Cultivated tomato (Solanum lycopersicum L.) germplasm shows limited genetic variation. Many DNA marker systems have been used for genetic diversity studies in wild and cultivated tomatoes, but their usefulness for characterizing phenotypic differences among very closely related cultivars remains uncertain. We have used 19 selected simple sequence repeat (SSR) markers and 7 amplified fragment length polymorphism (AFLP) primer combinations to characterize 48 cultivars of tomato, mainly traditional cultivars from the south-east of Spain. The main types were Solanum lycopersicum L. 'Muchamiel', 'De la pera', and 'Moruno'. The robustness of the dendrograms and the discrimination power reached with each marker type were similar. Unique fingerprinting even of the most closely related tomato cultivars could be obtained using a combination of some SSR and AFLP markers. A better grouping of the 'Muchamiel' cultivars was observed with SSR markers, whereas the grouping of cultivars of 'De la pera' type was best achieved with AFLPs. However, both types of markers adequately grouped cultivars of the main types, confirming the utility of SSR and AFLP markers for the identification of traditional cultivars of tomato.
Digital PCR (dPCR) is a breakthrough technology that able to provide sensitive and absolute nucleic acid quantification. It is a third-generation technology in the field of nucleic acid amplification. A unique feature of the technique is that of dividing the sample into numerous separate compartments, in each of which an independent amplification reaction takes place. Several instrumental platforms have been developed for this purpose, and different statistical approaches are available for reading the digital output data. The dPCR assays developed so far in the plant science sector were identified in the literature, and the major applications, advantages, disadvantages, and applicative perspectives of the technique are presented and discussed in this review.
Digital polymerase chain reaction (dPCR) is a breakthrough technology based on the partitioning of the analytical sample and detection of individual end-point amplifications into the separate compartments. Among the numerous applications of this technology, its suitability in mutation detection is relevant and characterized by unprecedented levels of precision. The actual applicability of this analytical technique to quantify the presence of a specific plant genotype, in both raw materials and transformed products, by exploiting a point polymorphism has been evaluated. As proof of concept, an Italian premium pasta production chain was considered and a dPCR assay based on a durum wheat target variety private point mutation was designed and evaluated in supply-chain samples. From the results obtained, the assay can be applied to confirm the presence of a target variety and to quantify it in raw materials and transformed products, such as commercial grain lots and pasta. The performance, costs, and applicability of the assay has been compared to analytical alternatives, namely simple sequence repeats (SSRs) and genotype-by-sequencing based on Diversity Arrays Technology sequencing (DArTseqTM).
A traditional Italian sweet pepper landrace, 'Peperone di Voghera', which faced the risk of extinction, was analyzed for its genetic, phenological, morphological, agronomic and biochemical traits. An extant population was compared with cultivars 'Quadrato d'Asti', 'Cuneo', and 'Giallo d'Asti', cultivated in the same area, in order to evaluate the chance of the landrace recovery. Amplified fragment length polymorphism (AFLP) analysis demonstrated that Voghera landrace is distinct with respect to reference cultivars, thus excluding extensive genetic contamination and providing a molecular basis of both phenological and biochemical differences. Leaf chlorophyll content is lower, fruits start ripening earlier than controls, and yield (1,100-1,300 g plant -1 ) does not significantly differ. Sensitivity to root pathogens, the main factor that led to the decline in the past, does not seem to compromise the future chance of recovery. Vitamin C concentration is high (200-240 mg/100 g) and preserved by cold storage; more than 25% of vitamin C is also kept in pickled fruits. 'Voghera' landrace has nutritional and gastronomic properties that can be appreciated by consumers. The high internal genetic variability shown by AFLP analysis indicates that future selection work is necessary to fully maintain the original traits of the landrace and to improve it.Additional key words: AFLP; biodiversity; Capsicum annuum; extinction; nutraceutical properties; root mycosis. ResumenCaracterización de la variedad de pimiento tradicional 'Voghera', rescatada en el norte de ItaliaEn este trabajo se han analizado las características genéticas, morfológicas, fenológicas, agronómicas y bioquími-cas de una variedad tradicional italiana de pimiento dulce, 'Peperone di Voghera', en peligro de extinción. Se comparó una población existente con cultivares de la misma zona, 'Quadrato d'Asti', 'Cuneo', y 'Giallo d'Asti', a fin de evaluar la posibilidad de recuperarla. Análisis AFLPs (polimorfismos en la longitud de fragmentos amplificados) demostraron que 'Voghera' es distinta con respecto a los cultivares de referencia, lo que excluye que haya una amplia contaminación genética y proporciona una base molecular de las diferencias, tanto fenológicas como bioquímicas. Su contenido en clorofila es menor y los frutos comienzan a madurar antes que los controles, pero el rendimiento (1.100-1.300 g planta -1 ) no es significativamente diferente. Su sensibilidad a patógenos de la raíz, el principal factor que llevó a su declive en el pasado, no parece poner en peligro la posibilidad de una futura recuperación. La concentración en vitamina C es alta (200-240 mg/100 g) y se preserva en el fruto conservado en cámara frigorífica; también se detecta una buena cantidad (más del 25%) de vitamina C en las frutas encurtidas. 'Voghera' tiene propiedades nutritivas y gastronómicas que pueden ser apreciadas por los consumidores. La alta variabilidad genética interna detectada en los análisis AFLP indica que el trabajo de selección es necesario para mantener plena...
Several food products, made from hulled wheats, are now offered by the market, ranging from grains and pasta to flour and bakery products. The possibility of verifying the authenticity of wheat species used at any point in the production chain is relevant, in defense of both producers and consumers. A chip digital PCR assay has been developed to detect and quantify percentages of hulless (i.e., common and durum wheat) and hulled (i.e., einkorn, emmer and spelt) wheats in grains, flours and food products. The assay has been designed on a polymorphism in the miRNA172 target site of the AP2-5 transcription factor localized on chromosome 5A and involved in wheat spike morphogenesis and grain threshability. The assay has been evaluated even in a real-time PCR system to assess its applicability and to compare the analytical costs between dPCR and real-time PCR approaches.
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