RESUMEN
Wheat is the predominant crop worldwide, contributing approximately 20% of protein and calories to the human diet. However, the yield potential of wheat faces limitations due to pests, diseases, and abiotic stresses. Although conventional breeding has improved desirable traits, the use of modern transgenesis technologies has been limited in wheat in comparison to other crops such as maize and soybean. Recent advances in wheat gene cloning and transformation technology now enable the development of a super wheat consistent with the One Health goals of sustainability, food security, and environmental stewardship. This variety combines traits to enhance pest and disease resistance, elevate grain nutritional value, and improve resilience to climate change. In this review, we explore ways to leverage current technologies to combine and transform useful traits into wheat. We also address the requirements of breeders and legal considerations such as patents and regulatory issues.
Asunto(s)
Plantas Modificadas Genéticamente , Triticum , Triticum/genética , Productos Agrícolas/genética , Fitomejoramiento , Ingeniería Genética , Enfermedades de las Plantas/parasitología , Enfermedades de las Plantas/prevención & control , Resistencia a la Enfermedad/genéticaRESUMEN
There is a worldwide concern to achieve food security with a sustainable approach, including the generation and implementation of techniques for the production of high-quality chemical-free crops. This food revolution has promoted the development and consolidation of programmes for integrated pest management. Some of those programmes include the use of diverse organisms (biological control agents) to suppress populations of pests potentially harmful to the crops. Among these biological control agents are entomopathogenic fungi that are highly effective in suppressing a diversity of insects and have, therefore, been produced and marketed throughout the world. However, the bottleneck for applying entomopathogenic fungi is the production of propagules (blastospores and conidia) with resistance to environment conditions and abiotic factors, maintaining high quality in terms of virulence. Therefore, this manuscript presents recent studies related to increasing resistance and quality using different bioreactors to produce conidia. The above presents a global panorama related to current developments that contribute to improving the resistance, quality, and production of entomopathogenic fungal propagules.
Asunto(s)
Agentes de Control Biológico , Control Biológico de Vectores , Animales , Productos Agrícolas , Hongos , Insectos/microbiología , Control Biológico de Vectores/métodos , Esporas FúngicasRESUMEN
Over the past decades, advances in plant biotechnology have allowed the development of genetically modified maize varieties that have significantly impacted agricultural management and improved the grain yield worldwide. To date, genetically modified varieties represent 30% of the world's maize cultivated area and incorporate traits such as herbicide, insect and disease resistance, abiotic stress tolerance, high yield, and improved nutritional quality. Maize transformation, which is a prerequisite for genetically modified maize development, is no longer a major bottleneck. Protocols using morphogenic regulators have evolved significantly towards increasing transformation frequency and genotype independence. Emerging technologies using either stable or transient expression and tissue culture-independent methods, such as direct genome editing using RNA-guided endonuclease system as an in vivo desired-target mutator, simultaneous double haploid production and editing/haploid-inducer-mediated genome editing, and pollen transformation, are expected to lead significant progress in maize biotechnology. This review summarises the significant advances in maize transformation protocols, technologies, and applications and discusses the current status, including a pipeline for trait development and regulatory issues related to current and future genetically modified and genetically edited maize varieties.
RESUMEN
Mesenchymal stem/stromal cells (MSCs) have the ability to secrete bioactive molecules, exerting multiple biological effects, such as tissue regeneration, reduction of inflammation, and neovascularization. The therapeutic potential of MSCs can be increased by genetic modification to overexpress cytokines and growth factors. Here we produced mouse MSCs overexpressing human leukemia inhibitory factor (LIF) to assess their proangiogenic potential in vitro and in vivo. Mouse bone marrow-derived MSCs were transduced by using a second-generation lentiviral system to express human LIF. Leukemia inhibitory factor expression was confirmed by RT-qPCR and by ELISA, allowing the quantification of the transcript and secreted protein, respectively. Flow cytometry analysis and trilineage differentiation assay showed that the MSC_LIF cell line maintained the immunophenotype and a multipotency characteristic of MSCs. The immunosuppressive activity of MSC_LIF was confirmed using a lymphoproliferation assay. Moreover, gene expression analysis demonstrated upregulation of genes coding for strategic factors in the neovascularization process, such as angiogenin, IL-8, MCP-1, and VEGF, and for the perivascular cell markers αSMA, Col4a1, SM22, and NG2. To evaluate the pro-angiogenic potential of MSC_LIF, we first tested its effects on endothelial cells obtained from umbilical vein in a scratch wound healing assay. Conditioned medium (CM) from MSC_LIF promoted a significant increase in cell migration compared to CM from control MSC. Additionally, in vitro tube formation of endothelial cells was increased by the presence of MSC_LIF, as shown in microvessel sprouting in aortic ring cultures. Finally, an in vivo Matrigel plug assay was performed, showing that MSC_LIF were more potent in promoting in vivo angiogenesis and tissue vascularization than control MSCs. In conclusion, LIF overexpression is a promising strategy to increase the proangiogenic potential of MSCs and sets precedents for future investigations of their potential applications for the treatment of ischemic diseases and tissue repair.
RESUMEN
[This corrects the article DOI: 10.3389/fmicb.2019.00357.].
RESUMEN
Species from the genus Candida are among the most important human fungal pathogens. Several of them are frequent commensals of the human microbiota but are also able to cause a variety of opportunistic infections, especially when the human host becomes immunocompromised. By far, most of the research to understand the molecular underpinnings of the pathogenesis of these species has focused on Candida albicans, the most virulent member of the genus. However, epidemiological data indicates that related Candida species are also clinically important. Here, we describe the generation of a set of strains and plasmids to genetically modify C. dubliniensis and C. tropicalis, the two pathogenic species most closely related to C. albicans. C. dubliniensis is an ideal model to understand C. albicans pathogenesis since it is the closest species to C. albicans but considerably less virulent. On the other hand, C. tropicalis is ranked among the four most common causes of infections by Candida species. Given that C. dubliniensis and C. tropicalis are obligate diploids with no known conventional sexual cycle, we generated strains that are auxotrophic for at least two amino acids which allows the tandem deletion of both alleles of a gene by complementing the two auxotrophies. The strains were generated in two different genetic backgrounds for each species - one for which the genomic sequence is available and a second clinically important one. In addition, we have adapted plasmids developed to delete genes and epitope/fluorophore tag proteins in C. albicans so that they can be employed in C. tropicalis. The tools generated here allow for efficient genetic modification of C. dubliniensis and C. tropicalis, and thus facilitate the study of the molecular basis of pathogenesis in these medically relevant fungi.
RESUMEN
In Brazil, bioethanol is produced by sucrose fermentation from sugarcane by Saccharomyces cerevisiae in a fed-batch process that uses high density of yeast cells (15-25 % of wet weight/v) and high sugar concentration (18-22 % of total sugars). Several research efforts have been employed to improve the efficiency of this process through the isolation of yeasts better adapted to the Brazilian fermentation conditions. Two important wild strains named CAT-1 and PE-2 were isolated during the fermentation process and were responsible for almost 60 % of the total ethanol production in Brazil. However, in the last decade the fermentative substrate composition was much modified, since new sugar cane crops were developed, the use of molasses instead of sugar cane juice increase and with the prohibition of burning of sugarcane prior harvest. As consequence, these previously isolated strains are being replaced by new wild yeasts in most of ethanol plants. In this new scenario the isolation of novel better adapted yeasts with improved fermentative characteristics is still a big challenge. Here, we discuss the main aspects of Brazilian ethanol production and the efforts for the selection, characterization and genetic modifications of new strains with important phenotypic traits such as thermotolerance.
Asunto(s)
Biocombustibles , Etanol/metabolismo , Saccharomyces cerevisiae/crecimiento & desarrollo , Saccharomyces cerevisiae/genética , Técnicas de Cultivo Celular por Lotes , Brasil , Fermentación , Ingeniería Genética , Microbiología Industrial , Saccharomyces cerevisiae/aislamiento & purificación , Saccharomyces cerevisiae/metabolismo , Saccharum , Selección GenéticaRESUMEN
Ethylene response factor 1 (ERF1) is an essential integrator of the jasmonate and ethylene signalling pathways coordinating a large number of genes involved in plant defences. Its orthologue in Hevea brasiliensis, HbERF-IXc5, has been assumed to play a major role in laticifer metabolism and tolerance to harvesting stress for better latex production. This study sets out to establish and characterize rubber transgenic lines overexpressing HbERF-IXc5. Overexpression of HbERF-IXc5 dramatically enhanced plant growth and enabled plants to maintain some ecophysiological parameters in response to abiotic stress such as water deficit, cold and salt treatments. This study revealed that HbERF-IXc5 has rubber-specific functions compared to Arabidopsis ERF1 as transgenic plants overexpressing HbERF-IXc5 accumulated more starch and differentiated more latex cells at the histological level. The role of HbERF-IXc5 in driving the expression of some target genes involved in laticifer differentiation is discussed.
Asunto(s)
Hevea/metabolismo , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Hevea/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
The lethality of infectious diseases has decreased due to the implementation of crucial sanitary procedures such as vaccination. However, the resurgence of pathogenic diseases in different parts of the world has revealed the importance of identifying novel, rapid, and concrete solutions for control and prevention. Edible vaccines pose an interesting alternative that could overcome some of the constraints of traditional vaccines. The term "edible vaccine" refers to the use of edible parts of a plant that has been genetically modified to produce specific components of a particular pathogen to generate protection against a disease. The aim of this review is to present and critically examine "edible vaccines" as an option for global immunization against pathogenic diseases and their outbreaks and to discuss the necessary steps for their production and control and the list of plants that may already be used as edible vaccines. Additionally, this review discusses the required standards and ethical regulations as well as the advantages and disadvantages associated with this powerful biotechnology tool.
RESUMEN
Gene editing technologies are a group of recent innovations in plant breeding using molecular biology, which have in common the capability of introducing a site-directed mutation or deletion in the genome. The first cases of crops improved with these technologies are approaching the market; this has raised an international debate regarding if they should be regulated as genetically modified crops or just as another form of mutagenesis under conventional breeding. This dilemma for policymakers not only entails issues pertaining safety information and legal/regulatory definitions. It also demands borrowing tools developed in the field of social studies of science and technology, as an additional basis for sound decision making.
Asunto(s)
Productos Agrícolas/genética , Edición Génica/métodos , Genoma de Planta/genética , Regulación Gubernamental , Fitomejoramiento/métodos , Biotecnología/legislación & jurisprudencia , Biotecnología/métodos , Inocuidad de los Alimentos , Alimentos Modificados Genéticamente/normas , Edición Génica/legislación & jurisprudencia , Ingeniería Genética/legislación & jurisprudencia , Ingeniería Genética/métodos , Fitomejoramiento/legislación & jurisprudencia , Plantas Modificadas Genéticamente , Investigación , Factores SocioeconómicosRESUMEN
The association between inflammatory bowel diseases and colorectal cancer is well documented. The genetic modification of lactic acid bacteria as a tool to increase the anti-inflammatory potential of these microorganisms has also been demonstrated. Thus the aim of the present work was to evaluate the anti-cancer potential of different genetically modified lactic acid bacteria (GM-LAB) producing antioxidant enzymes (catalase or superoxide dismutase) or the anti-inflammatory cytokine IL-10 (protein or DNA delivery) using a chemical induced colon cancer murine model. Dimethilhydrazine was used to induce colorectal cancer in mice. The animals received GM-LAB producing anti-oxidant enzymes, IL-10 or a mixture of different GM-LAB. Intestinal damage, enzyme activities and cytokines were evaluated and compared to the results obtained from mice that received the wild type strains from which derived the GM-LAB. All the GM-LAB assayed showed beneficial effects against colon cancer even though they exerted different mechanisms of action. The importance to select LAB with innate beneficial properties as the progenitor strain was demonstrated with the GM-LAB producing anti-oxidant enzymes. In addition, the best effects for the mixtures GM-LAB that combine different anti-inflammatory mechanism. Results indicate that mixtures of selected LAB and GM-LAB could be used as an adjunct treatment to decrease the inflammatory harmful environment associated to colorectal cancer, especially for patients with chronic intestinal inflammation who have an increased risk to develop colorectal cancer.
Asunto(s)
Antineoplásicos/uso terapéutico , Catalasa/genética , Neoplasias Colorrectales/terapia , Enfermedades Inflamatorias del Intestino/terapia , Interleucina-10/genética , Lactococcus lactis/fisiología , Superóxido Dismutasa/genética , Animales , Antioxidantes/metabolismo , Catalasa/metabolismo , Neoplasias Colorrectales/inducido químicamente , Modelos Animales de Enfermedad , Femenino , Humanos , Inmunidad Mucosa/genética , Interleucina-10/uso terapéutico , Ácido Láctico/metabolismo , Ratones , Ratones Endogámicos BALB C , Organismos Modificados Genéticamente , Superóxido Dismutasa/metabolismoRESUMEN
The discovery of novel plant resistance (R) genes (including their homologs and analogs) opened interesting possibilities for controlling plant diseases caused by several pathogens. However, due to environmental pressure and high selection operated by pathogens, several crop plants have lost specificity, broad-spectrum or durability of resistance. On the other hand, the advances in plant genome sequencing and biotechnological approaches, combined with the increasing knowledge on Rgenes have provided new insights on their applications for plant genetic breeding, allowing the identification and implementation of novel and efficient strategies that enhance or optimize their use for efficiently controlling plant diseases. The present review focuses on main perspectives of application of R-genes and its co-players for the acquisition of resistance to pathogens in cultivated plants, with emphasis on biotechnological inferences, including transgenesis, cisgenesis, directed mutagenesis and gene editing, with examples of success and challenges to be faced.
Asunto(s)
Proteínas de Arabidopsis/inmunología , Resistencia a la Enfermedad/genética , Regulación de la Expresión Génica de las Plantas/inmunología , Enfermedades de las Plantas/inmunología , Proteínas de Plantas/inmunología , Plantas/genética , Proteínas Serina-Treonina Quinasas/inmunología , Proteínas de Arabidopsis/genética , Biotecnología/métodos , Sistemas CRISPR-Cas , Edición Génica/métodos , Interacciones Huésped-Patógeno/genética , Interacciones Huésped-Patógeno/inmunología , Mutagénesis Sitio-Dirigida , Fitomejoramiento/métodos , Enfermedades de las Plantas/genética , Inmunidad de la Planta/genética , Proteínas de Plantas/genética , Plantas/inmunología , Plantas/microbiología , Plantas/virología , Plantas Modificadas Genéticamente , Isoformas de Proteínas/genética , Isoformas de Proteínas/inmunología , Proteínas Serina-Treonina Quinasas/genética , Transducción de SeñalRESUMEN
"New Breeding Techniques" (NBTs) are a group of recent innovations in plant breeding using molecular biology tools. It is becoming evident that NBTs can introduce advantageous traits for agriculture that could be commercially available very soon However, there is still a need of clarifying its regulatory status, particularly in regards to worldwide regulations on Genetically Modified Organisms (GMOs). This article reviews the meaning of the NBTs concept, performs an overall regulatory analysis of these technologies and reports the first regulation in the world that is applied to these technologies, which was issued by the Argentine Government.
Asunto(s)
Agricultura/legislación & jurisprudencia , Cruzamiento/legislación & jurisprudencia , Ingeniería Genética/legislación & jurisprudencia , Organismos Modificados Genéticamente , Agricultura/métodos , Agricultura/tendencias , Argentina , Cruzamiento/métodos , Ingeniería Genética/métodos , Variación Genética , Regulación Gubernamental , SeguridadRESUMEN
SRY-related high-mobility-group box 9 (Sox9) gene is a cartilage-specific transcription factor that plays essential roles in chondrocyte differentiation and cartilage formation. The aim of this study was to investigate the feasibility of genetic delivery of Sox9 to enhance chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUC-MSCs). After they were isolated from human umbilical cord blood within 24 h after delivery of neonates, hUC-MSCs were untreated or transfected with a human Sox9-expressing plasmid or an empty vector. The cells were assessed for morphology and chondrogenic differentiation. The isolated cells with a fibroblast-like morphology in monolayer culture were positive for the MSC markers CD44, CD105, CD73, and CD90, but negative for the differentiation markers CD34, CD45, CD19, CD14, or major histocompatibility complex class II. Sox9 overexpression induced accumulation of sulfated proteoglycans, without altering the cellular morphology. Immunocytochemistry demonstrated that genetic delivery of Sox9 markedly enhanced the expression of aggrecan and type II collagen in hUC-MSCs compared with empty vector-transfected counterparts. Reverse transcription-polymerase chain reaction analysis further confirmed the elevation of aggrecan and type II collagen at the mRNA level in Sox9-transfected cells. Taken together, short-term Sox9 overexpression facilitates chondrogenesis of hUC-MSCs and may thus have potential implications in cartilage tissue engineering.
Asunto(s)
Humanos , Diferenciación Celular/genética , Condrogénesis/genética , Sangre Fetal/citología , Células Madre Mesenquimatosas/citología , Factor de Transcripción SOX9/genética , Agrecanos/biosíntesis , Western Blotting , Cartílago/metabolismo , Proliferación Celular/genética , Condrocitos/metabolismo , Colágeno Tipo II/biosíntesis , Citometría de Flujo , Proteínas Fluorescentes Verdes , Regulación de la Expresión Génica/fisiología , Células Endoteliales de la Vena Umbilical Humana/citología , Inmunohistoquímica , Inmunofenotipificación , Cultivo Primario de Células , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Ingeniería de Tejidos , TransfecciónRESUMEN
Considerando que las actitudes respecto a la biotecnología dependen del organismo sobre el cual se aplica y del tipo de modificación realizada se determinó la importancia del tipo de modificación genética (MG) en la compra de carne bovina en consumidores de Temuco (Región de La Araucanía, Chile) en diferentes segmentos de mercado, mediante una encuesta a 400 personas. Utilizando análisis de conjunto se determinó que el tipo de MG fue más importante que el precio en la compra (66,2 y 33,8%; respectivamente). Mediante análisis de conglomerados jerárquicos se distinguieron dos segmentos principales con diferencias significativas en la importancia de estos atributos. El segmento mayoritario (63,8%) dio alta importancia al tipo de MG, el segundo segmento (28,2%) atribuyó mayor importancia al precio, pero los consumidores de ambos segmentos prefirieron carne de un animal sin MG o de un animal con MG para disminuir el contenido de colesterol y rechazaron la carne de un animal alimentado con granos MG o de animales con MG para reducir el costo de alimentación. Los segmentos presentaron diferencias según el tamaño del grupo familiar, frecuencia de consumo de carne bovina y estilo de vida. Es posible concluir que la ausencia de MG en carne bovina es una condición deseable para los consumidores de Temuco, sin embargo, la totalidad de la muestra encuestada se muestra positiva hacia la carne de un animal MG que presente beneficios para la salud y el 8% acepta carne de un animal alimentado con granos MG.
Considering that attitudes to biotechnology depend on the organism to which it is applied and the type of modification made, a survey was carried out on 400 persons to determine the importance of the type of genetic modification (GM) in the purchase of beef by consumers in Temuco (La Araucanía Region, Chile) in different market segments. Using conjoint analysis it was determined that the type of GM was more important than the price in the purchase decision (66.2 and 33.8%, respectively). Two principal segments were identified by analysis of hierarchical conglomerates, with significant differences in the importance of these attributes. The majority segment (63.8%) attached great importance to the type of GM, the second segment (28.2%) attributed more importance to the price, however consumers of both segments preferred meat from an animal without GM or one with GM to reduce the cholesterol content, and rejected meat from an animal fed with GM grain or subjected to GM to reduce feeding costs. The segments presented differences related to the size of the family group, frequency of beef consumption and life-style. It may be concluded that the absence of GM in beef is a desirable condition for the consumers of Temuco, however the whole sample responded positively to meat from a GM animal which presents health benefits and 8% accept meat from an animal which has been fed with GM grain.