A preliminary pharmacophylogenetic study of Solanaceae medicinal plants containing tropane alkaloids / 中国中药杂志

The Solanaceae plants distributed in China belong to 105 species and 35 varietas of 24 genera. Some medicinal plants of Solanaceae are rich in tropane alkaloids(TAs), which have significant pharmacological activities. In this paper, the geographical distribution, chemical components, traditional therapeutic effect, pharmacological activities, and biosynthetic pathways of TAs in Solanaceous plants were summarized. Besides, the phylogeny of medicinal plants belonging to Solanaceae was visualized by network diagram. Fourteen genera of Solanaceae plants in China contain TAs and have medical records. TAs mainly exist in Datura, Anisodus, Atropa, Physochlaina, and Hyoscyamus. The TAs-containing species were mainly concentrated in Southwest China, and the content of TAs was closely related to plant distribution area and altitude. The Solanaceae plants containing TAs mainly have antispasmodic, analgesic, antiasthmatic, and antitussive effects. Modern pharmacological studies have proved the central sedative, pupil dilating, glandular secretion-inhibiting, and anti-asthma activities of TAs. These pharmacological activities provide a reasonable explanation for the traditional therapeutic efficacy of tropane drugs. In this paper, the geographical distribution, chemical components, traditional therapeutic effect, and modern pharmacological activities of TAs-containing species in Solanaceae were analyzed for the first time. Based on these data, the genetic relationship of TAs-containing Solanaceae species was preliminarily discussed, which provided a scientific basis for the basic research on TAs-containing solanaceous species and was of great significance for the development of natural medicinal plant resources containing TAs.

Efecto del termofosfato sobre el crecimiento y producción de papa (Solanum tuberosum L.) / Effect of thermophosphate on the growth and production of potatoes (Solanum tuberosum L.)

RESUMEN El cultivo de papa es uno de los más representativos de los Andes colombianos. El cultivo tiene altos costos de producción, entre los cuales, la fertilización representa un 36% del total. Los avances tecnológicos en nutrición vegetal han permitido el desarrollo de nuevos compuestos, que generan una mayor asimilación y disponibilidad de los elementos nutritivos, durante los periodos productivos. Dentro de las diferentes fuentes de fósforo empleadas en Colombia, está como alternativa, el termofosfato o fosfato térmico. Por esto, el objetivo del presente trabajo fue evaluar la aplicación del termofosfato en el cultivo de papa variedad 'Superior', en condiciones de campo, en el municipio de Villa Pinzón, Cundinamarca. Se empleó un diseño completamente al azar, con 4 tratamientos, que correspondieron a diferentes porcentajes de fosfato térmico 100% fosfato diamónico (DAP) (manejo convencional); 80% DAP, 20% fosfato térmico (FT); 40% DAP, 60% FT y 20% DAP, 80% FT, (0, 300, 600 y 900kg ha-1), con tres réplicas, para un total de 12 unidades experimentales. Las variables evaluadas fueron pesos fresco y seco, de parte área y raíz, concentración de fósforo en tejido y rendimiento por calidades y total. Se observó que la aplicación de termofosfato en diferentes concentraciones generó un efecto positivo con diferencias estadísticas (P≤0,05), en todas las variables evaluadas. El tratamiento de 600kg ha-1 de termofosfato, se muestra como una alternativa para el aporte de fósforo dentro de los planes de fertilización, en busca de una mejor respuesta fisiológica y productiva del cultivo de papa.

ABSTRACT Potato cultivation is one of the most representative activities of the Colombian Andes. The crop has high production costs, among which fertilization represents 36% of the total Technological advances in plant nutrition have allowed the development of new compounds that generate greater assimilation and availability of nutritional elements during productive periods. Within the different phosphorus sources available in Colombia, thermophosphate exists as an alternative. For this reason, the objective of this research was to evaluate the application of thermophosphate in the potato crop, Superior 'variety, under field conditions in the municipality of Villa Pinzón-Cundinamarca. A completely randomized design was used with 4 treatments that corresponded to different percentages of thermal phosphate: 100% diammonium phosphate (DAP) (conventional management); 80% DAP, 20% thermal phosphate (TF); 40% DAP, 60% TF and 20% DAP, 80% TF (0, 300, 600 y 900kg ha-1), with three replicas for a total of 12 experimental units. The variables evaluated were fresh and dry weights of areal and root part, phosphorus concentration in tissue and yield by qualities and the total. It was observed that the application of thermophosphate at different concentrations generated a positive effect, statistically different (P≤0.05) in all the variables evaluated. Thermophosphate, at 600kg ha-1 was found to be an alternative for the contribution of phosphorus within the fertilization plans, in search of a better physiological and productive response of the potato crop.

Detección de virus de ARN en cultivos de tomate del Oriente Antioqueño mediante secuenciación de alto rendimiento y RT-qPCR / Detection of RNA viruses in tomato fields in eastern Antioquia using high-throughput sequencing (HTS) and RT-qPCR

RESUMEN En Antioquia, el cultivo de tomate (Solanum lycopersicum) se ve afectado por diversas enfermedades virales, que ocasionan la disminución en la calidad de los frutos y de los rendimientos; sin embargo, pocos estudios han identificado, a nivel de especie, los agentes causales de dichas enfermedades. En los últimos años, la secuenciación de alto rendimiento (HTS), se ha convertido en una herramienta eficiente de diagnóstico de fitopatógenos, permitiendo la detección y la caracterización genómica de un alto número de virus, en diferentes plantas. En este trabajo, se evaluó la presencia de virus de ARN infectando tomate var. Chonto del oriente Antioqueño, mediante HTS y RT-PCR, en tiempo real (RT-qPCR), en muestras de tejido foliar y en semillas. El análisis de HTS indicó la infección de los virus Potato virus S (PVS), Potato virus Y (PVY), Potato yellow vein virus (PYVV), Potato virus X (PVX), Southern tomato virus (STV) y Bell pepper endornavirus (BPEV), en los cultivos de tomate de esta región, obteniéndose los genomas completos de PYVV, STV y BPEV. Las pruebas de RT-qPCR indicaron la presencia de PYVV en el 100% de las muestras foliares analizadas, mientras que PVX, PVY, STV y PVS, se encontraron en niveles de 94,4, 77,8, 72,2 y 5,6%, respectivamente. La evaluación de estos virus en lotes de semilla sexual comercial y no comercial y en sus plantas derivadas evidenció la presencia de cinco virus en dicho material, con niveles de prevalencia del 13 al 93% e infecciones mixtas, que incluyeron combinaciones, desde dos a cinco virus.

ABSTRACT In Antioquia, the tomato crop (Solanum lycopersicum) is seriously affected by a wide range of viral diseases that affect yield and the quality of fruits. Despite of this, there are few studies aimed at identifying these viruses at the species level. With the advent of High-throughput sequencing (HTS) methods, it is now possible to achieve an efficient characterization of viruses infecting plant hosts. In this work, the presence of RNA viruses infecting tomato var. Chonto in eastern Antioquia was tested using HTS and Real-time RT-PCR (RT-qPCR) in leaf tissues and seeds. HTS revealed infection with Potato virus S (PVS), Potato virus Y (PVY), Potato yellow vein virus (PYVV), Potato virus X (PVX), Southern tomato virus (STV) and Bell pepper endornavirus (BPEV). Complete genome sequences were obtained for PYVV, STV and BPEV. RT-qPCR showed prevalence of 100%, 94.4%, 77.8%, 72.2% and 5.6% for PYVV, PVX, PVY, STV and PVS in leaf samples, respectively. These viruses were also found infecting commercial and informal seeds and in their seedlings with a prevalence between 13 and 93%. Mixed infections were found to combine a mixture of two to five viruses.

PRIMER REPORTE DE BEGOMOVIRUS INFECTANDO CULTIVOS DE AJÍ (Capsicum spp.) EN COLOMBIA / First report of begomoviruses infecting pepper (Capsicum spp.) crops in Colombia

RESUMEN Virus del género Begomovirus infectan cultivos de importancia económica en todo el mundo, incluyendo ají. A la fecha, en Colombia no hay reportes de la presencia de begomovirus infectando este cultivo, por lo que el objetivo de esta investigación fue identificar la presencia de virus de este género en ají empleando estrategias moleculares. Se colectaron 197 muestras de ají en diez municipios del Valle del Cauca. Se extrajo el DNA genómico total vegetal y mediante PCR se detectó la presencia de begomovirus. Para establecer la identidad molecular del virus se amplificaron fragmentos de 1,4 kb de muestras colectadas en Palmira y Vijes. Los fragmentos fueron clonados, secuenciados y analizados. Se encontró que el 85,7 % de las muestras de ají evaluadas fueron positivas para begomovirus. Los análisis de secuencia de los fragmentos virales de 1,4 kb arrojaron una identidad de 91,8 % entre ellos y los de secuencia de nucleótidos de los virus aislados en Vijes y Palmira mostró que éstos presentan los valores de identidad más altos (87,2 % y 86,6 %) con el virus de la distorsión de la hoja de maracuyá, un begomovirus aislado de maracuyá en Colombia. Estos análisis estarían indicando que este begomovirus aislado de ají podría ser una nueva especie. De acuerdo con la literatura, este es el primer reporte de un begomovirus infectando cultivos de ají en Colombia.

ABSTRACT Viruses of the genus Begomovirus infect crops of economic importance around the world, including pepper. To date, in Colombia there are no reports of the presence of begomoviruses infecting this crop; therefore, this research work aimed to identify the presence of viruses of this genus in pepper using molecular strategies. Around 197 pepper samples were collected in ten municipalities in Valle del Cauca. Total plant genomic DNA was extracted, and the presence of begomoviruses was detected by using PCR. In order to establish the molecular identity of the virus, fragments of 1.4 kb were amplified from samples collected in Palmira and Vijes municipalities. The fragments obtained were cloned, sequenced, and analyzed. The results show that about 85.7 % of the pepper samples evaluated were positive for begomoviruses. Sequence analysis of the viral fragments of 1.4 kb showed an identity of 91.8 % among them. The nucleotide sequence analysis of the begomoviruses isolated in Vijes and Palmira showed its highest identity values (87.2 % and 86.6 %) with the passion fruit leaf distortion virus, a begomovirus that is affecting passion fruit crops in Colombia. These sequences analyze would indicate that this begomovirus isolated from pepper could be a new species that has not been reported worldwide. To our knowledge, this is the first report of a begomovirus infecting pepper crops in Colombia.

Molecular characterization of a new Begomovirus isolated from five weeds species collected in tomato crops in Valle del Cauca / Caracterización molecular de un nuevo begomovirus aislado de cinco especies de arvenses colectadas en cultivos de tomate en valle del cauca

RESUMEN Las arvenses son hospederos alternos de begomovirus (Geminiviridae), los cuales facilitan su persistencia y propagación a cultivos de interés agronómico, como el tomate. El objetivo de esta investigación fue obtener el genoma completo de un begomovirus bipartita encontrado en Amaranthus dubius, Rivina humilis, Rhynchosia minima, Desmodium sp. y Caesalpinia sp., las cuales fueron colectadas en cultivos de tomate en Ginebra y Cerrito, Valle del Cauca. El genoma del begomovirus fue obtenido utilizando amplificación por círculo rodante y digestión con las enzimas EcoRI y EcoRV, las cuáles cortan el componente genómico A y B, respectivamente. Estos fragmentos fueron clonados, secuenciados y analizados. Finalmente, se verificó la presencia de este begomovirus en todas las arvenses mediante PCR específico. Se obtuvieron tres clonas EcoRI y cinco clonas EcoRV. Los fragmentos que portan los componentes A y B presentan un tamaño de 2 584 y 2 543 nt, respectivamente. El análisis de secuencia de nucleótidos del genoma begomoviral A con otros begomovirus previamente reportados, mostró la mayor identidad (90,9 %) con el virus del mosaico dorado de Rhynchosia de Yucatán. Tomando como base el criterio de demarcación actual para las especies de Begomovirus establecido por el Comité Internacional de Taxonomía de Virus, el geminivirus aislado de las arvenses A. dubius, R. humilis, R. minima, Desmodium sp. y Caesalpinia sp., constituye una nueva especie begomoviral. Con base en la sintomatología observada en campo, se propone el nombre de Virus del mosaico dorado de Rhynchosia de Colombia para designar a esta nueva especie.

ABSTRACT The weeds are alternative hosts of begomoviruses (Geminiviridae), which facilitate their persistence and propagation to crops of agronomic interest, such as tomatoes. The objective of this investigation was to obtain the complete genome of a bipartite begomovirus found in Amaranthus dubius, Rivina humilis, Rhynchosia minima, Desmodium sp. and Caesalpinia sp., which were collected in tomato crops in Ginebra and Cerrito, Valle del Cauca. The genome of the begomovirus was obtained using rolling circle amplification and digestion with the enzymes EcoRI and EcoRV, which cut the genomic component A and B, respectively. These fragments were cloned, sequenced, and analyzed. Finally, the presence of this begomovirus in all weeds was verified by specific PCR. Three EcoRI clones and five EcoRV clones were obtained. The fragments carrying components A and B have a size of 2 584 and 2 543 nt, respectively. The analysis of the nucleotide sequence of the begomoviral A genome with other previously reported begomoviruses showed the highest identity (90.9 %) with the Rhynchosia golden mosaic virus Yucatán. Based on the current demarcation criterion for the Begomovirus species established by the International Committee on Taxonomy of Viruses, the geminivirus isolated from the weeds A. dubius, R. humilis, R. minima, Desmodium sp. and Caesalpinia sp., constitutes a new begomoviral species. Based on the symptoms observed in the field, the name of the Rhynchosia golden mosaic Colombia virus is proposed to designate this new species.

COINFECCIÓN NATURAL DE VIRUS DE ARN EN CULTIVOS DE PAPA (Solanum tuberosum subsp. Andigena) EN ANTIOQUIA (COLOMBIA) / Natural coinfection of RNA viruses in potato (Solanum tuberosum subsp. Andigena) crops in Antioquia (Colombia)

RESUMEN Las enfermedades virales son uno de los principales problemas fitopatológicos de la papa. Con el fin de determinar los virus más prevalentes en cultivos de papa var. Diacol Capiro en el oriente Antioqueño (Colombia), se evaluó mediante RT-qPCR la presencia de diez virus de ARN (PVY, PVA, PVV, TaLMV, PVS, PLRV, PYVV, PVX, ToRSV y PMTV) en 36 muestras de tejido foliar. Los resultados indicaron la ocurrencia de cinco de los diez virus evaluados, con niveles de prevalencia de 88,9 %, 75 %, 75 %, 41,7 % y 25 % para PVY, PVX, PYVV, PLRV y PVS, respectivamente. Con fines comparativos, cuatro virus también se evaluaron mediante ELISA, siendo detectados PVS (80,5 %), PVY (55 %) y PLRV (5,5 %); mientras que PVX no fue encontrado con esta prueba. La comparación de estas técnicas mediante la razón de prevalencia (RP), indicó que la RT-qPCR ofrece niveles superiores de detección con valores de RP = 1,6 y RP = 7,5 para los virus PVY y PLRV; mientras que para PVS la ELISA detectó más muestras positivas que RT-qPCR (RP = 3,22), evidenciándose la necesidad de diseñar nuevos cebadores ajustados a la diversidad de este virus en Antioquia. La coinfección mixta más frecuente fue PVY-PYVV-PVX (22,2 %), mientras que los cinco virus se encontraron en el 11,1 % de las muestras. Finalmente, utilizando secuenciación Sanger de la cápside y NGS para los genomas completos, se confirmó la circulación de todos los virus detectados en los cultivos de papa del oriente Antioqueño. Estos resultados señalan la necesidad de fortalecer los programas de manejo integrado de enfermedades virales en Antioquia.

ABSTRACT Viral diseases are one of the main phytopathological problems affecting potato crops worldwide. To determine the most prevalent viruses in potato var. Diacol Capiro crops in Eastern Antioquia, 36 leaf samples were tested for the presence of PVY, PVA, PVV, TaLMV, PVS, PLRV, PYVV, PVX, ToRSV and PMTV using RT-qPCR. Detected viruses included PVY, PVX, PYVV, PLRV and PVS with prevalence levels of 88.9 %, 75.0 %, 75.0 %, 41.7 % and 25.0 %, respectively. PVS, PVY, PLRV and PVX were also tested by ELISA. PVS, PVY and PLRV tested positive in 80.5 %, 55.0 % and 5.5 % of samples; PVX was not detected. Prevalence Ratios (PR) suggests that detection is higher for PVY (PR = 1.6) and PLRV (PR = 7.5) using RT-qPCR. ELISA worked better for PVS with a PR of 3.2; this result suggests that the RT-qPCR primers used for PVS must be adjusted to reflect the genome diversity of virus in Antioquia. The most frequent coinfection was PVY-PYVV-PVX, which occurred in 22.2 % of samples; coinfection with PVY, PVX, PYVV, PLRV and PVS was present in 11.1 % of samples. The circulation of these viruses in Eastern Antioquia was further confirmed using Sanger and high-throughput sequence. This work highlights the need to strengthen integrated disease management programs of viruses in Antioquia.

Design of primers for specific detection of Potato Virus Y (PVY) by RT-PCR / Diseño de cebadores específicos para la detección por rt-pcr del potato virus y (pvy)

RESUMEN El Potato virus Y (PVY) es uno de los virus más limitantes para la producción de papa (Solanum tuberosum y S. phureja) en el mundo. Este virus es transmitido por tubérculo-semilla de papa y por diferentes especies de áfidos. Para su manejo es fundamental la siembra de tubérculos certificados por su sanidad viral, para lo que se requieren metodologías de detección altamente sensibles como ELISA y RT-PCR. Para éstas últimas pruebas, es necesario disponer de cebadores específicos que permitan el diagnóstico del virus en tejidos asintomáticos. En este estudio se reportan los cebadores PVY_Col para la detección del PVY en RT-PCR convencional y en tiempo real (RT-qPCR). Estos cebadores fueron diseñados con base en las secuencias de este virus que se han reportado en Colombia sobre diferentes hospedantes, así como de las diferentes variantes encontradas en el mundo. Una particularidad adicional de estos cebadores es que no presentan reacción cruzada con el genoma del Potato virus V (PVV), otro potyvirus que recientemente se ha encontrado afectando cultivos de papa en Colombia. Se espera que los cebadores PVY_Col sean utilizados para apoyar los programas de certificación de material de siembra de papa, así como para adelantar estudios epidemiológicos y de manejo fitosanitario de este virus.

ABSTRACT Potato virus Y (PVY) is one of the most limiting viruses in the production of potato (Solanum tuberosum and S. phureja) worldwide. This virus is transmitted by aphids and infected tuber-seeds, and for this reason, disease management of PVY requires, among others, using certified planting material through highly sensitive techniques such as ELISA and RT-PCR. However, RT-PCR-based methods require primers of high specificity to allow the unequivocal detection of viruses in asymptomatic tissues. In this work, we report a new set of primers (PVY_Col) for detection of PVY by RT-PCR and real-time RT-PCR (RT-qPCR). These primers were designed using sequences of PVY isolates from Colombia and the rest of the world. An essential feature of these primers is their specificity, since they don't amplify templates from Potato virus V (PVV), a close PVY relative that also infects potato crops in Colombia. It is expected that the PVY_Col primer set will be useful to support potato seed certification programs, epidemiological studies, and PVY disease management programs.

Ciclo de vida y enemigos naturales de Mechanitis menapis (Lepidoptera: Ithomiini)

Las mariposas de la tribu Ithomiini son uno de los modelos biológicos más estudiados en años recientes en trabajos de biogeografía, taxonomía y evolución. Sin embargo, aun cuando la biología y distribución de sus especies es mejor conocida que la de otros grupos de mariposas, existen aspectos de la historia natural poco estudiados que permitirían un mejor entendimiento del comportamiento de sus poblaciones y las interacciones con su ambiente. En este trabajo, se estudió la historia natural de la mariposa, Mechanitis menapis mantineus Hewitson (Nymphalidae: Ithomiini) y sus enemigos naturales en el oeste de Ecuador. Para ello, se realizó la identificación de las plantas hospederas, se estudió el ciclo de vida y los factores de mortalidad de los estados inmaduros. Adicionalmente, se identificaron los parasitoides asociados a la especie en la zona de estudio. Se identificaron tres plantas hospederas, las cuales son nuevos registros para la especie en la región. Asimismo, se describió la morfología de los estados inmaduros y su tiempo de desarrollo. Finalmente, se identificaron los principales enemigos naturales y se describieron aspectos de la historia natural y comportamiento de Hyposoter sp. (Ichneumonidae: Campopleginae), principal parasitoide de larvas. Los resultados de este trabajo, proporcionan información esencial para la revisión taxonómica del género Mechanitis F. y para continuar el estudio de la interacción de esta especie con sus plantas hospederas y enemigos naturales.

Life cycle and natural enemies of Mechanitis menapis (Lepidoptera: Ithomiini). The butterflies of the Ithomiini tribe are one of the most-studied biological models of recent years in terms of biogeography, taxonomy, and evolution. However, even though their biology and distribution is better known than many other groups of butterflies, there are unknown aspects of their natural history that would improve our understanding of their behavior, population dynamics, and interactions with their environment. In this work, we studied the natural history of the butterfly Mechanitis menapis mantineus Hewitson (Nymphalidae: Ithomiini), and its natural enemies, in Western Ecuador. We identified three host plants, which are new records for the species in this region. We documented the life cycle and described the morphology of the immature stages, their development time, and studied the factors associated with mortality of these immature stages. Additionally, we identified the parasitoids associated with the species in the study area. In particular, we documented aspects of the natural history and behavior of Hyposoter sp. (Ichneumonidae: Campopleginae), the main parasitoid of the larvae. The results of this work provide essential information for the taxonomic revision of the genus Mechanitis F. as well as the continued study of the interactions between the butterfly, its host plants, and natural enemies.

Cloning and molecular characterization of a putative habanero pepper SERK1 cDNA expressed during somatic and zygotic embryogenesis

Background: Plant gene homologs that control cell differentiation can be used as biotechnological tools to study the in vitro cell proliferation competence of tissue culture-recalcitrant species such as peppers. It has been demonstrated that SERK1 homologs enhance embryogenic competence when overexpressed in transformed tissues; therefore, cloning of a pepper SERK1 homolog was performed to further evaluate its biotechnological potential. Results: A Capsicum chinense SERK full-length cDNA (CchSERK1) was cloned and characterized at the molecular level. Its deduced amino acid sequence exhibits high identity with sequences annotated as SERK1 and predicted-SERK2 homologs in the genomes of the Capsicum annuum CM-334 and Zunla-1 varieties, respectively, and with SERK1 homologs from members of the Solanaceae family. Transcription of CchSERK1 in plant tissues, measured by quantitative RT-PCR, was higher in stems, flowers, and roots but lower in leaves and floral primordia. During seed development, CchSERK1 was transcribed in all zygotic stages, with higher expression at 14 days post anthesis. During somatic embryogenesis, CchSERK1 was transcribed at all differentiation stages, with a high increment in the heart stage and lower levels at the torpedo/cotyledonal stages. Conclusion: DNA sequence alignments and gene expression patterns suggest that CchSERK1 is the C. chinense SERK1 homolog. Significant levels of CchSERK1 transcripts were found in tissues with cell differentiation activities such as vascular axes and during the development of zygotic and somatic embryos. These results suggest that CchSERK1 might have regulatory functions in cell differentiation and could be used as a biotechnological tool to study the recalcitrance of peppers to proliferate in vitro.

Nucleotide sequence polymorphism in the RFL-PPR genes of potato

Cytoplasmic male sterility (CMS) is widely used for hybrid seed production in cultivated Solanaceae species. However, there is very limited information about CMS-Rf genetic systems in potato (Solanum tuberosum). Studying the CMS-Rf systems in potato is both of theoretical and practical significance due to the emergence of a new revolutionary strategy of reinventing potato as adiploid inbred line-based crop to develop F1 hybrid seed potato breeding (Lindhout et al. 2011; Jansky et al. 2016). To search for potato Rf gene candidates, the comparative genetic approach was applied. Based on similarity to petunia Rf-PPR592 gene, 38 fragments were identified in five loci of the whole-genome nucleotide sequence of the accession DM 1-3 516 R44 S. tuberosum Phureja group (https://blast.ncbi.nlm.nih.gov/Blast.cgi). The putative encoded mitochondrial proteins have 589–597 amino acid residues, similarto RF-PPR proteins of petunia and chili pepper and contain 14 or 15 PPR motifs. Primers have been developed flanking the most variable 782–865 bp regions of the selected loci, and polymorphism of the cloned fragments has been investigated in a subset of nine potato genotypes. The amplified fragments included seven or eight PPR motifs and lacked introns. The SNP frequencies ranged from 7.0 to 19.8% depending on the locus, while the ratio of nonsynonymous to synonymous substitutions varied between 0.9 and 2.1.Positions 1, 3 and 6 were the most variable in the studied PPR motifs. Our results demonstrated that the analysed sequences belong to the RFL-PPR gene subfamily and may be considered as Rf gene candidates in potato.

Diversidad genética de tres poblaciones de Physalis peruviana a partir del fraccionamiento y patrón electroforético de proteínas de reserva seminal / Genetic diversity in three populations of Physalis peruvianausing fractionation and electrophoretic patterns of seed storage protein

En el presente trabajo, es estudiada la diversidad genética de tres poblaciones atribuidas a ecotipos de aguaymanto, Physalis peruaviana. Las tres poblaciones eran atribuidas a los ecotipos Agroandino (provincia de San Pablo), Celendino (provincia de Celendín) y Cajabamba (provincia de Cajabamba) del departamento de Cajamarca. Se realizó la cuantificación proteica y evaluó el polimorfismo de las proteínas de reserva seminal (SSPs) mediante electroforesis en gel de poliacrilamida denaturante (SDS-PAGE). Además, se identificaron características bioquímicas de las proteínas seminales en esta especie. No se hallaron diferencias entre las tres poblaciones basados en la cuantificación proteica. Las globulinas (82.4%) fueron la fracción mayoritaria seguida por las albuminas (13.9%), glutelinas (3.7%) y prolaminas (0.7%). Sólo las albuminas mostraron polimorfismo, hallándose 21 proteínas entre ~ 6.5 a ~45 kDa y tres perfiles electroforéticos diferentes, los cuales fueron compartidos entre las poblaciones. Se identificaron las leguminas y vicilinas en la fracción globulina. Las glutelinas mostraron proteínas de mismo peso molecular (PM) a las leguminas; y las prolaminas sólo una banda de bajo PM. La población de San Pablo fue completamente homogénea a diferencia de la población de Cajabamba que mostró la mayor diversidad genética seguida de Celendín. No fue posible diferenciar las poblaciones designadas como ecotipos Agroandino, Cajabamba y Celendino basados en el análisis de proteínas seminales.

The genetic diversity of three populations designated as ecotypes of golden berry (Physalis peruaviana) is studied using protein quantification and polymorphism of seed storage proteins (SSPs) by denaturating polyacrylamide gel electrophoresis (SDS-PAGE). As well, biochemical characteristics of seed proteins were identified. The populations were from San Pablo province (Agroandino ecotype), Celendín province (Celendino ecotype) and Cajabamba province (Cajabamba ecotype), all from Cajamarca Department. There was not difference among the three populations based on protein quantification. Globulins (82.4%) were the majority fraction followed for albumins (13.9%), glutelins (3.7%) and prolamins (0.7%). Only albumins showed polymorphism, showing 21 proteins between ~6.5 to ~45 kDa and three different electrophoretic profiles, which were share among the three populations. Legumins and vicilins were identified in globulin fraction. Glutelins showed proteins of same molecular weight (MW) to legumins; and prolamins only a band of low MW. San Pablo province population (Agroandino ecotype) was completely uniform, while Cajabamba population showed higher genetic diversity followed by Celendin population. Our results shows that, based on seed proteins analyses is not possible to distinguish the three populations designated as Agroandino, Cajabamba and Celendino ecotypes.

Development of a direct transformation method by GFP screening and in vitro whole plant regeneration of Capsicum frutescens L

Background: Capsicum is a genus of an important spice crop that belongs to the chili lineage. However, many Capsicum species (family Solanaceae) are known to be recalcitrant to genetic transformation and in vitro regeneration, thus hampering the effort in using Capsicum species for detailed biological investigation. In this study, we have developed an optimized protocol for the direct transformation of Capsicum frutescens L. cv. Hot Lava using a biolistic particle delivery system. In addition, a procedure for in vitro whole plant regeneration from the hypocotyl explants of C. frutescens was established. Results: In this study on the biolistic system, explant target distance, bombardment helium (He) pressure, and the size of the microcarrier were the key parameters to be investigated. The optimized parameters based on the screening of GFP expression were determined to have a target distance of 6 cm, helium pressure of 1350 psi, and gold particle (microcarrier) size of 1.6 µm. The greatest number of shoots was obtained from hypocotyls as explants using Murashige and Skoog medium supplemented with 5.0-mg/L 6-benzylaminopurine and 0.1-mg/L 1-naphthaleneacetic acid. On an average, five shoots per explant were formed, and of them, one shoot managed to form the root and developed into a whole plant. Conclusions: We obtained an optimized protocol for the biolistic transformation of chili and in vitro regeneration of chili plantlets. The establishment of the protocols will provide a platform for molecular breeding and biological studies of chili plants.

Different leaf extracts from Brunfelsia uniflora in mice / Diferentes extratos das folhas de Brunfelsia uniflora em camundongos

ABSTRACT: In Brazil, at least 16 poisonous plant species can affect the central nervous system of livestock. Recently in the state of Piauí, Northeastern Brazil, Brunfelsia uniflora was reported as a cause of nervous signs in donkeys, cattle, and small ruminants. In order to assess the toxicity of B. uniflora extracts, 20 Swiss mice were distributed into four groups (n=5) receiving by gavage alkaloids, flavonoids, saponins, or saline (control group). After administration of the extracts in a single dose (5g kg-1), all mice were observed daily for clinical signs. Mice that received the extracts showed moderate to severe clinical signs, including piloerection, vocalization, and seizures. All mice dosed with saponins died between 10 and 20min after administration. Serum biochemical evaluation of animals that received saponins revealed slight increases in total protein levels and decreased magnesium and chlorite levels. In conclusion, saponins of B. uniflora leaves induced acute toxic neurological effects and death in mice.

RESUMO: No Brasil, pelo menos 16 espécies de plantas tóxicas podem afetar o sistema nervoso central dos bovinos. Recentemente no estado do Piauí, no Nordeste do Brasil, Brunfelsia uniflora foi relatada como causadora de sinais nervosos em jumentos, bovinos e pequenos ruminantes. Para avaliar a toxicidade de B. uniflora, 20 camundongos foram distribuídos em quatro grupos de cinco animais recebendo diferentes extratos da planta. Os extratos de alcalóides, flavonóides, saponinas ou solução salina (grupo controle) foram administrados por gavagem. Após a administração dos extratos, em dose única (5g kg-1), todos os animais foram observados diariamente. Os camundongos que receberam os extratos de B. uniflora apresentaram sinais clínicos moderados a graves, incluindo piloeração, vocalização e convulsões. Todos os camundongos que receberam extrato de saponinas morreram entre 10 e 20 minutos após. A avaliação bioquímica sérica dos animais que receberam saponinas, revelou discretos aumentos nos níveis de proteína total e diminuição nos níveis de magnésio e cloro. Conclui-se que, as saponinas presentes nas folhas de B. uniflora induzem efeitos neurológicos tóxicos agudos e, morte em camundongos.

Yield analysis of Physalis ixocarpa Brot. ex Hornem varieties under greenhouse and field conditions / Análise de rendimento das variedades Physalis ixocarpa Brot. ex Hornem sob condições de estufa e de campo

ABSTRACT: Physalis ixocarpa (husk tomato) is traditionally cultivated in Mexico, and nowadays the yield is low. In this study, four cultivars of husk tomato were evaluated, under greenhouse and field conditions. It was carried out a split-plot experimental design in a 4x2 factorial arrangement. The large plot was the growth conditions and the small plot was the variety. The microclimate, soil and yield variables were recorded. In the field, the Integrated Photosynthetic Active Radiation (IPAR) was higher 12mol m-2 d-1 than in the greenhouse. The average temperature was slightly higher 1.18°C in the greenhouse and the average relative humidity was slightly higher 0.89% in the field. Plants grown in the greenhouse had lower negative values for the matric potential of the soil. Water consumption and water use efficiency were 10.31 and 53.43% higher in the greenhouse that in the field, respectively. The number of fruits and yield increased significantly in all varieties grown under greenhouse conditions.

RESUMO: A Physalis ixocarpa (Tomate de cáscara) é cultivada tradicionalmente no México. Entretanto, sua produtividade de frutos é considerada baixa. Neste estudo, avaliou-se quatro cultivares de Physalis ixocarpa cultivadas em ambiente protegido e a céu aberto. O delineamento experimental utilizado foi em blocos ao acaso no esquema de parcelas subdivididas com arranjo fatorial 2x4. Foi alocado na parcela principal o fator ambiente de cultivo e na subparcela o fator cultivar. Foram analisadas variáveis climáticas de solo e a produtividade de frutos. A radiação fotossinteticamente ativa integrada (RFAI) no ambiente a céu aberto foi maior em 12mol m-2 dia-1, em relação ao ambiente protegido. A temperatura média foi ligeiramente maior em 1,18oC no ambiente protegido e a umidade relativa foi ligeiramente maior em 0,89% a céu aberto. Os valores negativos do potencial mátrico do solo foram mais baixos no ambiente protegido. O consumo e a eficiência do uso da água foram, respectivamente, 10,31 e 54,43% maiores no ambiente protegido, em relação ao ambiente a céu aberto. O número e a produtividade de frutos incrementaram, significativamente, em todas as variedades avaliadas nas condições de ambiente protegido.

Chemical constituents from roots of Datura metel / 中国中药杂志

The chemical constituents from the n-butanol fraction of the 70% ethanol extract of Datura metel roots were separated by silica gel and ODS chromatogram columns as well as preparative HPLC. On the basis of spectral data analysis, their structures were elucidated. Twenty-one compounds were obtained and their structures were identified as citroside A (1), coniferin (2), paeoniflorin (3), (6R,7E,9R)-9-hydroxy-4,7-megastigmadien-3-one 9-O-[α-L-arabin-opyranosyl-(l→6)-β-D-glucopyranoside] (4), (1R,7R,10R,11R)-12-hydroxyl anhuienosol (5), kaurane acid glycoside A (6), ent-2-oxo-15,16-dihydroxypimar-8(14)-en-16-O-β-glucopyranoside (7), ginsenoside Rg₁(8), ginsenoside Re (9), notoginsenosides R₁(10), N-butyl-O-β-D-fructofuranoside (11), salidroside (12), hexyl β-sophoroside (13), 2,6-dimethoxy-4-hydroxyphenol 1-glucoside (14), benzyl-O-β-D-xylopyranoxyl(1→6)-β-D-glucopyranoside (15), (Z)-3-hexenyl-O-α-L-arabinopyranosyl-(1→6)-β-D-glucopyranoside (16), N-[2-(3,4-dihydro-xyphenyl)-2-hydroxyethyl]-3-(4-methoxyphenyl) prop-2-enamide (17), cannabisin D (18), cannabisin E (19), melongenamide B (20), paprazine (21). Compounds 2-17 and 20-21 were isolated from the Solanaceae family for the first time.

SECUENCIACIÓN DEL GENOMA COMPLETO DEL Potato yellow vein virus (PYVV) EN TOMATE (Solanum lycopersicum) EN COLOMBIA / Genome Sequencing of Potato yellow vein virus (PYVV) Strain Infecting Solanum lycopersicum in Colombia

RESUMEN Potato yellow vein virus (PYVV), es uno de los fitopatógenos más limitantes para la producción de papa en la región de Los Andes. A pesar que se le ha detectado infectando tomate en Colombia, el conocimiento de las características biológicas de las cepas presentes en este hospedante es muy limitado. En este estudio, utilizando secuenciación masiva de nueva generación (NGS), se obtuvo la secuencia completa de los tres segmentos genómicos del PYVV en plantas de tomate en Marinilla (Antioquia) y se evalúo la utilidad de tres juegos de cebadores para su detección mediante pruebas de RT-PCR convencional y en tiempo real (RT-qPCR). El genoma de la secuencia consenso presentó tamaños de 8043 nt (ARN1), 5346 nt (ARN2) y 3896 nt (ARN3) y se identificaron los diez ORF previamente reportados en este virus, aunque, en general, éstos presentaron menores niveles de identidad que los registrados entre cepas de PYVV de papa. Análisis de variación y de selección identificaron dos regiones en los ORF MET/HEL y CPm que presentan selección positiva, lo que podría estar asociado a la adaptación por hospedante. Los tres juegos de cebadores amplificaron las regiones esperadas de la cápside de PYVV, siendo posible identificar, por diferencias en valores de temperatura de fusión (Tm) y por secuenciación Sanger, la ocurrencia de al menos dos variantes principales de este virus en el Oriente Antioqueño, lo que concuerda con los niveles moderados de polimorfismos encontrados en las secuencias obtenidas por NGS.

ABSTRACT Potato yellow vein virus (PYVV) is one of the most important pathogens of potato in the Andean region. In spite of having been detected in tomato crops in Colombia, knowledge on the biological characteristics of PYVV is limited on this host. In this study, next-generation sequencing (NGS) of a PYVV strain infecting tomato in Marinilla District (Antioquia) was performed; additionally, three primer set useful in RT-PCR and RT-qPCR detection were also tested. The consensus genome consisted of three RNA segments of 8043 nt (RNA1), 5346 nt (RNA2) and 3896 nt (RNA3) encoding ten ORF with slight lower sequence identity in relation to PYVV isolates from potato. Sequence analysis suggests the presence of regions potentially undergoing positive selection in the ORFs coding for MET/HEL and CPm possibly as a result of host adaptation. Experimental validation of primers resulted in amplicon with the expected size while melting temperature analysis and sequencing suggest the presence of at least two PYVV infecting S. lycopersicum in east Antioquia in agreement with the NGS data.

Research progress on physalis pubescens / 中草药

Physalis pubescens is used as both food and medicine over 1 200 years. From the phytochemical view, the second metabolites such as flavonoids, sterols, phenylpropanoids, alkaloids, fatty acid and so on have been reported from this species. And this plant shows the activities of antitumor, antimicrobial, anti-oxidant, diuresis, immunosuppressive and so on in the recent studies. This paper reviews the researches on the chemical constituents and pharmacological activities of this plant in recent 30 years, by which the prospect of development and utilization of P. pubescens could be prospected. It provides the reference for the further rational development and comprehensive utilization of the resources of P. pubescens.

Phenylpropanoids from roots of Daturae Flos / 中草药

Objective To study the phenylpropanoids from the roots of Datura metel. Methods The separations and purifications were taken by silica gel and ODS chromatogram columns as well as preparative HPLC, and the structural identification based on physicochemical property, 1H-NMR and 13C-NMR as well as HR-MS data. Results Nineteen compounds obtained from the butanol fraction of the 70% ethanol extract of D. metel roots, which were identified as icariside E5 (1), alangisesquin A (2), Glycopentoside F (3), conicaoside (4), (7S,8R) dehydrodiconiferyl alcohol 9’-O-β-glucopyranoside (5), iariciresinol-4’-O-β-D-glucopyranoside (6), 7R,8R-threo-4,7,9-trihy-droxy-3,3’-dimethoxy-8-O-4’-neolignan-9’-O-β-D-glucopyranoside (7), vitrifol A (8), leptolepisol D (9), thero-2,3-bis-(4-hydroxy-3-methoxypheyl)-3-methoxy-propanol (10), hyuganoside IIIb (11), officinalioside (12), iariciresinol-9-O- β-D-glucopyranoside (13), stroside A (14), erythro-buddlenol B (15), sargentodoside D (16), (+)-(7S,8S)-4-hydroxy-3,3’,5’- trimethoxy-8’,9’-dinor-8,4’-oxyneolignan-7,9-diol-7’-oic acid (17), 5’-methoxy lariciresinol (18), and dehydrodiconiferyl alcohol 4-O-β-D-glucopyranoside (19). Conclusion Compounds 2-19 are isolated from Solanaceae family for the first time and compound 1 is firstly isolated from genus Datura L.

Detección serológica y molecular del Potato virus X (PVX) en tubérculos-semilla de papa (Solanum tuberosum L. y Solanum phureja Juz. & Bukasov) en Antioquia, Colombia / Serological and molecular detection of Potato virus X (PVX) in seed potato tubers (Solanum tuberosum L. and Solanum phureja Juz. & Bukasov) from Antioquia, Colombia

El Potato virus X (PVX) es uno de los virus más limitantes del cultivo de la papa en el mundo. Es transmitido solamente por contacto y por tubérculo-semilla. Su control se fundamenta en la siembra de tubérculos certificados por su sanidad viral y en la disponibilidad de metodologías de diagnóstico altamente sensibles. En este trabajo se evaluó la prevalencia del PVX en cuatro diferentes tejidos de tubérculos de Solanum tuberosum subsp. andigena var. Diacol-Capiro y S. phureja var. Criolla Colombia utilizando pruebas de DAS-ELISA para 128 submuestras y de RT-qPCR para 32 grupos de submuestras (4 submuestras/grupo). Los resultados de las pruebas serológicas indicaron la presencia de PVX en el 6,25 y 50% de las submuestras analizadas para la variedad Diacol-Capiro y Criolla Colombia, respectivamente; mientras que los niveles de prevalencia del PVX utilizando la detección por RT-qPCR fueron del 93,75%, independientemente de la variedad de papa y del tejido evaluado. Los valores promedio del ciclo umbral (Ct) en las RT-qPCR fueron de 25,6 (Ct=18,02 a 34,49) y el análisis de las curvas de desnaturalización permitió identificar dos variantes del virus con valores de Tm de 79,5±1°C y 83,7±1°C. La secuenciación de los amplicones obtenidos por RT-qPCR para los controles positivos y para dos de las muestras, confirmó su naturaleza viral. Estos resultados señalan unos muy altos niveles de prevalencia de PVX en el material de siembra de papa en Antioquia y la necesidad de fortalecer los programas de certificación de semilla con pruebas de detección como RT-qPCR.

Potato virus X (PVX) is one of the most important virus affecting potato crops worldwide. The virus is only transmitted mechanically and through tuber-seeds. Control of PVX is based on the usage of certified tubers, which in turn depends on the availability of sensitive diagnostic tests that allow its direct detection on seeds. In this work, the prevalence of PVX in four different tuber tissues of Solanum tuberosum subsp. andigena var. Diacol-Capiro and S. phureja var. Criolla was evaluated using DAS-ELISA (128 subsamples) and RT-qPCR (4 subsamples per group). DAS-ELISA revealed the presence of PVX in 6.25 and 50% of Diacol-Capiro and Criolla Colombia subsamples; in contrast, RT-qPCR detected PVX in 93.75% of the samples independent of the potato variety or type of tissue. Ct values were in the 18.02 to 34.49 range with a mean value of 25.6. Melting curve analysis allowed the identification of two virus variants with Tm values of 79.5±1°C and 83.7±1°C. Sanger sequencing of the positive controls and two of the samples confirmed RT-qPCR amplicons to be PVX. These results reveal a high level of prevalence of PVX in potato tuber seeds used in Antioquia and the need to strengthen seed certification programs in Colombia through RT-qPCR detection assays.

BIOATIVIDADE DE Solanum melongena L. E Capsicum annuum L. SOBRE Callosobruchus maculatus (COLEOPTERA: BRUCHIDAE) / Bioactivity of Solanum melongena L. and Capsicum annuum L. on Callosobruchus maculatus (Coleoptera: Bruchidae) / Bioactividad de Solanum melongena L. y Capsicum annuum L. en Callosobruchus maculatus (Coleoptera: Bruchidae)

O uso contínuo e indiscriminado de produtos químicos na agricultura pode trazer sérios prejuízos à saúde humana e ao meio ambiente. Uma opção é o emprego de plantas com ação inseticida. Diante do exposto, o objetivo desse trabalho foi avaliar a atividade inseticida do pó de folhas de Solanum melongena L. e Capsicum annuum L. contra Callosobruchus maculatus. O experimento foi conduzido no Laboratório de Entomologia da Universidade Federal de Campina Grande (UFCG), Campus de Pombal, Paraíba, Brasil. Os grãos de feijão-caupi foram tratados com os pós nas concentrações 0,0; 2,5; 5,0 e 10,0 % [100*(massa do pó/massa de grãos)] e realizados testes de sobrevivência e repelência contra C. maculatus. Os dados da sobrevivência foram analisados utilizando o teste de Log-rank (p ≤ 0,05), pelo método de D-collet e para a repelência utilizou-se o teste do Qui-Quadrado (p ≤ 0,05). Todos os pós e concentrações avaliadas foram repelentes contra C. maculatus, com exceção do pó de C. annuum na concentração de 2,5 %. No que se refere à sobrevivência, ambas as espécies vegetais causaram elevada mortalidade em C. maculatus, com morte total dos insetos em até 120 h.

The continuous and indiscriminate use of chemicals in agriculture can bring serious problems to human health and the environment. One option is the use of plants with insecticidal action. Given the above, the aim of this work was to evaluate the insecticide activity powder of leaves of Solanum melongena L. and Capsicum annuum L. against Callosobruchus maculatus in three concentrations. The experiment was conducted at the Laboratory of Entomology, Federal University of Campina Grande (UFCG), Campus de Pombal, Paraiba, Brazil. The grains of cowpea were treated with the powders in concentrations 0.0, 2.5, 5.0 and 10.0 % 6#91;100*(mass of powder/ mass of grains)] and performed tests of survival and repellency against C. maculatus. Survival data were analyzed using the Log-rank test (p ≤ 0.05) by D-collet method and the repellence we used the Chi-square test (p ≤ 0.05). All post and concentrations were repellent C. maculatus except the C. annuum powder at a concentration of 2.5 % that was not statistically significant difference. With respect to survival, both plant species caused high mortality on C. maculatus, with total kill insects up to 120 h.

El uso continuo y indiscriminado de productos químicos en la agricultura puede causar graves danos a la salud humana y al medio ambiente. Una opción es el uso de plantas con acción insecticida. Teniendo en cuenta lo anterior, el objetivo de este trabajo fue evaluar la actividad insecticida del polvo de las hojas de Solanum melongena L. y Capsicum annuum L. contra Callosobruchus maculatus en tres concentraciones. El experimento se realizó en el Laboratorio de Entomología de la Universidad Federal de Campina Grande (UFCG), Campus de Pombal, Paraiba, Brasil. Los granos de caupí fueron tratados con los polvos, en las concentraciones 0,0; 2,5; 5,0 a 10,0 % [100*(masa de polvo/masa de granos)] y se realizaron ensayos de supervivencia y repelencia contra C. maculatus. Los datos de supervivencia se analizaron mediante el análisis de Log-rank (p ≤ 0.05) por el método de D-Collet y para la repelencia se utilizó el análisis de Chi-cuadrado (p ≤ 0,05). Todos los polvos y las dosis evaluadas fueron repelentes contra C. maculatus, con la excepción del polvo de C. annuum a una dosis de 2,5 % que se observó ninguna diferencia estadística. Con respecto a la supervivencia, ambas especies de plantas causaron alta mortalidad en C. maculatus, con muerte total de los insectos hasta 120 h.