Population Structure of Pseudocercospora fijiensis in Costa Rica Reveals Shared Haplotype Diversity with Southeast Asian Populations.

Pseudocercospora fijiensis is the causal pathogen of black Sigatoka, a devastating disease of banana that can cause 20 to 80% yield loss in the absence of fungicides in banana crops. The genetic structure of populations of P. fijiensis in Costa Rica was examined and compared with Honduran and global populations to better understand migration patterns and inform management strategies. In total, 118 isolates of P. fijiensis collected from Costa Rica and Honduras from 2010 to 2014 were analyzed using multilocus genotyping of six loci and compared with a previously published global dataset of populations of P. fijiensis. The Costa Rican and Honduran populations shared haplotype diversity with haplotypes from Southeast Asia, Oceania, and the Americas but not Africa for all but one of the six loci studied. Gene flow and shared haplotype diversity was found in Honduran and Costa Rican populations of the pathogen. The data indicate that the haplotypic diversity observed in Costa Rican populations of P. fijiensis is derived from dispersal from initial outbreak sources in Honduras and admixtures between genetically differentiated sources from Southeast Asia, Oceania, and the Americas.

Micromorfología de los foliolos de pejibaye Bactris gasipaes (Arecaceae) var. Diamantes-10 / Micromorphology of pejibaye leaflets Bactris gasipaes (Arecaceae) var. diamonds-10

ResumenBactris gasipaes se cultiva ampliamente para el aprovechamiento del palmito y el consumo de los frutos En este trabajo se realizó una descripción de las características micro morfológicas de los foliolos de plantas adultas de la variedad sin espinas Diamantes-10 de B. gasipaes, recolectadas en la Estación Experimental Diamantes en Guápiles, Costa Rica. Se utilizó microscopía de luz, electrónica de barrido y de transmisión. Para el estudio se recolectaron 25 foliolos y se evidenció abundante cera epicuticular en la superficie adaxial y abaxial. La epidermis está compuesta de células isodiamétricas y las células de la hipodermis tienen forma rectangular y son de mayor tamaño que las de la epidermis, también se observaron células buliformes. Los foliolos son anfiestomáticos en las áreas intercostales y alternan con bandas de células epidérmicas. El mayor número de estomas se encuentra en la superficie abaxial. Estos foliolos presentan tres tipos de tricomas. El parénquima presenta varios estratos no muy bien definidos con presencia de astroesclereidas. También se observan haces de fibras entre el parénquima que forman grupos de células muy compactas. La vena central presenta varios haces vasculares, algunos dispuestos de forma continua y todos ellos se encuentran rodeados de una vaina de tejido esclerotizado, algunas de estas fibras presentaron protoplastos vivos. Todas las venas menores presentan la misma anatomía de los haces vasculares de la vena central. Los vasos del protoxilema y metaxilema tienen paredes laterales con ornamentaciones escalariformes. El floema se ubica hacia la superficie adaxial de la vena y en el corte se observaron las células cribosas y células compañeras rodeadas de células de parénquima y fibras. Las células compañeras presentaron plasmodesmatas ramificados unidos a un elemento criboso y en ellos se evidenció cuerpos de proteína, llamada proteína-P. La diferencia principal en la anatomía de los foliolos de la var. Diamantes-10 radica en la ausencia de espinas, pero en el resto de la morfología no parece presentar cambios morfológicos con el resto de las variedades de B. gasipaes K.

Abstract:Bactris gasipaes is widely cultivated for the consumption of palm hearts and fruits. The present work describes the micro morphological characteristics of leaflets from adult plants of B. gasipaes, thornless variety Diamantes-10, collected in the Diamantes Experimental Station in Guápiles, Costa Rica. We collected 25 leaflets and analyses were performed with a combination of microscopy techniques: light microscopy, scanning electron microscopy and transmission electron microscopy to study their structure. Our results showed that leaflets have abundant epicuticular wax on adaxial and abaxial surfaces. Analyses from the epidermis indicated that it is composed of isodiametric cells, and it is also evident that hypodermis cells have rectangular shape and are larger than the other epidermal cells. We observed stomata on both surfaces, but they were more abundant in the abaxial surface. On the other hand, the epidermis showed the presence of trichomes with three different morphologies. In the parenchyma, cells are large and not well defined, and we observed the presence of astroesclereids, and compact groups of fiber bundles between parenchyma cells. The central vein has several vascular bundles, arranged in a continuous manner, and they are surrounded by sclerotic tissue; some of these fibers presented live protoplasts. All minor veins showed the same anatomy as the central vein. In these veins, the vessel elements of protoxylem and metaxylem showed scalariform ornaments on their walls. Phloem is located towards the adaxial surface of the vein and we observed sieve and companion cells surrounded by fibers and parenchyma cells. The companion cells presented branched plasmodesmata attached to a sieve element, and in these elements we found protein bodies called P-protein. The main anatomical difference in the leaflets of the var. Diamantes-10, compared to the other varieties of B. gasipaes K, is the lack of thorns; the other morphological features seem to be conserved. Rev. Biol. Trop. 64 (3): 1273-1285. Epub 2016 September 01.

[Micromorphology of pejibaye leaflets Bactris gasipaes (Arecaceae) var. diamonds-10]. / Micromorfología de los foliolos de pejibaye Bactris gasipaes (Arecaceae) var. Diamantes-10.

Bactris gasipaes is widely cultivated for the consumption of palm hearts and fruits. The present work describes the micro morphological characteristics of leaflets from adult plants of B. gasipaes, thornless variety Diamantes-10, collected in the Diamantes Experimental Station in Guápiles, Costa Rica. We collected 25 leaflets and analyses were performed with a combination of microscopy techniques: light microscopy, scanning electron microscopy and transmission electron microscopy to study their structure. Our results showed that leaflets have abundant epicuticular wax on adaxial and abaxial surfaces. Analyses from the epidermis indicated that it is composed of isodiametric cells, and it is also evident that hypodermis cells have rectangular shape and are larger than the other epidermal cells. We observed stomata on both surfaces, but they were more abundant in the abaxial surface. On the other hand, the epidermis showed the presence of trichomes with three different morphologies. In the parenchyma, cells are large and not well defined, and we observed the presence of astroesclereids, and compact groups of fiber bundles between parenchyma cells. The central vein has several vascular bundles, arranged in a continuous manner, and they are surrounded by sclerotic tissue; some of these fibers presented live protoplasts. All minor veins showed the same anatomy as the central vein. In these veins, the vessel elements of protoxylem and metaxylem showed scalariform ornaments on their walls. Phloem is located towards the adaxial surface of the vein and we observed sieve and companion cells surrounded by fibers and parenchyma cells. The companion cells presented branched plasmodesmata attached to a sieve element, and in these elements we found protein bodies called P-protein. The main anatomical difference in the leaflets of the var. Diamantes-10, compared to the other varieties of B. gasipaes K, is the lack of thorns; the other morphological features seem to be conserved.

Primer caso de onicomicosis causada por Cylindrocarpon lichenicola, en un paciente inmunosuprimido en Costa Rica / First case of onychomycosis caused by Cylindrocarpon lichenicola in an immunosuppressed patient un Costa Rica

Antecedentes: la onicomicosis es una infección fúngica de las uñas, que puede ser causada por dermatofitos, levaduras y hongos filamentosos no dermatofitos. El objetivo del presente trabajo fue identificar-morfológica y molecularmente- un aislamiento realizado a partir de la uña del dedo medio de la mano derecha de un paciente inmunosuprimido con trasplante renal. Métodos: a la muestra se le realizó un examen microscópico directo en KOH 40 por ciento, y las características de la colonia fueron evaluadas en medio de cultivo Agar Sabouraud Glucosado y Agar Mycosel. El hongo aislado se identificó molecularmente mediante la amplificación, por reacción en cadena de la polimerasa (PCR) y secuenciación de la región espaciadora transcrita (ITS) de los genes ribosomales y de una porción del gen del factor de elongación 1 alfa (EF-1). La secuencia de nucleótidos de cada región fue comparada con las secuencias publicadas en la base de datos del Anco de Genes del Centro Nacional para la Información Biotecnológica (NCBI). Resultados: las características macro y microscópicas de la colonia concordaron con las directrices para Cylindrocarpon lichenicola (sinónimo Fusarium lichenicola). La secuencia de nucleótidos, tanto de la región ITS (544 pb) como de la porción del gen EF-1alfa (710 pb), presentó un 100 por ciento de similitud con secuencias de F. lichenicola. Conclusión: se identificó por primera vez en Costa Rica C. lichenicola asociado con onicomicosis en un paciente inmunosuprimidos. Este tipo de pacientes deben ser instruidos sobre las posibles fuentes de agentes infecciosos, particularmente los riesgos que pueden generar la jardinería y la agricultura...

Phylogenetic relationships of Phytophthora andina, a new species from the highlands of Ecuador that is closely related to the Irish potato famine pathogen Phytophthora infestans.

Phylogenetic relationships of Phytophthora infestans sensu lato in the Andean highlands of South America were examined. Three clonal lineages (US-1, EC-1, EC-3) and one heterogeneous lineage (EC-2) were found in association with different host species in genus Solanum. The EC-2 lineage includes two mitochondrial (mtDNA) haplotypes, Ia and Ic. Isolates of P. infestans sensu lato EC-2 fit the morphological description of P. infestans but are different from any genotypes of P. infestans described to date. All isolates of P. infestans sensu lato from Ecuador were amplified by a P. infestans specific primer (PINF), and restriction fragment length patterns were identical in isolates amplified with ITS primers 4 and 5. The EC-1 clonal lineage of P. infestans sensu lato from S. andreanum, S. columbianum, S. paucijugum, S. phureja, S. regularifolium, S. tuberosum and S. tuquerense was confirmed to be P. infestans based on sequences of the cytochrome oxidase I (cox I) gene and intron 1 of ras gene. The EC-2 isolates with the Ic haplotype formed a distinct branch in the same clade with P. infestans and P. mirabilis, P. phaseoli and P. ipomoeae for both cox I and ras intron 1 phylogenies and were identified as the newly described species P. andina. Ras intron 1 sequence data suggests that P. andina might have arisen via hybridization between P. infestans and P. mirabilis.

An Andean origin of Phytophthora infestans inferred from mitochondrial and nuclear gene genealogies.

Phytophthora infestans (Mont.) de Bary caused the 19th century Irish Potato Famine. We assessed the genealogical history of P. infestans using sequences from portions of two nuclear genes (beta-tubulin and Ras) and several mitochondrial loci P3, (rpl14, rpl5, tRNA) and P4 (Cox1) from 94 isolates from South, Central, and North America, as well as Ireland. Summary statistics, migration analyses and the genealogy of current populations of P. infestans for both nuclear and mitochondrial loci are consistent with an "out of South America" origin for P. infestans. Mexican populations of P. infestans from the putative center of origin in Toluca Mexico harbored less nucleotide and haplotype diversity than Andean populations. Coalescent-based genealogies of all loci were congruent and demonstrate the existence of two lineages leading to present day haplotypes of P. infestans on potatoes. The oldest lineage associated with isolates from the section Anarrhichomenun including Solanum tetrapetalum from Ecuador was identified as Phytophthora andina and evolved from a common ancestor of P. infestans. Nuclear and mitochondrial haplotypes found in Toluca Mexico were derived from only one of the two lineages, whereas haplotypes from Andean populations in Peru and Ecuador were derived from both lineages. Haplotypes found in populations from the U.S. and Ireland was derived from both ancestral lineages that occur in South America suggesting a common ancestry among these populations. The geographic distribution of mutations on the rooted gene genealogies demonstrate that the oldest mutations in P. infestans originated in South America and are consistent with a South American origin.

Mitochondrial genome sequences and molecular evolution of the Irish potato famine pathogen, Phytophthora infestans.

The mitochondrial genomes of haplotypes of the Irish potato famine pathogen, Phytophthora infestans, were sequenced. The genome sizes were 37,922, 39,870 and 39,840 bp for the type Ia, IIa and IIb mitochondrial DNA (mtDNA) haplotypes, respectively. The mitochondrial genome size for the type Ib haplotype, previously sequenced by others, was 37,957 bp. More than 90% of the genome contained coding regions. The GC content was 22.3%. A total of 18 genes involved in electron transport, 2 RNA-encoding genes, 16 ribosomal protein genes and 25 transfer RNA genes were coded on both strands with a conserved arrangement among the haplotypes. The type I haplotypes contained six unique open reading frames (ORFs) of unknown function while the type II haplotypes contained 13 ORFs of unknown function. Polymorphisms were observed in both coding and non-coding regions although the highest variation was in non-coding regions. The type I haplotypes (Ia and Ib) differed by only 14 polymorphic sites, whereas the type II haplotypes (IIa and IIb) differed by 50 polymorphic sites. The largest number (152) of polymorphic sites was found between the type IIb and Ia haplotypes. A large spacer flanked by the genes coding for tRNA-Tyr (trnY) and the small subunit RNA (rns) contained the largest number of polymorphic sites and corresponds to the region where a large indel that differentiates type II from type I haplotypes is located. The size of this region was 785, 2,666 and 2,670 bp in type Ia, IIa and IIb haplotypes, respectively. Among the four haplotypes, 81 mutations were identified. Phylogenetic and coalescent analysis revealed that although the type I and II haplotypes shared a common ancestor, they clearly formed two independent lineages that evolved independently. The type II haplotypes diverged earlier than the type I haplotypes. Thus our data do not support the previous hypothesis that the type II lineages evolved from the type I lineages. The type I haplotypes diverged more recently and the mutations associated with the evolution of the Ia and Ib types were identified.