Generating Genetic Resources for \u3cem\u3ePhytophthora capsici\u3c/em\u3e (L.) and Studying \u3cem\u3eP. capsici\u3c/em\u3e and \u3cem\u3ePhytophthora\u3c/em\u3e Hybrids in Peru

The genus Phytophthora includes more than 90 described species infecting over 1000 plant species. Population studies were conducted to investigate the survival and spread of P. capsici in the Peruvian coastal region. A total of 227 P. capsici isolates, recovered at widely distant localities from 2005-2007, were fingerprinted with AFLPs and SNP genotyping. A clonal population (PcPE-1) represented by 221 isolates was found to be distributed throughout the country. Atypical isolates of P. nicotianae were isolated from loquat trees in Peru and nuclear (internal transcribed spacer [ITS], the phenol acid carboxylase gene, and AFLPs) and mitochondrial genotyping (cytochrome oxidase gene [coxI]) identified this species as a hybrid between P. nicotianae and P. cactorum A comparison of five Phytophthora hybrid isolates from Peru and Taiwan (also infecting loquat trees) suggested that isolates from Peru likely originated from a single hybridization event and that the two isolates from Taiwan originated through different hybridization events. The generation of genetic resources for the study of complex genetic traits in P. capsici was initiated by studying its inbreeding up to the sixth generation. A total of 692 oospore-derived isolates were fingerprinted and a subset was characterized for pathogenicity in cucumber and jalapeno fruits and for segregation of the mating type. The traits tested revealed no-Mendelian segregation, and apomixis were observed to be more prevalent (100%) in deep (fifth generation) inbreeding crosses. Inbreeding was measured by studying the segregation of 20 AFLP markers, which indicated a loss of heterozygosity of ~75% by the sixth generation. The seminal cross from this study was used as a mapping population (F1) for generating a genetic linkage framework with 189 AFLP and 18 SNP markers. A total of 18 linkage groups were produced for each parental isolate using 65 and 42 markers for CBS121657 and CBS121656 isolates respectively covering 409 cM. SNP markers FL5 and FL6 were used for estimating the genome size of P. capsici and precision of the genome assembly. In order to conduct functional studies in P. capsici, we tested the efficacy of the polyethylene glycol mediated transformation. We regenerated up to 30 antibiotic resistant isolates and 53% of them were stable after three months of subculturing

Marketing de contenido para incrementar las ventas de la Empresa Mar y Copas, Chiclayo

La presente investigación tuvo como objetivo general proponer estrategias de marketing de contenidos para incrementar las ventas en la empresa Mar y Copas, Chiclayo; se realizó un diagnóstico de la situación actual del marketing de contenidos, se analizó el nivel de las ventas y se diseñó las estrategias de marketing de contenidos que permita el incrementar las ventas en la empresa Mar y Copas, Chiclayo. La investigación fue cuantitativa, tipo aplicada, descriptiva, propositiva de corte transversal; se aplicó una encuesta a 334 clientes. Los resultados señalan que la situación del uso de marketing de contenido en el restaurante es baja (38%), sus dimensiones de implementación de estrategias son medio (35%) y el análisis para medir el impacto es bajo (34%); el nivel de ventas oscila entre bajo y medio (34%), sus dimensiones como la demanda es bajo (34%), la oferta es baja (46%), la decisión de compra es bajo (36%) y la fidelización de clientes es baja (35%). Conclusión que la propuesta de estrategias de marketing de contenido va lograr incrementar las ventas en el restaurante Mar y Copas

Genetics and mapping of a new anthracnose resistance locus in Andean common bean Paloma

Background: The Andean cultivar Paloma is resistant to Mesoamerican and Andean races of Colletotrichum lindemuthianum, the fungal pathogen that causes the destructive anthracnose disease in common bean. Remarkably, Paloma is resistant to Mesoamerican races 2047 and 3481, which are among the most virulent races of the anthracnose pathogen. Most genes conferring anthracnose resistance in common bean are overcome by these races. The genetic mapping and the relationship between the resistant Co-Pa gene of Paloma and previously characterized anthracnose resistance genes can be a great contribution for breeding programs. Results: The inheritance of resistance studies for Paloma was performed in F2 population from the cross Paloma (resistant) × Cornell 49–242 (susceptible) inoculated with race 2047, and in F2 and F2:3 generations from the cross Paloma (resistant) × PI 207262 (susceptible) inoculated with race 3481. The results of these studies demonstrated that a single dominant gene confers the resistance in Paloma. Allelism tests performed with multiple races of C. lindemuthianum showed that the resistance gene in Paloma, provisionally named Co-Pa, is independent from the anthracnose resistance genes Co-1, Co-2, Co-3, Co-4, Co-5, Co-6, Co-12, Co-13, Co-14, Co-15 and Co-16. Bulk segregant analysis using the SNP chip BARCBean6K_3 positioned the approximate location of Co-Pa in the lower arm of chromosome Pv01. Further mapping analysis located the Co-Pa gene at a 390 kb region of Pv01 flanked by SNP markers SS82 and SS83 at a distance of 1.3 and 2.1 cM, respectively. Conclusions: The results presented here showed that Paloma cultivar has a new dominant gene conferring resistance to anthracnose, which is independent from those genes previously described. The linkage between the Co-Pa gene and the SS82 and SS83 SNP markers will be extremely important for marker-assisted introgression of the gene into elite cultivars in order to enhance resistance.EEA SaltaFil: Castro, Sandra Aparecida de Lima. Universidade Estadual de Maringá. Departamento de Agronomia; BrasilFil: Gonçalves-Vidigal, Maria Celeste. Universidade Estadual de Maringá. Departamento de Agronomia; BrasilFil: Gilio, Thiago Alexandre Santana. Universidade Estadual de Maringá. Departamento de Agronomia; BrasilFil: Lacanallo, Giselly Figueiredo. Universidade Estadual de Maringá. Departamento de Agronomia; BrasilFil: Valentini, Giseli. Universidade Estadual de Maringá. Departamento de Agronomia; BrasilFil: Martins, Vanusa da Silva Ramos. Universidade Estadual de Maringá. Departamento de Agronomia; BrasilFil: Qijian, Song. United States Department of Agriculture. Agricultural Research Service. Soybean Genomics and Improvement Laboratory; Estados UnidosFil: Galvan, Marta Zulema. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta; Argentina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Salta; ArgentinaFil: Hurtado-Gonzales, Oscar P. United States Department of Agriculture. Agricultural Research Service. Soybean Genomics and Improvement Laboratory; Estados UnidosFil: Pastor-Corrales, Marcial Antonio. United States Department of Agriculture. Agricultural Research Service. Soybean Genomics and Improvement Laboratory; Estados Unido

RESISTANT REACTION OF ANDEAN COMMON BEAN LANDRACE G19833, REFERENCE GENOME, TO 13 RACES OF \u3ci\u3eUROMYCES APPENDICULATUS\u3c/i\u3e SUGGESTS BROAD SPECTRUM RUST RESISTANCE

INTRODUCTION- The Andean common bean landrace G19833 (Chaucha Chuga) was recently used to obtain the first reference genome of Phaseolus vulgaris (Schmutz et al, 2014). A large quantity of sequence information is available for this landrace which includes BAC libraries, cDNA libraries, SNP databases, gene expression profiles from various tissues (RNAseq), etc. Additionally, G19833 has been used to generate RIL populations for mapping traits such as phosphorous acquisition, agronomic performance, and disease resistance. G19833 has also been reported as resistant to the anthracnose, angular leaf spot, and Ascochyta blight pathogens and susceptible to the bean golden mosaic and bean common mosaic, viruses (Reviewed by Broughton et al, 2003). However, little is known about the reaction of G19833 to the bean rust pathogen. We report here the reaction of G19833 to a set of 13 races of the rust pathogen, 10 Mesoamerican and three Andean. Together, these races overcome all known rust resistance genes in common bean. Rust resistance in the under-utilized Andean beans lags behind that of Mesoamerican beans. The objective of this study was to determine the spectrum of resistance of Andean G19833 to 13 different virulent races of the bean rust pathogen. MATERIALS AND METHODS - All disease evaluations were conducted under greenhouse conditions using published methodologies (Stavely, 1983). Landrace G19833 was inoculated with races 15-1 (41), 6-10 (44), 15-3 (47), 22-6 (49), 31-1 (53), 31-7 (58), 31-22 (67), 21-0 (72), 73 (6-15), 84 (37-1), 85 (6-31), 102 (29-0), and 108 (22-52). Rust resistance controls included Pinto 114, Aurora (Ur-3), Mexico 235 (Ur-3+), Early Gallatin (Ur-4), Mexico 309 (Ur-5), Golden Gate Wax (Ur-6), Great Northern 1140 (Ur-7), Pompadour Checa 50 (Ur-9/Ur-12), PI181996 (Ur-11), Redlands Pioneer (Ur-13), Ouro Negro (Ur-14), PI260418 (Ur-Unnamed), and PI310762 (Ur-Unnamed), and Compuesto Negro de Chimaltenango, CNC (Ur-Unnamed). Rust phenotypic evaluations were conducted twelve days after inoculations

FINE MAPPING THE BROAD SPECTRUM ANTHRACNOSE RESISTANCE GENE IN AMENDOIM CAVALO

INTRODUCTION: The Andean common bean landrace Amendoim Cavalo (AC) is resistant to races 2, 7, 9, 19, 23, 39, 55, 65, 73, 89, 1545, 2047 and 3481 of Colletotrichum lindemuthianum (Nanami et al, 2014). None of the common bean anthracnose resistance genes known to date, were resistant to all 13 races mentioned above, to which AC was resistant. The resistance in AC is conferred by a single and dominant gene (Co-AC) that is independent of the other known genes (Nanami et al., 2014 and Gilio et al, 2016). The AC locus has been located in the lower arm of chromosome Pv01 of common bean (Gilio et al., 2016). The objective of this study was to use fine mapping to locate the position Co-AC locus in the common bean genome

Thermotherapy Followed by Shoot Tip Cryotherapy Eradicates Latent Viruses and Apple Hammerhead Viroid from In Vitro Apple Rootstocks

Virus and viroid-free apple rootstocks are necessary for large-scale nursery propagation of apple (Malus domestica) trees. Apple stem grooving virus (ASGV) and Apple chlorotic leaf spot virus (ACLSV) are among the most serious apple viruses that are prevalent in most apple growing regions. In addition to these viruses, a new infectious agent named Apple hammerhead viroid (AHVd) has been identified. We investigated whether thermotherapy or cryotherapy alone or a combination of both could effectively eradicate ACLSV, ASGV, and AHVd from in vitro cultures of four apple rootstocks developed in the Cornell-Geneva apple rootstock breeding program (CG 2034, CG 4213, CG 5257, and CG 6006). For thermotherapy treatments, in vitro plants were treated for four weeks at 36 °C (day) and 32 °C (night). Plant vitrification solution 2 (PVS2) and cryotherapy treatments included a shoot tip preculture in 2 M glycerol + 0.8 M sucrose for one day followed by exposure to PVS2 for 60 or 75 min at 22 °C, either without or with liquid nitrogen (LN, cryotherapy) exposure. Combinations of thermotherapy and PVS2/cryotherapy treatments were also performed. Following treatments, shoot tips were warmed, recovered on growth medium, transferred to the greenhouse, grown, placed in dormancy inducing conditions, and then grown again prior to sampling leaves for the presence of viruses and viroids. Overall, thermotherapy combined with cryotherapy treatment resulted in the highest percentage of virus- and viroid-free plants, suggesting great potential for producing virus- and viroid-free planting materials for the apple industry. Furthermore, it could also be a valuable tool to support the global exchange of apple germplasm

Fine Mapping of Ur-3, a Historically Important Rust Resistance Locus in Common Bean

Bean rust, caused by Uromyces appendiculatus, is a devastating disease of common bean (Phaseolus vulgaris) in the Americas and Africa. The historically important Ur-3 gene confers resistance to many races of the highly variable bean rust pathogen that overcome other rust resistance genes. Existing molecular markers tagging Ur-3 for use in marker-assisted selection produce false results. Here, we describe the fine mapping of the Ur-3 locus for the development of highly accurate markers linked to Ur-3. An F2 population from the cross Pinto 114 (susceptible) × Aurora (resistant with Ur-3) was evaluated for its reaction to four different races of U. appendiculatus. A bulked segregant analysis using the SNP chip BARCBEAN6K_3 placed the approximate location of Ur-3 in the lower arm of chromosome Pv11. Specific SSR and SNP markers and haplotype analysis of 18 sequenced bean varieties positioned Ur-3 in a 46.5 kb genomic region from 46.96 to 47.01 Mb on Pv11. We discovered in this region the SS68 KASP marker that was tightly linked to Ur-3. Validation of SS68 on a panel of 130 diverse common bean cultivars containing all known rust resistance genes revealed that SS68 was highly accurate and produced no false results. The SS68 marker will be of great value in pyramiding Ur-3 with other rust resistance genes. It will also significantly reduce time and labor associated with the current phenotypic detection of Ur-3. This is the first utilization of fine mapping to discover markers linked to rust resistance in common bean

HTS-Based Diagnostics of Sugarcane Viruses: Seasonal Variation and Its Implications for Accurate Detection

Rapid global germplasm trade has increased concern about the spread of plant pathogens and pests across borders that could become established, affecting agriculture and environment systems. Viral pathogens are of particular concern due to their difficulty to control once established. A comprehensive diagnostic platform that accurately detects both known and unknown virus species, as well as unreported variants, is playing a pivotal role across plant germplasm quarantine programs. Here we propose the addition of high-throughput sequencing (HTS) from total RNA to the routine quarantine diagnostic workflow of sugarcane viruses. We evaluated the impact of sequencing depth needed for the HTS-based identification of seven regulated sugarcane RNA/DNA viruses across two different growing seasons (spring and fall). Our HTS analysis revealed that viral normalized read counts (RPKM) was up to 23-times higher in spring than in the fall season for six out of the seven viruses. Random read subsampling analyses suggested that the minimum number of reads required for reliable detection of RNA viruses was 0.5 million, with a viral genome coverage of at least 92%. Using an HTS-based total RNA metagenomics approach, we identified all targeted viruses independent of the time of the year, highlighting that higher sequencing depth is needed for the identification of DNA viruses