Prevalencia de virus de ARN en tubérculos-semilla certificados de Solanum tuberosum cv. Diacol Capiro de la sabana Cundiboyacense

La papa (Solanum tuberosum) Diacol Capiro es uno de los cultivares con mayor producción y consumo interno en Colombia, siendo los departamentos de Cundinamarca y Boyacá, los principales productores. Este cultivo, se ve afectado por un complejo de virus, que disminuye la calidad de los tubérculos y los rendimientos. En este trabajo, se evaluó la prevalencia de los virus de ARN: PLRV, PVY, PVX, PVS, PVV, PYVV, PMTV y PVB, en brotes de tubérculos-semilla certificados, provenientes de la sabana Cundiboyacense, mediante RT-qPCR. Los resultados revelan la ocurrencia de siete de los ochos virus en las muestras, con niveles de infección de 100 % (PVS, PVX y PYVV), 75 % (PLRV), 50 % (PVY), 37,5 % (PMTV) y 12,5 % (PVB). Adicionalmente, con el fin de obtener información de los genomas de los virus detectados, se utilizó secuenciación de alto rendimiento (HTS), de una muestra compuesta (bulk) de brotes, siendo posible obtener el genoma completo del PLRV y el genoma parcial del PVY. Los análisis filogenéticos realizados con dichas secuencias ubicaron a los virus PLRV y PVY en clados, conformados por aislamientos colombianos, con niveles de identidad superiores al 97 %. Estos hallazgos evidencian la necesidad de fortalecer los programas de certificación de tubérculos-semilla de papa en el país, mediante la utilización de pruebas moleculares de detección viral.

Diacol-Capiro is one of the most important potato (Solanum tuberosum) cultivars in Colombia with most production concentrated in the provinces of Cundinamarca and Boyacá. Unfortunately, this crop is seriously affected by several viruses that compromise the quality of tubers and yields. In this work, it was evaluated the prevalence of the RNA viruses: PLRV, PVY, PVX, PVS, PVV, PYVV, PMTV, and PVB in certified tuber-seed sprouts produced in the highlands of Cundinamarca and Boyacá by RT-qPCR. Results revealed a prevalence of 100 % for PVS, PVX, and PYVV; 75 % for PLRV, 50 % for PVY, 37.5 % for PMTV, and 12.5 % for PVB. Additionally, high-throughput sequencing from a sprout´s bulk sample was used to gather genomic information of infecting viruses, which resulted in a partial PVY sequence, and a complete PLRV genome. Phylogenetic analysis revealed that both assemblies cluster within clades comprising other Colombian isolates with more than 97 % nucleotide sequence identity. These findings highlight the need to update potato seed-tuber certification programs in Colombia with the implementation of more sensitive molecular tests.

Expression of Pinellia ternata leaf agglutinin under rolC promoter confers resistance against a phytophagous sap sucking aphid, Myzus persicae

BACKGROUND: Piercing/sucking insect pests in the order Hemiptera causes substantial crop losses by removing photoassimilates and transmitting viruses to their host plants. Cloning and heterologous expression of plantderived insect resistance genes is a promising approach to control aphids and other sap-sucking insect pests. While expression from the constitutive 35S promoter provides broad protection, the phloem-specific rolC promoter provides better defense against sap sucking insects. The selection of plant-derived insect resistance genes for expression in crop species will minimize bio-safety concerns. RESULTS: Pinellia ternata leaf agglutinin gene (pta), encodes an insecticidal lectin, was isolated and cloned under the 35S and rolC promoters in the pGA482 plant transformation vector for Agrobacterium-mediated tobacco transformation. Integration and expression of the transgene was validated by Southern blotting and qRT-PCR, respectively. Insect bioassays data of transgenic tobacco plants showed that expression of pta under rolC promoter caused 100% aphid mortality and reduced aphid fecundity up to 70% in transgenic tobacco line LRP9. These results highlight the better effectivity of pta under rolC promoter to control phloem feeders, aphids. CONCLUSIONS: These findings suggested the potential of PTA against aphids and other sap sucking insect pests. Evaluation of gene in tobacco under two different promoters; 35S constitutive promoter and rolC phloemspecific promoter could be successfully use for other crop plants particularly in cotton. Development of transgenic cotton plants using plant-derived insecticidal, PTA, would be key step towards commercialization of environmentally safe insect-resistant crops.

A systematic review of conventional and advanced approaches for the control of plant viruses

Viruses are the obligatory intracellular parasites infecting microbes, plants, animals, and humans. They aredead outside host cell but can take-over the host’s cell machinery as soon as they are into it. Several studies oninhibitor compounds have been done for animal viruses including those that are affecting humans, but thereis inadequacy in terms of research and literature for plant viruses that are responsible for losses in crop yieldand quality loss all across the globe. This could be focal point to study plant viruses, their transmission andpathogenicity, and to establish widely used, effective, and advanced approaches for their control. The purposeof this review is to discuss various approaches to control plant viruses that have been developed and applied tocombat plant viral infections. We have divided these approaches into two categories conventional (meristemtip culture, cryotherapy, thermotherapy, and chemotherapy) and advanced (nucleic acid-based approacheslike RNA Silencing, cross-protection, transgenic plants, gene pyramiding, and protein-protein interaction).Moreover, we have discussed and compared the principles, methodologies, advantages, and disadvantages ofeach technique. The approaches have been explored to promote their application in best suited way on variousplants to control viral diseases and to improve food crops quality with increase in production.

Molecular Identification of Potyviruses Infecting Potato Plots in Eastern Antioquia (Colombia) / Identificación molecular de potyvirus infectando cultivos de papa en el oriente de antioquia (colombia)

RESUMEN Los potyvirus son uno de los grupos de virus más limitantes en los cultivos de papa (Solanum tuberosum y S. phureja) en el mundo, siendo PVY, PVV y PVA las especies más prevalentes. En este trabajo se evaluó la presencia de estos potyvirus en cuatro lotes de S. tuberosum cv. Diacol-Capiro y cuatro lotes de S. phureja cv. Criolla-Colombia ubicados en el oriente de Antioquia, analizando la cápside viral mediante RT-PCR/secuenciación Sanger y secuenciación de nueva generación (NGS) para S. tuberosum. Los resultados indicaron la ocurrencia de los potyvirus PVY y PVV en las muestras de S. tuberosum y S. phureja, respectivamente; siendo detectadas mediante cebadores específicos la presencia de tres diferentes cepas de PVY (PVYN, PVYNTN y PVY°) en la región de estudio. Este hallazgo fue confirmado por NGS, obteniendo las secuencias completas de los genomas de estas tres cepas, lo que representa el primer reporte de PVYO en Colombia. Por su parte, los análisis de secuencias de la región CP de PVV indicaron niveles de identidad superiores a 99% con respecto a aislamientos del linaje PVVPhu reportado previamente en Antioquia. Estos hallazgos evidencian la necesidad de ajustar los sistemas de detección de virus en los programas de certificación de tubérculo-semilla de papa adelantados en el país.

ABSTRACT Potyviruses are one of the most limiting viruses for the potato (Solanum tuberosum and S. phureja) production worldwide, being PVY, PVV and PVA the most prevalent species. In this work, we tested the presence of these viruses in four commercial S. tuberosum cv. Diacol-Capiro and S. phureja cv. Criolla-Colombia plots in eastern Antioquia using RT-PCR and Sanger sequencing of the coat protein (CP) region, as well as next generation sequencing (NGS) for S. tuberosum samples. Our results revealed the presence of a PVV strain infecting S. phureja with high sequence identity (>99%) to lineage PVVPhu previously reported in Antioquia. Three strains of PVY (PVYN, PVYNTN and PVY°) were found to infect S. tuberosum. The presence of these three PVY strains was confirmed by NGS, allowing the assembly of their complete genomes. This is the first report of the full genome sequence of PVYo strain in Colombia. These findings highlight the need to adjust the tuber-seed certification programs currently implemented in the country.

Efeito da aplicação de maravilha (Mirabilis jalapa L. ), primavera (Bougainvillea spectabilis L. ) e isolados de Trichoderma na produção de alface / Effect of four o'clock flowers (Mirabilis jalapa L. ), bougainvillea (Bougainvillea spectabilis L. ) and Trichoderma spp. isolates in production of lettuce

Objetivou-se com esse trabalho avaliar o efeito de extratos foliares de primavera (Bougainvillea spetabilis L.) e maravilha (Mirabilis jalapa L.) e de Trichoderma spp., isoladamente ou em combinação, no manejo de viroses e na produção de alface. Os ensaios foram conduzidos em área de produção de hortaliças folhosas no município de Pinhalzinho/SP. O delineamento experimental foi de blocos casualizados, com dez tratamentos comparados com controle químico convencional. Foram observadas reduções de 18 por cento e 32 por cento na população de bactérias da rizosfera nas plantas de alface tratadas com o isolado IB18/22 e extrato de primavera + isolados de Trichoderma spp., respectivamente. Verificou-se que o extrato de maravilha estimulou o aumento do número de UFC/g da população de fungos. Entretanto, não houve diferença significativa na massa fresca da parte aérea e, apesar da pressão de inóculo e da alta densidade populacional de insetos vetores de vírus no local, não foi constatada a ocorrência de viroses, com exceção do espessamento de nervuras, que ocorreu em menor número em plantas tratadas, comparadas as do controle. Economicamente, houve 27,8 por cento de lucratividade e margem bruta de 38,5 por cento, indicando a viabilidade do uso de ferramentas de base ecológica no cultivo de alface. Além disso, os produtos usados são de fácil aquisição, seguros em termos de aplicação, meio ambiente e para o consumidor.

The aim of the work was to evaluate the effect of leaf extracts of bougainvillea (Bougainvillea spetabilis L.) and of four o'clock flowers (Mirabilis jalapa L.) plants and Trichoderma spp., alone or in combination, on viruses management and lettuce production. The tests were conducted in a producing area in Pinhalzinho-SP. The experimental design was randomized blocks with ten treatments in comparison with conventional chemical control. Reductions of 18 percent and 32 percent in the bacterial population of the lettuce rhizosphere treated with IB18/22 isolate and bougainvillea extract + Trichoderma spp. isolates, respectivelly, were observed. Four o'clock plant leaf extract stimulated growth of CFU / g number of fungi population. However, no significant change in fresh mass and size of lettuce was observed, and despite the inoculum pressure and high population density of virus vector insects in place, the occurrence of viruses was not found, except for big-vein syndrome which was higher in control than in treated plants. Economically, 27.8 percent net profit and 38.5 percent gross profit margin were verified, indicating that the use of ecological tools in lettuce crops is feasible. Moreover, the products are easy to obtain, safe to apply and safe for environment and human consumption.

Importância, problemas e perspectivas do melhoramento visando resistência a viroses em plantas / Importance, problems and perspectives of plant breeding concerning resistance to viruses

As viroses são um sério problema para a agricultura, podendo se tomar um fator limitante para o desenvolvimento de determinadas espécies. Medidas de controle, como a eliminação dos vetores, o uso de material sadio, a rotação de culturas e a erradicação de plantas infectadas são apenas soluções temporárias. A mais eficiente estratégia de controle envolve o uso de cultivares melhoradas para resistência ao vírus ou a seu vetor. A reduzida disponibilidade de fontes de resistência pode ser aumentada através da tecnologia do DNA recombinante, que traz novas perspectivas para o melhoramento de plantas resistentes a viroses.

Virus diseases are a serious problem to agricuiture, can be a limitant factor to normal development of some crops. Control measures, like vectors elimination, healthy material use, culture rotation and infected plants eradication, are only transient solutions. The more efficient approach for control involves plant breeding resistant to virus or its vector. Reduced availability of resistance source can be increased through recombinant DNA technology, which brings new breeding perspectives to virus resistant crops.

Similarities in the genomic sequence and coat protein structure of plant virsuses.

The genomic sequences of several RNA plant viruses including cucumber mosaic virus, brome mosaic virus, alfalfa mosaic virus and tobacco mosaic virus have become available recently. The former two viruses are icosahedral while the latter two are bullet and rod shaped, respectively in particle morphology. The non-structural 3a proteins of cucumber mosaic virus and brome mosaic virus have an amino acid sequence homology of 35% and hence are evolutionarily related. In contrast, the coat proteins exhibit little homology, although the circular dichroism spectrum of these viruses are similar. The non-coding regions of the genome also exhibit variable but extensive homology. Comparison of the brome mosaic virus and alfalfa mosaic virus sequences reveals that they are probably related although with a much larger evolutionary distance. The polypeptide folds of the coat protein of three biologically distinct isometric plant viruses, tomato bushy stunt virus, southern bean mosaic virus and satellite tobacco necrosis virus have been shown to display a striking resemblance. All of them consist of a topologically similar 8-standard β-barrel. The implications of these studies to the understanding of the evolution of plant viruses will be discussed.