Insights into the molecular mechanism of the responses for Cyperus alternifolius to PhACs stress in constructed wetlands.

Cyperus alternifolius has been widely reported to be an effective phytoremediation plant in constructed wetland systems (CWs). In this context, an integrated biochemical and proteomic analysis of C. alternifolius leaves exposed to pharmaceutically active compounds (PhACs) in CWs was conducted to understand the mechanism of phytoremediation. The obtained results showed the antioxidant enzyme activities were induced throughout the experiment; however over time, the malondialdehyde content is not significantly different from the control and the photosynthetic pigment contents in plant were subsequently slowly recovered. Therefore, we concluded that reactive oxygen species could be effectively counteracted by the enhanced antioxidant enzyme activities, and therefore the photosynthetic pigments were ultimately restored. Leaf extract proteome maps were obtained through 2-DE, and an average of 55, 49, and 24 spots were significantly altered by 30, 100, and 500 µg/L of PhACs over the control, respectively. Protein expression patterns showed that proteins in C. alternifolius leaves are associated with photosynthesis, energy metabolism, defense, and protein synthesis. Moreover, the most relevant pathways modulated by PhACs were photosynthesis and energy metabolism. The protein expression involved in antioxidant defense and stress response generally increased in all the PhAC treatments. The regulated proteins may favor PhAC degradation in CWs; however, the role of these proteins in degrading PhACs remains unknown; further biochemical studies should be conducted. This study indicated that C. alternifolius can tolerate multiple PhACs.

Dynamic high pressure microfluidization-assisted extraction and bioactivities of Cyperus esculentus (C. esculentus L.) leaves flavonoids.

The aim of this work was to study the effect of dynamic high pressure microfluidization (DHPM) on extracting total flavonoids from Cyperus esculentus L. (C. esculentus L.) leaves and to evaluate the antioxidant activity and antibacterial property of these flavonoids. In all the assays, pretreatment with DHPM was found to not only efficiently improve the yield of total flavonoids but also strengthen the antioxidant activity of the total flavonoids. C. esculentus L. leaves flavonoids had pronounced antioxidant activity in vivo that could significantly elevate the content of superoxide dismutase (SOD) without increasing the malondialdehyde (MDA) levels, and could also improve total antioxidant capacity in mice with a dose-dependent fashion. C. esculentus L. leaves flavonoids inhibited the growth of both Gram positive and Gram negative bacteria while no obvious inhibitory effect on Penicillium and Aspergillus could be observed. Our studies indicate that flavonoids from C. esculentus L. leaves can be taken as a natural antioxidant and bacteriostatic substance in food and pharmaceutical industry.

First report of resistance to acetolactate-synthase-inhibiting herbicides in yellow nutsedge (Cyperus esculentus): confirmation and characterization.

BACKGROUND: Yellow nutsedge is one of the most problematic sedges in Arkansas rice, requiring the frequent use of halosulfuron (sulfonylurea) for its control. In the summer of 2012, halosulfuron at 53 g ha(-1) (labeled field rate) failed to control yellow nutsedge. The level of resistance to halosulfuron was determined in the putative resistant biotype, and its cross-resistance to other acetolactate synthase (ALS) inhibitors from four different herbicide families. ALS enzyme assays and analysis of the ALS gene were used to ascertain the resistance mechanism. RESULTS: None of the resistant plants was killed by halosulfuron at a dose of 13 568 g ha(-1) (256× the field dose), indicating a high level of resistance. Based on the whole-plant bioassay, the resistant biotype was not controlled by any of the ALS-inhibiting herbicides (imazamox, imazethapyr, penoxsulam, bispyribac, pyrithiobac-sodium, bensulfuron and halosulfuron) tested at the labeled field rate. The ALS enzyme from the resistant biotype was 2540 times less responsive to halosulfuron than the susceptible biotype, and a Trp574 -to-Leu substitution was detected by ALS gene sequencing using the Illumina HiSeq. CONCLUSION: The results suggest a target-site alteration as the mechanism of resistance in yellow nutsedge, which accounts for the cross-resistance to other ALS-inhibiting herbicide families.

Cross-resistance to herbicides of five ALS-inhibiting groups and sequencing of the ALS gene in Cyperus difformis L.

Resistance to ALS-inhibiting herbicides in Cyperus difformis has evolved rapidly in many rice areas worldwide. This study identified the mechanism of resistance, assessed cross-resistance patterns to all five chemical groups of ALS-inhibiting herbicides in four C. difformis biotypes, and attempted to sequence the ALS gene. Whole-plant and ALS enzyme activity dose-response assays indicated that the WA biotype was resistant to all ALS-inhibiting herbicides evaluated. The IR biotype was resistant to bensulfuron-methyl, orthosulfamuron, imazethapyr, and propoxycarbazone-sodium and less resistant to bispyribac-sodium and halosulfuron-methyl, and susceptible to penoxsulam. ALS enzyme activity assays indicated that resistance is due to an altered target site yet mutations previously found to endow target-site resistance in weeds were not detected in the sequences obtained. The inability to detect resistance mutations in C. difformis may result from the presence of additional ALS genes, which were not amplified by the primers used. This study reports the first ALS gene sequence from Cyperus difformis. Certain ALS-inhibiting herbicides can still be used to control some resistant C. difformis biotypes. However, because cross-resistance to all five classes of ALS-inhibitors was detected in other resistant biotypes, these herbicides should only be used within an integrated weed management program designed to delay the evolution of herbicide resistance.

Adaptation to flooding in upland and lowland ecotypes of Cyperus rotundus, a troublesome sedge weed of rice: tuber morphology and carbohydrate metabolism.

BACKGROUND AND AIMS: In recent years, Cyperus rotundus has become a problem weed in lowland rice (Oryza sativa) grown in rotation with vegetables in the Philippines. As the growth of C. rotundus is commonly suppressed by prolonged flooding, the ability of the weed to grow vigorously in flooded as well as upland conditions suggests that adapted ecotypes occur in these rotations. Studies were conducted to elucidate the mechanisms that permit C. rotundus to tolerate flooded soil conditions. METHODS: Upland and lowland ecotypes of C. rotundus were compared in terms of growth habit, carbohydrate reserves and metabolism, and activities of enzymes involved in alcoholic fermentation - alcohol dehydrogenase (ADH) and pyruvate decarboxylase (PDC). KEY RESULTS: The lowland ecotype has much larger tubers than the upland ecotype. Prior to germination, the amylase activity and total non-structural carbohydrate content in the form of soluble sugars were greater in the tubers of lowland plants than in those of upland C. rotundus. At 24 h after germination in hypoxic conditions, PDC and ADH activities in the lowland plants increased, before decreasing at 48 h following germination. In contrast, ADH and PDC activities in the upland plants increased from 24 to 48 h after germination. CONCLUSIONS: Tolerance of lowland C. rotundus of flooding may be attributed to large carbohydrate content and amylase activity, and the ability to maintain high levels of soluble sugars in the tubers during germination and early growth. This is coupled with the modulation of ADH and PDC activities during germination, possibly to control the use of carbohydrate reserves and sustain substrate supply in order to avoid starvation and death of seedlings with prolonged flooding.

Contaminated soil phytoremediation by Cyperus laxus Lam. cytochrome p450 EROD-activity induced by hydrocarbons in roots.

Laboratory and greenhouse experiments with Cyperus laxus Lam were conducted to determine the rate and extent of phytoremediation and the effect of hydrocarbons on the cytochrome P450 EROD (7-ethoxyresorufin-O-deethylase) enzymatic activity in roots. Plants were cultivated on hydrocarbon-contaminated soil (HCS) and spiked perlite. Phytoremediation was evaluated using 6.5 kg HCS (173 +/- 15 mg total petroleum hydrocarbons [TPH] g(-1) of dry soil) pots at different moisture contents; the average removal rate was 3.46-0.25 mg TPH g(-1) dry soil month(-1) and 48% was removed when moisture was kept at 60%. The aromatic hydrocarbon fraction was the mostly removed, 60%; aliphatic, 51%; and polar 24% after 24-month experiments. In unplanted pots, TPH concentration did not exhibit significant differences with respect to the initial concentration. We confirmed that the presence of hydrocarbons induced ERODactivity up to 6.5-fold. Moreover, short-term experiments (up to 13 d) with spiked perlite demonstrated that two EROD activities in roots contributed to the total detected; 60% was found in the cytosolic and 40% in the microsomal fraction. To our knowledge, this is the first work that tries to build links between the hydrocarbon-inducible character of ERODactivity in roots and the phytoremediation ability of C. laxus in highly contaminated soils.

Cross-resistance pattern and alternative herbicides for Cyperus difformis resistant to sulfonylurea herbicides in Korea.

A Cyperus difformis L accession from Chonnam province, Korea was tested for resistance to the sulfonylurea herbicide, imazosulfuron. The accession was confirmed to be resistant (R) and was cross-resistant to other sulfonylurea herbicides, bensulfuron-methyl, cyclosulfamuron and pyrazosulfuron-ethyl, the pyrimidinyl thiobenzoate herbicide, bispyribac-sodium, and the imidazolinone herbicide imazapyr, but not to imazaquin. Multiple resistance was tested using twelve herbicides with target sites other than acetolactate synthase (ALS). The R biotype could be controlled by other herbicides with different modes of action such as butachlor, carfentrazone-ethyl, clomeprop, dithiopyr, esprocarb, mefenacet, oxadiazon, pretilachlor, pyrazolate and thiobencarb, applied to soil at recommended rates. Several sulfonylurea herbicide-based mixtures can control both the R and S biotypes of C difformis, except sulfonylurea plus dimepiperate, molinate or pyriftalid, and pyrazolate plus butachlor. Although mixtures of sulfonylurea herbicides might be more effective, they should be avoided and used only in special cases. In terms of in vitro ALS activity, the R biotype was 1139-, 3583-, 1482-, 416-, 5- and 9-fold more resistant to bensulfuron-methyl, cyclosulfamuron, imazosulfuron, pyrazosulfuron-ethyl, bispyribac-sodium and imazapyr, respectively, than the S biotype. The in vivo ALS activity of the R biotype was also less affected by the sulfonylurea herbicides, imazosulfuron and pyrazosulfuron-ethyl, than the S biotype. Results of in vitro and in vivo ALS assays indicated that the resistance mechanism of C difformis to ALS inhibitor herbicides was primarily due to an alteration in the target enzyme, ALS. Greenhouse experiments showed delayed flowering and reduced seed production of the R biotype, which could possibly result in reduced fitness. This unusual observation needs to be confirmed in field situations.

Evaluation of resistance in Cyperus difformis populations to ALS inhibiting herbicides.

Due to the intensification of rice cultivation in Spain and the repeated use of herbicides as an effective tool to control weeds, one important fact is the appearance of resistant populations of weeds, which are no longer controlled effectively at field doses. Therefore, it is necessary to increase doses to such an extent as to produce phytotoxicity in the rice crop. In paddy fields in Badajoz (Spain), a few years ago, populations of Cyperus difformis resistant to bensulfuron-methyl appeared. In order to characterize this resistance, whole plant assays were carried out in fourteen populations, eight of them originating from treated Spanish paddy fields. The results were expressed as resistance factor (RF) = ED50(R)/ED50(S). Two populations of Cyperus difformis, resistant (R) and tolerant (T) to bensulfuron-methyl, displayed resistance factors greater than 30 and 12.5, respectively. Furthermore, in order to evaluate the existence of a possible cross-resistance, the recommended field doses of 3 other herbicides (imazamox, ethoxysulfuron and bentazone + MCPA) were applied to the plants. All populations were susceptible to both imazamox and bentazone + MCPA and two populations showed cross-resistance to ethoxysulfuron. ALS activity was assayed in vitro in four biotypes, the R/S ratios of the I50 values calculated indicate that resistance involves a reduction in the sensitivity of the ALS mainly in the R population (biotype 4) to bensulfuron-methyl and ethoxysulfuron, displaying R/S ratios of 37500 and 142857, respectively, which may be attributed to inherent differences in the ALS protein itself and/or in the stability of the enzyme.