Globe fringerush (Fimbristylis miliacea) cross resistance to als-inhibitor herbicides under field conditions in irrigated rice in the south of Brazil

ALS-inhibiting herbicides usually provide adequate weed control in irrigated rice fields. After consecutive years of use, the Cyperaceae species, globe fringerush (Fimbristylis miliacea) began to show resistance to ALS (acetolactate synthase) inhibitors. Globe fringerush is one of the most problematic herbicide-resistant weeds in irrigated rice in the state of Santa Catarina in the South of Brazil. The objective of this research was to examine cross resistance of globe fringerush to ALS inhibitors, under field conditions. Two experiments were conducted in a rice field naturally infested with ALS-resistant globe fringerush in Santa Catarina, in the 2008/09 and 2009/10 cropping seasons. The experimental units were arranged in randomized complete block design, with five replicates, consisting of two factors (herbicide and dose) in a 4 x 5 factorial arrangement. ALS herbicides included bispyribac-sodium, ethoxysulfuron, pyrazosulfuron-ethyl and penoxsulam. Six-leaf globe fringerush was sprayed with herbicide doses of 0, 0.5, 1, 2 and 4X the recommended doses in a spray volume of 200 L ha-1. The number of rice culm, filled and sterile grains, plant height, dry shoot biomass and grain yield were recorded. Globe fringerush control was evaluated 28 and 70 days after herbicide application (DAA); shoots were harvested at 13 weeks after herbicide application and dry weight recorded. Competition with globe fringerush reduced the number of culm and rice grain yield. The globe fringerush biotype in this field was resistant to all ALS herbicides tested. Penoxsulam had the highest level of activity among treatments at 28 and 70 DAA, but the control level was only 50% and 42%, respectively, in the second year of assessment. This was not enough to prevent rice yield loss. Alternative herbicides and weed control strategies are necessary to avoid yield losses in rice fields infested with ALS-resistant biotypes of globe fringerush.

Herbicide selectivity in alfalfa crops

This study aimed to determine the selectivity of herbicides applied in pre- and post-emergence for alfalfa crops. Three separate experiments were carried out under greenhouse conditions. The first experiment was arranged in a completely randomized design with three replications in a 4 x 11 + 1 factorial scheme , with eleven herbicides (bentazon, chlorimuron-ethyl, fomesafen, fluazifop-p-butyl, saflufenacil, imazethapyr, clethodim, nicosulfuron, imazaquin, haloxyfop-methyl and MSMA), four doses of each herbicide (0.5 D, 0.75 D, 1.0 D and 1.25 D, where D = recommended dose), plus an untreated control. The products were applied to alfalfa plants at the stage of 4 to 5 leaf pairs. In the second experiment, the effect of pre-emergent herbicides on early alfalfa development was observed through a completely randomized design with five replications in a 3 x 4 x 2 factorial scheme, with three herbicides (hexazinone, atrazine + simazine, S-metolachlor), four doses (0.5 D, 0.75 D, 1.0 D and 1.25 D), and two types of soil texture (loamy and clay soil), plus an untreated control. The third experiment evaluated the action of atrazine, 2,550 g ha-1; clomazone - 600 g ha-1; diclosulam - 25 g ha-1; diuron+hexazinone - 936 + 264 g ha-1 and diuron+hexazinone +sulfometuron - 1,386 + 391 + 33.35 g ha-1 on alfalfa sown at different times after herbicide application. The effects of the treatments on alfalfa were evaluated according to visual phytotoxicity symptoms, plant height, and biomass of roots and shoots. Among the herbicides applied at post-emergence, imazethapyr, clethodim, haloxyfop-p-methyl and MSMA were selective for alfalfa, while among those applied at pre-emergence, none were selective, regardless of soil texture. The results of the third experiment showed that the herbicides diclosulam, hexazinone + diuron and atrazine caused less toxicity in alfalfa plants.

Effect of selectivity of herbicides and plant growth regulators used in sugarcane crops on immature stages of Trichogramma galloi (Hymenoptera: Trichogrammatidae)

Herbicides and plant growth regulators are often used in sugarcane management. However, the use of non-selective pesticides can cause adverse effects on the efficiency of beneficial insects in integrated pest management. Within this context, this study aimed to evaluate the effect of such products on the immature stages of the parasitoid Trichogramma galloi. Eggs of Diatraea saccharalis containing the parasitoid at the egg-larva stage and at the prepupal and pupal stages were immersed in test solutions of the following pesticides (maximum recommended doses for sugarcane): herbicides clomazone and diuron + hexazinone, and plant growth regulators trinexapac-ethyl and sulfometuron-methyl. The biological properties evaluated were emergence (F1 and F2) and number of eggs parasitized by T. galloi (F1). The products were classified according to percentage of reduction in emergence and parasitism: harmless (<30%), slightly harmful (30-79%), moderately harmful (80-99%) and harmful (>99%). The pesticides evaluated were considered to be harmless or slightly harmful to immature T. galloi and, thus, their use should be preferred for preserving this parasitoid species in sugarcane management programs.

Multiple resistance of Conyza sumatrensis to Chlorimuron­ethyl and to Glyphosate

Weed resistance to herbicides has been a major issue in Brazil, mainly due to the inefficiency of the herbicides used in no-till areas and to the high cost of these herbicide treatments. Failures in controlling the weed Conyza have been reported in Western and Northern grain crop areas in Paraná (Brazil). This work aimed to evaluate the potential occurrence of C. sumatrensis biotypes resistant to the herbicides chlorimuron-ethyl and glyphosate. Experiments were carried out under greenhouse conditions with four biotypes (Cascavel-2, Toledo-4, Tupãssi-6, and Assis Chateaubriand-7) possibly resistant to, as well as a population considered susceptible to chlorimuron-ethyl and glyphosate. To obtain dose-response curves, eight herbicide doses of chlorimuron-ethyl (0, 2.5, 5, 10, 20, 40, 80 and 160 g ha-1) and glyphosate (0, 90, 180, 360, 720, 1,440, 2,880 and 5,760 g e.a. ha-1) were applied and weed control and shoot biomass evaluations were made. Results provided evidence that two biotypes (Cascavel-2 and Tupãssi-6) were resistant to glyphosate and four biotypes (Cascavel-2, Toledo-4, Tupãssi-6 and Assis Chateaubriand-7) were resistant to chlorimuron­ethyl. Multiple resistance to glyphosate and chlorimuron was confirmed for biotypes Cascavel­2 and Tupãssi 6. This is the first report on multiple resistance in Conyza sumatrensis, worldwide.

PHYSIOLOGICAL QUALITY OF SOYBEAN SEEDS DEPENDING ON THE PREHARVEST DESICCATION

ABSTRACTDepending on the cultivar, the use of desiccants in the preharvest can favor maintenance of physiological quality. The objective of the study was to assess the physiological quality of soybean seeds as due to the use of apreharvest desiccant and desiccation time in two harvests (2011/12 and 2012/13). The treatments were five soybean cultivars, two growth stages of application, a control (without desiccant application), and three desiccants (glufosinate-ammonium, carfentrazone-ethyl and paraquat) (2011/12 harvest). In the 2012/13 harvest the carfentrazone-ethyl desiccant was replaced by diquat. The physiological quality of seeds was assessed by the percentage of viability and vigor (cold test, tetrazolium test and accelerated aging test). In 2011/12 harvest, there was an early harvest in six days with the use of glufosinate-ammonium and paraquat desiccants, when desiccation was done in stage R7.1, with maintenance of seed quality; however it was dependent on the cultivar. In the 2012/13 harvest there was no early harvesting due to the presence of rain in the preharvest and the use of desiccation did not affect the physiological quality of the seeds either. Cultivar NA5909 RG was more tolerant to remaining in the cultivation environment and maintained higher viability than 90% and greater vigor of 71% by the cold test compared to cultivar BMX Turbo (2011/12 harvest). It is concluded that desiccation can be a viable alternative to the soybean early harvesting, but it depends on the cultivar, the time of desiccation, the active principle of the desiccant and the absence of rain in preharvest.

SELECTION OF HERBICIDES TARGETING THE USE IN CROP SYSTEMS CULTIVATED WITH SHOWY CROTALARIA

ABSTRACT The increase in the area planted with Crotalaria spectabilishas occurred by several factors, highlighting the potential to reduce the nematodes, nitrogen fixation and the high production of biomass. By becoming a species sown as a crop, it is necessary to control the weeds that coexist with showy crotalaria. This change in the use of this crop creates the possibility of this specie becoming a weed. The aim of this study was to assess the potential use of herbicides applied in preemergence and postemergence of C.spectabilisfor different purposes (control of volunteer and selectivity plants). Three experiments were installed in a greenhouse (two with herbicides applied in preemergence - in soils with distinct textural categories; and one experiment with herbicides applied in postemergence). The results of the experiments with herbicides applied in preemergence showed that: amicarbazone, atrazine, diuron, metribuzin, prometryn, fomesafen and sulfentrazone showed effectiveness for control of C.spectabilis in clayey soil. Besides these, flumioxazin and isoxaflutole also showed potential to be used in the control of showy crotalaria in soils with loam texture. In relation to the postemergence herbicides, atrazine, diuron, prometryn, flumioxazin, fomesafen, lactofen, saflufenacil, amonio-glufosinate and glyphosate can be used aiming the chemical control of C.spectabilis. Herbicides chlorimuron-ethyl, diclosulan, imazethapyr, pyrithiobac-sodium, trifloxysulfuron-sodium, clomazone, pendimethalin, S-metolachlor and trifluralin applied in preemergence, and imazethapyr, pyrithiobac-sodium, flumiclorac, bentazon and clethodim applied in postemergence caused low levels of injury to C.spectabilis plants, making necessary the development of new searches to ensure the selectivity of these products.

WEED MANAGEMENT WITH HERBICIDES APPLIED IN PRE AND POSTEMERGENCE ON CASTOR CROP

ABSTRACT The aim of this work was to evaluate the weed management with herbicides applied in pre and postemergence on castor crop. The treatments were constituted by management systems using herbicides in pre (PRE) and postemergence (POS): 1) S-metolachlor at 576 g ha-1 (PRE 1) and chlorimuron-methyl at 15 g ha-1 (POS 1); 2) PRE 1 + clomazone at 650 g ha-1(PRE 2) and POS 1; 3) PRE 1, POS 1 and halosulfuron-methyl at 112.5 g ha-1(POS 2); 4) PRE 1, POS 1 and ethoxysulfuron at 60 g ha-1 (POS 3); 5) PRE 1, POS 1 and ethoxysulfuron at 120 g ha-1 (POS 4); 6) PRE 1 + PRE 2, POS 1 and POS 2; 7) PRE 1 + PRE 2, POS 1 and POS 3; 8) PRE 1 + PRE 2, POS 1 and POS 4; 9) hand-hoeing control and 10) non-weeded control. The experimental design was randomized blocks with four replications. The association of the herbicides in PRE combined to sequence applications of chlorimuron-ethyl and halosulfuron-methyl or ethoxysulfuron in POS resulted in the highest control levels of Richardia scabraand Cyperus ferax. Mild phytointoxication symptoms to the crop were more persistent with ethoxysulfuron at 120 g ha-1. The acceptable control of the weed species and similar yield to the hand-hoeing control permitted to conclude that S-metolachlor + clomazone PRE, followed by chlorimuron-ethyl and halosulfuron-methyl POS proved to be more adequate as a weed management strategy on castor crop.

CHEMICAL CONTROL OF WHITE CLOVER IN SOYBEAN CROPS

ABSTRACTWhite clover is tolerant to many herbicides, making difficult a chemical control of this species during soybean crop establishments. The objective of this research was to select herbicides applied postemergence to control white clover in soybean and know the effects of this control on soybean yield. Seven herbicides were assessed, applied postemergence, with or without sequential application of glyphosate, and two control treatments (no control and total control of white clover). Glyphosate (with two sequential applications), fomesafen (with a sequential application of glyphosate), chlorimuron-ethyl and lactofen have shown a satisfactory control of white clover (above 80%). The lower control efficiency has resulted in lower production of soybeans.

REGULADOR DE CRESCIMENTO NA PRODUTIVIDADE E QUALIDADE DE SEMENTES DE AVEIA-BRANCA

RESUMO A aveia-branca (Avena sativa) é uma importante alternativa para cultivos de inverno no sul do Brasil. Contudo, elevados percentuais de acamamento limitam sua produtividade. Nesse contexto, a utilização de reguladores de crescimento pode ser uma opção para evitar o acamamento e obter maior produtividade. O objetivo deste trabalho foi avaliar o efeito de doses do regulador de crescimento trinexapac-ethyl sobre as características agronômicas da aveia-branca, bem como sua influência na qualidade fisiológica e sanitária das sementes. Um experimento foi conduzido em campo, onde foi avaliada a influência de quatro doses de trinexapac-ethyl (0, 50, 100 e 150 g i.a. ha-1) sobre a estatura de planta, o diâmetro de colmo, o percentual de acamamento e os componentes do rendimento. Um segundo experimento foi conduzido em laboratório, onde sementes provenientes do estudo em campo foram avaliadas quanto a qualidade fisiológica a partir da mensuração da germinação, primeira contagem da germinação, condutividade elétrica, comprimento e massa de plântulas, envelhecimento acelerado, teste de frio, índice de velocidade de germinação, índice de velocidade de emergência, massa seca de plântulas em campo e sanidade pelo método de Blotter Test. A utilização de doses crescentes de trinexapac-ethyl reduziu de forma quadrática a estatura, bem como o acamamento de plantas de aveia-branca. A dose de 100 g i.a ha-1 de regulador de crescimento proporcionou os melhores resultados para os componentes de produtividade em A. sativa. O uso do trinexapac-ethyl afeta negativamente a germinação e o vigor de sementes de aveiabranca, assim como a sua sanidade, reduzindo a capacidade de estabelecimento e desenvolvimento inicial da cultura.

Chemical composition, acetylcholinesterase inhibitory and antifungal activities of Pera glabrata (Schott) Baill. (Euphorbiaceae)

Pera glabrata (Schott) Baill. was selected for this study after showing a preliminary positive result in a screening of Atlantic Forest plant species in the search for acetylcholinesterase inhibitors and antifungal compounds. The bioassays were conducted with crude ethanol extract of the leaves using direct bioautography method for acetylcholinesterase and antifungal activities. This extract was partitioned with hexane, chloroform and ethyl acetate solvents. The active chloroform fraction was submitted to silica gel chromatography column affording 12 groups. Caffeine, an alkaloid, which showed detection limits of 0.1 and 1.0 µg for anticholinesterasic and antifungal activities, respectively, was isolated from group nine. After microplate analyses, only groups four, nine, 10, 11 and 12 showed acetylcholinesterase inhibitory activity of 40% or higher. The group 12 was purified by preparative layer chromatography affording four sub-fractions. Two sub-fractions from this group were analyzed by gas chromatography-mass spectrometry and gas chromatography-flame ionization detector. The first sub-fraction showed anticholinesterasic activity and contained two major compounds: 9-hydroxy-4-megastigmen-3-one (84%) and caffeine (6%). The second sub-fraction presented five major compounds identified as 9-hydroxy-4-megastigmen-3-one, isololiolide, (-) loliolide, palmitic acid and lupeol and did not show activity.

Alcohol dependence induced in rats by semivoluntary intermittent intake

The objective of the present experiment was to assess ethyl alcohol (ETOH) dependence brought about by a semivoluntary intermittent intake regimen in rats. Male Wistar rats weighing 150-250 g at the onset of the experiment were assigned to the following groups: 0% ETOH (N = 11), 5% ETOH (N = 20), 20% ETOH (N = 20) and 40% ETOH (N = 18). ETOH solutions were offered at the end of the day and overnight from Monday to Friday, and throughout weekends, for 90 days. The concentration of the ETOH solutions was increased in a stepwise fashion allowing the rats to get used to the taste of alcohol. Reposition of pure water was permitted during 1-h water drinking periods in the morning. Daily volume intake (± SEM) averaged 25.4 ± 0.4 ml (0% ETOH), 23.8 ± 0.6 ml (5% ETOH), 17.6 ± 0.7 ml (20% ETOH) and 17.5 ± 0.6 ml (40% ETOH). ETOH consumption differed significantly (P<0.05) among groups, averaging 4.4 ± 0.2 g kg-1 day-1 (5% ETOH), 10.3 ± 0.3 g kg-1 day-1 (20% ETOH) and 26 ± 1.2 g kg-1 day-1 (40% ETOH). Furthermore, ETOH detection in plasma 10-12 h after offering the solution indicated that its consumption in the 40% ETOH group was sufficient to override its metabolism. Overt signs of ETOH dependence, such as increased thirst, hyperactivity, puffing, hair ruffling and startle responsiveness as well as reduced drowsiness, were significantly increased in the 20% and 40% ETOH groups compared to the 0% and 5% groups. Accordingly, the model described here proved to be a useful tool for the evaluation of subtle or moderate behavioral and physical consequences of long-term ETOH intake

Somaclonal variation: a morphogenetic and biochemical analysis of Mandevilla velutina cultured cells

Cell cultures of Mandevilla velutina have proved to be an interesting production system for biomass and secondary metabolites able to inhibit the hypotensive activity of bradykinin, a nonapeptide generated in plasma during tissue trauma. The crude ethyl acetate extract of cultured cells contains about 31- to 79-fold more potent anti-bradykinin compounds (e.g., velutinol A) than that obtained with equivalent extracts of tubers. Somaclonal variation may be an explanation for the wide range of inhibitor activity found in the cell cultures. The heterogeneity concerning morphology, differentiation, carbon dissimilation, and velutinol A production in M. velutina cell cultures is reported. Cell cultures showed an asynchronous growth and cells in distinct developmental stages. Meristematic cells were found as the major type, with several morphological variations. Cell aggregates consisting only of meristematic cells, differentiated cells containing specialized cell structures such as functional chloroplasts (cytodifferentiation) and cells with embryogenetic characteristics were observed. The time course for sucrose metabolism indicated cell populations with significant differences in growth and metabolic rates, with the highest biomass-producing cell line showing a cell cycle 60% shorter and a metabolic rate 33.6% higher than the control (F2 cell population). MALDI-TOF mass spectrometric analysis of velutinol A in selected cell lines demonstrated the existence of velutinol A producing and nonproducing somaclones. These results point to a high genetic heterogeneity in general and also in terms of secondary metabolite content.

Sulfated fucan as support for antibiotic immobilization

Xylofucoglucuronan from Spatoglossum schröederi algae was tested as a support for antibiotic immobilization. The polysaccharide (20 mg in 6 ml) was first activated using carbodiimide, 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide methiodide (20 mg in 2 ml), under stirring for 1 h at 25ºC and pH from 4.5 to 5.0. After adjusting the pH to 8.0, either gentamicin or amikacin (62.5 mg in 1.25 ml) was then immobilized on this chemically modified polysaccharide with shaking for 24 h in a cold room. Infrared spectra of the activated carbodiimide xylofucoglucuronan showed two bands to carbonyl (C = O at 1647.9 and 1700.7 cm-1) and to amide (CÝ-NH2) groups (1662.8 and 1714.0 cm-1). Microbial characterization of the derivatives was carried out by the disk diffusion method using Staphylococcus aureus or Klebsiella pneumoniae incorporated in Müller Hinton medium. Inhibition halos of bacterial growth were observed for the antibiotics immobilized on this sulfated heteropolysaccharide before and after dialysis. However, the halos resulting from the samples after dialysis were much smaller, suggesting that dialysis removed either non-covalently bound antibiotic or other small molecules. In contrast, bacterial growth was not inhibited by either xylofucoglucuronan or its activated form or by gentamicin or amikacin after dialysis. An additional experiment was carried out which demonstrated that the sulfated heteropolysaccharide was hydrolyzed by the microorganism. Therefore, the antibiotic immobilized on xylofucoglucuronan can be proposed as a controlled drug delivery system. Furthermore, this sulfated heteropolysaccharide can be extracted easily from sea algae Spatoglossum schröederi.

Skin morphology of the mutant hairless USP mouse

The morphology of the skin of the mutant hairless USP mouse was studied by histological, histochemical and immunohistochemical methods and compared to the skin of BALB/c mice. Representative sections of the dorsal skin from mice of both strains aged 18 days, and 1, 3, 6, and 8 months were studied. Sections stained with hematoxylin and eosin showed cystic formations called utricles and dermal cysts in the dermis that increased in size and number during growth. Skin thickness increased significantly at 8 months. Sections stained with picrosirius and examined with polarized light, displayed different colors, suggesting different thicknesses of dermal collagen fibers (probably types I and III). Weigert, Verhoeff and resorcin-fuchsin stains revealed fibers of the elastic system. The PAS and Alcian blue methods revealed neutral and acid glycosaminoglycans in the skin ground substance of both mouse strains. Immunohistochemical staining for fibronectin and laminin did not show differences between the mutant and BALB/c mice. Mast cells stained by the Gomori method and macrophages positive for HAM 56 antibodies were observed in both mouse strains. Except for the presence of enlarged cysts in the hairless strain, no qualitative differences were found during development of the skin of BALB/c and the mutant hairless mice.

Electrical parameters and water permeability properties of monolayers formed by T84 cells cultured on permeable supports

T84 is an established cell line expressing an enterocyte phenotype whose permeability properties have been widely explored. Osmotic permeability (P OSM), hydraulic permeability (P HYDR) and transport-associated net water fluxes (J W-transp), as well as short-circuit current (I SC), transepithelial resistance (R T), and potential difference (deltaV T) were measured in T84 monolayers with the following results: P OSM 1.3 ± 0.1 cm.s-1 x 10-3; P HYDR 0.27 ± 0.02 cm.s-1; R T 2426 ± 109 omega.cm², and deltaV T 1.31 ± 0.38 mV. The effect of 50 µM 5,6-dichloro-1-ethyl-1,3-dihydro-2H-benzimidazol-2-one (DCEBIO), a "net Cl- secretory agent", on T84 cells was also studied. We confirm the reported important increase in I SC induced by DCEBIO which was associated here with a modest secretory deltaJ W-transp. The present results were compared with those reported using the same experimental approach applied to established cell lines originating from intestinal and renal epithelial cells (Caco-2, LLC-PK1 and RCCD-1). No clear association between P HYDR and R T could be demonstrated and high P HYDR values were observed in an electrically tight epithelium, supporting the view that a "water leaky" barrier is not necessarily an "electrically leaky" one. Furthermore, the modest secretory deltaJ W-transp was not consistent with previous results obtained with RCCD-1 cells stimulated with vasopressin (absorptive fluxes) or with T84 cells secreting water under the action of Escherichia coli heat stable enterotoxin. We conclude that, while the presence of aquaporins is necessary to dissipate an external osmotic gradient, coupling between water and ion transport cannot be explained by a simple and common underlying mechanism.