schreb.) with spring applications, but summer applications may excessively injure tall fescue. The objective of this research was to investigate physiological effects of temperature on amicarbazone efficacy, absorption, translocation and metabolism in annual bluegrass, bermudagrass and tall fescue. RESULTS: At 25/20 °C (day/night), annual bluegrass absorbed 58 and 40% more foliar-applied amicarbazone than bermudagrass and tall fescue, respectively, after 72 h. Foliar absorption increased at 40/35 °C in all species, compared with 25/20 °C, and tall fescue had similar absorption to annual bluegrass at 40/35 °C. At 6 days after treatment, annual bluegrass metabolized 54% of foliar-applied amicarbazone, while bermudagrass and tall fescue metabolized 67 and 64% respectively. CONCLUSION: Tall fescue is more tolerant to amicarbazone than annual bluegrass at moderate temperatures (≈25/20 °C) owing to less absorption and greater metabolism. However, tall fescue susceptibility to amicarbazone injury at high temperatures (40/35 °C) results from enhanced herbicide absorption compared with lower temperatures (25/20 °C). Bermudagrass is more tolerant to amicarbazone than annual bluegrass and tall fescue owing to less herbicide absorption, regardless of temperature.

Methiozolin sorption and mobility in sand-based root zones.

BACKGROUND: Methiozolin is a herbicide currently used for annual bluegrass control in golf course putting greens. Previous research indicates that maximum weed control efficacy requires root exposure; however, soil sorption and mobility of methiozolin have not been established. Research was conducted to investigate soil sorption and subsequent desorption by dilution of methiozolin, as well as soil mobility using batch equilibrium experiments and thin-layer chromatography in nine root zones. Evaluations focused on sand-based systems typical of many golf course putting greens. RESULTS: Sorption coefficients (Kd values) ranged from 0.4 to 29.4 mL g(-1) and averaged 13.8 mL g(-1) . Sorption was most influenced by organic matter content; conversely, soil pH had a negligible effect. Methiozolin desorption did not occur with a 0.01 M CaCl2 dilution. Methiozolin mobility was low; retardation factors (Rf values) were <0.05 for all media with ≥0.3% organic matter. Sand (0.1% organic matter) resulted in an Rf value of 0.46. CONCLUSION: Approximately 24% of applied methiozolin is available for root uptake, and mobility is limited, suggesting resistance to loss through leaching displacement.