Plant characterization of insect-protected soybean.

Insect-protected soybean (SIP) that produces the Cry1A.105 and Cry2Ab2 insecticidal crystal proteins has been developed to provide protection from feeding damage caused by targeted lepidopteran insect pests. Typically, as part of environmental risk assessment (ERA), plant characterization is conducted, and the data submitted to regulatory agencies prior to commercialization of genetically modified (GM) crops. The objectives of this research were to: (a) compare soybean with and without the SIP trait in plant characterization field trials designed to fulfill requirements for submissions to global regulatory agencies and address China-specific considerations and (b) compare risk assessment conclusions across regions and the methodologies used in the field trials. The soybean with and without the SIP trait in temperate, tropical, and subtropical germplasm were planted in replicated multi-location trials in the USA (in 2012 and 2018) and Brazil (in 2013/2014 and 2017/2018). Agronomic, phenotypic, plant competitiveness, and survival characteristics were assessed for soybean entries with and without the SIP trait. Regardless of genetic background, growing region, season, or testing methodology, the risk assessment conclusions were the same: the evaluated insect-protected soybean did not differ from conventional soybean in evaluated agronomic, phenotypic, competitiveness, and survival characteristics indicating no change in plant pest/weed potential. These results reinforce the concept of data transportability across global regions, different seasons, germplasm, and methodologies that should be considered when assessing environmental risks of GM crops.

Glyphosate hormesis induced by treatment via seed stimulates the growth and biomass accumulation in soybean seedlings.

Glyphosate hormesis, identified as a potential means to enhance crop yields, encounters practical constraints because it is typically assessed through foliar applications. The expression and extend of hormesis in this approach are influenced by unpredictable environmental conditions, highlighting the need to explore alternative glyphosate application methods, such as seed treatment. This study aimed to assess glyphosate hormesis on growth rates and biomass accumulation in seedlings soybean cultivars. Two dose-response experiments [doses from 0 to 2880 g acid equivalent (ae) ha-1], one via foliar and one via seed, were conducted on three soybean cultivars [one non-glyphosate-resistant (NGR) and two glyphosate-resistant (GR, one RR and one RR2)]. In a subsequent experiment, three safe glyphosate doses (0, 90 and 180 g ae ha-1) applied via seed were evaluated on four soybean cultivars (two RR and two RR2). For foliar applications, the range of glyphosate doses increasing growth rates and dry biomass by 12-28 % were 5.6-45 g ae ha-1 for the NGR cultivar, of 45-720 g ae ha-1 for RR and of 11.25-180 g ae ha-1 for RR2. In the seed treatment, biomass increases of 16-60 % occurred at 45-180 g ae ha-1 for the NGR and RR cultivars, and 90-360 g ae ha-1 for RR2. Glyphosate doses of 90 and 180 g ae ha-1, applied via seeds, provided greater growth and biomass accumulation for the RR and RR2 soybean cultivars. Both foliar and seed applications of glyphosate increased growth and biomass accumulation in soybean cultivars, with seed treatments showing greater and more consistent enhancements. These findings propose practical and viable alternative for harnessing glyphosate hormesis to facilitate the early development of soybeans and potentially enhance crop yield.

Bt Soybean Cry1Ac Does Not Affect Development, Reproduction, or Feeding Behavior of Red-Banded Stink Bug Piezodorus guildinii (Hemiptera: Pentatomidae).

Bt soybean cultivation is increasing worldwide. The Cry1Ac protein expressed in Bt soybean efficiently controls several lepidopteran pests. The stink bug, Piezodorus guildinii (Westwood), a major pest for soybean in the Americas, is not controlled by Bt crops, although possible sub-lethal effects may occur. Even if there were no negative effects for sting bug, ingesting toxins could affect its bio-controllers. We tested through ELISA detection if P. guildinii ingests Cry1Ac from Bt soybean and possible effects on its development, reproduction, survival, and feeding behavior. Biological traits were evaluated under controlled conditions of nymphs and adults feeding on pods of near-isogenic cultivars DM5958iPRO (Bt) and DM59i (non-Bt). Feeding behavior was recorded using an AC-DC electropenetrography (EPG) device. Results indicated that P. guildinii ingested the Cry1Ac protein; however, nymphal period and accumulated survival percentage did not differ between cultivars. Feeding on Bt soybean pods did not affect fecundity (i.e., number of egg masses and eggs/female) nor egg viability. Different feeding behaviors were only detected on the pathway phase (stylet penetration into plant tissue), which was more pronounced in the Bt cultivar. However, the total duration of the feeding activities on seeds was numerically higher (ca. 2X) on Bt plants compared to non-Bt. This is the first study to demonstrate that P. guildinii does ingest the Cry1Ac protein and excrete it without being absorbed, probably explaining the lack of direct adverse effects on its biological parameters. EPG could indicate that Bt soybean plants might be less palatable than non-Bt to red-banded stink bug.

Risk assessment of the antifungal and insecticidal peptide Jaburetox and its parental protein the Jack bean (Canavalia ensiformis) urease.

Jaburetox (JBTX) is an insecticidal and antifungal peptide derived from jack bean (Canavalia ensiformis) urease that has been considered a candidate for developing genetically modified crops. This study aimed to perform the risk assessment of the peptide JBTX following the general recommendations of the two-tiered, weight-of-evidence approach proposed by International Life Sciences Institute. The urease of C. ensiformis (JBU) and its isoform JBURE IIb (the JBTX parental protein) were assessed. The history of safe use revealed no hazard reports for the studied proteins. The available information shows that JBTX possesses selective activity against insects and fungi. JBTX and JBU primary amino acids sequences showed no relevant similarity to toxic, antinutritional or allergenic proteins. Additionally, JBTX and JBU were susceptible to in vitro digestibility, and JBU was also susceptible to heat treatment. The results did not identify potential risks of adverse effects and reactions associated to JBTX. However, further allergen (e.g. serum IgE binding test) and toxicity (e.g. rodent toxicity tests) experimentation can be done to gather additional safety information on JBTX, and to meet regulatory inquiries for commercial approval of transgenic cultivars expressing this peptide.

Quality of silage produced from intercropped corn RR and soybeans RR / Qualidade de silagem produzida a partir de milho rr consorciado com soja RR

The objective of this study was to produce silage from intercropped RR glyphosate tolerant corn (RR - Roundup Ready) and RR soybeans under Brazilian Cerrado conditions. The research was carried out in off-season between February and June 2015, in Rio Verde, Goiás (Brazil). In the experimental period the rainfall was 865 mm and the average temperature of 23.7°C. Two trials were performed. The first had corn rows spaced in 1 m with a row of soybeans in the interrow. The second had double corn rows interspersed with one row of soybeans, all spaced in 0.50 m. In both trials, we used a randomized block design, in a 2x2+3 factorial scheme, with four replications. Primary factor consisted of two types of pre-sowing fertilization: one only in corn rows and the other over the total area. The secondary factor corresponded to two soybean varieties: medium and late cycle variety. In addition, three control treatments were made: one of corn and two of soybeans. The corn cutoff point was at the hard farinaceous stage when the milk line reaches half the grain, and in the sole crop soybean at the R5.5 final phase of grain filling. Silage was stored in PVC experimental silos for 60 days. The intercropped treatments, in both spatial arrangements, promoted increases in neutral detergent fiber and acid detergent fiber contents. Soybean monoculture presented high values of crude protein and ethereal extract. There were higher effluent losses in medium-cycle soybean variety silage when it was grown in single cropping. However, the lowest effluent losses were recorded for silages of medium-cycle soybean intercropped under a double row pre-sowing corn fertilization system. The soybeans mixed silages produced together with corn under off-season conditions did not yield satisfactory results.

Objetivou-se neste trabalho adequar o sistema de consórcio entre milho com tolerância ao herbicida glifosato (RR - Roundup Ready) e soja RR para obtenção de silagem de qualidade em condições do Cerrado brasileiro. A pesquisa foi conduzida na safrinha entre fevereiro e junho de 2015, em Rio Verde, Goiás (Brasil). A pluviosidade do período experimental foi de 865mm e a temperatura média de 23,7°C. Realizou-se dois ensaios, adequando formas de adubação de semeadura, arranjo de plantas e cultivares. Utilizou-se delineamento em blocos casualizados, em fatorial 2x2+3, com quatro repetições, sendo o fator primário duas modalidades de adubação de semeadura na linha: somente no milho, e em área total; o fator secundário correspondeu a duas variedades de soja: ciclo médio e tardio; mais três testemunhas: uma de milho e duas de soja. O ponto de corte do milho foi no estádio farináceo duro quando a linha do leite atinge metade do grão, e na soja em monocultivo no estádio fenológico R5.5, fase final de enchimento de grãos. Armazenou-se a silagem em silos experimentais de PVC por 60 dias. Os tratamentos de consórcio em ambos os arranjos aumentaram os teores de fibra em detergente neutro e fibra em detergente ácido. Os monocultivos de soja obtiveram valores elevados de proteína bruta e extrato etéreo. Ocorreu maior perda por efluente na silagem da variedade de soja de ciclo médio cultivada em monocultivo. No entanto, as menores perdas por efluente foram registradas para silagens de culturas de soja de ciclo médio consorciadas sob um sistema de fertilização pré-semeadura de linha de milho, no arranjo de linha dupla. A inserção de soja para produzir silagens mista juntamente com o milho em condições de safrinha não teve resultados satisfatórios.

Dynamics of glyphosate and AMPA in the soil surface layer of glyphosate-resistant crop cultivations in the loess Pampas of Argentina.

This study investigates the dynamics of glyphosate and AMPA in the soil surface layer of two fields growing glyphosate-resistant crops in the loess Pampas of Córdoba Province, Argentina. Glyphosate decay and AMPA formation/decay were studied after a single application, using decay kinetic models. Furthermore, glyphosate and AMPA concentrations were investigated in runoff to evaluate their off-site risk. During a 2.5-month study, cultivations of glyphosate-resistant soybean and maize received an application of 1.0 and 0.81 kg a.e. ha-1, respectively, of Roundup UltraMax©. Topsoil samples (0-1, 1-2 cm) were collected weekly (including before application) and analysed for glyphosate, AMPA and soil moisture (SM) contents. Runoff was collected from runoff plots (3 m2) and weirs after 2 erosive rainfall events, and analysed for glyphosate and AMPA contents (water, eroded-sediment). Under both cultivations, background residues in soil before application were 0.27-0.42 mg kg-1 for glyphosate and 1.3-1.7 mg kg-1 for AMPA. In the soybean area, the single-first-order (SFO) model performed best for glyphosate decay. In the maize area, the bi-phasic Hockey-Stick (HS) model performed best for glyphosate decay, due to an abrupt change in SM regimes after high rainfall. Glyphosate half-life and DT90 were 6.0 and 19.8 days, respectively, in the soybean area, and 11.1 and 15.4 days, respectively, in the maize area. In the soybean area, 24% of the glyphosate was degraded to AMPA. In the maize area, it was only 5%. AMPA half-life and DT90 were 54.7 and 182 days, respectively, in the soybean area, and 71.0 and 236 days, respectively, in the maize area. Glyphosate and AMPA contents were 1.1-17.5 times higher in water-eroded sediment than in soil. We conclude that AMPA persists and may accumulate in soil, whereas both glyphosate and AMPA are prone to off-site transport with water erosion, representing a contamination risk for surface waters and adjacent fields.

Bt Proteins Have No Detrimental Effects on Larvae of the Green Lacewing, Chrysopa pallens (Rambur) (Neuroptera: Chrysopidae).

Biosafety of a genetically modified crop is required to be assessed prior to its commercialization. For this, a suitable artificial diet was developed and used to establish a dietary exposure test for assessing the toxicity of midgut-active Bt insecticidal proteins on Chrysopa pallens (Rambur). Subsequently, this dietary exposure test was used to evaluate the toxicity of the proteins Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, and Vip3Aa on C. pallens larvae. Temporal stability, bioactivity, and the intake of the insecticidal proteins were confirmed by enzyme-linked immunosorbent assay and a sensitive-insect bioassay. The life history characteristics, such as survival, pupation, adult emergence, 7-day larval weight, larval developmental time, and emerged male and female fresh weights remained unaffected, when C. pallens were fed the pure artificial diet (negative control) and the artificial diets containing 200 µg/g of each purified protein: Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, or Vip3Aa. On the contrary, all of the life history characteristics of C. pallens larvae were adversely affected when fed artificial diet containing boric acid (positive control). The results demonstrate that diets containing the tested concentrations of Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, and Vip3Aa have null effects on C. pallens larvae. The outcome indicates that genetically modified crops expressing the tested Bt proteins are safe for the lacewing, C. pallens.

Outcrossing potential between 11 important genetically modified crops and the Chilean vascular flora.

The potential impact of genetically modified (GM) crops on biodiversity is one of the main concerns in an environmental risk assessment (ERA). The likelihood of outcrossing and pollen-mediated gene flow from GM crops and non-GM crops are explained by the same principles and depend primarily on the biology of the species. We conducted a national-scale study of the likelihood of outcrossing between 11 GM crops and vascular plants in Chile by use of a systematized database that included cultivated, introduced and native plant species in Chile. The database included geographical distributions and key biological and agronomical characteristics for 3505 introduced, 4993 native and 257 cultivated (of which 11 were native and 246 were introduced) plant species. Out of the considered GM crops (cotton, soya bean, maize, grape, wheat, rice, sugar beet, alfalfa, canola, tomato and potato), only potato and tomato presented native relatives (66 species total). Introduced relative species showed that three GM groups were formed having: a) up to one introduced relative (cotton and soya bean), b) up to two (rice, grape, maize and wheat) and c) from two to seven (sugar beet, alfalfa, canola, tomato and potato). In particular, GM crops presenting introduced noncultivated relative species were canola (1 relative species), alfalfa (up to 4), rice (1), tomato (up to 2) and potato (up to 2). The outcrossing potential between species [OP; scaled from 'very low' (1) to 'very high' (5)] was developed, showing medium OPs (3) for GM-native relative interactions when they occurred, low (2) for GMs and introduced noncultivated and high (4) for the grape-Vitis vinifera GM-introduced cultivated interaction. This analytical tool might be useful for future ERA for unconfined GM crop release in Chile.

Cultivos genéticamente modificados: las promesas y las buenas intenciones no bastan

Genetically modified crops: promises and good intentions are not enough (refutation to Espinoza et al. 2004, Rev. Biol. Trop. 52 (3): 727-732). The arguments presented by Espinoza et al. in their paper "Relationship of genetically modified crops with the environment and health of the Costa Rican human population" published in this journal (Rev. Biol. Trop. 52: 727-732, 2004) are questioned and refuted. The arguments are confronted with evidence offered by scientists and national and international independent organizations around the world (e.g. World Health Organization, Consumers international, Physicians and Scientists for Responsible Application of Science and Technology, international Union for Conservation of Nature and Natural Resources, the Council of the University of Costa Rica, and the independent Science Panel) showing the current uncertainty and limitations of science in this area, as well as those of proposed and applied biosafety approaches. Environment, biodiversity and food security are so important and basic matters, that there is need of serious testing, particularly when promises seem to be based on environmentally dangerous ideas promoted half a century ago by the so called "green revolution". Debate should continue, based on a holistic analysis of facts and with ethical reasoning, avoiding emotional positions that can confuse virtual reality with reality. Rev. Biol. Trop. 55 (2): 347-364. Epub 2007 June, 29.

Se refutan y cuestionan los razonamientos expuestos por las autoras en el artículo "Relación de los cultivos modificados genéticamente con el ambiente y la salud de la población costarricense" publicado en esta revista por Espinoza et al. (52: 727-732, 2004). Para ello se contraponen éstos con las evidencias expuestas y analizadas tanto por científicos como por organizaciones nacionales e internacionales independientes de diverso tipo en diferentes lugares del mundo (e.g. Organización Mundial de la Salud, internacional de Consumidores, Médicos y Científicos por una Aplicación Responsable de la Ciencia y la Tecnología, Unión internacional para la Conservación de la Naturaleza, Consejo Universitario de la Universidad de Costa Rica y el Panel de Ciencias independiente), que confirman la incertidumbre y limitaciones actuales de la ciencia en esta área, así como de los mecanismos de bioseguridad propuestos y puestos en vigor. En materias tan importantes y básicas para la supervivencia como las que nos ocupan -el ambiente, la biodiversidad y la seguridad alimentaria- no es prudente apostar a promesas basadas en la misma lógica promocionada desde hace poco más de cinco décadas por la denominada "revolución verde". Es necesario continuar este debate basado en el análisis holístico de los hechos, así como en razonamientos éticos, evitando caer en posiciones emocionales que llegan a confundir la realidad virtual con la verdadera realidad.