Memoria para optar al título profesional de Ingeniero Agrónomo Mención Fruticultura / El almendro es una especie predominantemente autoincompatible entre algunas de sus variedades, lo que obliga a los productores a establecer huertos con al menos dos variedades intercompatibles. Esta incompatibilidad actúa para prevenir la autofecundación, la que es del tipo gametofítica, por la expresión de proteínas específicas dentro de los estilos. Es por esto que es de suma importancia conocer la compatibilidad entre las variedades, existiendo diversos métodos para determinarla. En los últimos años, se ha establecido la compatibilidad a través de evaluaciones moleculares, que utilizan técnicas específicas para determinar el genotipo S de cada variedad, y han existido avances importantes en el desarrollo de técnicas biológicas para la determinación de la compatibilidad. Durante la floración del año 2011, con las variedades de almendro Desmayo Largueta, Desmayo Rojo, Nonpareil, Fritz, Wood Colony, Carmel, Thompsom, Sonora, Solano, Masbovera, Francolí, Price, Ne plus Ultra, Glorieta y Marcona, se utilizó la técnica propuesta por Mori et al. (2006), basada en el protocolo del laboratorio Daphne Preuss, para determinar la compatibilidad entre ‘Marcona’ y 14 variedades de almendro. Esta técnica está basada en la tinción de la callosa de los tubos polínicos con anilina azul, permitiendo la observación del crecimiento, utilizando microscopio de fluorescencia. Además, se realizó un seguimiento de los estados fenológicos de las variedades analizadas, en campo, determinando que Desmayo Rojo, Thompson, Glorieta, Masbovera y Francolí coincidieron en menos de 5 días con la floración de ‘Marcona’. Se determinó la compatibilidad de todas las variedades con Marcona, resultando compatible con las 14 variedades ensayadas. Los resultados concuerdan con los análisis genéticos moleculares encontrados en diversos estudios, validando a este método como una herramienta para determinar compatibilidad, sobre todo en variedades donde aún los alelos S no han sido identificados. / Almond cultivars are predominantly self-incompatible which requires to plant at least two inter-compatible varieties. This incompatibility acts to prevent self-fertilization, which is the gametophytic type given by specific proteins produced in the styles. Because of this, knowing the compatibility between cultivars is required and different methods to determine it have been established in recent years through molecular or biological techniques. During the 2011 flowering, the compatibility between Marcona and the cultivars Desmayo Largueta, Desmayo Rojo, Nonpareil, Fritz, Wood Colony, Carmel, Thompsom, Sonora, Solano, Masbovera, Francolí, Price, Ne plus Ultra and Glorieta, were analyzed using the technique developed by Mori et al. (2006), based on Daphne Preuss laboratory protocol. This technique stains callose using aniline blue, allowing pollen tube growth to be observed with flourescence microscopy. Furthermore, flowering of analized cultivars was monitored in the field, showing that Desmayo Rojo, Thompson, Glorieta, Masbovera and Francolí cultivars coincided in less than five days with Marcona cultivar. Marcona was compatible with all studied cultivars, in accordance with genetic molecular analysis found in different studies. Also, these results validated this method as useful to determine genetic compatibility, especially for cultivars where the S alleles have not been identified yet.

Almond (Prunus dulcis) is the most important nut crop in terms of world production. Due to its health benefit and high nutritional value the consumption and world supply of almond is increasing. To remain competitive in the world market, the Australian almond breeding program was established to produce cultivars with better adaptation to Australian conditions. As part of this program an almond mapping population consisting of 93 F₁ progeny derived from a cross between the American cultivar ‘Nonpareil’ (NP) and the European self-compatible cultivar ‘Lauranne’ (LA) was produced to construct the genetic linkage maps. The first almond linkage map developed prior to the commencement of this project failed to produce the eight linkage groups similar to the basic chromosome number of almond (x = 8) and many large gaps were also observed on the linkage groups. Therefore, more markers were needed to saturate the maps. Microsatellite markers are considered one of the best choices for mapping studies. 195 microsatellite markers isolated from Prunus species were obtained from published papers or by personal communication. Polymorphism was revealed by three different methods, and in general, polyacrylamide gel electrophoresis (PAGE) compared to the fluorescent labelled marker detection using an automated DNA sequencer or agarose gel electrophoresis, showed the most efficient and cost effective method of genotyping. A subset of 54 markers which produced reliable and easily interpretable polymorphic bands was selected to screen the whole mapping population. Microsatellites originally isolated from almond species showed the highest rate of amplification and polymorphism followed by peach microsatellites and the least informative markers were isolated from cherry. It seems that the level of transportability and usefulness of microsatellite markers is related to the genetic distance of the closely related species. Almond and peach belong to the same subgenus (Amygdalus) and other Prunus species are classified in Prunophora subgenus. The nut, or kernel, is the commercial part of the almond tree, thus to improve the quality of fruit an understanding of environmental influence, heritability and correlation of traits is required. Pomological and quality characters such as: shell hardness, kernel size, shape, taste, pubescence, colour, and percentage of doubles were measured during three consecutive years (2005-2007) on the total mapping population, but data analysis (ANOVA) was performed only on trees that survived for all three years. Most of the traits showed high broad-sense heritability and kernel shape showed the highest heritability of H² = 0.92 suggesting high genetic control of this trait. Occasionally larger kernels than either parent were found in the progeny indicating potential for improvement of this trait even with smaller kernel size parent that encompass many desirable characters. High correlation was also found between the in-shell and kernel weight (r = 0.74), kernel length / kernel width (r = 0.67), kernel weight to kernel length (r = 0.78) and kernel width (r = 0.80). This correlation estimation pointed out in this study indicates that the improvement of one character may result the progress in another trait. Neither of the parents in the mapping population had bitter or obvious slightly bitter taste but slightly bitter kernels were observed among the progeny. Amygdalin was assumed to be responsible for bitter taste in almond; therefore we measured the amount of amygdalin in sweet and slightly bitter kernel progeny by HPLC. However, the results showed that amygdalin exists in sweet kernels as well. Although the average amount of amygdalin in slightly bitter kernels (20.34 mg kg⁻¹ FW) was higher than sweet kernels (3.67 mg kg⁻¹ FW), some sweet kernels had higher amounts of amygdalin suggesting the impact of other components on slightly bitter kernel. The highest variability within the traits was observed in the percentage of double kernel, which showed the highest standard error. Strong environmental effects, particularly low temperature at pre-blossom time is speculated to produce much higher double kernels. Three genetic linkage maps, one for each parent and an integrated map were constructed by the addition of 54 new microsatellite markers to the previous dataset. All the data was scored and coded according to the coding system necessary by JoinMap3 which was used for map construction. 131 markers including microsatellite, ISSR, RAPD, SCAR and S-allele markers were placed on the integrated map covering 590.7 cM with the average density of 4.5 cM/marker. The minimum number of six microsatellite markers was placed on linkage group 8 and the linkage group 1 which is the longest linkage group has 14 microsatellite markers. Comparative mapping study with other Prunus maps, especially with the highly saturated reference map showed complete synteny and minor changes in the order of four markers on linkage groups compared with Prunus reference map. The conservation of molecular marker order observed in this study supports the idea of looking at Prunus genome as a single genetic system and practical application of this similarity would be in cross-transportability of microsatellite markers from well developed linkage maps to the less studied species in Prunus. Ten microsatellite loci placed on our map have not been reported before and could be used to improve the density of other Prunus maps, especially the reference map. This study contributed to the better understanding of the mode of inheritance and environmental effect on morphological traits and the effect of amygdalin on kernel taste. The most saturated microsatellite based almond linkage map developed in this study can serve as a framework for future almond breeding program in Australia and benefit Prunus improvement programs internationally. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1348850 / Thesis (Ph.D.) - University of Adelaide, School of Agriculture, Food and Wine, 2008

Objetivo: Determinar el efecto neuroprotector de la administración de la semilla de Prunus dulcis “almendra” sobre el tejido nervioso en ratones inducidos a estrés por desorientación motora. Diseño: Estudio analítico, transversal, experimental y prospectivo. Lugar: Laboratorios del Centro de Investigación de Bioquímica y Nutrición Alberto Guzmán Barrón, Facultad de Medicina, UNMSM, Lima, Perú. Materiales: Ratones albinos BALB/c (Mus musculus) machos y Prunus dulcis “almendra”. Métodos: Se utilizó 42 ratones, según expertos, de 3 meses de edad y 31 ± 4,4 g de peso, distribuidos aleatoriamente en seis grupos (n=7). Todos los grupos recibieron la misma dieta balanceada y agua ad libitum durante 5 días. Recibieron los siguientes tratamientos, por cinco días, vía peroral: grupo I y II: suero fisiológico (NaCl 0,9g% 10mL/kg), grupo III: vitamina E 400mg/kg, grupo IV: almendra 100 mg/kg, grupo V: almendra 500 mg/kg y grupo VI: almendra 1000 mg/kg; 12 horas antes de finalizar el Tto. se cortaron los bigotes de los ratones, excepto al grupo I; y luego de 12 horas se realizó el sacrificio. Principales medidas de los resultados: Nivel de lipoperoxidación expresado en sustancias reactivas al ácido tiobarbitúrico (TBARs) y nivel de Grupos sulfhídrilos no proteicos (GS-NP), además de cambios histopatológicos de tejido de cerebro y cerebelo. Resultados: La administración de Prunus dulcis “almendra” aumenta significativamente (p<0.05) los niveles de GS-NP en todos los grupos (excepto G VI) en comparación con el G II en cerebro; los niveles de TBARs disminuyen significativamente (p<0.05) en el grupo V y VI comparado con el grupo II, y en relación a los cambios histológicos se observa una mejora leve en el G V en comparación con el GII. Conclusiones: La administración de la suspensión de la semilla del Prunus dulcis “almendra” expreso un efecto neuroprotector en los indicadores bioquímicos (TBARs y GS-NP), sobre el tejido nervioso en ratones inducidos a estrés por desorientación motora. / Objetive: Determine the neuroprotective effect of administration of Prunus dulcis seed "almond" on the nervous tissue in motor stress induced disorientation mice. Design: Analytical, transverse, experimental and prospective study. Location: Laboratories of the Research Center of Biochemistry and Nutrition Alberto Guzman Barron, Faculty of Medicine, UNMSM, Lima, Peru. Materials: Mice albino BALB / c (Mus musculus) males and Prunus dulcis "almond". Methods: 42 mice was used, experts say, 3 months and 31 ± 4,4 g in weight, randomized into six groups (n = 7). All groups received the same balanced diet and water ad libitum for 5 days. They received the following treatments for five days, perorally: group I and II: saline (NaCl 0.9g% 10 mL / kg), group III: Vitamin E 400 mg / kg, group IV: almond 100 mg / kg, group V : almond 500 mg / kg and group VI: almond 1000 mg / kg; 12 hours before the end of Tto. Cut whiskers of mice, except the group I; and after 12 hours they were sacrificed. Main outcome measures: Level of lipid peroxidation expressed in thiobarbituric acid (TBARS) and level of non-protein sulfhydryl groups (GS-NP) substances in addition to histopathological changes of brain tissue and cerebellum. Results: Administration of Prunus dulcis "almond" significantly increased (p <0.05) levels of GS-NP in all groups (except G VI) compared to the G II in brain; TBARS levels decreased significantly (p <0.05) in the V and VI group compared with group II, and in relation to the histological changes seen a slight improvement in the GV compared to GII. Conclusions: The administration of the suspension of Prunus dulcis seed "almond" demonstrated the neuroprotective effect in biochemical (TBARs y GS-NP) on the nervous tissue in mice induced to stress motor disorientation. KEYWORDS: Prunus dulcis "almond" non-protein sulfhydryl groups (GS-NP), lipid peroxidation (TBARS), histopathological changes, disorientation motor stress.