Quick bioassay test from tillers for detecting ALS herbicide resistance of weedy rice and barnyardgrass

Resistance to acetolactate synthase (ALS) inhibitors have increased recently in South Brazil where the major weeds of flooded rice (barnyardgrass and weedy rice) have evolved resistance to imazapyr+imazapic. The aim of this research was to evaluate a growth medium for tissue regeneration of tillers in barnyardgrass, as well as an agar-based bioassays test (also from tillers) to detect susceptible and resistant biotypes of weedy rice and barnyardgrass to imazapyr+imazapic in vitro. Greenhouse experiments were conducted to detect ALS-resistant (R) and susceptible (S) weedy rice and barnyardgrass biotypes, and bioassays were carried out to evaluate an adequate growth medium for barnyardgrass tiller regeneration and determine the concentration of herbicide to distinguish R and S plants. The culture medium that provided a suitable barnyardgrass growth was MS 50% with the addition of benzylamino-purine. The tissue regeneration in vitro with the growth medium containing imazapyr+imazapic allowed to discriminate between R and S barnyardgrass and weedy rice plants. The concentration required for satisfactory control of susceptible barnyardgrass and weedy rice explants grown in vitro was 0.9 µM and 1.3 µM of imazapyr+imazapic herbicide, respectively. The bioassay in vitro using tiller regeneration provides an opportunity to predict effectively imazapyr+imazapic resistance in barnyardgrass and weedy rice.

Effect of different herbicides on Bidens pilosa and Euphorbia heterophylla biotypes resistant to ALS inhibitors

Select herbicides with different mechanism of action is a satisfactory option for resistant weed control. Then, the present work aimed to study the efficiency of different herbicides and their mixtures on Bidens pilosa (blackjack) and Euphorbia heterophylla (wild poinsettia) biotypes, resistant to ALS herbicides in two development stages. The trials we arranged in a completely randomized design with four replications. The treatments tested were (g a.i/a.e ha-1): imazethapyr at 70 and 140 (WG formulation) + 1.0% Assist; imazethapyr at 57.6 and 72 (SL formulation) + 1.0% Assist; imazapic + imazethapyr at 56 and 70 + 1.0% Assist; glyphosate + imazethapyr (596); saflufenacil + glyphosate at 35 + 720 + 0.5% Dash in tank mix, glyphosate at 720 and, a control without herbicide application. Control efficiency was evaluated, as well as dry matter accumulation at the end of the studies. Plants of both species were more susceptible to herbicides at the early stage of development (2 to 4 leaves). The treatments with saflufenacil + glyphosate, (imazethapyr + glyphosate) and glyphosate promoted the best controls, regardless of the species studied and the application stage. The mixture with saflufenacil provided the highest control speed, and the mixture (imazethapyr + glyphosate) was less efficient among three excellent treatments when applied to plants in the 4-6 leaf stage. The treatments (imazethapyr, in both formulations) and (imazethapyr + imazapic) were ineffective in controlling the studied biotypes, regardless of dose and developmental stage studied.

Herbicide performance in the control of Conyza spp. where three plant heights

Intensive use of the herbicide glyphosate has led to herbicide resistant Conyza spp. populations. Thus, there is a need to indicate alternative herbicides and the appropriate developmental stage for controlling these populations. This study identifies alternatives for controlling glyphosate-resistant horseweed, with treatment applications at different plant heights. For this purpose, field experiments were conducted in the 2016/17 and 2017/18 crop years. The evaluated treatments were: glyphosate (540 g ae ha-1), glyphosate (1080 g ae ha-1), glyphosate (2160 g ae ha-1), glyphosate (3240 g ae ha-1), glyphosate + 2.4-D (1080 + 1005 g ae ha-1), glyphosate + saflufenacil (1080 + 49 g ae/ai ha-1), paraquat (400 g ai ha-1), diquat (400 g ai ha-1), ammonium glufosinate (600 g ai ha-1), and control (without application). These treatments were applied to plants with a maximum of 5 cm; plants between 6 and 15 cm; and plants between 16 and 25 cm. The results showed that glyphosate did not control weeds, regardless of rate. With the exception of 2,4-D, which needs complementation with sequential application of another contact herbicide, all alternatives were viable for the control of Conyza spp. plants with a maximum height of 5 cm.

Estimation of sulfentrazone sorption in isolated application and mixed with glyphosate / Estimativa da sorção do sulfentrazone na aplicação isolada e em mistura com glifosato

The use of herbicides belonging to a single mechanism of action for several consecutive years in the same area may lead to the selection of weed biotypes resistant to herbicides. Weeds resistant to glyphosate have been problem worldwide. As an alternative control, farmers have used glyphosate mixed with sulfentrazone. When two herbicides are mixed, the parameters that govern the behaviour of herbicides in the soil can be altered. In this study, the sorption of sulfentrazone was estimated when applied in isolation with glyphosate formulations. For this, increasing doses of sulfentrazone were applied using Red-Yellow Latosol substrate and washed sand. Each dose of sulfentrazone was applied in a mixture with 1080 g a. e. ha-1 Roundup Ready®, Roundup Ultra® and Zapp Qi®. To evaluate the presence of sulfentrazone in the soil solution, Sorghum bicolor was used as an indicator species. Symptoms of intoxication were evaluated at 7, 14 and 21 days after sowing. At 21 days after sowing, the plants were collected and dried in an oven at 70 ± 10 °C to determine the dry matter. Based on the intoxication of the indicator plants, it was verified that, regardless of the formulation, the presence of glyphosate in the mixture increased the sorption of sulfentrazone in the Red-Yellow Latosol. However, based on dry matter accumulation, sulfentrazone sorption increased because of the mixture with Roundup Ultra®, but decreased due to the presence of Roundup Ready® and Zapp Qi®. Therefore, the application of sulfentrazone in mixture with glyphosate alters the sorptive forces of sulfentrazone in a Red-Yellow Latosol.

A utilização de herbicidas pertencentes a um único mecanismo de ação por vários anos consecutivos na mesma área pode levar a seleção de biótipos de plantas daninhas resistentes a herbicidas. Plantas daninhas resistentes ao glyphosate têm sido problema no mundo inteiro. Como alternativa de controle, os agricultores têm utilizado glyphosate em mistura com sulfentrazone. Quando se misturam duas moléculas herbicidas, os parâmetros que governam o comportamento de herbicidas no solo podem ser alterados. Neste estudo foi estimada a sorção do sulfentrazone quando aplicado de forma isolada e em mistura com formulações de glyphosate. Para isso, doses crescentes de sulfentrazone foram aplicadas utilizando-se como substrato Latossolo Vermelho-Amarelo e areia lavada. Cada uma das doses de sulfentrazone foi aplicada em mistura com 1080 g ha-1 de glyphosate comercializado nas formulações Roundup Ready®, Roundup Ultra® e Zapp Qi®. Para avaliar a presença do sulfentrazone na solução do solo utilizou-se o Sorghum bicolor como espécie indicadora. Os sintomas de intoxicação foram avaliados aos 7, 14 e 21 dias após a semeadura. Aos 21 dias após a semeadura, as plantas foram coletadas e secadas em estufa a 70 ±10C para determinação da matéria seca. Com base na intoxicação das plantas indicadoras, verificou-se que, independente da formulação, a presença glyphosate na mistura, aumentou a sorção do sulfentrazone no Latossolo Vermelho-Amarelo. No entanto com base no acúmulo de matéria seca, constatou-se que a sorção do sulfentrazone aumentou em função da mistura com Roundup Ultra®, mas diminuiu devido a presença de Roundup Ready® e Zapp Qi®. Conclui-se que a aplicação de sulfentrazone em mistura com glyphosate altera as forças sortivas do sulfentrazone no Latossolo Vermelho-Amarelo.

Performance of RR soybean submitted to postemergence application of glyphosate with a foliar elicitor product / Desempenho da soja RR submetida a aplicação de glifosato associado a produto elicitor foliar em pós-emergência

With the advent of glyphosate-resistant soybean, postemergence applications of the herbicide have become routine. In this way, problems related to the selectivity of glyphosate have been shown to be increasingly intense, due to the use of increased dosages and applications of the herbicide at unrecommended moments. In order to evaluate the agronomic performance of soybean plants subjected to the postemergence application of glyphosate ­ both in isolation and together with Crop+ ­ at different stages and dosage levels, a randomized block design experiment was carried out with two adjacent checks and four repetitions. The treatments consisted of: glyphosate (1296 g·ha-1); glyphosate + Crop+ (1296 + 0.25 g or L·ha-1); glyphosate + Crop+ (1296 + 0.5 g or L·ha-1); glyphosate (2592 g·ha-1); glyphosate + Crop+ (2592 + 0.25 g or L·ha-1); glyphosate + Crop+ (2592 + 0.5 g or L·ha-1); 2 × glyphosate (2 × 1296 g·ha-1); 2 × glyphosate + Crop+ (2 × 1296 + 0.25 g or L·ha-1); 2 × glyphosate + Crop+ (2 × 1296 + 0.5 g or L·ha-1) and Crop+ (0.5 g·ha-1). The phytotoxicity, soil plant analysis development (SPAD) index, height, stand, mass of 100 grains, and yield of the soybean plants were evaluated. The use of Crop+ on soybean plants led to an increase in the SPAD index; the use of Crop+ in association with glyphosate made possible the attenuation of visible symptoms of damage, preventing reductions in crop yield.(AU)

Com o advento da soja resistente ao glifosato, aplicações em pós-emergência da cultura tornaram-se corriqueiras. Dessa forma, problemas relacionados à seletividade desse herbicida mostraram-se cada vez mais intensos, devido ao aumento da dose utilizada e à aplicação em momentos não recomendados. Para avaliar o desempenho agronômico da soja submetida à aplicação de glifosato isolado em pós-emergência e em associação ao Crop+ em diferentes estádios e doses, um experimento foi realizado em delineamento de blocos ao acaso, em esquema de testemunhas duplas adjacentes, adotando-se quatro repetições. Os tratamentos foram constituídos por: glifosato (1296 g·ha-1); glifosato + Crop+ (1296 + 0,25 g ou L·ha-1); glifosato + Crop+ (1296 + 0,5 g ou L·ha-1);glifosato (2592 g·ha-1); glifosato + Crop+ (2592 + 0,25 g ou L·ha-1);glifosato + Crop+ (2592 + 0,5 g ou L·ha-1); 2 × glifosato (2 × 1296 g·ha-1); 2 × glifosato + Crop+ (2 × 1296 + 0,25 g ou L·ha-1); 2 × glifosato + Crop+ (2 × 1296 + 0,5 g ou L·ha-1) e Crop+ (0,5 g·ha-1). Foram avaliados fitointoxicação das plantas de soja, índice SPAD (soil plant analysis development), altura, estande, massa de cem grãos e produtividade. Nos resultados, a utilização de Crop+ em plantas de soja proporcionou o aumento no índice SPAD e, quando o Crop+ foi associado ao glifosato, possibilitou a atenuação dos sintomas visuais de injúrias, prevenindo reduções na produtividade da cultura.(AU)

The glyphosate controversy: an update / A controvérsia do glifosato: uma atualização

The demand for food in the world grows year after year due in part to population growth, but also to the improvement of emerging markets. Brazil is one of the largest food producers in the world. In 2017, its cereal, legume and oilseed crops totaled 238.6 million tons, 29.2% more than the year before. Much of the great increase in productivity is due to the incorporation of transgenic seeds, especially cotton, maize and soybean, which possess genes that will increase plant's adaptability to harsh soil and water conditions and, resistance to pests, but also tolerance to herbicides. Virtually, all transgenic soybeans planted in Brazil are resistant to glyphosate, the herbicide initially launched on the market by Monsanto in the 1970s under the trade name Roundup. Due to the existence of several transgenic crops tolerant to glyphosate, such as soy, wheat, corn and canola, this product is the most commercialized herbicide in the world. The use of glyphosate allows the sowing of transgenic crops immediately after application, making the planting and maintenance processes very practical. Soybeans, such as other transgenics, have biological safety already well defined, but the use of glyphosate is still an extremely controversial subject. This review presents some historical aspects of the binomial Roundup Ready soybean/glyphosate and discusses the most recent controversies about the use of glyphosate in Brazil and worldwide.(AU)

A demanda por alimentos no mundo cresce ano após ano devido em parte pelo aumento populacional, mas também pela melhoria dos mercados emergentes. O Brasil é um dos maiores produtores de alimentos do mundo. Em 2017, a safra nacional de cereais, leguminosas e oleaginosas totalizou 238,6 milhões de toneladas, 29,2% superior à obtida em 2016. Boa parte do grande aumento da produtividade se dá pela incorporação de sementes transgênicas, especialmente de algodão, milho e soja, que apresentam genes que conferem maior adaptabilidade da planta ao solo, pragas, condições hídricas, mas também conferem resistência a herbicidas. Praticamente toda a soja transgênica plantada no país é resistente ao glifosato, inicialmente lançado no mercado pela empresa Monsanto nos anos 1970 pelo nome comercial Roundup, que é hoje o herbicida mais utilizado no mundo, principalmente pela disseminação da soja e outros produtos transgênicos como trigo, milho e canola. A utilização do glifosato permite a semeadura de culturas transgênicas imediatamente após a aplicação, tornando o processo de plantio muito prático. A soja, como outros transgênicos, apresenta segurança biológica já definida, mas a utilização do glifosato ainda é um assunto extremamente controverso. Esta revisão apresenta alguns aspectos históricos do binômio soja transgênica Roundup Ready/glifosato e discute as controvérsias mais recentes sobre o uso do glifosato no Brasil e no mundo.(AU)

Glyphosate, isolated or in associations, at agronomic performance and seed quality of the RR®2 soybean / Glifosato, isolado ou em associações, no desempenho agronômico e qualidade de sementes de soja RR ® 2

The "second generation" of glyphosate-tolerant soybean (RR®2 soybean) was developed through a different technique of insertion of the glyphosate-insensitive EPSPs gene. Information on the selectivity of glyphosate, alone or in combination, in RR2 soybean is lacking. This study evaluated the effects of glyphosate, isolated or in associations, applied at post-emergence (V4), at agronomic performance and seed quality of soybean cultivar NS 6700 IPRO (RR2). The experimental design was randomized block with four replications and seven treatments, conducted in the field for two growing seasons. The treatments consisted of glyphosate herbicide, alone or in combination with clethodim, cloransulam, chlorimuron, lactofen and fluazifop, besides the control without application. Analysis was performed for crop injury, Soil and Plant Analyzer Development (SPAD) index, as well as variables related to agronomic performance (height, number of pods per plant, yield and 1,000-seed weight) and seed quality (vigor, germination, abnormal seedlings and dead seeds). An additional test was conducted with the same cultivar and treatments in a greenhouse in a completely randomized design with four replications. The herbicides did not affect agronomic performance and seed quality of RR2 soybean. Thus, the soybean cultivar NS 6700 IPRO (RR2) was tolerant to glyphosate, alone or combined with other herbicides applied in post-emergence (V4).(AU)

A "segunda geração" de soja tolerante ao glifosato (soja RR®2) foi desenvolvida por meio de uma diferente inserção do gene EPSPs insensível ao referido herbicida. Informações sobre a seletividade de glifosato, isolado ou em associação, são faltantes em soja RR2. Este estudo avaliou os efeitos do glifosato, isolado ou em associações, aplicado em pós-emergência (V4), no desempenho agronômico e na qualidade de sementes do cultivar de soja NS 6700 IPRO (RR2). Adotou-se o delineamento experimental de blocos ao acaso com quatro repetições e sete tratamentos, realizados no campo em duas safras. Os tratamentos consistiram no herbicida glifosato, isolado ou em associação com clethodim, cloransulam, chlorimuron, lactofen e fluazifop, além da testemunha sem aplicação. Foi realizada a avaliação dos sintomas de injúria e índice SPAD (Soil and Plant Analyzer Development), bem como de variáveis relacionadas ao desempenho agronômico (altura, número de vagens por planta, produtividade e massa de 1.000 sementes) e à qualidade das sementes (vigor, germinação, plântulas anormais e sementes mortas). Experimento complementar foi realizado com o mesmo cultivar e tratamentos em casa de vegetação em delineamento inteiramente casualizado com quatro repetições. Os herbicidas não afetaram o desempenho agronômico e a qualidade de sementes da soja RR2. Assim, o cultivar NS 6700 IPRO (RR2) foi tolerante ao glifosato, isolado ou em associação com outros herbicidas aplicados em pós-emergência (V4).(AU)

Construction of an RNAi vector for knockdown of GM-ACS genes in the cotyledonary nodes of soybean

Background: Ethylene plays an important role in the regulation of floral organ development in soybean, and 1-aminocyclopropane-1-carboxylate synthase (ACS) is a rate-limiting enzyme for ethylene biosynthesis. However, whether ACS also regulates floral organ differentiation in soybean remains unknown. To address this, we constructed an RNAi vector to inhibit ACS expression in cotyledonary nodes. Linear DNA cassettes of RNAi-ACS obtained by PCR were used to transform soybean cotyledonary nodes. Results: In total, 131 of 139 transiently transformed plants acquired herbicide resistance and displayed GUS activities in the new buds. In comparison to untransformed seedling controls, a greater number of flower buds were differentiated at the cotyledonary node; GM-ACS1 mRNA expression levels and ethylene emission in the transformed buds were reduced. Conclusion: These results indicate that the cotyledonary node transient transformation system may be suitable for stable transformation and that the inhibition of ACS expression may be an effective strategy for promoting floral organ differentiation in soybean.

Soybean yield loss and economic thresholds due to glyphosate resistant hairy fleabane interference / Perda de rendimento da soja e nível de dano econômico pela interferência de buva resistente ao glifosato

The interference of glyphosate resistant hairy fleabane (Conyza bonariensis) has caused yield losses in soybean crop. Knowledge of the economic threshold is an important aspect for the economic management of resistant weeds. Because of it, the interference of the glyphosate resistant hairy fleabane density on the soybean yield loss was evaluated, as well as to estimate the economic threshold. Two experiments were conducted to verify the effect of different weed densities on the yield of two soybean cultivars (BRS Estância and BMX Turbo). Weed density ranged from 0 to 124 plants m-2. The yield of the BRS Estância decreased by 1.4% to the increase of one single plant, while for the BMX Turbo the loss was 25.9%. Soybean yield and economic threshold were lower for the BMX Turbo cultivar compared to the cultivar BRS Estância, which can be attributed to the genotype, competitive ability, weed development stage and/or environment. Increases in crop yield expectation, soybean price and herbicide efficiency reduce the economic threshold, indicating that the adoption of control measures should be taken when the weed density is low. Due to the high cost of hairy fleabane control, it is only feasible at high densities.(AU)

A interferência de plantas de buva (Conyza bonariensis) resistente ao herbicida glyphosate tem causado perdas de produtividade na cultura da soja. O conhecimento do nível de dano econômico é um importante fator para o manejo econômico de plantas daninhas resistentes na cultura da soja. Para isso, foi avaliada a interferência da densidade de buva resistente ao glyphosate sobre a perda de rendimento da soja, bem como estimado o nível de dano econômico. Dois experimentos foram conduzidos para verificar o efeito de diferentes densidades da planta daninha sobre a produtividade de dois cultivares de soja (BRS Estância e BMX Turbo). As densidades de buva variaram de 0 a 124 plantas m-2. A produtividade do cultivar BRS Estância reduziu 1,4% ao aumento de uma planta, enquanto para o cultivar BMX Turbo a perda foi de 25,9%. A produtividade da soja e o nível de dano econômico foram menores para o cultivar BMX Turbo, em comparação com o cultivar BRS Estância, o que pode ser atribuído ao genótipo, à habilidade competitiva, ao estádio de desenvolvimento da buva e/ou a ambiente. Aumentos na expectativa de produtividade da cultura, no preço da soja e na eficiência dos herbicidas reduzem o nível de dano econômico, indicando que a adoção de medidas de controle deve ser tomada quando a população de buva é baixa. Em função do elevado custo de controle da buva, este somente é viável em altas densidades.(AU)

Alternatives for chemical management of sourgrass / Controle químico de capim-amargoso

Intensive use of Glyphosate has been selected multiple resistant sourgrass biotypes in the areas of intensive agriculture in Brazil. Thus, it is important that viable management alternatives are evaluated for this invasion, which currently is regarded as the main problem of Brazilian agriculture. The study aimed to evaluate strategies for chemical management of sourgrass. Three trials divided according to the weed development stages were performed: Post- Initial (up to tillering), Post-Delayed (one to four tillers) and Adult-Plant (adult stage characterized by the beginning of the reproductive phase). The sourgrass seeds were collected from biotypes with evidence of Glyphosate tolerance in the municipality of Rio Brilhante ­ MS. In each trial were evaluated thirteen treatments and one witness, where the treatments were herbicide mixtures applications with subsequent sequential applications at different dates (7, 10 and 14 days after the first application) and the herbicides application without mixtures. Sourgrass plants are tolerant to Glyphosate and Clethodim herbicides. Formulated mixtures of Paraquat + Diuron and Haloxifop are the most suitable for controlling sourgrass plants derived from seeds, regardless of the control period. Treatments that have the Haloxyfop herbicide in the first application provided better control when compared to the other treatments with mixtures and sequential applications at all stages of applications.

O uso intensivo do herbicida glyphosate tem selecionado vários biótipos resistentes de capimamargoso nas áreas de agricultura intensiva no Brasil. Dessa forma, é importante que sejam estudadas alternativas viáveis de manejo para esta invasora, que hoje é considerada como um dos principais problemas da agricultura brasileira. O trabalho teve como objetivo a avaliação de estratégias para o manejo químico de capim-amargoso. Foram realizados três experimentos divididos em função dos estágios de desenvolvimento da planta daninha: Pós-inicial (até o perfilhamento), Pós-tardio (um a quatro perfilhos) e Planta Adulta de capim-amargoso (estágio adulto, caracterizado pelo inicio da fase reprodutiva). As sementes de Capim-amargoso foram coletadas de biótipos com indícios de tolerância ao herbicida Glyphosate no município de Rio Brilhante ­ MS. Em cada experimento foram avaliados treze tratamentos e uma testemunha, onde os tratamentos foram constituídos por aplicações de misturas de herbicidas com posteriores aplicações sequenciais com diferentes datas de aplicações da sequencial (7, 10 e 14 dias após a primeira aplicação) e pela aplicação de herbicidas sem misturas. As plantas de Capim-amargoso apresentam tolerância aos herbicidas Glyphosate e Clethodim. A mistura formulada dos herbicidas Paraquat + Diuron e o herbicida Haloxifop foram os melhores para o controle de plantas de capim-amargoso provenientes de sementes, independente da época de controle. Os tratamentos que possuem o herbicida Haloxifop na primeira aplicação proporcionaram melhores controles quando comparados aos demais tratamentos com misturas e aplicações sequenciais em todos os estágios de aplicações.

Analysis of exogenous gene and protein digestion and absorption of SD rats (Rattus norvegicus) fed roundup ready soybean meal / 生物工程学报

Metabolism and deposition of exogenous gene and protein from transgenic glyphosate herbicide-tolerant soybean meal in SD rats were studied in the experiment. The transgenic soybean GTS40-3-2 meal and its non-transgenic counterpart (parent A5403) were fed to the generation and the second generation Sprague-Dawley (SD) rats (Rattus norvegicus). The study added the genetically modified (GM) soybean meal and its non-transgenic control soybean meal (parent A5403) in a ratio of 20% respectively to the feeds. By using qualitative, quantitative PCR and ELISA methods to detect transgenic soybean residues of metabolism ingredients in rats, the safety and influence of GM soybean were evaluated. The results showed that the intestinal fecal and cecum contents of rats were detected with residues of GM ingredients, intestinal flora and organs were not found related genes and protein. These results indicated that transgenic glyphosate herbicide-tolerant soybean GTS40-3-2 meal was as safe as its non-GM soybean meal in long-term feeding study.

Characterization of the acetohydroxyacid synthase multigene family in the tetraploide plant Chenopodium quinoa

Background Currently, the technology called Clearfield® is used in the development of crops resistant to herbicides that inhibit the enzyme acetohydroxy acid synthase (AHAS, EC 2.2.1.6). AHAS is the first enzyme of the biosynthetic pathway that produces the branched-chain of the essential amino acids valine, leucine, and isoleucine. Therefore, multiple copies of the AHAS gene might be of interest for breeding programs targeting herbicide resistance. In this work, the characterization of the AHAS gene was accomplished for the Chenopodium quinoa Regalona-Baer cultivar. Cloning, sequencing, and Southern blotting were conducted to determine the number of gene copies. Results The presence of multiple copies of the AHAS gene as has been shown previously in several other species is described. Six copies of the AHAS gene were confirmed with Southern blot analyses. CqHAS1 and CqAHAS2 variants showed the highest homology with AHAS mRNA sequences found in the NR Database. A third copy, CqAHAS3, shared similar fragments with both CqAHAS1 and CqAHAS2, suggesting duplication through homeologous chromosomes pairing. Conclusions The presence of multiple copies of the gene AHAS shows that gene duplication is a common feature in polyploid species during evolution. In addition, to our knowledge, this is the first report of the interaction of sub-genomes in quinoa.

Habilidade competitiva relativa de arroz irrigado com arroz-vermelho suscetível ou resistente ao herbicida imazapyr + imazapic / Relative competitive ability of irrigated rice with red rice susceptible or resistant to the herbicide imazapyr + imazapic / Relative competitive ability of irrigated rice with red rice susceptible or resistant to the herbicide imazapyr + imazapic

O uso intenso de herbicidas com o mesmo mecanismo de ação tem selecionado espécies de plantas daninhas resistentes como, por exemplo, o arroz-vermelho (Oryza sativa L.) com resistência à mistura formulada dos herbicidas imazapyr + imazapic. A competição de plantas daninhas ocasiona perda de produtividade das culturas, geralmente crescente, quanto mais semelhantes forem suas características morfofisiológicas. Ainda, a capacidade competitiva do biótipo geralmente é alterada pela característica de resistência, e seu conhecimento é fundamental para a compreensão da dinâmica da população resistente. Diante disso, o objetivo da pesquisa foi estudar a habilidade competitiva entre biótipos de arroz-vermelho resistente e suscetível ao herbicida imazapyr + imazapic com arroz irrigado, utilizando o método de série de substituição. Para isso, foram conduzidos experimentos com a cultivar Puitá INTA CL, competindo com o biótipo de arroz-vermelho suscetível ou resistente. As populações de plantas por vaso totalizaram 24, o equivalente a 942 plantas m-2. Para o arroz-vermelho suscetível ou resistente ao herbicida, a competição interespecífica é favorável em relação à competição intraespecífica, enquanto para a cultura do arroz irrigado o competidor independe. A cultivar Puitá INTA CL, em geral, apresenta habilidade competitiva equivalente aos biótipos de arroz-vermelho suscetível ou resistente ao herbicida imazapyr + imazapic, competindo pelos mesmos recursos do ambiente.(AU)

The intensive use of herbicides with the same mechanism of action has selected resistant weeds, among which is the red rice (Oryza sativa L.) after the use of the formulated herbicide mixture of imazapyr and imazapic. The competition of weeds leads to usually increasing yield losses of crops, the more similar the morphological and physiological characteristics between weed and crop. Moreover, the competitive ability of the biotype is generally modified by the resistance characteristic; besides, the knowledge of competitive ability is fundamental to understand the dynamics of the resistant population. Therefore, the aim of this research was to study the competitive ability of red rice biotypes that are resistant and susceptible to imazapyr and imazapic herbicide with rice, using the series substitution method. Experiments were conducted with Puitá INTA CL cultivar competing with susceptible or resistant biotypes of red rice. The total plant populations per pot were of 24, similar to 942 plants m-2. For red rice that is resistant or susceptible to herbicide, the interspecific competition is favorable with regard to intraspecific competition, while for the irrigated rice the competitor is independent. Generally, the Puitá INTA CL cultivar presents competitive ability equivalent to red rice biotypes that are susceptible or resistant to imazapyr and imazapic herbicide, competing for the same resources from the environment.(AU)

One-step, codominant detection of imidazolinone resistance mutations in weedy rice (Oryza sativa L. )

Background Weedy rice (Oryza sativa L.) is a noxious form of cultivated rice (O. sativa L.) associated with intensive rice production and dry seeding. A cost-efficient strategy to control this weed is the Clearfield rice production system, which combines imidazolinone herbicides with mutant imidazolinone-resistant rice varieties. However, imidazolinone resistance mutations can be introgressed in weedy rice populations by natural outcrossing, reducing the life span of the Clearfield technology. Timely and accurate detection of imidazolinone resistance mutations in weedy rice may contribute to avoiding the multiplication and dispersion of resistant weeds and to protect the Clearfield system. Thus, highly sensitive and specific methods with high throughput and low cost are needed. KBioscience's Allele Specific PCR (KASP) is a codominant, competitive allele-specific PCR-based genotyping method. KASP enables both alleles to be detected in a single reaction in a closed-tube format. The aim of this work is to assess the suitability and validity of the KASP method for detection in weedy rice of the three imidazolinone resistance mutations reported to date in rice. Results Validation was carried out by determining the analytical performance of the new method and comparing it with conventional allele-specific PCR, when genotyping sets of cultivated and weedy rice samples. The conventional technique had a specificity of 0.97 and a sensibility of 0.95, whereas for the KASP method, both parameters were 1.00. Conclusions The new method has equal accuracy while being more informative and saving time and resources compared with conventional methods, which make it suitable for monitoring imidazolinone-resistant weedy rice in Clearfield rice fields.

Avaliação do sistema integrado de controle parasitário em uma criação semi-intensiva de caprinos na região de Santa Catarina / Evaluation of the integrated parasite control in a semi-intensive goat farming in the region of Santa Catarina

O objetivo deste trabalho foi subsidiar um programa sustentável de controle parasitário em um rebanho caprino em São Francisco do Sul, SC, aplicando o Sistema Integrado de Controle Parasitário (SICOPA). Foram utilizados 63 caprinos, distribuídos em nove grupos para teste de eficácia de anti-helmínticos, exames de contagem de ovos por grama de fezes (OPG), coprocultura, grau Famacha, micro-hematócrito, escore corporal e contagem de larvas no pasto. A redução da OPG 15 dias pós-tratamento com closantel, albendazole, nitroxinil, levamisole, ivermectina+levamisole+albendazol, moxidectina, ivermectina, abamectina e sulfóxido de albendazol foi de 89, 83, 65, 63, 57, 37, 31, 0 e 0%, respectivamente. A média de graus Famacha 1 e 2 no estudo foi de 81%, e de graus 3, 4 e 5 foi de 19%, e não se observou correlação com os valores do micro-hematócrito em razão da prevalência de Trichostrongylus sp. (92%). A contagem de larvas infectantes (L3) na pastagem apresentou valores abaixo de 1000 L3/kg/MS, predominando Trichostrongylus sp. Nenhuma das drogas testadas foi considerada eficaz, evidenciando resistência parasitária múltipla. A aplicação de ferramentas do SICOPA e a adoção de estratégias de manejo e nutrição adequados são fundamentais para estabelecer um programa sanitário sustentável.

The objective of this study was to subsidize a sustainable parasite control program in a flock of goats in São Francisco do Sul, SC, Brazil, applying the integrated system for parasite control. Sixty three adult animals were used in nine groups to perform an anthelmintic efficacy test, faecal egg count (FEC), Famacha method, haematocrit, body condition score, coproculture, and the presence of larvae on pasture. Drug efficacy measured by FEC 15 days post-treatment with closantel, ivermectin + Levamisole + albendazole, albendazole, nitroxinyl, levamisol, abamectin, ivermectin, moxidectin and albendazole was 85, 57, 83, 65, 63, 31, 28, 24 and 0%, respectively. The Famacha score 1 or 2 was given to 81% and scores 3, 4 or 5 were given to 19%, without significant correlation with haematocrit values since the predominant was Trichostrongylus sp. (92%). The count of infective larvae levels on pasture revealed values below 1000 L3/kg/DM predominantly Trichostrongylus sp. None of the tested drugs was effective, showing multiple parasitic resistance. The correct application of the SICOPA and adoption of management strategies and proper nutrition are essential to establish a sustainable health program.

Intellectual property rights related to the genetically modified glyphosate tolerant soybeans in Brazil

The present work analyzes the different modalities of protection of the intellectual creations in the biotechnology agricultural field. Regarding the Brazilian legislations related to the theme (the Industrial Property Law - no. 9. 279/96 and the Plant Variety Protection Law - no. 9. 456/97), and based in the international treaties signed by Brazil, the present work points to the inclusions of each of them, as well as to their interfaces using as reference the case study of glyphosate tolerant genetically modified soybean. For this case study, Monsanto's pipelines patents were searched and used to analyze the limits of patent protection in respect to others related to the Intellectual Property (IP) laws. Thus, it was possible to elucidate the complex scenario of the Intellectual Property of the glyphosate tolerant soybeans, since for the farmer it is hard to correlate the royalties payment with the IP enterprise's rights.

O presente trabalho analisa as diferentes modalidades de proteção das criações intelectuais no campo da biotecnologia agrícola. A partir das leis Brasileiras relacionadas ao tema (Lei da Propriedade Industrial - nº 9.279/96 e Lei da Proteção de Cultivares - nº 9.456/97), e com base nos tratados internacionais assinados pelo Brasil, o presente trabalho aponta as inclusões de cada uma, assim como, suas interfaces usando como referência o estudo de caso da soja geneticamente modificada para tolerância ao glifosato. Para este caso, patentes pipelines da Monsanto foram buscadas e usadas para analisar os limites de proteção das patentes frente às outras leis de Propriedade Intelectual (PI) relacionadas. Assim, foi possível elucidar o cenário complexo da Propriedade Intelectual das sojas tolerantes ao glifosato, já que para o agricultor não é fácil correlacionar o pagamento dos royalties com os direitos de PI da empresa.

In vitro effect of the herbicide glyphosate on human blood platelet aggregation and coagulation / Efeito in vitro do herbicida glifosato na agregação plaquetária e coagulação sanguínea em humanos

Glyphosate [N-(phosphonomethyl)-glycine] is a broad-spectrum, non-selective, post-emergence herbicide that is extensively used in agriculture. Published data referring to the effects of this product on human health are contradictory. We showed previously that long-term treatment of rats with low doses of Glyphosate-Biocarb® may induce hepatic histological changes and bleeding without decreasing platelet counts. The aim of the current study was to investigate, in vitro, the effect of glyphosate on human blood platelet aggregation and coagulation. Materials and methods: Platelet aggregation was determined in the platelet-rich plasma using the agents: 6µM-adenosine diphosphate, 6µM-epinephrine and 4µg/mL-collagen. Pretreatment with 500µg/mL glyphosate showed significant hypofunction of the three aggregating agents. The inhibitory effect was dose-dependent at concentrations from 50 to 500 µg/mL. The release of ATP was lower for glyphosate-treated platelets after stimulation by collagen. On the other hand, glyphosate did not promote any inhibitory effects on prothrombin time, thromboplastin time and thrombin time. In conclusion, the results demonstrate that glyphosate promotes changes in the platelet metabolism with an inhibitory effect on primary hemostasis.

O glifosato [N-(phosphonomethyl)-glycine] é um herbicida pós-emergente não seletivo de amplo espectro muito utilizado na agricultura. Dados da literatura referentes aos efeitos desse produto na saúde humana são contraditórios. Em estudos prévios demonstramos que ratos previamente tratados com glifosato apresentavam lesões hepáticas e sangramento sem alterações quantitativas de plaquetas. O objetivo do presente estudo é investigar os efeitos in vitro do glifosato (GP) na agregação plaquetária e coagulação sanguínea em humanos. A agregação plaquetária foi determinada em plasma rico em plaquetas (PRP) usando os agentes adenosina difosfato (ADP) 6µM, epinefrina 6µM e colágeno 4µg/mL. Pré-tratamento com GP 500µg/mL demonstrou significativa hipofunção dos três agentes agregantes. O efeito inibitório foi dose dependente em concentrações de 50-500 µg/mL. Utilizando-se a quantificação de ATP como um índice da capacidade de secreção plaquetária, foi observado diminuição da liberação das plaquetas tratadas com GP. Por outro lado, o GP não promoveu efeito inibidor no tempo de protrombina (TP), tempo de tromboplastina parcial ativada (ATTP) e tempo de trombina (TT). Em conclusão, os resultados demonstram que o GP promove mudanças no metabolismo plaquetário com efeito inibitório na hemostasia primária.

Avaliação de equivalência substancial e potencial de alergenicidade de cultivares de soja tolerantes ao herbicida glifosato / Evaluation of substantial equivalence and potential of allergenic reactions of soybean cultivars tolerant to the glyphosate herbicide

Os parâmetros de avaliação de segurança de alimentos geneticamente modificados fundamentam-se na comparação de equivalência substançial entre as variedades e pela inocuidade de proteínas da planta GM com as proteínas encontradas nas plantas convencionais. O objetivo deste trabalho foi avaliar a segurança alimentar de três cultivares de sojas geneticamente modificadas para tolerarem o herbicida glifosato através da determinação da equivalência substancial e do potencial alergênico das mesmas quando comparadas às suas respectivas parentais isogênicas. Seis amostras de soja foram analisadas, sendo três convencionais parentais e três GM, referentes ao cultivo de 2004-2005, em Goiás. Para a composição química foram realizadas análises em triplicata de proteínas, lipídeos, umidade, minerais e fibra alimentar. Análises complementares para determinação de aminoácidos, ácidos graxos, isoflavonas e ácido fítico também foram realizadas. O potencial de alergenicidade foi avaliado em extratos protéicos brutos de três cultivares convencionais e suas correspondentes GM. Os mesmos extratos protéicos foram fracionados para obter as globulinas 7S e 11S por precipitação e posterior purificação em coluna de bioafinidade Sepharose 4B...

Análise comparativa de mapas protéicos de amostras de soja convencionais e tolerantes ao herbicida glifosato visando à inocuidade alimentar / Comparative analysis of maps soy protein samples of conventional and tolerant to the herbicide glyphosate for food safety

A soja geneticamente modificada tolerante ao herbicida glifosato tem sido a cultura derivada da engenharia genética mais cultivada atualmente no mundo. Como todo alimento GM a soja tem sido alvo de investigação em relação a sua Biossegurança. Novas estratégias têm sido desenvolvidas e aplicadas neste campo de pesquisa, sendo que métodos rápidos e eficientes de análise proteômica têm sido utilizados para avaliação e monitoramento da segurança e inocuidade alimentar, indicando mudanças no perfil protéico entre variedades convencionais e GM. O objetivo do presente trabalho foi avaliar os mapas protéicos de amostras de soja convencionais e suas derivadas geneticamente modificadas tolerantes ao herbicida glifosato, utilizando técnicas de análise proteômica com ênfase para inocuidade alimentar. Foram utilizadas seis amostras de soja, sendo três convencionais parentais e três derivadas GM, cultivadas entre 2004-2005, em Goiás. O extrato bruto protéico foi submetido à análise por eletroforese unidimensional e bidimensional. A eletroforese 2D, foi realizada utilizando tiras com gradiente de pH de 3-10 e 4-7. As imagens dos mapas protéicos das seis variedades, produzidas em replicatas, foram analisadas pelo software ImageMaster 2D Platinum. O potencial alergênico do extrato protéico bruto foi avaliado para todas as variedades utilizando soro de pacientes alérgicos à soja através de immunoblotting. Nos resultados obtidos observou-se a presença das principais frações protéicas da soja pela eletroforese unidimensional sem alteração significativa entre as amostras parentais e GM, exceto para uma banda de 115 kDa presente nas amostras parentais, mas ausente nas amostras GM. A partir da análise por eletroforese 2D foram identificadas as formas peptídicas correspondentes às frações de β-conglicinina e glicinina bem como diversas outras proteínas encontradas na soja como o inibidor de tripsina e a lipoxigenase. Através do software foi possível observar que um spot apresentou...

Genetically modified soya-tolerant to the herbicide glyphosate culture has been derived from the more cultivated genetic engineering in the world today. As GM soya beans whole food has been investigated in relation to your biosafety. New strategies have been developed and applied research in this field, and fast and efficient methods of analysis proteomics have been used for assessment and monitoring of food security and safety, indicating changes in own protein profile between conventional and GM varieties. The aim of this work was to assess the maps soy protein samples of conventional and genetically modified their derived to the herbicide glyphosate-tolerant, using Proteomics analysis techniques with emphasis on food safety. Six samples were used for conventional soya, three and three derived from GM parental, grown between 2004-2005. The crude protein extract own was subjected to analysis by electrophoresis one-dimensional and two-dimensional. 2D electrophoresis using Strip was held with pH gradient of 3-10 and 4-7. Protein maps images of six varieties produced in replicates have been analysed by the 2D Platinum software ImageMaster. The potential allergenic in crude protein extracts was evaluated for all varieties using allergic patient serum soya by immunoblotting. In the results obtained noted the presence of the main protein fractions of soya by one-dimensional electrophoresis without significant change between parental and GM samples, except for a band of 115 parental kDa present in the sample, but absent in GM samples. From the analysis by 2D electrophoresis peptides forms were identified corresponding to fractions of β-conglicinina and glicinina as well as several other proteins found in soy as trypsin inhibitor and lipoxygenase. Through the software has been possible to observe that a spot presented statistical difference between the samples tested, expressed in greater concentration in the samples GM in parenting. In tests of allergenicity, GM...

Expression of acetohydroxyacid synthase isozyme genes ilvBN, ilvGM, ilvIH and their resistance to AHAS-inhibitor herbicides / 生物工程学报

Acetohydroxyacid synthase (AHAS) catalyses the first reaction in the pathway for synthesis of the branched-chain amino acids. AHAS is the target for sulfonylurea, imidazolinone and other AHAS-inhibitor herbicides. Herbicides-resistant AHAS genes have potential application in plant transgenetic engineering and development of new generation herbicide. The AHAS isozyme genes ilvBN, ilvGM and ilvIH were cloned from metsulfuron-methyl resistant strain Klebsiella sp. HR11 and metsulfuron-methyl sensitive strain Klebsiella pneumoniae MGH 78578. Homologous sequences comparison indicated that the differences in AHAS isozyme genes at amino acid levels between strain HR11 and strain MGH 78578 were mainly on the large subunits of ilvBN and ilvGM. The three AHAS isozyme genes from HR11 and MGH 78578 were ligated into the expression vector pET29a(+) and expressed in Escherichia coli BL21, respectively. The results of enzyme inhibition assay showed that only ilvBN and ilvGM from strain HR11 showed strong resistance to AHAS-inhibitor herbicides, while ilvIH from strain HR11 and ilvBN, ilvGM and ilvIH from strain MGH78578 were sensitive to AHAS-inhibitor herbicides.