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.

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.

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.

Biologia e ecofisiologia de buva (Conyza bonariensis e Conyza canadensis): [revisão] / Biology and ecophysiology of hairy fleabane (Conyza bonariensis) and horseweed (Conyza canadensis)

Conyza bonariensis (L.) Cronquist e Conyza canadensis (L.) Cronquist são duas espécies de plantas daninhas pertencentes à família Asteraceae. Com freqüência, elas infestam pomares, vinhedos e outras culturas, como milho, soja e algodão e, também, culturas forrageiras, pastagens e áreas não-cultivadas. Esta revisão de literatura tem por objetivos descrever e discutir as características botânicas, biológicas e ecofisiológicas daquelas espécies com a finalidade de facilitar a adoção de estratégias para seu manejo em culturas ou limitar seu estabelecimento. Conyza canadensis é uma espécie anual ou bienal, nativa da América do Norte, enquanto C. bonariensis é uma espécie anual, nativa da América do Sul. A propagação destas espécies dá-se através de sementes que são facilmente dispersas através do vento e da água. As sementes não apresentam dormência e podem germinar prontamente em condições de temperatura e umidade favoráveis. Em vários países, inclusive no Brasil, foi constatada resistência aos herbicidas em biótipos destas espécies. Adoção de práticas culturais diversificadas, integradas ao controle químico, pode minimizar os efeitos negativos provocados por buva e, também, prevenir casos de resistência.

Conyza bonariensis (L.) Cronquist and Conyza canadensis (L.) Cronquist are two weed species belonging to Asteraceae family. Frequently, they occur in orchards, vineyards, and in other crops, such as corn, soybean, and cotton and, also, in forage crops and pastures, as well as in fallow areas. This literature review has as objectives to describe and discuss botanical, biological, and ecopysiological characteristics of Conyza species that may facilitate adoption of strategies for their management in crops or to limit their establishment. Conyza canadensis is an annual or biennial species, native of North America; whereas, C. bonariensis is an annual species, native of South America. Spreading of both species occurs by seeds, which are easily dispersed through wind and water. Seeds do not present dormancy and can germinate promptly under favorable conditions of temperature and humidity. In various countries, including Brazil, Conyza biotypes resistant to herbicides were found. Adoption of diversified cultural practices, integrated to were found chemical control, may minimize the negative effects of Conyza, as well as, prevent cases of resistance.

Expression of bacterial genes in transgenic tobacco: methods, applications and future prospects

Tobacco is the most commonly used plant for expression of transgenes from a variety of organisms, because it is easily grown and transformed, it provides abundant amounts of fresh tissue and has a well-established cell culture system. Many bacterial proteins involved in the synthesis of commercial products are currently engineered for production in tobacco. Bacterial enzymes synthesized in tobacco can enhance protection against abiotic stresses and diseases, and provide a system to test applied strategies such as phytoremediation. Examples of bacterial gene expression in tobacco include production of antigen proteins from several human bacterial pathogens as vaccines, bacterial proteins for enhancing resistance against insects, pathogens and herbicides, and bacterial enzymes for the production of polymers, sugars, and bioethanol. Further improvements in the expression of recombinant proteins and their recovery from tobacco will enhance production and commercial use of these proteins. This review highlights the dynamic use of tobacco in bacterial protein production by examining the most relevant research in this field.