Conservação genética de populações de Cryptocarya moschata Nees (Lauraceae) na Mata Atlântica do estado de São Paulo

Através da análise de 18 locos isoenzimáticos polimórficos, foram estimadas as freqüências alélicas referentes a 214 indivíduos adultos de quatro populações naturais de Cryptocarya moschata de duas regiões do estado de São Paulo. Com base nas heterozigosidades observadas e esperadas, foram obtidas estimativas das estatísticas F de Wright. Para fins de comparação, utilizou-se também o método da análise da variância para estimação dos parâmetros correspondentes F = FIT, CE="Symbol">qP= FST e f = FIS. Os dois métodos forneceram resultados bastante concordantes: c="http:/img/fbpe/rbb/v22s2/f.gif" alt="f.gif (76 bytes)">IT = 0,142; c="http:/img/fbpe/rbb/v22s2/f.gif" alt="f.gif (76 bytes)">ST = 0,140; c="http:/img/fbpe/rbb/v22s2/f.gif" alt="f.gif (76 bytes)">IS = 0,002 e c="http:/img/fbpe/rbb/v22s2/f.gif" alt="f.gif (76 bytes)">= 0,116; c="http:/img/fbpe/rbb/v22s2/a3form05.gif" alt="a3form05.gif (151 bytes)">P = 0,123 e c="http:/img/fbpe/rbb/v22s2/a3form06.gif" alt="a3form06.gif (144 bytes)">= ­0,008, indicando que os indivíduos dentro das populações devem ser panmíticos e que a diversidade entre populações é bastante alta, sendo similar à que se espera para famílias com estruturação de meios-irmãos. Calculando c="http:/img/fbpe/rbb/v22s2/a3form05.gif" alt="a3form05.gif (151 bytes)">P com as populações tomadas duas a duas, notou-se tendência de c="http:/img/fbpe/rbb/v22s2/a3form05.gif" alt="a3form05.gif (151 bytes)">P crescer com a distância geográfica o que sugere tendência de isolamento pela distância. O fluxo gênico foi estimado em 0,9 indivíduos por geração, o que corrobora a pronunciada diferenciação populacional encontrada. Devido ao valor negligível encontrado de c="http:/img/fbpe/rbb/v22s2/a3form07.gif" alt="a3form07.gif (245 bytes)">IS, o tamanho efetivo de variância de cada população é equivalente ao número de indivíduos amostrados. As estratégias de manejo e conservação necessárias para a preservação da alta variabilidade genética intrapopulacional de C. moschata implicam na manutenção de populações com número grande de indivíduos. Além disso, para a preservação da espécie como um todo, a manutenção de muitas populações provavelmente é necessária.

Morfologia e estabelecimento de plântulas de Cryptocarya moschata Nees, Ocotea catharinensis Mez e Endlicheria paniculata (Spreng.) MacBride - Lauraceae

Foram descritos aspectos morfológicos de plântulas de Cryptocarya moschata Nees, Ocotea catharinensis Mez e Endlicheria paniculata (Spreng.) MacBride em desenvolvimento, em condições naturais e em casa de vegetação, de material coletado no Parque Estadual de Carlos Botelho-SP (24º44' - 24º03'S e 47º46' - 48º10'W). C. moschata apresentou um padrão de desenvolvimento de plântulas diferente das demais, com pec��olos cotiledonares alongados e soldados na região hipocotilar, ausência de catáfilos, torção na região do hipoc��tilo, e emissão de um par de eófilos simultâneos. O. catharinensis e E. paniculata apresentaram plântulas com emissão simultânea de três a cinco eófilos espiralados, em média. Na natureza, as plântulas de C. moschata caracterizaram-se por permanecer latentes por períodos longos, até o surgimento de clareiras ou incidência maior de luz, evidenciado pela ausência de indivíduos jovens em condições de sub-bosque da mata. As plântulas de O. catharinensis se estabeleceram na sombra, independentemente da formação de clareiras para o seu desenvolvimento, apresentando inúmeros juvenis no sub-bosque. Para E. paniculata, houve indicações de que a espécie apresenta plasticidade alta, desenvolvendo-se tanto em condições sombreadas como a pleno sol, em áreas perturbadas de borda ou de grandes clareiras, podendo ser colonizadora de áreas degradadas.

Estrutura genética intrapopulacional em Cryptocarya moschata Nees (Lauraceae)

Pela análise de seis sistemas enzimáticos (ACP, ALP, CAT, GOT, PPO e PO), com 18 locos polimórficos, foram calculadas as freqüências alélicas de 24 alelos referentes a 141 indivíduos adultos de uma população natural de Cryptocarya moschata (noz-moscada-do-Brasil) do Parque Estadual Carlos Botelho (24º03'21" S, 47º59'36" W), São Miguel Arcanjo, SP, amostrados em uma área de cerca de 647 ha. A estrutura genética espacial foi investigada através do método de autocorrelação espacial, utilizando-se coeficientes I de Moran estimados para 10 classes de distância. Adicionalmente, para as mesmas classes de distância, estimaram-se os coeficientes r através de uma análise multivariada. Ambas as abordagens mostraram-se similares qualitativamente, sendo que 15 correlogramas de coeficientes I foram significativos ao nível de 95% de probabilidade. O correlograma total gerado pelos coeficientes r indicou estruturação espacial significativa para a classe de distância de 0 a 150 m. Pela subdivisão desta classe em intervalos de 10 m, detectou-se autocorrelação espacial positiva na maioria das classes, indicando que os agrupamentos de árvores de C. moschata, que se encontram separadas a distâncias menores que 50 m, são constituídos por indivíduos semelhantes geneticamente e provavelmente aparentados. Contudo, a maior similaridade genética ocorreu entre indivíduos distanciados em torno de 105 m, podendo estar associada à dispersão de sementes promovida por Brachyteles arachnoides (Primates - Cebidae). Pela análise dos resultados, conclui-se que para uma amostragem adequada, visando a detec��ão de diversidade genética, a coleta de indivíduos distanciados acima de 750 m seria recomendável.

Morfologia e estabelecimento de plântulas de Cryptocarya moschata Nees, Ocotea catharinensis Mez e Endlicheria paniculata (Spreng.) MacBride - Lauraceae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Conservação genética de populações de Cryptocarya moschata Nees (Lauraceae) na Mata Atlântica do estado de São Paulo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estrutura genética intrapopulacional em Cryptocarya moschata Nees (Lauraceae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Herança de formato e produção de frutos imaturos em abobrinha (Cucurbita moschata)

O objetivo deste trabalho foi determinar a herança de formato e produção de frutos imaturos de 2 novas linhagens de C. moschata. Para tal obtiveram-se as gerações Fl, F2 e retrocruzamentos para ambos os progenitores. Para a avaliação das diferentes gerações utilizou-se o delineamento em blocos ao acaso com 4 repetições e número variável de plantas por repetição. As estimativas dos parâmetros genéticos foram obtidas segundo o modelo de MATHER & JINKS. Foram avaliados o peso, comprimento e diâmetros próximos das extremidades peduncular e estilar dos frutos. Observou-se dominância para maior velocidade de crescimento dos frutos, caracterizada pelos maiores comprimentos, diâmetros e peso médio na geração F1. Observou-se também dominancia para frutos mais bojudos e com menor relação entre o comprimento e a média dos diâmetros. Devido ao maior peso médio de frutos, e não ao número, obteve-se heterose significativa (20,5%) para produção total por planta.

Seed dispersal and germination of Cryptocarya moschata Nees & Martius Ex Nees, Ocotea catharinensis Mez and Endlicheria paniculata (Sprengel) Macbride (Lauraceae)

Data of seed dispersal and germination of the studied species are presented, comparing both observations from the wild and nursery. Fruits were collected from Carlos Botelho State Park (24 degrees 44' to 24 degrees 03'S, 47 degrees 46' to 48 degrees 10'W), south of São Paulo State, Brazil. O. catharinensis had low germination percentage, both in the wild and in nursery, and did not tolerate seed storage at low temperature. In nursery, diaspores with removed mesocarp of E. paniculata presented greater germination than those in entire fruits. The high levels of seedling mortality beneath mother-trees of C. moschata, when in comparison to those observed to established seedlings from diaspores dispersed by muriquis (Brachyteles arachnoides E. Geoffroy 1806, Cebidae, Primates), allied to the absence of juveniles in the understory, are in accord with the escape model of Janzen-Connel. In nursery, diaspores of C. moschata dispersed by the primates had greater germination, in smaller time, than those collected from mother-trees.

Evaluation of micronuclei frequency in Tradescantia pallida pollen mother cells treated with ethanolic extracts isolated from Cryptocarya mandioccana, Cryptocarya moschata and Pterogyne nitens

Although herbal extracts contain several classes of compounds with pharmacological activity, they also present toxic substances with mutagenic effects. The aim of the present study was to verify the mutagenicity of Cryptocarya moschata, Cryptocarya mandioccana and Pterogyne nitens using micronucleus assay in pollen mother cells (tetrads) in Tradescantia pallida (Trad-MCN). T. pallida inflorescences were treated with different concentrations of ethanolic extracts from the selected plant species. For C. mandioccana C. moschata and P. nitens, Trad-MCN assays were carried out simultaneoulsly, followed by positive control (formaldehyde 10000 ppm), negative control (Hoagland's solution), and vehicle control (Tween 20 20% or DMSO 3%). MCN present in tetrads were quantified in 300 tetrads/inflorescence and the mean (%) and standard error (SE) were established for at least 10 inflorescences per treatment. The extracts demonstrated dose response mutagenicity (clastogenic/aneugenic effects), respectively, C. mandioccana (0.5, 1.0 and 2.0 mg/mL) and P. nitens (1.0 and 2.0 mg/mL) However, no mutagenic effect was observed to C. moschata at the concentrations evaluated in the present study. We can conclude that the C. mandioccana and P. nitens extracts demonstrated clastogenic/aneugenic effects in highest concentrations whereas C. moschata extract did not demonstrate the same effect. © 2006 Sociedade Brasileira de Toxicologia.

Assessment of carotenoids in pumpkins after different home cooking conditions

Carotenoids have antioxidant activity, but few are converted by the body into retinol, the active form of vitamin A. Among the 600 carotenoids with pro-vitamin A activity, the most common are α- and β-carotene. These carotenoids are susceptible to degradation (e.g., isomerization and oxidation) during cooking. The aim of this study was to assess the total carotenoid, α- and β-carotene, and 9 and 13-Z- β-carotene isomer contents in C. moschata after different cooking processes. The raw pumpkin samples contained 236.10, 172.20, 39.95, 3.64 and 0.8610 µg.g- 1 of total carotenoids, β-carotene, α-carotene, 13-cis-β-carotene, and 9-Z-β-carotene, respectively. The samples cooked in boiling water contained 258.50, 184.80, 43.97, 6.80, and 0.77 µg.g- 1 of total carotenoids, β-carotene, α-carotene, 13-Z-β-carotene, and 9-Z-β-carotene, respectively. The steamed samples contained 280.77, 202.00, 47.09, 8.23, and 1.247 µg.g- 1 of total carotenoids, β-carotene, α-carotene,13-Z-β-carotene, and 9-Z-β-carotene, respectively. The samples cooked with added sugar contained 259.90, 168.80, 45.68, 8.31, and 2.03 µg.g- 1 of total carotenoid, β-carotene, α-carotene, 13-Z- β-carotene, and 9-Z- β-carotene, respectively. These results are promising considering that E- β-carotene has 100% pro-vitamin A activity. The total carotenoid and carotenoid isomers increased after the cooking methods, most likely as a result of a higher availability induced by the cooking processes.

Ecological niche modeling and geographical distribution of pollinator and plants: A case study of Peponapis fervens (Smith, 1879) (Eucerini: Apidae) and Cucurbita species (Cucurbitaceae)

The bees of the Peponapes genus (Eucerini, Apidae) have a Neotropical distribution with the center of species diversity located in Mexico and are specialized in Cucurbita plants. which have many species of economic importance. such as squashes and pumpkins Peponapis fervens is the only species of the genus known from southern South America The Cucurbita species occurring in the same area as P fervens Include four domesticated species (C ficifolia, C maxima maxima, C moschata and C pepo) and one non-domesticated species (Cucurbita maxima andreana) It was suggested that C. in andreana was the original pollen source to P fervens, and this bee expanded its geographical range due to the domestication of Cucurbita The potential geographical areas of these species were determined and compared using ecological niche modeling that was performed with the computational system openModeller and GARP with best subsets algorithm The climatic variables obtained through modeling were compared using Cluster Analysis Results show that the potential areas of domesticated species practically spread all over South America The potential area of P fervens Includes the areas of C m andreana but reaches a larger area, where the domesticated species of Cucurbita also Occur The Cluster Analysis shows a high climatic similarity between P fervens and C. m. andreana Nevertheless. P fervens presents the ability to occupy areas with wider ranges of climatic variables and to exploit resources provided by domesticated species (C) 2009 Elsevier B V All rights reserved

Epiderme e padrão de venação foliar de espécies de Lauraceae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

El injerto en pepino corto tipo español (Cucumis sativus L.) recomencaciones para su empleo en la zona central española

El injerto en hortalizas es uno de los temas de más actualidad en el panorama hortícola, no solo español, sino occidental, y recalcamos occidental, pues en muchos países que no corresponden a ese ámbito, sobre todo asiáticos: Japón, Corea, China, Filipinas, etc., esta es una técnica que cuenta con una gran difusión desde hace décadas, siendo, por ejemplo en Japón, la mayoría de sus cultivos de cucurbitáceas y solanáceas realizados con planta injertada. A finales de los noventa quedó claro que el empleo de bromuro de metilo tenía una fecha de caducidad y que las zonas que tenían una fuerte dependencia de este desinfectante de suelo debían de buscar alternativas a un plazo lo más corto posible, con un punto añadido sobre etapas anteriores, debían ser alternativas lo más respetuosas posible con el medio ambiente y que no incrementaran, de forma importante, los costes de producción. En la zona centro y concretamente en los invernaderos de la Comunidad de Madrid y zonas cercanas de Toledo y Guadalajara el pepino era y es el cultivo predominante, los horticultores empleaban el bromuro de metilo de forma sistemática para desinfectar sus suelos y la desaparición de este producto les planteaba una gran incertidumbre, lo que llevó a que desde diferentes instancias se buscaran diferentes alternativas. Tras analizar las posibilidades que se podían implementar y conocido el buen resultado que había dado el injerto en sandía en Almería, se decidió acometer los trabajos que conforman esta Tesis Doctoral, planteando en la zona, diferentes ensayos con la idea de conocer, si el injerto en pepino, con los cultivares empleados habitualmente, podía ser una alternativa real para los horticultores, tanto de Madrid, como los de las zonas cercanas de Toledo y Guadalajara. Se pretendía conocer sobre todo las repercusiones agronómicas y si esta técnica podría emplearse en solitario o era necesario complementarla con otras alternativas: desinfectantes químicos, solarización, biofumigación e incluso desinfección con vapor de agua. Los ensayos fueron realizados de forma secuencial entre el año 1999 y el 2011, comprobándose en primer lugar que el empleo de portainjertos híbridos de calabaza era posible con los cultivares de pepino corto tipo español, mayoritariamente empleados en los últimos años del siglo XX y primeros del XXI, fundamentalmente: Serena. Tras los primeros ensayos, Shintoza parec��a el portainjerto híbrido de calabaza (Cucurbita maxima x C. moschata) con mejores perspectivas de empleo, pues presentaba la ventaja adicional de ser bien conocido por los semilleros que producen planta injertada al ser, en esos momentos, el portainjerto más empleado en sandía, lo que garantizaba por su lado, su empleo en pepino, y que los horticultores pudiesen disponer de planta injertada. Más adelante los trabajos se encaminaron hacia la determinación de la densidad y tipo de poda más adecuado para la planta injertada, realizándose múltiples ensayos en esta dirección, que culminaron con la conclusión de que el extravigor que los portainjertos conferían a las plantas permitía conducir estas a dos o más brazos (se suelen emplear dos, por mejor adaptación a los trabajos de manejo de la planta por parte de los agricultores), con lo que se podría disminuir la densidad de planta y por tanto ahorrar en este capítulo, cosa que preocupaba y preocupa a los agricultores. Se llegó a determinar que es posible reducir la densidad de plantación en alrededor de un 25%, estando la densidad de brazos más adecuada entre 3 y 3.5 br•m-2. Tras las primeras decisiones tomadas sobre el portainjerto y la densidad más adecuada, se continuó con el estudio de adaptación de estas propuestas a los nuevos cultivares que las empresas de semillas iban proponiendo y los agricultores adoptando. Estas acciones se complementaron con la introducción de nuevos portainjertos susceptibles de sustituir a Shintoza o rotar con él para cambiar de sistema radicular, lo que es conveniente cuando se emplean, como es el caso, portainjertos que no son resistentes a nematodos, principalmente de la especie Meloidogyne incognita, el mayor problema en la zona, debido al suelo. Cultivares como Trópico, en un primer momento, y Urano y Motril más recientemente, se adaptaron muy bien a esta técnica. Entre los portainjertos que mostraron buena adaptación a la técnica de injerto y suficientemente buena compatibilidad con la mayoría de los cultivares ensayados destacan: RS-841, Strongtosa y Camel. Azman también mostró un comportamiento relevante, pero este portainjerto no podrá ser empleado, al ser recientemente retirado del mercado por la empresa que lo obtuvo y comercializó Aunque no era el objetivo principal de esta Tesis Doctoral, se ha comprobado que puede ser interesante combinar el empleo del injerto con otras técnicas alternativas al bromuro de metilo para superar los problemas debidos a enfermedades del suelo o nematodos, pero debe seguirse trabajando pues este es un tema en continua evolución, tanto si se trata de desinfectantes, a la mayoría de los cuales les está siendo retirado el permiso para su comercialización, como de otros métodos como la biofumigación o el empleo de vapor de agua. Queda muy claro que el injerto puede considerarse entre los métodos respetuosos con el medio ambiente, si no el que más, en lo que alternativas al bromuro de metilo se refiere. También en otro momento, se comprobó que con plantas injertadas es posible reducir el aporte de nutrientes, sobre todo nitrógeno, lo que además de un ahorro supone una mejora más desde el punto de vista medioambiental. En definitiva, queda demostrado que es factible el empleo del injerto en pepino corto tipo español, que las selecciones de los híbridos entre Cucurbita maxima y C. moschata que habitualmente se están empleando en sandía son también de aplicación en estos pepinos y que su empleo puede llevarnos a producciones suficientemente remuneradoras, alargándose en muchos casos el ciclo y no modificando, de forma apreciable, la calidad. Queda también demostrado que aunque los portainjertos no sean resistentes a nematodos, su extravigor les hace desarrollarse, desde el punto de vista productivo, suficientemente, llegando por tanto, a “convivir” con ese problema. Al no ser resistentes los portainjertos, y permanecer e incluso agravarse el problema de nematodos es conveniente poder contar con diferentes portainjertos que nos permitan rotar entre ellos y utilizar diferentes sistemas radiculares que harán menos fácil el parasitismo de los nematodos, como recomiendan los nematólogos que se haga. ABSTRACT Vegetable grafting is one of the most current practices in horticulture, not only in Spain, but also in other Western and Asian countries, such as Japan, South Korea, China, the Philippines, etc. This is a decades-old, widespread technique: In fact, most cucurbit and solanaceous crops in Japan and Korea are grafted. At the end of the 1990s, it was clear that methyl bromide had an expiry date. Consequently, the areas strongly dependant on this soil disinfectant had to look for alternatives as quickly as possible. Besides, these had to be as environmentally friendly as possible and should not increase production costs significantly. The cucumber has been and still is the most important crop in greenhouses of the Comunidad de Madrid and in areas near Toledo and Guadalajara. Cucumber growers used methyl bromide systematically to disinfect the soil. The banning of this chemical product brought about uncertainty, which encouraged the search for different alternatives. After analyzing the different possibilities and taking into account the good results of watermelon grafting in Almería, it was decided to carry out the works that make up this doctoral thesis. Different trials were made in order to know if the cultivars used in cucumber grafting might be a real alternative for farmers, not only in Madrid, but also in the areas near Toledo and Guadalajara. The main aim was to assess the agronomic repercussions and whether that technique could be used alone, or if other complementary alternatives, such as chemical disinfectants, solarisation, biofumigation, or even steam disinfection, were necessary. Trials were carried out sequentially from 1999 to 2011. It was observed that the use of pumpkin hybrid rootstocks could be applied to cultivars of Spanish short cucumbers, mainly grown in the late 20th and early 21st centuries eg Serena. After the early trials, Shintoza (Cucurbita maxima x C. moschata), a pumpkin hybrid rootstock, seemed to be the best option, as it had the additional advantage of being well known by nurseries growing grafting plants. Bearing this in mind, Shintoza was then the hybrid rootstock to be used in cucumbers and consequently growers could have grafted plants at their disposal. Later on, research was focused on density and the most adequate type of pruning, by carrying out several trials. These experiments showed that, the extra vigour the rootstocks gave to the plants, allowed them to have two or three stems, (normally nurserymen use two, as it is easier for them to manage the plants). These findings would lead to the lessening the density of the plant and thus reduce costs, something which worried and still worries farmers. It was stated that it would be possible to reduce the density of the plants by about 25%, the optimum density of the stems ranging from 3 to 3.5 stem-m-2. Once decisions were taken both on the rootstock and the appropriate density, we went on to study how to apply these proposals to the new cultivars which the seed companies were proposing and the farmers were applying. These measures were complemented with the introduction of new rootstocks capable of replacing Shintoza, or rotating with it in order to change the root system. This is particularly necessary when rootstocks, non-resistant to nematodes, mainly of the species Meloidogyne incognita, are used. This is the main problem due to the soil of that area. Cultivars such as Trópico, at first, and Urano and Motril, more recently, adapted quite well to this technique. Among the rootstocks which adapted well to grafting and which were compatible with most tested cultivars, were, in particular, RS-841 Strongtosa and Camel. The behaviour of Azman was worth studying, but this rootstock was removed from the market by the company which had bought and commercialized it. Although not the main purpose of the research, it was observed that combining grafting with other alternatives to methyl bromide in order to overcome problems due to soil diseases or nematodes may be worthwhile. However, further research is needed, as this topic is in constant evolution, not only when we come to disinfectants, most of which are being affected by the removal of the permit for commercialization, but also when we refer to other techniques such as biofumigation or the use of steam. Results also showed that grafted plants may reduce the amount of fertilizers, particularly nitrogen, used: This means both saving money and the protection of the environment. We may conclude by saying that grafting Spanish short cucumbers is feasible, that the selections of the hybrids between Cucurbita maxima and C. moschata, habitually used in watermelon grafting, can also be applied to these cucumbers. It can also be concluded that the use of these grafting techniques may lead to profitable yields, by lengthening the growing cycle in many cases and by maintaining the quality to a large extent. Although these rootstocks are not resistant to nematodes, the results showed that their extra vigour enables them to develop in terms of production, and thus they cope with this problem. Since these rootstocks are not resistant to nematodes and the problem with these nematodes may even worsen, it is recommended that different types of rootstocks should be available to enable both the rotation and the use of different root systems, which will encourage the parasitism of nematodes.

Correction of the misclassification of species in the Portuguese collection of Cucurbita pepo L. using DNA markers

In this study, the genetic variability among 130 accessions of the Portuguese germplasm collection of Cucurbita pepo L. maintained at the Banco Portugues de Germoplasma Vegetal was assessed using AFLP (amplified fragment length polymorphism) and RAPD (random amplified polymorphic DNA) techniques for the identification of a genetically diverse core group of accessions for field phenotypic analysis. The surprisingly completely different molecular patterns exhibited by multiple accessions was later confirmed in the distribution of the putative C. pepo plants into two clusters drastically separated at a very low level of genetic similarity (DICE coefficient = 0.37). Additional analyses with RAPD and ISSR (inter single sequence repeat) markers and the introduction of standard genotypes of C. maxima L. and C. moschata L. into the analyses allowed the identification of multiple accessions of the last species wrongly included in the C. pepo collection. This study is a good example of the usefulness of DNA markers in the establishment and management of plant germplasm collections.

Correction of the misclassification of species in the Portuguese collection of Cucurbita pepo L. using DNA markers

In this study, the genetic variability among 130 accessions of the Portuguese germplasm collection of Cucurbita pepo L. maintained at the Banco Portugues de Germoplasma Vegetal was assessed using AFLP (amplified fragment length polymorphism) and RAPD (random amplified polymorphic DNA) techniques for the identification of a genetically diverse core group of accessions for field phenotypic analysis. The surprisingly completely different molecular patterns exhibited by multiple accessions was later confirmed in the distribution of the putative C. pepo plants into two clusters drastically separated at a very low level of genetic similarity (DICE coefficient = 0.37). Additional analyses with RAPD and ISSR (inter single sequence repeat) markers and the introduction of standard genotypes of C. maxima L. and C. moschata L. into the analyses allowed the identification of multiple accessions of the last species wrongly included in the C. pepo collection. This study is a good example of the usefulness of DNA markers in the establishment and management of plant germplasm collections.