937 resultados para Solanum lycopersicum L


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Structural differences such as abnormalities, damage and free spaces in seeds may affect germination. The aim of this study was to study the relationship between eggplant seed morphology and seed germination. Ten seed lots of the eggplant cultivar Embu were evaluated by X-ray image analysis and the germination test. Seed image analysis was performed by Image Pro Plus® software and the whole seed area and free space between the embryo and endosperm were measured. The internal seed area filled by the embryo and endosperm was calculated from the difference between the whole seed and free space areas. Based on these results and visual seed analysis, seeds were classified into three categories and information on germination was obtained for each one. X-ray image analysis provides a perfect view of the internal seed parts and for seed morphology studies. An increase in seed area filled by the endosperm and embryo does not improve seed germination. Mechanical seed damage and deteriorated tissues can adversely affect seed germination.

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O presente trabalho teve como objetivo avaliar dois sistemas de tutoramento e condução de plantas na produtividade e seus componentes em híbridos de tomateiro dos segmentos Salada Longa Vida e Santa Cruz de crescimento indeterminado, visando ao consumo in natura. Adotou-se o delineamento em blocos ao acaso com os tratamentos em esquema fatorial 5x2, quatro repetições, sendo avaliados cinco híbridos comerciais, três pertencentes ao grupo Salada Longa Vida ('Alambra', 'Paron' e 'Forty') e dois do grupo Santa Cruz ('Débora Pto' e 'Ellus'), em dois sistemas de condução de plantas (bambu e fitilho). O experimento foi conduzido em condições de campo, em Itatiba (SP), de julho de 2007 a janeiro de 2008. Foram consideradas todas as plantas de cada parcela e avaliaram-se a produtividade total, produtividade comercial, produção por planta, massa média de frutos, número total de frutos, número de frutos comercializáveis, número de frutos das classes '3A', '2A' e '1A', número de frutos manchado e com lóculo aberto. Não houve interação entre cultivar e sistema de condução de planta para maioria das características avaliadas, exceto número total de frutos, número de frutos comercializáveis e número de frutos da classe '2A'. O sistema de condução de plantas com uma haste tutorada com fitilho proporcionou incremento na massa média de frutos e número de frutos da classe '3A', e decréscimo do número de frutos da classe '1A', sem reduzir a produtividade total e comercial quando comparado ao método de condução tradicional no bambu com duas hastes por planta.

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The general combining ability (GCA), specific combining ability (SCA), and heterosis were studied in a complete diallel cross among fresh market tomato breeding lines with reciprocal excluded. Fifteen genotypes (five parents and ten hybrids) were tested using a randomized complete block design, with three replications, and the experiments were conducted in Itatiba, São Paulo state, Brazil, in 2005/06. The yield components evaluated were fruit yield per plant (FP), fruit number per plant (FN), average fruit weight (FW); cluster number per plant (CN); fruit number per cluster (FC), fruit wall thickness (FT) and number of locules per fruit (NL). Fruit quality components evaluated were total soluble solids (SS); total titratable acidity (TA); SS/TA ratio, fruit length (FL); fruit width (WI); length to width ratio (FL/WI). The data for each trait was first subjected to analysis of variance. Griffing's method 2, model 1 was employed to estimate the general (GCA) and specific (SCA) combining abilities. Parental and hybrid data for each trait were used to estimate of mid-parent heterosis. For plant fruit yield, IAC-2 was the best parental line with the highest GCA followed by IAC-4 and IAC-1 lines. The hybrids IAC-1 x IAC-2, IAC-1 x IAC-4 and IAC-2 x IAC-4 showed the highest effects of SCA. High heterotic responses were found for fruit yield and plant fruit number with values up to 49.72% and 47.19%, respectively. The best hybrids for fruit yield and plant fruit number were IAC-1 x IAC-2, IAC-1 x IAC-4 and IAC-2 x IAC-5, for fruit yield and plant fruit number, the main yield components.

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Variedades ou híbridos de tomateiro utilizados para produção de molhos e ketchups costumam ter hábito determinado, enquanto a maioria dos genótipos utilizados na produção para mesa (consumo in natura) possuem hábito indeterminado. Além de influenciar no manejo da cultura, o hábito de crescimento pode alterar parâmetros de produtividade, como o teor de sólidos solúveis totais (SST) nos frutos. O hábito de crescimento é controlado principalmente pelo gene SELF-PRUNING (SP), que é um dos componentes de uma pequena família gênica da qual faz parte também o gene SINGLE FLOWER TRUSS (SFT), atualmente considerado um dos componentes do tão buscado "florígeno". O entendimento da função bioquímica e o efeito fisiológico de tais genes em interação com o ambiente e outros genes (epistasia) possibilita a manipulação de parâmetros como precocidade e SST. Além disso, fornece subsídios para compreender a base genética do crescimento semideterminado, que combina vantagens do hábito determinado e indeterminado, podendo ser usado pelos melhoristas de plantas para o desenvolvimento de novas cultivares.

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Máster en Oceanografía

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Tomato (Lycopersicon esculentum Mill., Solanum lycopersicon L.) is one of the most popular vegetable throughout the world, and the importance of its cultivation is threatened by a wide array of pathogens. In the last twenty years this plant has been successfully used as a model plant to investigate the induction of defense pathways after exposure to fungal, bacterial and abiotic molecules, showing triggering of different mechanisms of resistance. Understanding these mechanisms in order to improve crop protection is a main goal for Plant Pathology. The aim of this study was to search for general or race-specific molecules able to determine in Solanum lycopersicon immune responses attributable to the main systems of plant defense: non-host, host-specific and induced resistance. Exopolysaccharides extracted by three fungal species (Aureobasidium pullulans, Cryphonectria parasitica and Epicoccum purpurascens), were able to induce transcription of pathogenesis-related (PR) proteins and accumulation of enzymes related to defense in tomato plants cv Money Maker,using the chemical inducer Bion® as a positive control. During the thesis, several Pseudomonas spp. strains were also isolated and tested for their antimicrobial activity and ability to produce antibiotics. Using as a positive control jasmonic acid, one of the selected strain was shown to induce a form of systemic resistance in tomato. Transcription of PRs and reduction of disease severity against the leaf pathogen Pseduomonas syringae pv. tomato was determined in tomato plants cv Money Maker and cv Perfect Peel, ensuring no direct contact between the selected rhizobacteria and the aerial part of the plant. To conclude this work, race-specific resistance of tomato against the leaf mold Cladosporium fulvum is also deepened, describing the project followed at the Phytopathology Laboratory of Wageningen (NL) in 2007, dealing with localization of a specific R-Avr interaction in transfected tomato protoplast cultures through fluorescence microscopy.

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In a majority of species, leaf development is thought to proceed in a bilaterally symmetric fashion without systematic asymmetries. This is despite the left and right sides of an initiating primordium occupying niches that differ in their distance from sinks and sources of auxin. Here, we revisit an existing model of auxin transport sufficient to recreate spiral phyllotactic patterns and find previously overlooked asymmetries between auxin distribution and the centers of leaf primordia. We show that it is the direction of the phyllotactic spiral that determines the side of the leaf these asymmetries fall on. We empirically confirm the presence of an asymmetric auxin response using a DR5 reporter and observe morphological asymmetries in young leaf primordia. Notably, these morphological asymmetries persist in mature leaves, and we observe left-right asymmetries in the superficially bilaterally symmetric leaves of tomato (Solanum lycopersicum) and Arabidopsis thaliana that are consistent with modeled predictions. We further demonstrate that auxin application to a single side of a leaf primordium is sufficient to recapitulate the asymmetries we observe. Our results provide a framework to study a previously overlooked developmental axis and provide insights into the developmental constraints imposed upon leaf morphology by auxin-dependent phyllotactic patterning.

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The spatial arrangement of leaves and flowers around the stem, known as phyllotaxis, is controlled by an auxin-dependent reiterative mechanism that leads to regular spacing of the organs and thereby to remarkably precise phyllotactic patterns. The mechanism is based on the active cellular transport of the phytohormone auxin by cellular influx and efflux carriers, such as AUX1 and PIN1. Their important role in phyllotaxis is evident from mutant phenotypes, but their exact roles in space and time are difficult to address due to the strong pleiotropic phenotypes of most mutants in phyllotaxis. Models of phyllotaxis invoke the accumulation of auxin at leaf initials and removal of auxin through their developing vascular strand, the midvein. We have developed a precise microsurgical tool to ablate the midvein at high spatial and temporal resolution in order to test its function in leaf formation and phyllotaxis. Using amplified femtosecond laser pulses, we ablated the internal tissues in young leaf primordia of tomato (Solanum lycopersicum) without damaging the overlying L1 and L2 layers. Our results show that ablation of the future midvein leads to a transient accumulation of auxin in the primordia and to an increase in their width. Phyllotaxis was transiently affected after midvein ablations, but readjusted after two plastochrons. These results indicate that the developing midvein is involved in the basipetal transport of auxin through young primordia, which contributes to phyllotactic spacing and stability.

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Summary Potato cells (Solanum tuberosum L.), cultivated in original Murashige-Skoog (MS) medium for 5 days were subsequently transferred into {MS} media containing nitrate or ammonium as sole inorganic N source and incubated under anoxia for 24 h. With regard to lipid stability, these cells behaved differently. Although lipid hydrolysis occurred in both cases by the same mechanism, nitrate was able to postpone free fatty acid release for about 6 h compared with ammonium within the 24 h anoxia treatment. The increased membrane lipid stability of nitrate-treated cells under anoxia was correlated with a higher nitrate reduction capability and an improved energy status.

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Durante la madurez del fruto se producen cambios morfológicos, fisiológicos y bioquímicos provocados por la expresión regulada de diferentes genes. El objetivo de este trabajo fue verificar si la presencia de polipéptidos totales del pericarpio en los estados verde maduro (VM) y rojo maduro (RM) permite caracterizar la madurez del tomate. Se analizaron 18 líneas endocriadas recombinantes obtenidas por selección antagónica-divergente de un cruzamiento entre la cv. Caimanta (Solanum lycopersicum) y la entrada LA722 (S. pimpinellifolium), que fueron incluidas junto a la F1 como testigos experimentales. Los extractos proteicos se obtuvieron de dos muestras independientes de cada estado según el protocolo estándar y se resolvieron en SDS-PAGE. Se analizó la presencia/ausencia de bandas por genotipos y por estado, detectándose 26 en VM y 29 en RM. Algunas bandas fueron comunes entre estados, mientras que otras resultaron propias de VM o RM, respectivamente. Se calcularon las distancias de Jaccard y se realizó un análisis de conglomerados según el método UPGMA. En el dendrograma (correlación cofenética = 0,43) se distinguieron dos grandes grupos definidos por el estado de madurez. Se concluye que los perfiles proteicos del pericarpio son una herramienta postgenómica apropiada para identificar dos estados de madurez del fruto de tomate.

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Mulch materials of different origins have been introduced into the agricultural sector in recent years alternatively to the standard polyethylene due to its environmental impact. This study aimed to evaluate the multivariate response of mulch materials over three consecutive years in a processing tomato (Solanum lycopersicon L.) crop in Central Spain. Two biodegradable plastic mulches (BD1, BD2), one oxo-biodegradable material (OB), two types of paper (PP1, PP2), and one barley straw cover (BS) were compared using two control treatments (standard black polyethylene [PE] and manual weed control [MW]). A total of 17 variables relating to yield, fruit quality, and weed control were investigated. Several multivariate statistical techniques were applied, including principal component analysis, cluster analysis, and discriminant analysis. A group of mulch materials comprised of OB and BD2 was found to be comparable to black polyethylene regarding all the variables considered. The weed control variables were found to be an important source of discrimination. The two paper mulches tested did not share the same treatment group membership in any case: PP2 presented a multivariate response more similar to the biodegradable plastics, while PP1 was more similar to BS and MW. Based on our multivariate approach, the materials OB and BD2 can be used as an effective, more environmentally friendly alternative to polyethylene mulches.

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La resistencia genética mediada por los genes R es uno de los sistemas de defensa de las plantas frente a patógenos y se activa una vez que los patógenos han superado la defensa basal que otorgan la cutícula y pared celular. Los mecanismos de resistencia genética se inician a su vez, por el reconocimiento de productos derivados de genes de avirulencia de los patógenos (avr) por parte de las proteínas R. Tanto la respuesta de defensa basal como la respuesta de defensa por genes R están influenciadas por patrones de regulación hormonal, que incluye a las principales hormonas vegetales ácido salicílico (SA), ácido jasmónico (JA) y etileno (ET). En tomate (Solanum lycopersicum) uno de los genes R es el gen MiG1, que confiere resistencia a nematodos formadores de nódulos (Meloidogyne javanica, M. incognita y M. arenaria). Uno de los eventos más importantes que caracterizan a la respuesta de resistencia es la reacción hipersensible (HR), que está mediada por la activación temprana de una serie de sistemas enzimáticos, entre los que destaca el de las peroxidasas (PRXs) Clase III. Su función es importante tanto para limitar el establecimiento y expansión del nematodo, al generar ambientes altamente tóxicos por su contribución en la producción masiva de ROS, como por su implicación en la síntesis y depósito de lignina generando barreras estructurales en el sitio de infección. Además de estos mecanismos de defensa asociados a la resistencia constitutiva, las plantas pueden desarrollar resistencia sistémica adquirida (SAR) que en la naturaleza ocurre, en ocasiones, en una fase posterior a que la planta haya sufrido el ataque de un patógeno. Así mismo hay diferentes productos de origen químico como el benzotiadiazol o BTH (ácido S-metil benzol-(1,2,3)-tiadiozole-7-carbónico ester) que pueden generar esta misma respuesta SAR. Como resultado, la planta adquiere resistencia sistémica frente a nuevos ataques de patógenos. En este contexto, el presente trabajo aborda en primer lugar el análisis comparativo, mediante microarrays de oligonucleótidos, de los transcriptomas de los sistemas radicales de plantas de tomate de 8 semanas de edad de dos variedades, una portadora del gen de resistencia MiG1 (Motelle) y otra carente del mismo y, por tanto, susceptible (Moneymaker), antes y después de la infección por M. javanica. Previo a la infección se observó que la expresión de un gran número de transcritos era más acusada en la variedad resistente que en la susceptible, entre ellos el propio gen MiG1 o los genes PrG1 (o P4), LEJA1 y ER24, lo que indica que, en ausencia de infección, las rutas hormonales del SA, JA y ET están más activas en la raíz de la variedad resistente. Por el contrario, un número mucho menor de transcritos presentaban su expresión más reducida en Motelle que en Moneymaker, destacando un gen de señalización para sintetizar la hormona giberelina (GA). La infección por M. javanica causa importantes cambios transcripcionales en todo el sistema radical que modifican sustancialmente las diferencias basales entre plantas Motelle y Moneymaker, incluida la sobreexpresión en la variedad resistente de los transcritos de MiG1, que se reduce parcialmente, mientras que las rutas hormonales del SA y el JA continuan más activas que en la susceptible (evidente por los genes PrG1 y LEJA1). Además, los cambios asociados a la infección del nematodo se evidencian por las grandes diferencias entre los dos tiempos post-infección considerados, de tal forma que en la fase temprana (2 dpi) de la interacción compatible predomina la sobreexpresión de genes de pared celular y en la tardía (12 dpi) los relacionados con el ARN. En el análisis de la interacción incompatible, aunque también hay muchas diferencias entre ambas fases, hay que destacar la expresión diferencial común de los genes loxA y mcpi (sobrexpresados) y del gen loxD (reprimido) por su implicación en defensa en otras interacciones planta-patógeno. Cabe destacar que entre las interacciones compatible e incompatible hubo muy pocos genes en común. En la etapa temprana de la interacción compatible destacó la activación de genes de pared celular y la represión de la señalización; en cambio, en la interacción incompatible hubo proteínas principalmente implicadas en defensa. A los 12 días, en la interacción compatible los genes relacionados con el ARN y la pared celular se sobreexpresaban principalmente, y se reprimían los de proteínas y transporte, mientras que en la incompatible se sobreexpresaron los relacionados con el estrés, el metabolismo secundario y el de hormonas y se reprimieron los de ARN, señalización, metabolismo de hormonas y proteínas. Por otra parte, la técnica de silenciamiento génico VIGS reveló que el gen TGA 1a está implicado en la resistencia mediada por el gen MiG1a M. javanica. Así mismo se evaluó el transcriptoma de todo el sistema radical de la variedad susceptible tras la aplicación del inductor BTH, y se comparó con el transcriptoma de la resistente. Los resultados obtenidos revelan que el tratamiento con BTH en hojas de Moneymaker ejerce notables cambios transcripcionales en la raíz; entre otros, la activación de factores de transcripción Myb (THM16 y THM 27) y del gen ACC oxidasa. Las respuestas inducidas por el BTH parecen ser de corta duración ya que no hubo transcritos diferenciales comunes a las dos fases temporales de la infección comparadas (2 y 12 dpi). El transcriptoma de Moneymaker tratada con BTH resultó ser muy diferente al de la variedad resistente Motelle, ambas sin infectar, destacando la mayor expresión en el primero del gen LeEXP2, una expansina relacionada con defensa frente a nematodos. Las respuestas inducidas por los nematodos en Moneymaker-BTH también fueron muy distintas a las observadas previamente en la interacción incompatible mediada por MiG1, pues sólo se detectaron 2 genes sobreexpresados comunes a ambos eventos. Finalmente, se abordó el estudio de la expresión diferencial de genes que codifican PRXs y su relación con la resistencia en la interacción tomate/M. javanica. Para ello, se realizó en primer lugar el estudio del análisis del transcriptoma de tomate de la interacción compatible, obtenido en un estudio previo a partir de tejido radical infectado en distintos tiempos de infección. Se han identificado 16 unigenes de PRXs con expresión diferencial de los cuales 15 se relacionan por primera vez con la respuesta a la infección de nematodos. La mayoría de los genes de PRXs identificados, 11, aparecen fuertemente reprimidos en el sitio de alimentación, en las células gigantes (CG). Dada la implicación directa de las PRXs en la activación del mecanismo de producción de ROS, la supresión de la expresión génica local de genes de PRXs en el sitio de establecimiento y alimentación pone de manifiesto la capacidad del nematodo para modular y superar la respuesta de defensa de la planta de tomate en la interacción compatible. Posteriormente, de estos genes identificados se han elegido 4: SGN-U143455, SGN-U143841 y SGN-U144042 reprimidos en el sitio de infección y SGN-U144671 inducido, cuyos cambios de expresión se han determinado mediante análisis por qRT-PCR y de hibridación in situ en dos tiempos de infección (2 dpi y 4 dpi) y en distintos tejidos radicales de tomate resistente y susceptible. Los patrones de expresión obtenidos demuestran que en la interacción incompatible la transcripción global de los 4 genes estudiados se dispara en la etapa más temprana en el sitio de infección, detectándose la localización in situ de transcritos en el citoplasma de las células corticales de la zona meristemática afectadas por el nematodo. A 4 dpi se observó que los niveles de expresión en el sitio de infección cambian de tendencia y los genes SGN-U144671 y SGN-U144042 se reprimen significativamente. Los diferentes perfiles de expresión de los genes PRXs en los dos tiempos de infección sugieren que su inducción en las primeras 48 horas es crucial para la respuesta de defensa relacionada con la resistencia frente a la invasión del nematodo. Por último, al analizar el tejido radical sistémico, se detectó una inducción significativa de la expresión en la fase más tardía de la infección del gen SGN-U144042 en el genotipo susceptible y del SGN-U143841 en ambos genotipos. En este estudio se describe por primera vez la inducción de la expresión sistémica de genes de PRXs en tomate durante la interacción compatible e incompatible con M. javanica lo que sugiere su posible implicación funcional en la respuesta de defensa SAR activada por la infección previa del nematodo. ABSTRACT Plants defend themselves from pathogens by constitutive and/or induced defenses. A common type of induced defense involves plant resistance genes (R), which are normally activated in response to attack by specific pathogen species. Typically, a specific plant R protein recognizes a specific pathogen avirulence (avr) compound. This initiates a complex biochemical cascade inside the plant that results in synthesis of antipathogen compounds. This response can involve chemical signaling, transcription, translation, enzymes and metabolism, and numerous plant hormones such as salicylic acid (SA), jasmonates (JA) and ethylene (ET). Induced plant defense can also activate Class III peroxidases (PRXs), which produce reactive oxygen species (ROS), regulate extracellular H2O2, and play additional roles in plant defense. R-gene activation and the resulting induced defense often remain localized in the specific tissues invaded by the plant pathogen. In other cases, the plant responds by signaling the entire plant to produce defense compounds (systemic induction). Plant defense can also be induced by the exogenous application of natural or synthetic elicitors, such as benzol-(1,2,3)-thiadiazole-7-carbothionic acid. There is much current scientific interest in R-genes and elicitors, because they might be manipulated to increase agricultural yield. Scientists also are interested in systemic induction, because this allows the entire plant to be defended. In this context, one of the aims of this investigation was the transcriptoma analysis of the root systems of two varieties of tomato, the resistant variety (Motelle) that carrier MiG1 and the susceptible (Moneymaker) without MiG1, before and after infection with M. javanica. The overexpression was more pronounced in the transcriptoma of the resistant variety compared with susceptible, before infection, including the MiG1 gene, PrG1 (or P4) genes, LEJA1 and ER24, indicating that hormone SA, JA and ET are active in the resistant variety. Moreover, GA hormone presents an opposite behavior. M. javanica infection causes significant transcriptional changes in both compatible (Moneymaker-M. javanica) and incompatible (Motelle-M. javanica) interaction. In the incompatible transcriptome root system, was notably reduced the expression of the MiG1 gene, and a continuity in the expression of the hormonal pathways of SA and JA. In other hand, transcriptional profile changes during compatible interaction were associated with nematode infection. The large differences between the two times point infection considered (2 dpi and 12 dpi) indicates an overexpression of cell wall related genes in the first phase, and conversely an overexpression of RNA genes in the late phase. Transcriptoma analysis of incompatible interaction, although there were differences between the two phases, should be highlighted the common differential gene expression: loxA and mcpi (overexpressed) and loxD gene (suppressed), as they are involved in defenses in other plant-pathogen interactions. The VIGS tool has provided evidence that TGA 1a is involved in MiG1 mediated resistance to M. javanica. Likewise, the systemic application of BTH was assessed and compared with susceptible and resistant variety. Root system transcriptoma of BTH treatment on leaves showed the activation of Myb transcription factors (THM16 and THM27), the ACC oxidase gene. and the LeEXP2 gene, encoding for an expansin enzyme, related with defense against nematodes. The activation appears to be reduced by subsequent infection and establishment of nematodes. To assist in elucidate the role of tomato PRXs in plant defence against M. javanica, the transcriptome obtained previously from isolated giant cells (GC) and galls at 3 and 7 dpi from the compatible interaction was analysed. A total of 18 different probes corresponding to 16 PRX encoding genes were differentially expressed in infection site compared to the control uninfected root tissues. Most part of them (11) was down-regulated. These results yielded a first insight on 15 of the PRX genes responding to tomato–Meloidogyne interaction and confirm that repression of PRX genes might be crucial for feeding site formation at the initial stages of infection. To study the involvement of PRX genes in resistance response, four genes have been selected: SGN-U143455, SGN-U143841 and SGN-U144042 consistently down-regulated and SGN-U144671 consistently up-regulated at infection site in compatible interaction. The expression changes were determined by qRT-PCR and in situ location at 2 dpi and 4 dpi, and in different root tissues of resistant and susceptible plants. Early upon infection (2 dpi), the transcripts levels of the four genes were strongly increased in infected tissue of resistant genotype. In situ hybridization showed transcript accumulation of them in meristem cortical cells, where the nematode made injury. The results obtained provide strong evidence that early induction of PRX genes is important for defence response of the resistance against nematode invasion. Moreover, the induction patterns of SGN-U144042 gene observed at 4 dpi in distal noninfected root tissue into the susceptible genotype and of SGN-U143841 gene in both genotypes suggest a potential involvement of PRX in the systemic defence response.

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En este estudio evaluamos la calidad de los lodos de una depuradora urbana y su aplicación en el cultivo de sandía (Citrullus lanatus) y tomate (Solanum lycopersicum).

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Intact amyloplasts from potato (Solanum tuberosum L.) were used to study starch biosynthesis and phosphorylation. Assessed by the degree of intactness and by the level of cytosolic and vacuolar contamination, the best preparations were selected by searching for amyloplasts containing small starch grains. The isolated, small amyloplasts were 80% intact and were free from cytosolic and vacuolar contamination. Biosynthetic studies of the amyloplasts showed that [1-14C]glucose-6-phosphate (Glc-6-P) was an efficient precursor for starch synthesis in a manner highly dependent on amyloplast integrity. Starch biosynthesis from [1-14C]Glc-1-P in small, intact amyloplasts was 5-fold lower and largely independent of amyloplast intactness. When [33P]Glc-6-P was administered to the amyloplasts, radiophosphorylated starch was produced. Isoamylase treatment of the starch followed by high-performance anion-exchange chromatography with pulsed amperometric detection revealed the separated phosphorylated α-glucans. Acid hydrolysis of the phosphorylated α-glucans and high-performance anion-exchange chromatography analyses showed that the incorporated phosphate was preferentially positioned at C-6 of the Glc moiety. The incorporation of radiolabel from Glc-1-P into starch in preparations of amyloplasts containing large grains was independent of intactness and most likely catalyzed by starch phosphorylase bound to naked starch grains.