HKT2;2/1, a K+-permeable transporter identified in a salt tolerant rice cultivar through surveys of natural genetic polymorphism

We have investigated OsHKT2;1 natural variation in a collection of 49 cultivars with different levels of salt tolerance and geographical origins. The effect of identified polymorphism on OsHKT2;1 activity was analysed through heterologous expression of variants in Xenopus oocytes. OsHKT2;1 appeared to be a highly conserved protein with only five possible amino acid substitutions that have no substantial effect on functional properties. Our study, however, also identified a new HKT isoform, No-OsHKT2;2/1 in Nona Bokra, a highly salt-tolerant cultivar. No-OsHKT2;2/1 probably originated from a deletion in chromosome 6, producing a chimeric gene. Its 5¢ region corresponds to that of OsHKT2;2, whose full-length sequence is not present in Nipponbare but has been identified in Pokkali, a salt-tolerant rice cultivar. Its 3¢ region corresponds to that of OsHKT2;1. No-OsHKT2;2/1 is essentially expressed in roots and displays a significant level of expression at high Na+ concentrations, in contrast to OsHKT2;1. Expressed in Xenopus oocytes or in Saccharomyces cerevisiae, No-OsHKT2;2/1 exhibited a strong permeability to Na+ and K+, even at high external Na+ concentrations, like OsHKT2;2, and in contrast to OsHKT2;1. Our results suggest that No-OsHKT2;2/1 can contribute to Nona Bokra salt tolerance by enabling root K+ uptake under saline conditions.

Contribution to the improvement of communication services in nanosatellite constellations using delay and disruption tolerant networking based architectures

Esta tesis se desarrolla dentro del marco de las comunicaciones satelitales en el innovador campo de los pequeños satélites también llamados nanosatélites o cubesats, llamados así por su forma cubica. Estos nanosatélites se caracterizan por su bajo costo debido a que usan componentes comerciales llamados COTS (commercial off-the-shelf) y su pequeño tamaño como los Cubesats 1U (10cm*10 cm*10 cm) con masa aproximada a 1 kg. Este trabajo de tesis tiene como base una iniciativa propuesta por el autor de la tesis para poner en órbita el primer satélite peruano en mi país llamado chasqui I, actualmente puesto en órbita desde la Estación Espacial Internacional. La experiencia de este trabajo de investigación me llevo a proponer una constelación de pequeños satélites llamada Waposat para dar servicio de monitoreo de sensores de calidad de agua a nivel global, escenario que es usado en esta tesis. Es ente entorno y dadas las características limitadas de los pequeños satélites, tanto en potencia como en velocidad de datos, es que propongo investigar una nueva arquitectura de comunicaciones que permita resolver en forma óptima la problemática planteada por los nanosatélites en órbita LEO debido a su carácter disruptivo en sus comunicaciones poniendo énfasis en las capas de enlace y aplicación. Esta tesis presenta y evalúa una nueva arquitectura de comunicaciones para proveer servicio a una red de sensores terrestres usando una solución basada en DTN (Delay/Disruption Tolerant Networking) para comunicaciones espaciales. Adicionalmente, propongo un nuevo protocolo de acceso múltiple que usa una extensión del protocolo ALOHA no ranurado, el cual toma en cuenta la prioridad del trafico del Gateway (ALOHAGP) con un mecanismo de contienda adaptativo. Utiliza la realimentación del satélite para implementar el control de la congestión y adapta dinámicamente el rendimiento efectivo del canal de una manera óptima. Asumimos un modelo de población de sensores finito y una condición de tráfico saturado en el que cada sensor tiene siempre tramas que transmitir. El desempeño de la red se evaluó en términos de rendimiento efectivo, retardo y la equidad del sistema. Además, se ha definido una capa de convergencia DTN (ALOHAGP-CL) como un subconjunto del estándar TCP-CL (Transmission Control Protocol-Convergency Layer). Esta tesis muestra que ALOHAGP/CL soporta adecuadamente el escenario DTN propuesto, sobre todo cuando se utiliza la fragmentación reactiva. Finalmente, esta tesis investiga una transferencia óptima de mensajes DTN (Bundles) utilizando estrategias de fragmentación proactivas para dar servicio a una red de sensores terrestres utilizando un enlace de comunicaciones satelitales que utiliza el mecanismo de acceso múltiple con prioridad en el tráfico de enlace descendente (ALOHAGP). El rendimiento efectivo ha sido optimizado mediante la adaptación de los parámetros del protocolo como una función del número actual de los sensores activos recibidos desde el satélite. También, actualmente no existe un método para advertir o negociar el tamaño máximo de un “bundle” que puede ser aceptado por un agente DTN “bundle” en las comunicaciones por satélite tanto para el almacenamiento y la entrega, por lo que los “bundles” que son demasiado grandes son eliminados o demasiado pequeños son ineficientes. He caracterizado este tipo de escenario obteniendo una distribución de probabilidad de la llegada de tramas al nanosatélite así como una distribución de probabilidad del tiempo de visibilidad del nanosatélite, los cuales proveen una fragmentación proactiva óptima de los DTN “bundles”. He encontrado que el rendimiento efectivo (goodput) de la fragmentación proactiva alcanza un valor ligeramente inferior al de la fragmentación reactiva. Esta contribución permite utilizar la fragmentación activa de forma óptima con todas sus ventajas tales como permitir implantar el modelo de seguridad de DTN y la simplicidad al implementarlo en equipos con muchas limitaciones de CPU y memoria. La implementación de estas contribuciones se han contemplado inicialmente como parte de la carga útil del nanosatélite QBito, que forma parte de la constelación de 50 nanosatélites que se está llevando a cabo dentro del proyecto QB50. ABSTRACT This thesis is developed within the framework of satellite communications in the innovative field of small satellites also known as nanosatellites (<10 kg) or CubeSats, so called from their cubic form. These nanosatellites are characterized by their low cost because they use commercial components called COTS (commercial off-the-shelf), and their small size and mass, such as 1U Cubesats (10cm * 10cm * 10cm) with approximately 1 kg mass. This thesis is based on a proposal made by the author of the thesis to put into orbit the first Peruvian satellite in his country called Chasqui I, which was successfully launched into orbit from the International Space Station in 2014. The experience of this research work led me to propose a constellation of small satellites named Waposat to provide water quality monitoring sensors worldwide, scenario that is used in this thesis. In this scenario and given the limited features of nanosatellites, both power and data rate, I propose to investigate a new communications architecture that allows solving in an optimal manner the problems of nanosatellites in orbit LEO due to the disruptive nature of their communications by putting emphasis on the link and application layers. This thesis presents and evaluates a new communications architecture to provide services to terrestrial sensor networks using a space Delay/Disruption Tolerant Networking (DTN) based solution. In addition, I propose a new multiple access mechanism protocol based on extended unslotted ALOHA that takes into account the priority of gateway traffic, which we call ALOHA multiple access with gateway priority (ALOHAGP) with an adaptive contention mechanism. It uses satellite feedback to implement the congestion control, and to dynamically adapt the channel effective throughput in an optimal way. We assume a finite sensor population model and a saturated traffic condition where every sensor always has frames to transmit. The performance was evaluated in terms of effective throughput, delay and system fairness. In addition, a DTN convergence layer (ALOHAGP-CL) has been defined as a subset of the standard TCP-CL (Transmission Control Protocol-Convergence Layer). This thesis reveals that ALOHAGP/CL adequately supports the proposed DTN scenario, mainly when reactive fragmentation is used. Finally, this thesis investigates an optimal DTN message (bundles) transfer using proactive fragmentation strategies to give service to a ground sensor network using a nanosatellite communications link which uses a multi-access mechanism with priority in downlink traffic (ALOHAGP). The effective throughput has been optimized by adapting the protocol parameters as a function of the current number of active sensors received from satellite. Also, there is currently no method for advertising or negotiating the maximum size of a bundle which can be accepted by a bundle agent in satellite communications for storage and delivery, so that bundles which are too large can be dropped or which are too small are inefficient. We have characterized this kind of scenario obtaining a probability distribution for frame arrivals to nanosatellite and visibility time distribution that provide an optimal proactive fragmentation of DTN bundles. We have found that the proactive effective throughput (goodput) reaches a value slightly lower than reactive fragmentation approach. This contribution allows to use the proactive fragmentation optimally with all its advantages such as the incorporation of the security model of DTN and simplicity in protocol implementation for computers with many CPU and memory limitations. The implementation of these contributions was initially contemplated as part of the payload of the nanosatellite QBito, which is part of the constellation of 50 nanosatellites envisaged under the QB50 project.

Use of thermographic imaging to screen for drought-tolerant genotypes in Brachypodium distachyon

Thermal imaging has been used to evaluate the response to drought and warm temperatures in a collection of Brachypodium distachyon lines adapted to varied environmental conditions. Thermographic records were able to separate lines from contrasting rainfall regimes. Genotypes from dryer environments showed warmer leaves under water deficit, which suggested that decreased evapotranspiration was related to a more intense stomatal closure. When irrigated and under high temperature conditions, drought-adapted lines showed cooler leaves than lines from wetter zones. The consistent, inverse thermographic response of lines to water stress and heat validates the reliability of this method to assess drought tolerance in this model cereal. It additionally supports the hypothesis that stomatal-based mechanisms are involved in natural variation for drought tolerance in Brachypodium. The study further suggests that these mechanisms are not constitutive but likely related to a more efficient closing response to avoid dehydration in adapted genotypes. Higher leaf temperature under water deficit seems a dependable criterion of drought tolerance, not only in B. distachyon but also in the main cereal crops and related grasses where thermography can facilitate high-throughput preliminary screening of tolerant materials.

Seven Ulmus minor clones tolerant to Ophiostoma novo-ulmi registered as forest reproductive material in Spain

The Spanish elm programme began in 1986 in response to the devastating impact of Dutch elm disease on natural elm stands and urban trees. Its main objectives were to conserve remaining genetic resources and select and breed tolerant native elm genotypes. After 27 years of work conducting susceptibility trials on thousands of elm genotypes, the first seven tolerant Ulmus minor trees are now being registered by the Spanish Environmental Administration. This paper presents the results of the susceptibility tests on these clones and their distinctive genetic, morphological and phenological features. In all susceptibility trials the commercial ?Sapporo Autumn Gold? clone, which is highly tolerant to O. novo-ulmi, was used as a control. The registered clones were named ?Ademuz?, ?Dehesa de la Villa?, ?Majadahonda?, ?Toledo?, ?Dehesa de Amaniel?, ?Retiro? and ?Fuente Umbría?. The most tolerant clone was ?Dehesa de Amaniel?, as its wilting values were below 5% during the two consecutive inoculation trials performed in Madrid. ?Fuente Umbría?, tested over four consecutive years in Guadalajara and Palencia, was the Spanish clone with the most reliable tolerance level to O. novo-ulmi. The ?Ademuz? and ?Majadahonda? clones had the highest ornamental scores and are promising trees for use in urban environments and tree breeding for ornamental quality. These two genotypes showed a later bud burst phenology than the other U. minor clones, demonstrating suitability to areas with late frost events. The Spanish programme aims to substantially increase the range of tolerant native elms through new selections and crossings to gain a better understanding of the genetic basis of resistance.

Identification of novel drought-tolerant-associated SNPs in common bean (Phaseolus vulgaris)

Common bean (Phaseolus vulgaris L.) is a leguminous in high demand for human nutrition and a very important agricultural product. Production of common bean is constrained by environmental stresses such as drought. Although conventional plant selection has been used to increase production yield and stress tolerance, drought tolerance selection based on phenotype is complicated by associated physiological, anatomical, cellular, biochemical, and molecular changes. These changes are modulated by differential gene expression. A common method to identify genes associated with phenotypes of interest is the characterization of Single Nucleotide Polymorphims (SNPs) to link them to specific functions. In this work, we selected two drought-tolerant parental lines from Mesoamerica, Pinto Villa, and Pinto Saltillo. The parental lines were used to generate a population of 282 families (F3:5) and characterized by 169 SNPs. We associated the segregation of the molecular markers in our population with phenotypes including flowering time, physiological maturity, reproductive period, plant, seed and total biomass, reuse index, seed yield, weight of 100 seeds, and harvest index in three cultivation cycles. We observed 83 SNPs with significant association (p < 0.0003 after Bonferroni correction) with our quantified phenotypes. Phenotypes most associated were days to flowering and seed biomass with 58 and 44 associated SNPs, respectively. Thirty-seven out of the 83 SNPs were annotated to a gene with a potential function related to drought tolerance or relevant molecular/biochemical functions. Some SNPs such as SNP28 and SNP128 are related to starch biosynthesis, a common osmotic protector; and SNP18 is related to proline biosynthesis, another well-known osmotic protector.