Pre-Harvest Factors Optimization Using Genetic Algorithm for Lettuce

The agricultural sector is facing problems on crop development due to climate change and global warming. Crops such as rice, tomato, corn, lettuce, potato, wheat, soybeans and others are affected. Through analyzing the graphical representation of data, no optimum values are observed. In this study, the suitability of the genetic algorithm in finding the best condition for producing high quality lettuce crop was determined. The parameters that were optimized are the light intensity, temperature and CO2. These parameters were essential preharvest factors for lettuce. The system selected the 50 fittest individuals based on the fitness score and then proceeds to the recombination process. A mutation has been applied to test if the solution is the global one. When the iterations had reached the required number of generation, the system stopped and gave the best condition for lettuce. Critical design on GA was done and the best fitness plot was obtained. The GA results showed that the optimum conditions for a highquality lettuce crop needs a light intensity of 175.22296 μmol/m2/s, a temperature of 19.36228 ºC and a CO2 level of 803.01855 ppm

Precision Agriculture Technology for Crop Farming

This book provides a review of precision agriculture technology development, followed by a presentation of the state-of-the-art and future requirements of precision agriculture technology. It presents different styles of precision agriculture technologies suitable for large scale mechanized farming; highly automated community-based mechanized production; and fully mechanized farming practices commonly seen in emerging economic regions. The book emphasizes the introduction of core technical features of sensing, data processing and interpretation technologies, crop modeling and production control theory, intelligent machinery and field robots for precision agriculture production

Improvements to the maize (Zea mays L.) in vivo maternal doubled haploid system

The maize (Zea mays L.) in vivo maternal doubled haploid system has been widely applied to maize breeding and genetics in recent decades and is an important part of the majority of public and private maize breeding programs today. The principal advantage of the doubled haploid system is the ability to generate completely homozygous inbred lines in as little as two seasons. Other advantages to this system include more rapid integration of loci of interest and increased usefulness over traditional lines developed through self-pollination. In this dissertation, some of the major problems in the maternal doubled haploid system are addressed. Namely, improvement of maternal inducers, improved understanding of the genetics controlling inducibility, development of an automated system to sort haploid kernels, investigation and application of spontaneous chromosome doubling, and a proposal for the acceleration of the breeding cycle beyond doubled haploids through the in vitro nursery. This dissertation provides some new insight into these problems, as follows. The development and release of a new improved maternal haploid inducer for use in doubled haploid programs. Improved understanding of the quantitative nature of inducibility and the effects of misclassification are discussed. Successful automated discrimination of haploid and diploid kernels using optical and fluorescence methods is described. In an effort to make the doubled haploid system more efficient and safe, a bypass of the colchicine doubling step is proposed through the application and investigation of spontaneous chromosome doubling in haploid plants. Finally, as a proposal for what could be the next step in accelerating the breeding cycle, the in vitro nursery and its applications is discussed

Precision Agriculture Technology for Crop Farming

This book provides a review of precision agriculture technology development, followed by a presentation of the state-of-the-art and future requirements of precision agriculture technology. It presents different styles of precision agriculture technologies suitable for large scale mechanized farming; highly automated community-based mechanized production; and fully mechanized farming practices commonly seen in emerging economic regions. The book emphasizes the introduction of core technical features of sensing, data processing and interpretation technologies, crop modeling and production control theory, intelligent machinery and field robots for precision agriculture production

Two Zona Pellucida vaccines expressed in different plant expression systems for spaying mammals

This is a concept study of designing two contraceptive vaccines, expressing them in different plant expression systems und testing their efficacy. As antigens fertilization-relevant Zona Pellucida proteins were used. Stable transformed pea seeds, tobacco seeds, tobacco leaves and carrot cells show vaccine accumulation. A transient expression system was used for producing and characterizing the vaccines. The vaccines elicited a strong immune response, which reduced offspring of BALB/c mice up to 52 %.Diese Arbeit ist eine Konzeptstudie über das Design von zwei kontrazeptiven Impfstoffen, der Expression in verschiedenen pflanzlichen Systemen und dem Wirkungstest. Als Antigene dienen befruchtungsrelevante Zona Pellucida Proteine. In stabil transformierten Erbsen-, Tabaksamen, -blättern und Karottenzellen konnte eine Akkumulation nachgewiesen werden. Ein transientes Expressionssystem wurde zur Produktion und Charakterisierung der Impfstoffe verwendet. Die Impfstoffe lösten eine starke Immunantwort aus, welche die Nachkommenschaft bei BALB/c Mäusen um bis zu 52 % reduziert

Proceedings of the European Conference on Agricultural Engineering AgEng2021

This proceedings book results from the AgEng2021 Agricultural Engineering Conference under auspices of the European Society of Agricultural Engineers, held in an online format based on the University of Évora, Portugal, from 4 to 8 July 2021. This book contains the full papers of a selection of abstracts that were the base for the oral presentations and posters presented at the conference. Presentations were distributed in eleven thematic areas: Artificial Intelligence, data processing and management; Automation, robotics and sensor technology; Circular Economy; Education and Rural development; Energy and bioenergy; Integrated and sustainable Farming systems; New application technologies and mechanisation; Post-harvest technologies; Smart farming / Precision agriculture; Soil, land and water engineering; Sustainable production in Farm buildings

Development and characterization of two new tools for plant genetic engineering: A CRISPR/Cas12a-based mutagenesis system and a PhiC31-based gene switch

Tesis por compendio[ES] La mejora genética vegetal tiene como objetivo la obtención de plantas con rasgos mejorados o características novedosas que podrían ayudar a superar los objetivos de sostenibilidad. Para este fin, la biotecnología vegetal necesita incorporar nuevas herramientas de ingeniería genética que combinen una mayor precisión con una mayor capacidad de mejora. Las herramientas de edición genética recientemente descubiertas basadas en la tecnología CRISPR/Cas9 han abierto el camino para modificar los genomas de las plantas con una precisión sin precedentes. Por otro lado, los nuevos enfoques de biología sintética basados en la modularidad y la estandarización de los elementos genéticos han permitido la construcción de dispositivos genéticos cada vez más complejos y refinados aplicados a la mejora genética vegetal. Con el objetivo final de expandir la caja de herramientas biotecnológicas para la mejora vegetal, esta tesis describe el desarrollo y la adaptación de dos nuevas herramientas: una nueva endonucleasa específica de sitio (SSN) y un interruptor genético modular para la regulación de la expresión transgénica. En una primera parte, esta tesis describe la adaptación de CRISPR/Cas12a para la expresión en plantas y compara la eficiencia de las variantes de Acidaminococcus (As) y Lachnospiraceae (Lb) Cas12a con Streptococcus pyogens Cas9 (SpCas9) descritos anteriormente en ocho loci de Nicotiana benthamiana usando expresión transitoria. LbCas12a mostró la actividad de mutagénesis promedio más alta en los loci analizados. Esta actividad también se confirmó en experimentos de transformación estable realizados en tres plantas modelo diferentes, a saber, N. benthamiana, Solanum lycopersicum y Arabidopsis thaliana. Para este último, los efectos mutagénicos colaterales fueron analizados en líneas segregantes sin la endonucleasa Cas12a, mediante secuenciación del genoma descartándose efectos indiscriminados. En conjunto, los resultados muestran que LbCas12a es una alternativa viable a SpCas9 para la edición genética en plantas. En una segunda parte, este trabajo describe un interruptor genético reversible destinado a controlar la expresión génica en plantas con mayor precisión que los sistemas inducibles tradicionales. Este interruptor, basado en el sistema de recombinación del fago PhiC31, fue construido como un dispositivo modular hecho de partes de ADN estándar y diseñado para controlar el estado transcripcional (encendido o apagado) de dos genes de interés mediante la inversión alternativa de un elemento regulador central de ADN. El estado del interruptor puede ser operado externa y reversiblemente por la acción de los actuadores de recombinación y su cinética, memoria y reversibilidad fueron ampliamente caracterizados en experimentos de transformación transitoria y estable en N. benthamiana. En conjunto, esta tesis muestra el diseño y la caracterización funcional de herramientas para la ingeniería del genómica y biología sintética de plantas que ahora ha sido completada con el sistema de edición genética CRISPR/Cas12a y un interruptor genético reversible y biestable basado en el sistema de recombinación del fago PhiC31.[CA] La millora genètica vegetal té com a objectiu l'obtenció de plantes amb trets millorats o característiques noves que podrien ajudar a superar els objectius de sostenibilitat. Amb aquesta finalitat, la biotecnologia vegetal necessita incorporar noves eines d'enginyeria genètica que combinen una major precisió amb una major capacitat de millora. Les eines d'edició genètica recentment descobertes basades en la tecnologia CRISPR/Cas9 han obert el camí per modificar els genomes de les plantes amb una precisió sense precedents. D'altra banda, els nous enfocaments de biologia sintètica basats en la modularitat i l'estandardització dels elements genètics han permès la construcció de dispositius genètics cada vegada més complexos i sofisticats aplicats a la millora genètica vegetal. Amb l'objectiu final d'expandir la caixa d'eines biotecnològiques per a la millora vegetal, aquesta tesi descriu el desenvolupament i l'adaptació de dues noves eines: una nova endonucleasa específica de lloc (SSN) i un interruptor genètic modular per a la regulació de l'expressió transgènica . En una primera part, aquesta tesi descriu l'adaptació de CRISPR/Cas12a per a l'expressió en plantes i compara l'eficiència de les variants de Acidaminococcus (As) i Lachnospiraceae (Lb) Cas12a amb la ben establida Streptococcus pyogens Cas9 (SpCas9), en vuit loci de Nicotiana benthamiana usant expressió transitòria. LbCas12a va mostrar l'activitat de mutagènesi mitjana més alta en els loci analitzats. Aquesta activitat també es va confirmar en experiments de transformació estable realitzats en tres plantes model diferents, a saber, N. benthamiana, Solanum lycopersicum i Arabidopsis thaliana. Per a aquest últim, els efectes mutagènics col·laterals van ser analitzats en línies segregants sense l'endonucleasa Cas12a, mitjançant seqüenciació completa del genoma i descartant efectes indiscriminats. En conjunt, els resultats mostren que LbCas12a és una alternativa viable a SpCas9 per a l'edició genètica en plantes. En una segona part, aquest treball descriu un interruptor genètic reversible destinat a controlar l'expressió gènica en plantes amb major precisió que els sistemes induïbles tradicionals. Aquest interruptor, basat en el sistema de recombinació del bacteriòfag PhiC31, va ser construït com un dispositiu modular fet de parts d'ADN estàndard i dissenyat per controlar l'estat transcripcional (encès o apagat) de dos gens d'interès mitjançant la inversió alternativa d'un element regulador central d'ADN. L'estat de l'interruptor pot ser operat externa i reversiblement per acció dels actuadors de recombinació i la seva cinètica, memòria i reversibilitat van ser àmpliament caracteritzats en experiments de transformació transitòria i estable en N. benthamiana. En conjunt, aquesta tesi mostra el disseny i la caracterització funcional d'eines per a l'enginyeria del genòmica i biologia sintètica de plantes que ara ha sigut completat amb el sistema d'edició genètica CRISPR/Cas12a i un interruptor genètic biestable i reversible basat en el sistema de recombinació del bacteriòfag PhiC31.[EN] Plant breeding aims to provide plants with improved traits or novel features that could help to overcome sustainability goals. To this end, plant biotechnology needs to incorporate new genetic engineering tools that combine increased precision with higher breeding power. The recently discovered genome editing tools based on CRISPR/Cas9 technology have opened the way to modify plant¿s genomes with unprecedented precision. On the other hand, new synthetic biology approaches based on modularity and standardization of genetic elements have enabled the construction of increasingly complex and refined genetic devices applied to plant breeding. With the ultimate goal of expanding the toolbox of plant breeding techniques, this thesis describes the development and adaptation to plant systems of two new breeding tools: a site-specific nuclease (SSNs), and a modular gene switch for the regulation of transgene expression. In a first part, this thesis describes the adoption of the SSN CRISPR/Cas12a for plant expression and compares the efficiency of Acidaminococcus (As) and Lachnospiraceae (Lb) Cas12a variants with the previously described Streptococcus pyogens Cas9 (SpCas9) in eight Nicotiana benthamiana loci using transient expression experiments. LbCas12a showed highest average mutagenesis activity in the loci assayed. This activity was also confirmed in stable genome editing experiments performed in three different model plants, namely N. benthamiana, Solanum lycopersicum and Arabidopsis thaliana. For the latter, off-target effects in Cas12a-free segregating lines were discarded at genomic level by deep sequencing. Collectively, the results show that LbCas12a is a viable alternative to SpCas9 for plant genome engineering. In a second part, this work describes the engineering of a new reversible genetic switch aimed at controlling gene expression in plants with higher precision than traditional inducible systems. This switch, based on the bacteriophage PhiC31 recombination system, was built as a modular device made of standard DNA parts and designed to control the transcriptional state (on or off) of two genes of interest by alternative inversion of a central DNA regulatory element. The state of the switch can be externally and reversibly operated by the action of the recombination actuators and its kinetics, memory, and reversibility were extensively characterized in N. benthamiana using both transient expression and stable transgenics. Altogether, this thesis shows the design and functional characterization of refined tools for genome engineering and synthetic biology in plants that now has been expanded with the CRISPR/Cas12a gene editing system and the phage PhiC31-based toggle switch.Bernabé Orts, JM. (2019). Development and characterization of two new tools for plant genetic engineering: A CRISPR/Cas12a-based mutagenesis system and a PhiC31-based gene switch [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/133055TESISCompendi