Controlled expression of recombinant proteins in Physcomitrella patens by a conditional heat-shock promoter: a tool for plant research and biotechnology.

The ability to express tightly controlled amounts of endogenous and recombinant proteins in plant cells is an essential tool for research and biotechnology. Here, the inducibility of the soybean heat-shock Gmhsp17.3B promoter was addressed in the moss Physcomitrella patens, using beta-glucuronidase (GUS) and an F-actin marker (GFP-talin) as reporter proteins. In stably transformed moss lines, Gmhsp17.3B-driven GUS expression was extremely low at 25 degrees C. In contrast, a short non-damaging heat-treatment at 38 degrees C rapidly induced reporter expression over three orders of magnitude, enabling GUS accumulation and the labelling of F-actin cytoskeleton in all cell types and tissues. Induction levels were tightly proportional to the temperature and duration of the heat treatment, allowing fine-tuning of protein expression. Repeated heating/cooling cycles led to the massive GUS accumulation, up to 2.3% of the total soluble proteins. The anti-inflammatory drug acetyl salicylic acid (ASA) and the membrane-fluidiser benzyl alcohol (BA) also induced GUS expression at 25 degrees C, allowing the production of recombinant proteins without heat-treatment. The Gmhsp17.3B promoter thus provides a reliable versatile conditional promoter for the controlled expression of recombinant proteins in the moss P. patens

Regulação da embriogênese semática in vitro com ênfase do papel de hormonios endógenos

Different aspects of the in vitro somatic embryogenesis regulation are reviewed in this paper.work. A description of g General aspects, such as terminology, uses, stages of development and factors associated with the somatic embryogenesis, are described. is carried out. Although a brief description ofn the effects of the addition of different plant growth regulators to the culture medium wasis given, the article is centereds itself on the effect that the endogenous hormone concentrations in the initial explants and in the tissue cultures derived from them could play oin the induction and expression of somatic embryogenesis. It is significant that few to emphasize the low amount of systematic studies have been conducted, in this subject, in which different species and hormone groups were compared in cultures with and without embryogenic capacity. Moreover, the lack of correlation between the results presented in different studies the distinct works indicates that the hormone content of the cultures is not the only factor involved.Neste trabalho se faz uma revisão de diversos aspectos da regulação da embriogêneses somático in vitro. Vários aspectos gerais a este fenômeno tem sido discutidos, tais como a definição de terminologia, descrição de eventuais aplicações, seus estados de desenvolvimento e outros fatores associados com sua indução e expressão. Embora se faça uma breve descrição do efeito da adição de diferentes reguladores de crescimento ao meio de cultivo, o artigo está centrado no efeito que as concentrações hormonais endogênas nos explantes iniciais e nos cultivos in vitro derivados deles podem ter na indução e expressão da embriogênese somática. Tem de se fazer ênfase na pouca quantidade de estudos sistemáticos realizados neste tema que comparem em várias espécies e diferentes grupos hormonais em cultivos com e sem competência embriogênica. Finalmente, indica-se que a falta de correlação entre os resultados destes poucos trabalhos parece indicar que os conteúdos hormonais endôgenos não são os únicos fatores envolvidos neste fenômeno.Grman Academic Exchange Service//DAAD/AlemaniaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro para Investigaciones en Granos y Semillas (CIGRAS

Novel and emerging biotechnological crop protection approaches

Traditional breeding or genetically modified organisms (GMOs) have for a long time been the sole approaches to effectively cope with biotic and abiotic stresses and implement the quality traits of crops. However, emerging diseases as well as unpredictable climate changes affecting agriculture over the entire globe force scientists to find alternative solutions required to quickly overcome seasonal crises. In this review, we first focus on cisgenesis and genome editing as challenging biotechnological approaches for breeding crops more tolerant to biotic and abiotic stresses. In addition, we take into consideration a toolbox of new techniques based on applications of RNA interference and epigenome modifications, which can be adopted for improving plant resilience. Recent advances in these biotechnological applications are mainly reported for non‐model plants and woody crops in particular. Indeed, the characterization of RNAi machinery in plants is fundamental to transform available information into biologically or biotechnologically applicable knowledge. Finally, here we discuss how these innovative and environmentally friendly techniques combined with traditional breeding can sustain a modern agriculture and be of potential contribution to climate change mitigation