Features of a unique intronless cluster of class I small heat shock protein genes in tandem with box C/D snoRNA genes on chromosome 6 in tomato (Solanum lycopersicum)

Physical clustering of genes has been shown in plants; however, little is known about gene clusters that have different functions, particularly those expressed in the tomato fruit. A class I 17.6 small heat shock protein (Sl17.6 shsp) gene was cloned and used as a probe to screen a tomato (Solanum lycopersicum) genomic library. An 8.3-kb genomic fragment was isolated and its DNA sequence determined. Analysis of the genomic fragment identified intronless open reading frames of three class I shsp genes (Sl17.6, Sl20.0, and Sl20.1), the Sl17.6 gene flanked by Sl20.1 and Sl20.0, with complete 5' and 3' UTRs. Upstream of the Sl20.0 shsp, and within the shsp gene cluster, resides a box C/D snoRNA cluster made of SlsnoR12.1 and SlU24a. Characteristic C and D, and C' and D', boxes are conserved in SlsnoR12.1 and SlU24a while the upstream flanking region of SlsnoR12.1 carries TATA box 1, homol-E and homol-D box-like cis sequences, TM6 promoter, and an uncharacterized tomato EST. Molecular phylogenetic analysis revealed that this particular arrangement of shsps is conserved in tomato genome but is distinct from other species. The intronless genomic sequence is decorated with cis elements previously shown to be responsive to cues from plant hormones, dehydration, cold, heat, and MYC/MYB and WRKY71 transcription factors. Chromosomal mapping localized the tomato genomic sequence on the short arm of chromosome 6 in the introgression line (IL) 6-3. Quantitative polymerase chain reaction analysis of gene cluster members revealed differential expression during ripening of tomato fruit, and relatively different abundances in other plant parts.

Evaluacion de alternativas de manejo contra el complejo mosca blanca (Bemisia tabaci Gennadius) - Geminivirus en el cultivo de tomate [Solanum lycopersicum L.) (=Lycopersicum esculentum Mill.)] en Tisma, Masaya (2009) y Camoapa, Boaco (2010)

Mosca blanca (Bemisia tabaci) y los Geminivirus, son un serio problema para los productores de tomate en Nicaragua, este complejo afecta drásticamente los rendimientos del cultivo. Se han reportado diferentes tecnologías como solución al problema, tales como el uso de microinvernadero en la etapa semillero, uso de insecticidas sintéticos y prácticas culturales en la etapa de campo sin embargo, la problemática de la plaga aún persiste. Con el propósito de identificar alternativas que disminuyan el daño ocasionado al medio ambiente y que sean económicamente viables, se estableció está investigación en la que se evaluó el efecto de alternativas botánicas y químicas sobre la poblaciones de adultos B. tabaci, porcentaje de incidencia de virus, porcentaje de severidad de virus, incidencia de otros insectos plaga y su efecto sobre los enemigos naturales. El ensayo se estableció en dos ambientes, uno en Tisma (Masaya) en el periodo de Octubre a Diciembre del 2009 y el otro en Camoapa (Boaco) en los meses de Abril a Junio del 2010. El diseño utilizado fue un bloque completo al azar (BCA) en ambas localidades, los tratamientos evaluados fueron: Actara 25 WG ( Thiametoxam); Engeo 24,7 SC (Thiametoxam+Lambda-Cihalotrina); macerado de hojas de madero negro (Gliricidia sepium); aceite vegetal+jabón líquido, chile (Capsicum sp)+ ajo (Allium sativum)+jabón y un testigo. Los resultados encontrados fueron los siguientes: los tratamientos con engeo, aceite vegetal + jabón líquido y hojas de madero negro registraron los promedios más bajo de mosca blanca por planta tanto en Tisma como en Camoapa. Los menores porcentajes de incidencia de virus y porcentaje de severidad del daño del virus fueron en engeo, aceite vegetal+jabón líquidos y madero negro. Los mejores rendimientos fueron registrados en engeo y hojas de madero negro en ambas localidades. El análisis económico demostró que engeo presentó un a TRM de 457 % en Tisma (2009) y 696 % en Camoapa (2010).

Alternativas de manejo contra el complejo mosca blanca (Bemisia tabaci Gennadius)-Geminivirus en el cultivo de tomate (Solanum lycopersicum L.= (Lycopersicum esculentum Mill.) en Tisma, Masaya (2009) y Camoapa, Boaco (2010)

Mosca blanca (Bemisia tabaci) y los Geminivirus, son un serio problema para los productores de tomate en Nicaragua, este complejo afecta drásticamente los rendimientos del cultivo. Se han reportado diferentes tecnologías como solución al problema, tales como el uso de microinvernadero en la etapa semillero, uso de insecticidas sintéticos y prácticas culturales en la etapa de campo, sin embargo, la problemática de la plaga aún persiste. Con el propósito de identificar alternativas que disminuyan el daño ocasionado al medio ambiente y que sean económicamente viables, se estableció la investigación en la que se comparo el efecto de alternativas botánicas y químicas sobre la poblaciones de adultos B. tabaci, porcentaje de incidencia de virus, porcentaje de severidad de virus, rendimientos y análisis económico. El ensayo se estableció en dos ambientes, uno en Tisma (Masaya) en el periodo de Octubre a Diciembre del 2009 y el otro en Camoapa (Boaco) en los meses de Abril a Junio del 2010. El diseño utilizado fue un bloque completo al azar (BCA) en ambas localidades, los tratamientos evaluados fueron: Actara 25 WG (Thiametoxam); Engeo 24,7 SC (Thiametoxam+Lambda-Cihalotrina); macerado de hojas de madero negro (Gliricidia sepium); aceite vegetal+jabón líquido, chile (Capsicum sp)+ ajo (Allium sativum)+jabón y un testigo. Los resultados encontrados fueron los siguientes: los tratamientos con engeo, aceite vegetal + jabón líquido y hojas de madero negro registraron los promedios más bajo de mosca blanca por planta tanto en Tisma como en Camoapa. Los menores porcentajes de incidencia de virus y porcentaje de severidad del daño del virus fueron en engeo, aceite vegetal+jabón líquidos y madero negro. Los mejores rendimientos fueron registrados en engeo con promedios 26581-18953 kg ha -1 y hojas de madero negro con promedios 25277-18107 kg ha -1 en Tisma (2009) y Camoapa (2010) respectivamente. El análisis económico demostró que engeo presentó una TRM de 457 % en Tisma (2009) y 696 % en Camoapa (2010).

Evaluación de productos botánicos para el manejo del complejo mosca blanca (Bemisia tabaci, Gennadius)-Geminivirus y otros insectos plagas en el cultivo de tomate (Solanum lycopersicum L.), en Tisma, Masaya

La mosca blanca (Bemisia tabaci) y los Geminivirus son las principales plagas causantes de pérdidas económicas en el cultivo de tomate (Solanum lycopersicum, Mill) en el municipio de Tisma-Masaya. En base a esta situación se realizó un estudio para evaluar la efectividad que tiene para controlar plagas los tratamientos botánicos: Chile+Ajo+Jabón, Madero Negro, Crisantemo, Extracto alcohólico+Chile, Extracto alcohólico+ Chile+ Ajo y Testigo en el período comprendido entre Diciembre 2013 a Febrero 2014. Las variables evaluadas fueron: número de mosca blanca por planta, Incidencia del daño de virosis de mosca blanca por planta, severidad del daño de virosis de mosca blanca por planta, y otros organismos plagas en el rubro: Halticus sp por planta, áfidos (Aphis gossypii) por planta, y minador de la hoja (Liriomyza sp) por planta. De los tratamientos evaluados, el menor número de moscas blancas por planta y menor porcentaje de incidencia(47%) y severidad (51%) lo obtuvo el tratamiento Extracto Alcohólico + Chile+ Ajo. El tratamiento Madero Negro presentó la mayor efectividad en el manejo de poblaciones de Halticus sp. y Lyriomiza sp., mientras que el Extracto Alcohólico + Chile + Ajo presentó los mejores resultados para el manejo de Aphis gossypii. El análisis económico reveló que el mayor rendimiento lo obtuvo el Extracto Alcohólico+ Chile + Ajo con 34685.18 kg/ha seguido del tratamiento Chile+ Ajo+ Jabón (30614.28 kg/ha). En el análisis de retorno marginal resultó que el tratamiento Chile+ Ajo+ Jabón es el que obtuvo la mejor tasa de retorno marginal con 1476 %, es decir, 14.76 US$ por cada dólar invertido. El análisis realizado en la Universidad de Tucson Arizona en muestra de mosca blanca procedente de Tisma- Masaya determinó que de la muestra un 66.66% corresponde a la mosca blanca autóctona (biotipo A), y el 33.33% corresponde al biotipo B que es más agresivo que el A. Y el análisis de los begomovirus determinó: Enrollamiento severo de la hoja de tomate (ToSLCV) con 97.4% de identidad y el Virus del mosaico dorado de la chiltoma (PepGMV) con un 98% de identidad.

Amplicon metagenomics for elucidating microbial community changes associated to conventional and organic farming of Beta vulgaris and Solanum lycopersicum

Soil microbial community changes associated to conventional and organic farming of two relevant crops (Beta vulgaris and Solanum lycopersicum) were analysed through 16s rRNA amplicon sequencing. This study revealed microbial communities in the agricultural soils studied to be similar to other reported nutrient-rich microbiomes, and some significant differences between the microbial communities associated to the two farming practices were found. Some phyla (Chloroflexi and Thermi) were found to be present in different abundances according to soil treatment. As chloroplast interference can be a stumbling block in plant-associated 16s rRNA amplicon metagenomics analysis of aerial plant tissues, two protocols for bacterial cell detachment (orbital shaking and ultrasound treatment) and two protocols for microbial biomass recovery (centrifugation and filtration) were tested regarding their efficiency at excluding plant-DNA. An alternative method to the one proposed by Rastogi et al (2010) for evaluating the chloroplast-amplicon content in post-PCR samples was tested, and the method revealed that filtration was the most efficient protocol in minimising chloroplast interference.

Parámetros fisicoquímicos para la obtención de modelos de predicción en la calidad poscosecha del tomate (solanum lycopersicum).

Tesis (Maestría en Ciencias Agrícolas) UANL, 2011.

Evaluación de sustratos y enrenados en producción de tomate bola (Solanum lycopersicum L.) en invernadero.

Tesis (Maestro en Ciencias en Producción Agrícola) UANL, 2014.

Genetic variation of and environmental effects on inducibility of resistance in tomatoes (Solanum lycopersicum L.) to Phytophthora infestans (Mont) de Bary

Many plant strengtheners are promoted for their supposed effects on nutrient uptake and/or resistance induction (IR). In addition, many organic fertilizers are supposed to enhance plant health and several studies have shown that tomatoes grown organically are more resistant to late blight, caused by Phytophthora infestans to tomatoes grown conventionally. Much is known about the mechanisms underlying IR. In contrast, there is no systematic knowledge about genetic variation for IR. Therefore, the following questions were addressed in the presented dissertation: (i) Is there genetic variation among tomato genotypes for inducibility of resistance to P. infestans? (ii) How do different PS compare with the chemical inducer BABA in their ability to IR? (iii) Does IR interact with the inducer used and different organic fertilizers? A varietal screening showed that contrary to the commonly held belief IR in tomatoes is genotype and isolate specific. These results indicate that it should be possible to select for inducibility of resistance in tomato breeding. However, isolate specificity also suggests that there could be pathogen adaptation. The three tested PS as well as two of the three tested organic fertilisers all induced resistance in the tomatoes. Depending on PS or BABA variety and isolate effects varied. In contrast, there were no variety and isolate specific effects of the fertilisers and no interactions with the PS and fertilisers. This suggests that the different PS should work independent of the soil substrate used. In contrast the results were markedly different when isolate mixtures were used for challenge inoculations. Plants were generally less susceptible to isolate mixtures than to single isolates. In addition, the effectiveness of the PS was greater and more similar to BABA when isolate mixtures were used. The fact that the different PS and BABA differed in their ability to induce resistance in different host genotype -pathogen isolate combinations puts the usefulness of IR as a breeding goal in question. This would result in varieties depending on specific inducers. The results with the isolate mixtures are highly relevant. On the one hand they increase the effectiveness of the resistance inducers. On the other hand, measures that increase the pathogen diversity such as the use of diversified host populations will also increase the overall resistance of the hosts. For organic tomato production the results indicate that it is possible to enhance the tomato growing system with respect to plant health management by using optimal fertilisers, plant strengtheners and any measures that increase system diversity.

Determination of lipophilic and hydrophilic antioxidants in Solanum Lycopersicum L. tomato from Gordal variety by LL-USAE combined with UHPLC-PDA/FLR

Tomato (Lycopersicon esculentum L.) is one of the main constituents of the Mediterranean diet. Its consumption has been proposed to reduce the risk of cardiovascular diseases and certain types of cancer. It is therefore one of the most popular and extensively consumed vegetable crop worldwide. To gain insights on the potential of Lycopersicon esculentum L. as bioactive food, two analytical methodologies were developed to determine the levels of the lipophilic -tocopherol, α-tocopherol, β-carotene, lycopene; and hydrophilic antioxidants ascorbic acid. The quantification of total carotenoids (β-carotene and lycopene) was assessed through a liquid–liquid ultrasound assisted extraction (LL-USAE) in combination with ultraviolet-visible spectroscopy (UV-Vis), according to method of mean, for total carotenoids (λmáx = 450 nm. The ultra-high performance liquid chromatographic using both photodiode array and fluorescence detection (UHPLC-PDA/FLR), allows the identification and quantification of the target lipophilic and hydrophilic antioxidants. This methodology UHPLC-PDA/FLR is fast, simple and revealed a high sensitivity for the compounds under study. The limits of detection (LODs) and quantification (LOQs) obtained were much lower (about 10 times) than the reported in literature. The method LL-USAE/UV-Vis was validated and applied to different tomato foodstuffs. The results reveal a small increase of carotenoids content during maturation, reaching the maximum level when ripe. These results complement those obtained by the ORAC and TBARS assays that show an increase of antioxidant capacity during maturation. The LODs ans LOQs obtained were also about 10 times lower than reported in literature. The carotenoid content was also evaluated by LL-USAE/UV-Vis in different tomatoes varieties. Regional variety present the high carotenoid level, followed by campari and gordal, and at last grape. This methodology was also applied to different processed food samples containing tomatoes derivatives. Highest carotenoids content were obtained in concentrated tomato foodstuffs.

The Tomato (Solanum Lycopersicum cv. Micro-Tom) Natural Genetic Variation Rg1 and the DELLA Mutant Procera Control the Competence Necessary to Form Adventitious Roots and Shoots

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Interação entre o gene TKN2 (KNOX-type I) e o miR156 node durante a transição de fase vegetativa para reprodutiva em tomateiro (Solanum lycopersicum)

Pós-graduação em Ciências Biológicas (Genética) - IBB

Caracterização molecular do gene que codifica a TCTP (Translationally Controlled Tumor Protein) em tomate (Solanum lycopersicum cv. Santa Clara)

The gene encoding TCTP (Translationally Controlled Tumour Protein) is present in all eukaryotes and its product is involved in various cellular processes. Although well characterized in mammals, there are only few works available in the literature related to the analysis of this protein in plants. In this present work, the expression of the gene encoding TCTP was analyzed in different organs/tissues of tomato plants (Solanum lycopersicum cv. Santa Clara). A quantification performed by RT-qPCR revealed the presence of TCTP transcript in all tissues/organs analyzed, with the highest expression level found in leaves. With the exception of fruits in intermediate stage of maturation, for which a small increase on the expression was detected, there was minimal variation in the relative expression of TCTP in other organ/tissues. In parallel, the effects of the constitutive expression of TCTP were investigated using transgenic tobacco lines able to overexpress this protein at different levels (T1, T2 and T3). Seedlings of these lines, and of a non-transgenic control line, were grown in MS culture medium for 21 days. At the end of this period, the length of roots and leaves was taken and the seedlings were photographed. According to Tukey's test, the analysis of the mean root length revealed a significant difference between T1 and T3 lines when compared to the control, although the same was not observed for the T2 line. For leaves, according to Kruskal-Wallis test, there was a statistical difference between the averages of leaf growth obtained for the different lines evaluated. According to these results, we can conclude that TCTP shows an ubiquitous expression in tomato plants, with the highest expression detected in leaves, and also that its overexpression promoted a higher root and leaf development in two of three transgenic tobacco lines tested

Abordagem fotomorfogenética para explorar o estresse abiótico em tomateiro (Solanum lycopersicum L.)

Pós-graduação em Agronomia (Produção Vegetal) - FCAV

Photomorphogenic modulation of water stress in tomato (Solanum lycopersicum L.): the role of phytochromes A, B1, and B2

Phytochromes are red/far-red light photoreceptors that mediate a variety of photomorphogenic processes in plants, from germination to flowering. In addition, there is evidence that phytochromes are also part of the stress signalling response, especially in response to water deficit stress, which is the major abiotic factor limiting plant growth and crop productivity worldwide. In this study, we used the phyA (far red-insensitive; fri), phyB1 (temporary red-insensitive; tri) and phyB2 mutants of tomato (Solanum lycopersicum L.) to study the roles of these three phytochromes in drought stress responses. Compared to wild type (WT) plants grown under water-deficit stress conditions, the fri, tri, and phyB2 mutants did not exhibit altered dry weights, leaf areas, stomatal densities, or stomatal opening. The stomatal conductance of all three mutants was severely reduced under both fully-hydrated and water-deficit conditions. Although relative water contents did change after drought stress in each mutant, the most significant reduction in water potential during water stress was observed in the fri mutant. However, this mutant returned its water status to WT levels during rehydration. Although the phyB2 mutant lost more water from detached leaves during abscisic acid (ABA) treatment, phyB2 behaved like WT plants, indicating that this mutant was not insensitive to ABA. Overall, these results indicate that the phytochromes phyA, phyB1, and phyB2 modulate drought stress responses in tomato.

Caracterização funcional da via miR159/SlyGAMYB-like ao longo do processo de frutificação em tomateiro (Solanum lycopersicum L.)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)