Translocação do glyphosate em biótipos de azevém (Lolium multiflorum)

Foram avaliadas neste trabalho a absorção foliar e a translocação do glyphosate por biótipos de azevém (Lolium multiflorum) sensíveis e resistentes a esse herbicida. Para isso, aplicou-se 14C-glyphosate utilizando-se uma microsseringa de precisão, adicionando-se 10 µL da calda sobre a face adaxial da primeira folha com lígula totalmente visível, quando as plantas de azevém se apresentavam com três perfilhos. A quantidade de glyphosate absorvido e translocado foi avaliada em intervalos de tempo (2, 4, 8, 16, 32 e 64 horas após a aplicação), por meio da medição da radiação emitida pelo 14C-glyphosate, em espectrômetro de cintilação líquida. Foram analisadas a parte aérea e as raízes, bem como a folha onde foi feita a aplicação e a solução de lavagem desta folha. A velocidade de absorção do glyphosate foi semelhante em ambos os biótipos de azevém, observando-se mais de 50% de absorção desse herbicida nas primeiras oito horas após a aplicação. Maior retenção de glyphosate foi observada na folha tratada do biótipo resistente: 81,64% do total de glyphosate absorvido até as 64 horas. No biótipo sensível esse valor foi de 55% no mesmo período. No restante da parte aérea e nas raízes, a maior quantidade do glyphosate absorvido foi encontrada no biótipo sensível, mostrando sua maior capacidade de translocação. Após 64 horas da aplicação do glyphosate, apenas 6%, em média, do glyphosate se encontrava nas plantas, indicando que a maior parte do produto pode ter sido exsudada. Conclui-se que a sensibilidade do azevém ao glyphosate pode ser atribuída à maior capacidade de translocação desse herbicida pelo biótipo sensível.

Efeito de formulações na absorção e translocação do glyphosate em soja transgênica

Este trabalho teve como objetivo avaliar a absorção e translocação de glyphosate em diferentes formulações por plantas de soja (variedade CD 219RR). Para isso, aplicou-se o 14C-glyphosate misturado à calda em três formulações comerciais (Roundup Ready® e R. Transorb®, ambas contendo o sal de isopropilamina, e Zapp Qi®, formulado à base do sal potássico), quando as plantas apresentavam o segundo trifólio completamente expandido. Transcorridas 4, 16, 40 e 64 horas após a aplicação, as plantas foram coletadas e fracionadas, separando-se a folha de aplicação (trifólio), a parte aérea, as raízes e os nódulos radiculares. O 14C-glyphosate não-absorvido foi recuperado e contado por meio da lavagem da folha (metanol 80%). Entre as formulações foi observada variação na penetração e na translocação do 14C-glyphosate para as diferentes partes avaliadas. Todavia, em todas as formulações a maior absorção se deu nos intervalos posteriores a 16 horas da aplicação. Em relação ao total de herbicida encontrado nas plantas de soja, maior percentual na parte aérea foi observado quando se aplicou o Zapp Qi® (sal potássico) e, nas raízes, o R. Transorb® (sal de isopropilamina). Detectou-se a presença de 14C glyphosate nos nódulos radiculares das plantas em todos os tratamentos, sendo o maior percentual observado quando se utilizou R. Transorb®, 40 horas após a aplicação (0,13% do total medido ou 0,4% considerando somente o total presente na planta). Os resultados reforçam a hipótese de que o glyphosate pode prejudicar a simbiose entre rizóbio e soja, uma vez que o microssimbionte também apresenta em seu metabolismo a EPSPS, sensível a esse herbicida.

Distribuição de glyphosate e acúmulo de nutrientes em biótipos de azevém

Objetivou-se neste trabalho avaliar o acúmulo de nutrientes e a translocação de glyphosate em biótipos de azevém. Para isso, foram montados dois ensaios: no primeiro aplicou-se 14C-glyphosate, adicionando 10 µL da calda sobre a face adaxial da primeira folha com lígula totalmente visível, quando as plantas de azevém apresentavam três perfilhos. A quantidade de glyphosate absorvido, translocado e exsudado foi avaliada 64 horas após aplicação, por meio da medição da radiação emitida pelo 14C-glyphosate, em espectrômetro de cintilação líquida. O glyphosate foi quantificado em folha de aplicação, perfilhos, raízes e na solução nutritiva onde foram cultivados os biótipos de azevém. No segundo experimento, aplicou-se o glyphosate (480 g ha-1) tanto no biótipo sensível quanto no resistente. Após dez dias da aplicação, a parte área e as raízes das plantas foram coletadas e secas em estufa, sendo determinados os teores de macronutrientes. No primeiro ensaio, verificou-se exsudação radicular em ambos os biótipos, nos quais a quantidade de glyphosate exsudada foi semelhante, não ultrapassando 5% do total que penetrou na planta. No perfilho principal do biótipo sensível, comparado ao resistente, foi observada maior concentração do produto marcado. O biótipo resistente apresentou maior acúmulo de produto marcado na folha de aplicação; no sensível, a maior parte do glyphosate foi encontrada nas raízes. Com relação ao segundo ensaio, na presença de herbicida o biótipo sensível apresentou menor teor de fósforo tanto na parte aérea quanto na planta. Os biótipos resistente e sensível, sem aplicação de herbicida, tenderam a apresentar maiores teores de N total e N inorgânico na parte aérea e na planta como um todo, quando comparados aos tratamentos em que foi realizada a aplicação do produto. Ambos os biótipos mostraram a mesma capacidade de absorção e acúmulo de macronutrientes na ausência do produto.

Exsudação radicular de glyphosate por Brachiaria decumbens e seus efeitos em plantas de eucalipto

Plantas de eucalipto com sintomas de intoxicação por glyphosate são comuns em áreas em que esse herbicida é usado. Uma das possíveis formas de contato com glyphosate é por meio da exsudação radicular do produto, por plantas daninhas tratadas, e subseqüente absorção pelas plantas de eucalipto. Objetivou-se com este trabalho avaliar a exsudação de glyphosate por Brachiaria decumbens e seus efeitos sobre plantas de eucalipto, por meio da aplicação de 14C-glyphosate misturado à calda de pulverização do produto comercial. Mudas de dois clones de eucalipto (UFV05 e UFV06) foram cultivadas em consórcio com Brachiaria decumbens (capim-braquiária), em vasos contendo dois tipos de solo: um arenoso e outro argiloso. Aos 35 dias após o transplantio das mudas, foram aplicados na braquiária 50 µL da mistura de 14C-glyphosate com a formulação comercial de glyphosate Scout®, utilizando-se uma microsseringa de precisão. Aos 2, 8, 16 e 24 dias após aplicação, as plantas de eucalipto foram coletadas e fracionadas em ápice primário, ápices secundários, folhas e raízes, sendo processadas de acordo com metodologia usual para determinação da radioatividade. Não foram observados sintomas de intoxicação por glyphosate nas plantas de eucalipto, em nenhuma das avaliações realizadas. Entretanto, o 14C-glyphosate foi encontrado em todas as plantas de eucalipto avaliadas, independentemente do solo, do clone e da época de avaliação, em maior concentração em plantas cultivadas no solo arenoso. Os resultados evidenciam a exsudação radicular do glyphosate e/ou de seus metabólitos pela braquiária e subseqüente absorção, via raízes, pelas plantas de eucalipto, em concentrações inferiores às necessárias para causar intoxicação na cultura.

Glyphosate translocation in hairy fleabane (Conyza bonariensis) biotypes

The objective of this work was to evaluate the translocation of glyphosate in C. bonariensis plants resistant and susceptible to that herbicide. The 14C-glyphosate was mixed with commercial gyhphosate (800 g ha-1) and applied on the center of the adaxial face of a third node leaf, using a micro syringe, and adding 10 µL of a solution with specific activity of 1,400 Bq, 45 days after plant emergence. The concentration of the glyphosate translocated in the plant was evaluated at time intervals of 6, 12, 36 and 72 hours after being applied on the application leaf, stem, roots and leaves. Ten hours after treatment application, the distribution of the product in the application leaf, divided into base, center and apex, was also evaluated by measuring the radiation emitted by 14C-glyphosate in a liquid scintillation spectrometer. Greater glyphosate retention was observed in the resistant biotype leaf, approximately 90% of the total absorbed up to 72 hours. In the susceptible biotype, this value was close to 70% in the same period. Susceptible biotype leaves, stem and roots showed greater concentration of glyphosate, indicating greater translocation efficiency in this biotype. In the resistant biotype, the herbicide accumulated in greater quantity at the apex and center of the application leaf, while in the susceptible biotype greater accumulation was observed at the base and center leaf. Thus, it can be stated that the resistance mechanism is related to the differential translocation of this herbicide in the biotypes.

Absorção, translocação e exsudação radicular de glyphosate em clones de eucalipto: clones

Objetivou-se com este trabalho avaliar a absorção, translocação e exsudação radicular de glyphosate por dois clones de eucalipto: 2277 e 531. O 14C-glyphosate foi aplicado na concentração de 1.440 g ha-1, distribuída uniformemente no terceiro e no quarto limbo foliar a partir do ápice caulinar, com radioatividade aproximada de 0,030 μCi. A absorção, translocação e exsudação radicular foram avaliadas pela radioatividade do 14C-glyphosate nos diferentes tecidos da planta, bem como na água de lavagem e solução nutritiva, nos intervalos de 0, 2, 8, 32 e 72 horas após a aplicação - HAA. A concentração de 14C-glyphosate na folha aplicada foi semelhante para os dois clones nas avaliações a partir de 8 HAA. Todavia, considerando a planta inteira, ela foi superior no clone 2277 em todas as épocas de avaliação. Maior quantidade de 14C-glyphosate foi verificada na água de lavagem da folha aplicada do clone 531, indicando menor absorção do herbicida nesse clone em relação ao 2277. Na parte aérea e no sistema radicular, a concentração do 14C-glyphosate foi semelhante entre os clones em todos os intervalos de avaliação, porém com concentrações maiores nas raízes. Pequena parte do total aplicado foi exsudada para solução nutritiva (valores entre 0,78 e 1,16%), não havendo diferença entre os clones quanto à translocação na planta e na exsudação radicular do herbicida. A absorção diferencial entre os clones, atribuída na maioria dos casos a diferenças na estrutura e composição da cutícula, pode ser uma possível explicação para a tolerância diferencial entre os genótipos.

Biodegradation of glyphosate in rhizospheric soil cultivated with Glycine max, Canavalia ensiformis e Stizolobium aterrimum

Biodegradation of glyphosate was evaluated in rhizospheric soil cultivated with Glycine max (soybean, var. BRS245-RR), Canavalia ensiformis and Stizolobium aterrimum. After these species were cultivated for 60 days, soil samples were collected, placed in flasks and treated with 14C-glyphosate. After 30 days of incubation, the total release rate of C-CO2 was determined along with microbial biomass (MBC), metabolic quotient (qCO2), and degradation percentage of the radio-labeled glyphosate released as 14C-CO2. A higher mass of rhizosphere-associated microorganisms was verified in the soil samples from pots cultivated with soybean, regardless of glyphosate addition. However, in the presence of the herbicide, this characteristic was the most negatively affected. Microorganisms from the C. ensiformis rhizosphere released a lower amount of 14C-CO2, while for those originated from S. aterrimum, the amount released reached 1.3% more than the total carbon derived from the respiratory activity. The rhizospheric soil from S. aterrimum also presented higher glyphosate degradation efficiency per microbial biomass unit. However, considering qCO2, the microbiota of the rhizospheric soil cultivated with soybean was more efficient in herbicide degradation.

Glyphosate translocation in herbicide tolerant plants

The objective of this study was to evaluate glyphosate translocation in glyphosate-tolerant weed species (I. nil, T. procumbens and S. latifolia) compared to glyphosate-susceptible species (B. pilosa). The evaluations of 14C-glyphosate absorption and translocation were performed at 6, 12, 36 and 72 hours after treatment (HAT) in I. nil and B. pilosa, and only at 72 HAT in the species T. procumbens and S. latifolia. The plants were collected and fractionated into application leaf, other leaves, stems, and roots. In S. latifolia, approximately 88% of the glyphosate remained in the application leaf and a small amount was translocated to roots at 72 HAT. However, 75% of the herbicide applied on T. procumbens remained in the leaf that had received the treatment, with greater glyphosate translocation to the floral bud. It was concluded that the smaller amount of glyphosate observed in S. latifolia and T. procumbens may partly account for their higher tolerance to glyphosate. However, I. nil tolerance to glyphosate may be associated with other factors such as metabolization, root exudation or compartmentalization, because a large amount of the herbicide reached the roots of this species.

Sorption and desorption of Suflafenacil in two soils in the state of São Paulo with different physical and chemical attributes

A study was conducted to evaluate the sorption and desorption of 14C herbicide saflufenacil (pyrimidinedione) in two soils in the State of São Paulo, classified as Red Yellow Latosol with clayey texture (LVA-1) and medium texture (LVA-2), using the batch method through isotherms. The soils were air dried and sieved a 2 mm mesh. The radioactivity was determined by liquid scintillation spectrometry in acclimatized room (25 ± 2 °C). Sorption isotherms were conducted for 5 concentrations of saflufenacil (5.0; 2.5; 1.0; 0.5 and 0.05 μg mL-1) and the results were adjusted to the Freundlich equation, thus obtaining the parameters of sorption followed by two extractions with 0.01 M CaCl2 to determine desorption parameters similarly to sorption. The results showed that saflufenacil sorption was low for both soils studied, being greater for the LVA with higher organic matter content. The desorption coefficients were greater than their sorption coefficients, suggesting the occurrence of hysteresis. The sorption and desorption isotherms (classified as type C isotherms), hysteresis and the t-test between the angular coefficient of the respective isotherms showed that both the sorption and desorption occur with equal intensity.

Effects of straw decomposition degree on leaching and weed control efficacy of tebuthiuron and hexazinone in green sugarcane harvesting

Green sugarcane harvesting may promote great changes in the dynamics of herbicides in the environment. Our goal was to evaluate the influence of straw decomposition degree on leaching and weed (Ipomoea grandifolia) control efficacy by (14C) tebuthiuron and hexazinone. The presence of straw on the soil surface affected leaching, mainly for hexazinone (leaching reduced from 37 to 5% of the applied amount in the presence of straw). Overall, tebuthiuron showed more efficient control of Ipomoea than hexazinone. The straw decomposition degree affected only hexazinone efficacy that was lowest for the least decomposed straw. Further studies are needed to evaluate the effects of sugarcane straw on herbicides dissipation, particularly on volatilization and photolysis, to better predict their efficacy and environmental fate.

Shikimate accumulation, glyphosate absorption and translocation in horseweed biotypes

In Brazil, few research works on mechanisms of weed resistance to glyphosate have been conducted so far. Therefore, this research aimed to study analytical procedures determining the relation between the concentration of plant shikimate after glyphosate application and the plant resistance to this herbicide; and evaluate the glyphosate absorption and translocation into two resistant ® and susceptible (S) horseweed biotypes to glyphosate. Horseweed plants with nine true leaves received glyphosate (720 g a.e. ha-1), and 2, 3, 4, 7 and 10 days after application (DAA) the concentration of shikimic acid was measured by HPLC. In another experiment, plants were treated with radiolabeled glyphosate (14C) (1.456 MBq mmol-1 specific activity) and radioactivity was measured 4, 8, 24, 48 and 72 hours after treatment (HAT) by liquid scintillation spectrometry. The shikimate concentration in plants increased 16,351.14 and 7,892.25 mg kg-1 of dry weight, for R and S plants respectively, at seven DAA. Therefore, the procedure for quantification of shikimic acid was suitable for R and S plants differentiation to glyphosate, indicating that the R population is actually resistant to glyphosate. On average, 98% of glyphosate applied was absorbed by the studied biotypes, at 72 HAT. Around 68% of the absorbed radioactivity remained on the biotypes leaves treated, the S biotype showing the highest translocation. Therefore, the R biotype resistance mechanism studied is associated to the differential translocation.

Interaction between NO and oxytocin: Influence on LHRH release

Nitric oxide synthase (NOS)-containing neurons have been localized in various parts of the CNS. These neurons occur in the hypothalamus, mostly in the paraventricular and supraoptic nuclei and their axons project to the neural lobe of the pituitary gland. We have found that nitric oxide (NO) controls luteinizing hormone-releasing hormone (LHRH) release from the hypothalamus acting as a signal transducer in norepinephrine (NE)-induced LHRH release. LHRH not only releases LH from the pituitary but also induces sexual behavior. On the other hand, it is known that oxytocin also stimulates mating behavior and there is some evidence that oxytocin can increase NE release. Therefore, it occurred to us that oxytocin may also stimulate LHRH release via NE and NO. To test this hypothesis, we incubated medial basal hypothalamic (MBH) explants from adult male rats in vitro. Following a preincubation period of 30 min, MBH fragments were incubated in Krebs-Ringer bicarbonate buffer in the presence of various concentrations of oxytocin. Oxytocin released LHRH at concentrations ranging from 0.1 nM to 1 µM with a maximal stimulatory effect (P<0.001) at 0.1 µM, but with no stimulatory effect at 10 µM. That these effects were mediated by NO was shown by the fact that incubation of the tissues with NG-monomethyl-L-arginine (NMMA), a competitive inhibitor of NOS, blocked the stimulatory effects. Furthermore, the release of LHRH by oxytocin was also blocked by prazocin, an a1-adrenergic receptor antagonist, indicating that NE mediated this effect. Oxytocin at the same concentrations also increased the activity of NOS (P<0.01) as measured by the conversion of [14C]arginine to citrulline, which is produced in equimolar amounts with NO by the action of NOS. The release of LHRH induced by oxytocin was also accompanied by a significant (P<0.02) increase in the release of prostaglandin E2 (PGE2), a mediator of LHRH release that is released by NO. On the other hand, incubation of neural lobes with various concentrations of sodium nitroprusside (NP) (300 or 600 µM), a releaser of NO, revealed that NO acts to suppress (P<0.01) the release of oxytocin. Therefore, our results indicate that oxytocin releases LHRH by stimulating NOS via NE, resulting in an increased release of NO, which increases PGE2 release that in turn induces LHRH release. Furthermore, the released NO can act back on oxytocinergic terminals to suppress the release of oxytocin in an ultrashort-loop negative feedback

Effect of FSH and insulin on lipogenesis in cultures of Sertoli cells from immature rats

Follicle-stimulating hormone (FSH) and insulin regulate glycide metabolism in Sertoli cells, thus stimulating lactate production. These stimulatory effects of FSH and insulin do not require protein synthesis, suggesting a modulation of enzyme activity and/or regulation of glucose transport. The present investigation was performed to characterize the hormonal control of lipid metabolism in Sertoli cells. The data indicate that FSH and insulin have a regulatory effect on lipid metabolism in Sertoli cells. After 8 h of preincubation with insulin (5 µg/ml), the activity of the enzyme ATP-citrate lyase in cultured Sertoli cells was increased from 0.19 to 0.34 nmol NAD+ formed µg protein-1 min-1. FSH (100 ng/ml) had no effect on this enzyme. Glycerol phosphate dehydrogenase activity was not affected by any of the hormones tested. When Sertoli cells from 19-day old rats were incubated with [1,2­14C]acetate for 90 or 360 min, the [14C] label was present predominantly in triglyceride and phospholipid fractions with minor amounts in other lipids. In Sertoli cells pretreated for 16 h with insulin and FSH, an increase in acetate incorporation into lipids was observed. Most of the label was in esterified lipids and this percentage increased with the time of treatment; this increase was remarkable in triglycerides of control cells (18.8% to 30.6%). Since Sertoli cell triglycerides participate in the control of spermatogenesis, the present data suggest that the hormonal control of lipid metabolism in Sertoli cells is important not only for maintaining the energy of the cell itself, but also for the control of the spermatogenesis process.

Application of isotope-selective non-dispersive infrared spectrometry for the evaluation of the 13C-urea breath test: comparison with three concordant methods

The aim of this work was to compare the performance of isotope-selective non-dispersive infrared spectrometry (IRIS) for the 13C-urea breath test with the combination of the 14C-urea breath test (14C-UBT), urease test and histologic examination for the diagnosis of H. pylori (HP) infection. Fifty-three duodenal ulcer patients were studied. All patients were submitted to gastroscopy to detect HP by the urease test, histologic examination and 14C-UBT. To be included in the study the results of the 3 tests had to be concordant. Within one month after admission to the study the patients were submitted to IRIS with breath samples collected before and 30 min after the ingestion of 75 mg 13C-urea dissolved in 200 ml of orange juice. The samples were mailed and analyzed 11.5 (4-21) days after collection. Data were analyzed statistically by the chi-square and Mann-Whitney test and by the Spearman correlation coefficient. Twenty-six patients were HP positive and 27 negative. There was 100% agreement between the IRIS results and the HP status determined by the other three methods. Using a cutoff value of delta-over-baseline (DOB) above 4.0 the IRIS showed a mean value of 19.38 (minimum = 4.2, maximum = 41.3, SD = 10.9) for HP-positive patients and a mean value of 0.88 (minimum = 0.10, maximum = 2.5, SD = 0.71) for negative patients. Using a cutoff value corresponding to 0.800% CO2/weight (kg), the 14C-UBT showed a mean value of 2.78 (minimum = 0.89, maximum = 5.22, SD = 1.18) in HP-positive patients. HP-negative patients showed a mean value of 0.37 (minimum = 0.13, maximum = 0.77, SD = 0.17). IRIS is a low-cost, easy to manage, highly sensitive and specific test for H. pylori detection. Storing and mailing the samples did not interfere with the performance of the test.

Abnormalities of glucose metabolism in spontaneously hypertensive rats

Abnormalities in glucose metabolism and insulin action are frequently detected in patients with essential hypertension. Spontaneously hypertensive rats (SHR) have been used as an experimental model to understand this pathological condition. The objective of the present study was to assess glucose metabolism and insulin action in SHR and Wistar rats under fed and fasting conditions. Peripheral glucose utilization was estimated by kinetic studies with [6-³H]-glucose and gluconeogenetic activity was measured during continuous [14C]-bicarbonate infusion. Plasma glucose levels were higher in the SHR group. Plasma insulin levels in the fed state were higher in the SHR group (99.8 ± 6.5 µM) than in the control group (70.4 ± 3.6 µM). Muscle glycogen content was reduced in SHR compared to control under the various experimental conditions. Peripheral glucose utilization was slightly lower in the SHR group in the fed state (8.72 ± 0.55 vs 9.52 ± 0.80 mg kg-1 min-1 in controls). Serum free fatty acid levels, hepatic glycogen levels, hepatic phosphoenolpyruvate carboxykinase activity and gluconeogenetic activity were similar in the two groups. The presence of hyperglycemia and hyperinsulinemia and the slightly reduced peripheral glucose utilization suggest the presence of resistance to the action of insulin in peripheral tissues of SHR. Hepatic gluconeogenesis does not seem to contribute to the metabolic alterations detected in these animals.