Two genes encoding sucrose synthase (SUS), namely SUS2 (At5g49190) and SUS3 (At4g02280), are strongly and differentially expressed in Arabidopsis seed. Detailed biochemical analysis was carried out in developing seeds 9-21 days after flowering (DAF) of wild type and two knockouts. SUS2 and SUS3 are not redundant genes since single knockouts show a phenotype in developing seeds. The mutants had 30-50% less SUS activity and therefore accumulated 40% more sucrose and 50% less fructose at 15 DAF. This did not affect the hexose-P pool, but led to 30-70% less starch in embryo and seed coat. Lipids were 55% higher in both mutants at 9-15 DAF. It seems that sucrolysis via SUS is not required for oil or protein synthesis but rather for channeling carbon toward ADP-glucose and starch in seeds. Metabolite profiling with GC-TOF revealed specific downstream changes in primary metabolism as a consequence of signaling or regulatory fine-tuning. While sucrose increased, hexoses and specific amino acids decreased reciprocally. There was a developmental shift regarding an earlier timing of dry weight accumulation, germinative maturity, oil deposition, sugar levels, transient starch buildup, and protein storage. Nevertheless, final seed size and composition were unaltered due to an earlier cessation of growth, thus giving rise to an apparent silent phenotype of mature mutant seeds. We conclude that SUS is important for metabolite homeostasis and timing of seed development, and propose that an altered sucrose/hexose ratio can modify carbon partitioning and the pattern of storage compounds in Arabidopsis.

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Sucrose synthase (SUS) is a key enzyme of carbon metabolism in heterotrophic tissues of plants. The Arabidopsis genome contains six SUS genes. Two members of this family, namely AtSUS2 (At5g49190) and AtSUS3 (At4g02280) are strongly and differentially expressed in Arabidopsis seed. Expression analysis was carried out using SUS:promoter-GUS fusion lines in a wild-type genetic background or in a mutant carrying a lesion in the transcription factor LEAFY COTYLEDON 2 (LEC2; At1g28300). The accumulation patterns of mRNA, protein, and SUS activity were altered in the lec2 mutant during seed development 9-18 days after flowering. This indicates that LEC2 acts epistatically on the expression of AtSUS2 and AtSUS3. It appears that LEC2 is required for cotyledon-specific expression of both SUS genes but it is not responsible for expression in the radicle tip during embryo development. The AtSUS2 promoter was induced in planta by feeding of glucose but less so by sucrose and trehalose. Non-phosphorylable glucose analogs such as 3-O-methyl-glucose and 2-deoxyglucose also caused an induction, suggesting that sugar signaling proceeds by a hexokinase-independent pathway, possibly involving hexose sensing. Analysis of transgenic lines carrying of truncated versions of the AtSUS2:promoter fused to Beta-glucuronidase activity revealed an internal 421 bp region that was responsible for expression in seeds. Bioinformatic sequence analysis revealed regulatory cis-elements putatively responsible for the spatio-temporal pattern of AtSUS2 expression such as the SEF3 (aaccca) and W-box (ttgact) motifs. These findings are discussed in relation to the roles played by AtSUS2, AtSUS3 and LEC2 in the biosynthesis of seed storage products in Arabidopsis.

Para la exportación de nopal verdura, México debe cumplir con los requisitos sanitarios de cada país. Una forma de garantizar la inocuidad de este producto es implementar buenas prácticas agrícolas (BPA). Sin embargo, sólo unos pocos productores de nopal verdura poseen reconocimiento en BPA. El objetivo del presente trabajo fue evaluar la calidad sanitaria de nopal verdura en punto de embarque de productores que usan BPA. Se seleccionaron al azar 10 productores de Otumba, Estado de México pertenecientes a una sociedad rural con reconocimiento BPA, y se tomaron muestras combinadas de nopal en punto de embarque durante los meses de comercialización de marzo de 2008 a mayo de 2009. Se determinó la carga de bacterias mesófilas aerobias (BMA), Coliformes totales (CT), Salmonella sp. y Escherichia coli, así como residuos de plaguicidas. Sólo 8% y 3% de las muestras rebasaron los límites especificados por la ICMSF en el conteo de BMA y CT, respectivamente. Una de las muestras colectadas en mayo de 2009 estuvo contaminada con Escherichia coli, y ninguna con Salmonella sp. Cinco productores rebasaron los límites permisibles de BMA, y dos los de CT en una de sus muestras. Estos casos se relacionaron con escasa higiene de los contenedores y el medio de transporte. Adicionalmente, se detectaron residuos de uno o varios plaguicidas en 8% de las muestras. Indicando el cumplimiento parcial de las BPA por estos productores en particular. Los resultados indican que el nopal verdura producido con BPA cumple con los requisitos sanitarios para exportación.

ResumenLa sacarosa sintasa (SUS) es una enzima clave en el metabolismo de la sacarosa. Esta enzima cataliza la conversión reversible de sacarosa y UDP a UDP-Glucosa y fructosa. En Arabidopsis, la familia de genes SUS consta de 6 miembros. Uno de estos, el gen SUS1 (At5g20830) se expresa durante la fase de desarrollo de la morfogénesis de la semilla, periodo que corresponde a la fase de la acumulación masiva de almidón y de hexosas, a la fecha se desconoce la función metabólica que tiene esta isoforma-1 en la síntesis de estos compuestos carbonados. Con la finalidad de aportar elementos que contribuyan a determinar las funciones de esta enzima (SUS1) en el desarrollo de la semilla, se llevó a cabo un análisis funcional, el cual comprende la caracterización bioquímica de un alelo nulo de esta isoforma. Los resultados obtenidos muestran la importancia de esta enzima (SUS1) para el metabolismo de la sacarosa en la semilla de Arabidopsis en la etapa de desarrollo. Por otro lado, la mutación SUS1 afecta principalmente el cociente hexosas/ sacarosa; lo cual provoca una disminución importante en la acumulación de ácidos grasos y de almidón transitorio durante los estadios tempranos del desarrollo de la semilla.Palabras clave: almidón, ácidos grasos, isoforma-1 de sacarosa sintasa, semillas en desarrollo. AbstractSucrose synthase (SUS) is a key enzyme in the metabolism of sucrose. This enzyme catalyzes the reversible conversion of sucrose and UDP to UDP-glucose and fructose. In Arabidopsis, the family SUS of genes consists of 6 members. One of these, the SUS1 gene (At5g20830) is expressed during the development phase of the morphogenesis of the seed, which corresponds to the phase of the massive accumulation of starch and hexoses to date unknown metabolic function has this isoform-1 in the synthesis of these carbon compounds. In order to provide elements that contribute to determine the roles of this enzyme (SUS1) in seed development was carried out a functional analysis, which comprises the biochemical characterization of a null allele of this isoform. The results show the importance of this enzyme (SUS1) for sucrose metabolism in Arabidopsis seeds in the development stage. Moreover, the mutation affects mainly SUS1 hexoses/sucrose ratio; which results in a significant accumulation of fatty acids and of transitory starch in the early stages of seed development decreased.