An integrative approach reveals five new species of highland papayas (Caricaceae, Vasconcellea) from northern Peru.

The assignment of accurate species names is crucial, especially for those with confirmed agronomic potential such as highland papayas. The use of additional methodologies and data sets is recommended to establish well-supported boundaries among species of Vasconcellea. Accordingly, six chloroplast (trnL-trnF, rpl20-rps12, psbA-trnH intergenic spacers, matK and rbcL genes) and nuclear (ITS) markers were used to delimit species in the genus Vasconcellea using phylogeny and four DNA-based methods. Our results demonstrated congruence among different methodologies applied in this integrative study (i.e., morphology, multilocus phylogeny, genetic distance, coalescence methods). Genetic distance (ABGD, SPN), a coalescence method (BPP), and the multilocus phylogeny supported 22-25 different species of Vasconcellea, including the following five new species from northern Peru: V. badilloi sp. nov., V. carvalhoae sp. nov., V. chachapoyensis sp. nov., V. pentalobis sp. nov., and V. peruviensis sp. nov. Genetic markers that gave better resolution for distinguishing species were ITS and trnL-trnF. Phylogenetic diversity and DNA-species delimitation methods could be used to discover taxa within traditionally defined species.

The Production and Quality of Different Varieties of Papaya Grown under Greenhouse in Short Cycle in Continental Europe.

In Europe, papaya consumption is growing due to its nutritional properties. The proximity of consumer markets to Southeast Spain allows fruits to be harvested at a more advanced stage of maturity compared to exporting countries from outside Europe, a timeline which improves the quality of the papaya. Experiments have been carried out to assess the adaptation of papaya to protected cropping systems (under greenhouse) in the region. In this paper, we showed the results obtained in an experiment with five varieties, taking the most cultivated variety as control, which was grafted on its own female rootstock, in addition to another four new varieties that were introduced. Transplanting was made with early sex-identified plants in the nursery. Cultivation was developed in a 446-day cycle, almost 15 months and fruits were always harvested from the soil, due to the height that the plant reached in that period. The best yield parameters and fruit characteristics were obtained from hermaphrodite Intenzza papaya grafted on female papaya rootstock, although there were also other varieties which gave results that made possible its cultivation under this production system.

The high content of ß-carotene present in orange-pulp fruits of Carica papaya L. is not correlated with a high expression of the CpLCY-ß2 gene.

We investigated the transcriptional regulation of six genes involved in carotenoid biosynthesis, together with the carotenoid accumulation during postharvest ripening of three different papaya genotypes of contrasting pulp color. Red-pulp genotype (RPG) showed the lowest content of yellow pigments (YP), such as ß-cryptoxanthin, zeaxanthin, and violaxanthin, together with the lowest relative expression levels (REL) of CpLCY-ß2 and CpCHX-ß genes. On the contrary, the yellow-pulp genotype (YPG) showed the highest content of YP and the highest REL of CpLCY-ß2 and CpCHX-ß genes. Interestingly, the orange-pulp genotype (OPG) showed intermediate content of YP and intermediate REL of CpLCY-ß2 and CpCHX-ß genes. The highest content of ß-carotene shown by OPG despite having an intermediate REL of the CpLCY-ß2 genes, suggests a post-transcriptional regulation. Thus, the transcriptional level of the genes, directing the carotenoid biosynthesis pathway, can partially explain the accumulation of carotenoids during the postharvest ripening in C. papaya genotypes of contrasting pulp color.

A wild origin of the loss-of-function lycopene beta cyclase (CYC-b) allele in cultivated, red-fleshed papaya (Carica papaya).

PREMISE OF THE STUDY: The red flesh of some papaya cultivars is caused by a recessive loss-of-function mutation in the coding region of the chromoplast-specific lycopene beta cyclase gene (CYC-b). We performed an evolutionary genetic analysis of the CYC-b locus in wild and cultivated papaya to uncover the origin of this loss-of-function allele in cultivated papaya. METHODS: We analyzed the levels and patterns of genetic diversity at the CYC-b locus and six loci in a 100-kb region flanking CYC-b and compared these to genetic diversity levels at neutral autosomal loci. The evolutionary relationships of CYC-b haplotypes were assessed using haplotype network analysis of the CYC-b locus and the 100-kb CYC-b region. KEY RESULTS: Genetic diversity at the recessive CYC-b allele (y) was much lower relative to the dominant Y allele found in yellow-fleshed wild and cultivated papaya due to a strong selective sweep. Haplotype network analyses suggest the y allele most likely arose in the wild and was introduced into domesticated varieties after the first papaya domestication event. The shared haplotype structure between some wild, feral, and cultivated haplotypes around the y allele supports subsequent escape of this allele from red cultivars back into wild populations through feral intermediates. CONCLUSIONS: Our study supports a protracted domestication process of papaya through the introgression of wild-derived traits and gene flow from cultivars to wild populations. Evidence of gene flow from cultivars to wild populations through feral intermediates has implications for the introduction of transgenic papaya into Central American countries.

Efeito dos óleos essenciais sobre a antracnose in vitro e em frutos de mamoeiro / Effect of essential oils on in vitro anthracnose and in fruits of the papaya plant

RESUMO As doenças pós-colheita do mamão são as principais responsáveis pelas perdas que ocorrem durante esse processo. A antracnose é uma doença causada pelo fungo Colletotrichum gloeosporioides. Esse patógeno tem a capacidade de se estabelecer no fruto imaturo, permanecendo em estado latente até que as condições se tornem favoráveis ao seu desenvolvimento. O presente estudo teve como objetivo determinar o efeito fungistático dos óleos essenciais sobre o fungo C.gloeosporioides. Os experimentos foram realizados no laboratório de Fitossanidade da Universidade Estadual de Mato Grosso do Sul, Unidade Universitária de Cassilândia, foram três etapas, quais sejam: I efeito dos óleos essenciais sobre a germinação de conídios, II- ação in vitro dos óleos sobre micélios fúngicos e III- efeito dos óleos essenciais sobre o fungo em frutos do mamoeiro. Para as três fases o delineamento experimental utilizado foi inteiramente casualizado, em esquema fatorial 6 X 5, (óleos essenciais de alecrim, menta, capim-limão, anis, árvore-chá e canela) e cinco concentrações (0 μL, 10 μL, 30 μL, 50 μL, 100 μL), com 5 repetições. A germinação de conídios foi afetada drasticamente pelos óleos de menta e árvore-chá. O efeito fungitóxico do óleo de menta foi confirmado nos experimentos subsequentes, nos quais essa substância na concentração de 100 µL inibiu completamente o crescimento micelial invivo e in vitro do fungo C. gloeosporioides. Os óleos de alecrim e árvore-chá também afetam o crescimento micelial in vivo e in vitro desse fungo, embora em menor intensidade.

ABSTRACT Post-harvest papaya diseases are primarily responsible for the losses suffered during this process. Anthracnose is a disease caused by the Colletotrichum gloeosporioides fungus. This pathogen has the ability to settle in the immature fruit, remaining dormant until conditions become favorable for its development. The purpose of this study was to determine the fungicidal effect of essential oils on the C. gloeosporioides fungus. The experiments were undertaken in the Plant Health laboratory of the State University of Mato Grosso do Sul, at the University Unit of Cassilandia, where an evaluation was performed in three stages, namely: I) the effect of essential oils on spore germination, II) the in vitro action of oils on mycelium fungi, and III) the effect of essential oils on the fungus in papaya fruit. For the three phases, the experimental design was completely randomized in a 6 x 5 factorial arrangement (essential oils of rosemary, mint, lemongrass, anise, tea tree, and cinnamon) and five concentrations (0 uL, 10 uL, 30 uL, 50 uL, 100 uL) with five replications. The spore germination was drastically affected by the mint and tea tree oils. The fungitoxic effect of peppermint oil was confirmed in subsequent experiments in which this substance at a concentration of 100 uL completely inhibited the mycelial in vivo and in vitro growth of the C. gloeosporioides fungus. The rosemary and tea tree oils also affect the mycelial in vivo and in vitro growth of this fungus, albeit to a lesser degree.

Comparative evaluation of volatiles, phenolics, sugars, organic acids and antioxidant properties of Sel-42 and Tainung papaya varieties.

The present study was designed to determine the phenolic compounds, organic acids, sugars, aroma profiles and antioxidant properties of Sel-42 and Tainung papayas grown in Turkey. High-performance liquid chromatography/electrospray ionisation tandem mass spectrometry (HPLC-ESI-MS/MS) method was used for the phenolic compounds analysis. Twelve phenolic compounds were identified and quantified in the samples. The total phenolic content of Sel-42 was clearly higher than that of Tainung. Protocatechuic acid-hexoside, gallic acid-deoxyhexoside, ferulic acid and chlorogenic acids were the most abundant phenolics in both cultivars. Aroma composition of papaya was analysed by gas chromatography-mass spectrometry (GC-MS). A total of 46 and 42 aroma compounds, including esters, alcohols, terpenes, lactones, acids, carbonyl compounds, and volatile phenols were identified in the Sel-42 and Tainung, respectively. The significant linear correlation was confirmed between the values for the total phenolic content and antioxidant activity of papaya extracts.

Extratos vegetais no controle da antracnose e na conservação da qualidade em frutos de mamoeiro / Plant extracts in controlling anthracnose and quality conservation in papayas fruit

RESUMO O presente trabalho teve como objetivo avaliar a ação antifúngica dos extratos de Allamanda blanchetti A.DC. e Momordica charantia L. no controle de Colletotrichumgloesporioides e qualidade pós-colheita em frutos de mamoeiro da cultivar Sunrise solo. Os experimentos foram conduzidos nos Laboratório de Fitopatologia e Laboratório de Produtos de Origem Animal, CCA-UFPB, Campus.II, Areia-PB. Os frutos foram colhidos no estádio 1 de maturação, em seguida, sanitizados com hipoclorito de sódio a 1% por três minutos e pulverizados com os extratos etanólicos de A. blanchetti e M. charantia nas concentrações: 0, 10, 100, 500 e 1000 µg.mL-1. Os frutos foram distribuídos em bandejas de polipropileno expandido em temperatura ambiente (23±2 °C) e umidade relativa em torno de 70%. Após doze dias de armazenamento, os frutos foram avaliados quanto à incidência natural de C. gloesporioides, severidade, atividade da enzima peroxidase, sólidos solúveis, acidez titulável, relação SS/AT, perda de massa e aparência externa dos frutos. Foi realizada análise de regressão polinomial testando modelos até o terceiro grau para as concentrações dos extratos utilizando análise de variância pelo teste F enquanto as médias foram comparadas pelo teste de Tukey a 5% de probabilidade, através do programa SAS® 9.2. Não se observou diferença estatística quando avaliou-se a incidência natural de C. gloesporioides em frutos de mamoeiro utilizando os extratos de A. blanchetti e M. charantia. Já na avaliação da severidade da antracnose, verificou-se diferença para o tratamento com o extrato de A. blanchetti, que apresentou as menores escalas de notas em relação ao tratamento com a M. charantia. Observou-se atividade da peroxidase (POD) em frutos de mamoeiro nos extratos de A. blanchetti e M. charantia. Os frutos tratados com A. blanchetti apresentaram teor de acidez, com máximo de 0,12 g de ácido cítrico.100 g-1 e no de M. charantiaapresentou aumento para 0,15 g de ácido cítrico.100 g-1. A relação SS/AT diminuiu com o aumento das concentrações dos dois extratos. Os frutos tratados com A. blanchetti apresentaram maior perda de massa durante todo o armazenamento, sendo maior que 10% aos nove dias. Já a perda de massa dos frutos tratados com M. charantia apresentou comportamento linear, sendo superior a 12% aos 12 dias de armazenamento. A cor evoluiu de 1 (verde) para 6 (casca alaranjada) e a aparência dos frutos diminuiu com o decorrer do tempo de 5 (excelente) para 3 (regular) em todas concentrações dos extratos de A. blanchetti e M. charantia. Os extratos de A. blanchetti e M. charantia não reduziram a incidência natural, porém, só o extrato de A. blanchetti foi responsável por reduzir a severidade da antracnose. Ocorreu a atividade da enzima peroxidase em todos os tratamentos avaliados e conservaram a qualidade pós-colheita em frutos até os nove dias de armazenamento.

ABSTRACT This study aimed to evaluate the antifungal effect of the extracts Allamandablanchetti A.DC. and Momordica charantia L. in the control of Colletotrichum gloesporioides in papaya fruits of the Sunrise Solo cultivation, as well as their post-harvest quality. The trials were conducted in the Laboratory of Plant Pathology and Animal Products Laboratory, CCA-UFPB, Campus II, Areia-PB. The fruits were harvested at maturity stage 1, then sanitized with sodium hypochlorite 1% for three minutes and sprayed with ethanol extracts of A. blanchetti and M.charantia at concentrations of 0, 10, 100, 500 and 1000 μg.mL-1.The fruits were distributed in polypropylene trays expanded at room temperature (23±2 °C) and relative humidity around 70%. After twelve days of storage, the fruits were evaluated for natural incidence of C. gloesporioides, severity, peroxidase enzyme activity, soluble solids, titratable acidity, SS/TA ratio, weight loss and external appearance of the fruit. Polynomial regression testing models to the third degree were employed for the extracts of concentrations, using analysis of variance by F test. The averages were compared by Tukey test at 5% probability, through the SAS® 9.2 program. There was no statistical difference observed when we evaluated the natural incidence of C. gloesporioides in the papaya fruit, using extracts from A. blanchetti and M. charantia. In the assessment of the severity of the anthracnose, there was no statistical difference for the treatment with the extract of A. blanchetti, which had smaller scales of notes when compared with the treatment using the M. charantia. It was observed peroxidase activity (POD) in the papaya fruits treated with the extracts of A. blanchetti and M. charantia. The fruits treated with A. blanchetti presented a maximum dose of acidity of 0.12 g g-1 of citric acid.100 g-1. In the fruits treated with M. charantia this increase was of 0.15g of citric acid.100 g-1. The SS/TA ratio decreased with increasing concentrations of the two extracts. The fruits treated with A. blanchettipresented higher weight loss of approximately 10%, since the M. charantia was over 12%. The color evolved from 1 (green) to 6 (Orange Peel) and the appearance of the fruit got worse according to the time, from 5 (excellent) to 3 (regular) in all the concentrations of the extracts of A. blanchetti and M. charantia. The A. blanchetti and M. charantia extracts did not reduce the natural incidence, however, only the A. blanchetti extract was able to decrease the severity of the anthracnose. The peroxidase enzyme presented activity in all treatments and kept the postharvest quality of the fruits in the nine days of storage.

Análise sazonal da produção e da atividade enzimática de látex fresco coletado de frutos de plantas femininas e hermafroditas de mamão (Carica papaya L. ) / Seasonal analysis of yield and enzyme activity of fresh latex collected from fruits of female and hermaphrodite plants of papaya (Carica papaya L. )

RESUMO O látex obtido do fruto de Carica papaya L. (mamão) é de grande importância industrial e farmacêutica. Sua obtenção, através do cultivo dessa espécie, depende de vários fatores ainda pouco estudados. Nesse sentido, o presente trabalho tem por objetivo avaliar e comparar plantas femininas e hermafroditas de C. papaya em relação à produção de látex e sua atividade enzimática em coletas semanais e consecutivas durante o 12º mês de cultivo (verão) e no decorrer do segundo ano de desenvolvimento das plantas, e em mais três coletas abrangendo as demais estações do ano (outono, inverno e primavera). Os resultados mostraram que, em quatro coletas semanais e sucessivas durante o verão, a massa de látex da primeira coleta foi superior às demais para plantas femininas. Plantas hermafroditas tiveram comportamento oscilante para esta variável. Plantas femininas tiveram médias de produção superiores comparadas com as hermafroditas. As médias das atividades enzimáticas desse período se mostraram superiores para plantas hermafroditas. Nas coletas estacionais foi observado que plantas hermafroditas têm um comportamento mais oscilante em relação à produção de látex e atividade enzimática. Plantas femininas produzem, em média, maior massa de látex e são mais constantes na produção. Nas estações avaliadas, observaram-se semelhanças nas médias das atividades enzimáticas entre plantas femininas e hermafroditas.

ABSTRACT The latex obtained from the fruit of Carica papaya L. (papaya) is of great industrial and pharmaceutical importance. Its obtainment, through the cultivation of this specie, depends on several factors still poorly studied. In this sense, this study aims to evaluate and compare hermaphrodite and female plants of C. papaya for the production of latex and its enzymatic activity in weekly and consecutive collections of fruits during the 12th month of cultivation (summer) and during the second year of the plant´s development, in three collections covering the other seasons (autumn, winter and spring). The results showed that, in four successive weekly tapping during the summer, the latex mass of the first collection was superior to the others, for female plants. Hermaphrodite plants had oscillating behavior for this variable. Female plants had higher production averages, compared with the hermaphrodites ones. The averages of the enzymatic activities of this period were higher for hermaphrodite plants. The seasonal tapping showed that hermaphrodite plants had oscillating behavior in the production of latex and enzyme activity. Female plants produced, on average, greater mass and were constant in latex production. In the evaluated seasons, similarities in the averages of the enzymatic activities between female and hermaphrodite plants were observed.

Isolation and characterization of the papaya MADS-box E-class genes, CpMADS1 and CpMADS3, and a TM6 lineage gene CpMADS2.

Papaya (Carica papaya L.) plants are polygamous, with female, male, and hermaphroditic flowers. To understand the roles of MADS-box genes in flower development and sex determination, we cloned cDNAs of E-class genes CpMADS1 and CpMADS3 and a TM6 lineage of the B-class gene CpMADS2 from young flower buds of papaya. Reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR analyses revealed that CpMADS1 and CpMADS3 were preferentially expressed in the carpel and also in petals and stamens. CpMADS2 was expressed in both petals and stamens early during floral development. Comparison of 10 papaya genotypes of 5 different sex phenotypes - hermaphrodite, male, female, progeny-all-hermaphrodite, and progeny-all-male - by Southern blot analysis of genomic DNAs with probes of the 3 genes revealed similar restriction patterns and copy number, suggesting a low relationship of the 3 CpMADS genes with sex expression of papaya plants at the genomic level.

Clonagem e análise da expressão de genes de proteínas de mamão papaia com atividade inibitória sobre poligalacturonases fúngicas / Cloning and expression analysis of papaya genes encoding proteins with inhibitory activity against fungal polygalacturonases

As proteínas inibidoras de poligalacturonases (PGIPs) presentes na parede celular são capazes de limitar o potencial destrutivo da poligalacturonase (PG) fúngica e, assim, constituem um tipo importante dentre os diversos sistemas de defesa do tecido vegetal frente à infecção fúngica. No mamão, o ataque fitopatogênico é o principal causador de danos pós-colheita, e sua alta susceptibilidade pode estar relacionada com a baixa eficácia ou pouca abundância dos meios de defesa anti-fitopatogênica. Uma vez que isso pode estar relacionado com as PGIPs e nada se conhece sobre o papel dessas proteínas nesse fruto, o objetivo do trabalho foi clonar os genes das PGIPs de mamoeiro e definir seu padrão de expressão em diferentes órgãos e tecidos e ao longo do amadurecimento. Para tanto, foram identificadas no genoma do mamoeiro, a partir de critérios que definem a identidade de uma PGIP, duas prováveis sequências dentre 13 candidatas iniciais. Ambas foram clonadas a partir das sequências genômicas e de cDNA, sequenciadas e sua identidade confirmada, sendo denominadas Cppgip4 e Cppgip6. As análises de expressão relativa em diversos tecidos e idades fisiológicas do mamoeiro demonstraram que os dois genes apresentaram diminuição da expressão com o desenvolvimento dos frutos, sendo que com a polpa apresentou redução dos níveis de expressão relativa de Cppgip4 em até 18 vezes dos 30 dias pós-antese (DPA) ao 9 dias pós-colheita (DPC). Na casca também houve redução significativa da expressão com o desenvolvimento. Para a expressão absoluta, nos frutos, sementes, caules, raízes e folhas, o número de cópias de ambos os transcritos decresceu com o desenvolvimento, sendo cerca de cem mil vezes mais abundante para Cppgip6 que para Cppgip4. As tentativas de expressão de proteínas recombinantes em Pichia pastoris não geraram resultado positivo, provavelmente em virtude das condições ideais de indução ainda não terem sido estabelecidas corretamente para o ensaio. A atividade de PGIPs extraídas diretamente do tecido foi medida por análise de difusão em ágar empregando pectinase de Aspergillus niger e revelou uma tendência à diminuição da porcentagem de inibição à medida que os frutos se desenvolveram, em concordância com os resultados da análise por qPCR. O conjunto de resultados sugere que a expressão varia com o estádio de desenvolvimento do fruto e é tecido-específica, possivelmente em resposta à diferente susceptibilidade dos tecidos ao ataque fitopatogênico, indicando que menores níveis de transcritos e atividade no amadurecimento, período de maior susceptibilidade, poderiam sinalizar para a regulação do processo degradativo marcando o início da senescência

Polygalacturonase inhibiting proteins (PGIPs) present in plant cell walls are able to inhibit the destructive action of fungal polygalacturonase (PG). In this way, they constitute an important type of plant defense system against fungal infections. In papaya fruit, the pathogenic attack is the main cause of post harvesting loss, and its high susceptibility may be related to the low efficiency or low abundance of anti-phytopathogenic defense. Since this fact could be related to PGIPs expression and little is known about the response of these proteins in the fruit, the aim of the present work was to clone the genes of PGIPs papaya fruit and set their expression pattern in different organs and tissues throughout fruit ripening. Thus, two probable PGIP sequences among 13 initial candidates were identified in the papaya genome by using specific criteria. Both sequences were cloned from cDNA and genomic samples, sequenced and confirmed its identity, and then being named Cppgip4 and Cppgip6. Analysis of relative expression in various tissues at different physiological stages demonstrated that both genes were down regulated during fruit development. The relative expression levels of Cppgip4 in papaya pulp was reduced by 18 times from the 30 days post-anthesis (DPA) to the 9 days post-harvest (DPH). Similarly, gene expression in papaya peel was significant down regulated during fruit development. Absolute expression analysis revealed gene expressions in the fruit pulp, seed, stem, root and leaf were also down regulated within development. Moreover, Cppgip6 gene expression was a hundred thousand times more abundant than Cppgip4. The recombinant protein expression in Pichia pastoris did not result positive, probably because of the ideal conditions of induction have not been properly established the yet. The activity of PGIPs extracted directly from the tissue was measured by the agar diffusion assay using pectinase from Aspergillus niger and showed decrease of inhibition during fruit developed in accordance with the results of the qPCR analysis. Based on the results it is possible to suggest the expression of these genes varies temporally with the developmental stage of the fruit and is tissue-specific, possibly in response to the different susceptibility of tissues to pathogenic attack. In addition, the lowest levels of PGIP expression were achieved at the fruit ripening, when the susceptibility to fungal infection is high and could signal for regulating the degradation process characterized by the onset of senescence

Evolutionary relationship and structural characterization of the EPF/EPFL gene family.

EPF1-EPF2 and EPFL9/Stomagen act antagonistically in regulating leaf stomatal density. The aim of this study was to elucidate the evolutionary functional divergence of EPF/EPFL family genes. Phylogenetic analyses showed that AtEPFL9/Stomagen-like genes are conserved only in vascular plants and are closely related to AtEPF1/EPF2-like genes. Modeling showed that EPF/EPFL peptides share a common 3D structure that is constituted of a scaffold and loop. Molecular dynamics simulation suggested that AtEPF1/EPF2-like peptides form an additional disulfide bond in their loop regions and show greater flexibility in these regions than AtEPFL9/Stomagen-like peptides. This study uncovered the evolutionary relationship and the conformational divergence of proteins encoded by the EPF/EPFL family genes.

Use of morpho-agronomic traits and DNA profiling for classification of genetic diversity in papaya.

We examined the genetic diversity of papaya (Carica papaya) based on morpho-agronomic and molecular data. Twenty-seven genotypes grown in Brazil were analyzed with 11 AFLP primer combinations, 23 ISSR markers, 22 qualitative, and 30 quantitative descriptors. For the joint analyses, we used the Gower algorithm (Joint Gower) and the average value of the individual dissimilarity matrix for each type of data (Average-Joint Gower); 359 AFLP and 52 ISSR polymorphic bands were found. Approximately 29.2 and 7.7% of the AFLP and ISSR bands, respectively, were genotype-specific and may therefore be used for papaya variety protection. Although there was a significant correlation between the qualitative and quantitative descriptor dissimilarity matrices (r = 0.43), the morpho-agronomic data were not highly correlated with the molecular data. Moreover, correlation between AFLP and ISSR dissimilarity matrices was nearly null (r = -0.01). Joint Gower analysis of all data showed high correlations, especially for AFLP markers, most likely due to the larger number of bands, generating a strong bias in the diversity estimates. The Average-Joint Gower analysis allowed a better balance between the correlations for the continuous and the discrete variables. The results generated by clustering analysis distinguished 5 genetically distinct groups. While we found that papaya genotypes are significantly variable for many traits, we observed that Average-Joint Gower analysis allowed for genotype clustering based on the most widely used criterion for classifying papaya genotypes, which is fruit type ('Formosa' or 'Solo'). This information helps provide an accurate estimate of the genetic diversity and structure of papaya germplasm, which will be used for further breeding strategies.

A dated phylogeny of the papaya family (Caricaceae) reveals the crop's closest relatives and the family's biogeographic history.

Papaya (Carica papaya) is a crop of great economic importance, and the species was among the first plants to have its genome sequenced. However, there has never been a complete species-level phylogeny for the Caricaceae, and the crop's closest relatives are therefore unknown. We investigated the evolution of the Caricaceae based on sequences from all species and genera, the monospecific Carica, African Cylicomorpha with two species, South American Jacaratia and Vasconcellea with together c. 28 species, and Mexican/Guatemalan Jarilla and Horovitzia with four species. Most Caricaceae are trees or shrubs; the species of Jarilla, however, are herbaceous. We generated a matrix of 4711 nuclear and plastid DNA characters and used maximum likelihood (ML) and Bayesian analysis to infer species relationships, rooting trees on the Moringaceae. Divergence times were estimated under relaxed and strict molecular clocks, using different subsets of the data. Ancestral area reconstruction relied on a ML approach. The deepest split in the Caricaceae occurred during the Late Eocene, when the ancestor of the Neotropical clade arrived from Africa. In South America, major diversification events coincide with the Miocene northern Andean uplift and the initial phase of the tectonic collision between South America and Panama resulting in the Panamanian land bridge. Carica papaya is sister to Jarilla/Horovitzia, and all three diverged from South American Caricaceae in the Oligocene, 27 (22-33) Ma ago, coincident with the early stages of the formation of the Panamanian Isthmus. The discovery that C. papaya is closest to a clade of herbaceous or thin-stemmed species has implications for plant breeders who have so far tried to cross papaya only with woody highland papayas (Vasconcellea).

Towards improved reconstruction of ancestral gene order in angiosperm phylogeny.

Whole genome doubling (WGD), a frequent occurrence during the evolution of the angiosperms, complicates ancestral gene order reconstruction due to the multiplicity of solutions to the genome halving process. Using the genome of a related species (the outgroup) to guide the halving of a WGD descendant attenuates this problem. We investigate a battery of techniques for further improvement, including an unbiased version of the guided genome halving algorithm, reference to two related genomes instead of only one to guide the reconstruction, use of draft genome sequences in contig form only, incorporation of incomplete sets of homology correspondences among the genomes, and addition of large numbers of "singleton" correspondences. We make use of genomic distance, breakpoint reuse rate, dispersion of sets of alternate solutions, and other means to evaluate these techniques, and employ the papaya (Carica papaya) and grapevine (Vitis vinifera) genomes to reconstruct the pre-WGD ancestor of poplar (Populus trichocarpa), as well as an early rosid ancestor. A significant result is that the papaya genome has rearranged at a greater rate from the rosid ancestor than phylogenetic relationships would predict.

Estudio nutritivo, bioquímico y toxicológico del fruto de la carica stipulata V. M. Badillo (papayita olorosa) / Nutritional, biochemical and toxicological study of the fruit of carica stipulata V. M. Badillo (aromatic small papaya)

El estudio de Carica stipulata V. M. Badillo nos permitió determinar su composición química y nutritiva demostrando que es una fuente de minerales como el hierro (2.65 mg/100 g) y magnesio (217.22 mg/100 g), además de la vitamina C (45 mg/100 g) y la pectina (1 mg/100 g). Muestra un bajo valor calórico de 25,06 KCal/100 g de alimentos. Presenta además flavonoles y ácidos fenólicos principalmente ferúlico (25.26 ppm) y cafeico (15.49 ppm). El estudio bioquímico en ratas adultas que ingirieron la pulpa liofilizada del fruto mostró un aumento significativo de glucosa sanguínea en el grupo de la dosis de 600 y 300 mg/kg de peso con respecto al grupo control confirmando que la papaya aumenta los niveles séricos de glucosa. Se encontró además una disminución significativa en los niveles de colesterol sérico siendo mayor en el grupo de 300 mg/kg de peso. Los niveles de proteína subieron significativamente especialmente en el grupo de dosis de 600 mg/kg de peso, habiendo un aumento significativo de globulina especialmente en los grupos de la dosis de 600 y 300 mg/Kg de peso respectivamente. Los resultados de toxicidad mostraron que es prácticamente no tóxica con un valor de DL50 de 7629.4 mg/kg de peso corporal.

The study of Carica stipulata V. M. Badillo allowed us to determine its chemical and nutritional composition, demonstrating that it is a mineral source of iron (2.65 mg/100 g) and magnesium (217.22 mg/100 g), in addition to vitamin e (45 mg/100 g) and pectin (1 mg%), it Shows a low caloric value of 25,06 Kcal/100 grams of food. It presents flavonoids and phenolic acids mainly ferulic (25.26 ppm) and caffeic (15.49 ppm). The biochemical study in adult rats that ingested the freeze-dried pulp of the fruit showed a significant increase in blood glucose specially in the group of dose of 600 and 300 mg/kg of weight with respect to the control group confirming that papaya increases the serum levels of glucose. There was a significant decrease in cholesterol levels, being more significant in the group of dose of 300 mg/kg of weight. The protein levels raised significantly specially in the group of dose of 600 mg/kg, finding a significant increase of globulin, mainly in the group that received doses of 600 and 300 mg/kg respectively. The toxicity results showed that it is practically nontoxic with a value of DL50 of 7629.4 mg/kg of corporal weight.

Hybrid 'Sinta' papaya exhibits unique ACC synthase 1 cDNA isoforms.

Five ripening-related ACC synthase cDNA isoforms were cloned from 80% ripe papaya cv. 'Sinta' by reverse transcription-PCR using gene-specific primers. Clone 2 had the longest transcript and contained all common exons and three alternative exons. Clones 3 and 4 contained common exons and one alternative exon each, while clone 1, the most common transcript, contained only the common exons. Clone 5 could be due to cloning artifacts and might not be a unique cDNA fragment. Thus, there are only four isoforms of ACC synthase mRNA. Southern blot analysis indicates that all five clones came from only one gene existing as a single copy in the 'Sinta' papaya genome. Multiple sequence alignment indicates that the four isoforms arise from a single gene, possibly through alternative splicing mechanisms. All the putative alternative exons were present at the 5'-end of the gene comprising the N-terminal region of the protein. 'Sinta' ACC synthase cDNAs were of the capacs 1 type and are most closely related to a 1.4 kb capacs 1-type DNA (AJ277160) from Eksotika papaya. No capacs 2-type cDNAs were cloned from 'Sinta' by RT-PCR. This is the first report of possible alternative splicing mechanism in ripening-related ACC synthase genes in hybrid papaya, possibly to modulate or fine-tune gene expression relevant to fruit ripening.

Detection of genetically modified organisms by electrochemiluminescence PCR method.

With the development of biotechnology, more and more genetically modified organisms (GMOs) have entered commercial market. Because of the safety concerns, detection and characterization of GMOs have attracted much attention recently. In this study, electrochemiluminescence polymerase chain reaction (ECL-PCR) combined with hybridization technique was applied to detect the GMOs in genetically modified (GM) soybeans and papayas for the first time. Whether the soybeans and the papayas contain GM components was discriminated by detecting the Cauliflower mosaic virus 35S (CaMV35S) promoter. The experiment results show that the detection limit for CaMV35S promoter is 100 fmol, and the GM components can be clearly identified in GM soybeans and papayas. The technique may provide a new means in GMOs detection due to its simplicity and high efficiency.

Phylogenetic analysis of the highland papayas ( Vasconcellea) and allied genera (Caricaceae) using PCR-RFLP.

The chloroplast and mitochondrial DNA diversity of 61 genotypes belonging to 18 Vasconcellea species, the so-called highland papayas, was studied by PCR-RFLP analysis of two non-coding cpDNA regions ( trnM- rbcL and trnK1- trnK2) and one non-coding mtDNA region ( nad4/1- nad4/2). This sample set was supplemented with six genotypes belonging to three other Caricaceae genera: the monotypic genus Carica, including only the cultivated papaya, and the genera Jacaratia and Cylicomorpha. Moringa ovalifolia was added as an outgroup species. The PCR-amplified cpDNA regions were digested with 18 restriction endonucleases, the mtDNA region with 11. A total of 22 point mutations and four insertion/deletions were scored in the sample. A higher level of interspecific variation was detected in the two cpDNA regions in comparison to the analysis of the mtDNA. Wagner parsimony and Neighbor-Joining analysis resulted in dendrograms with similar topologies. PCR-RFLP analysis supported the monophyly of Caricaceae, but among the 26 mutations scored, an insufficient number of markers discriminated between the different Caricaceae genera included in this study. Hence the inference of the intergeneric relationships within Caricaceae was impossible. However, some conclusions can be noted at a lower taxonomic level. The Caricaceae species were divided into two lineages. One group included only Vasconcellea spp., whereas the second included the remaining Vasconcellea spp., together with the papaya genotypes and those from the other Caricaceae genera. This may indicate a higher level of inter-fertility for the Vasconcellea species from the latter clade in interspecific crossings with papaya. The putative progenitors of the natural sterile hybrid V. x heilbornii, i.e. V. stipulata and V. cundinamarcensis, were only distantly related to V. x heilbornii. This indicates that probably none of these species was involved as the maternal progenitor in the origin of V. x heilbornii. Surprisingly, V. x heilbornii had organellar genome patterns identical with V. weberbaueri, suggesting a possible involvement of this species in the origin of V. x heilbornii. On the basis of discrepancy between morphological traits and the cpDNA profiles of some pairs of Vasconcellea species, we believe that besides V. x heilbornii, some other species have originated through interspecific hybridization. A reticulate evolution for Vasconcellea has therefore been suggested. Finally, intraspecific cpDNA variation was detected in V. microcarpa, thus providing molecular evidence for the high diversity previously indicated by morphological observations.