Institute Investigaciones en Fruticultura Tropical

Havana, Cuba

Institute Investigaciones en Fruticultura Tropical

Havana, Cuba
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Hernandez-Rodriguez L.,Institute Investigaciones en Fruticultura Tropical | Bertalmio A.,Instituto Nacional Of Investigacion Agropecuaria | Arruabarrena A.,Instituto Nacional Of Investigacion Agropecuaria | Rubio L.,Instituto Nacional Of Investigacion Agropecuaria | And 5 more authors.
Plant Disease | Year: 2017

Citrus tristeza virus (CTV) (Closteroviridae: Closterovirus) has multiple isolates classified into six major CTV genotype groups or strains: T36, VT, T3, RB, T68, and T30 (Harper 2013). The reported circulating strains in Uruguay are VT, T3, T36, and NC, a new genetic lineage (Benítez-Galeano et al. 2015). To continue surveying CTV genotypes in Uruguay, single aphid (Toxoptera citricida) transmission (SAT) was performed to separate the virus population mixtures of several field isolates as described by Powell and Lin (2005). SAT was conducted by inoculating field isolates in Mexican lime (Citrus aurantifolia), challenging 28 plants per field isolate. CTV infection was confirmed by ELISA with 3DF1+3CA5 and MCA13 antibodies (PlantPrint Diagnostics), and RNA silencing suppressor genes p20 and p25 were amplified by RT-PCR (Benítez-Galeano et al. 2015). In 2015, nine subisolates were obtained by SAT from a field isolate named SNH-A, which was collected from a 23-year-old asymptomatic ‘Newhall’ sweet orange (C. sinensis) grafted onto Poncirus trifoliata in a commercial orchard at Paysandú, Uruguay, in January 2014. CTV was detected in one of these subisolates (SNH-9-sA) by 3DF1+3CA5 and MCA13 antibodies 30 days after SAT. RNA was extracted as recommended in Arruabarrena et al. (2016), and fragments with expected sizes of 561 and 677 bp were amplified after retrotranscription for the p20 and p25 genes, respectively. When comparing p20 and p25 sequences of SNH-A (KU928192, KU900357) and SNH-9-sA (KU928185, KU900356) with each other the nucleotide identities were 94.5 and 93.5%, respectively. These sequences also showed nucleotide identities ranging from 91 to 98% with reference isolates of the CTV strains (T36: AY340974, U16304, VT: EU857538, T3: KC525952, RB: FJ525431, T68: JQ965119, T30: AF260651, Y18420, and NC: GQ454870). The highest nucleotide identity for the two analyzed genes of SNH-9-sA sequences was with the resistance-breaking RB strain, and for SNH-A sequences with T3 (p20) and NC (p25) strains. The RB phenotype of subisolate SNH-9-sA was demonstrated by grafting three bark patches onto three P. trifoliate seedlings (rutaceous species where only RB strains can systemically move to cause an infection, Dawson and Mooney 2000). One of the three SNH-9-sA inoculated plants (SNH-9-sA-Tp3) was 3DF1+3CA5 and MCA13 positive 8 months after graft-inoculation. Amplicons for the p20 and p25 genes were amplified and the corresponding sequences (KY559506, KY367397) showed the highest nucleotide identity with the RB strain. Maximum likelihood phylogenetic trees of the two genes grouped the SNH-9-sA, SNH-9-sA-Tp3, and RB strain sequences in the same phylogenetic cluster, while SNH-A was grouped in different clades depending on the analyzed gene. To our knowledge, this is the first report of the presence of the CTV RB genotype in South America. Taking into consideration the widespread use of P. trifoliata as rootstock in Uruguay, the described findings highlight the urgent need to study the epidemiology of the RB genotype in the country, and enforce management strategies that consider the presence of this genotype for the implementation of control measures. © 2017, American Phytopathological Society. All rights reserved.


Arruabarrena A.,Instituto Nacional Of Investigacion Agropecuaria | Benitez-Galeano M.J.,University of the Republic of Uruguay | Giambiasi M.,Instituto Nacional Of Investigacion Agropecuaria | Bertalmio A.,Instituto Nacional Of Investigacion Agropecuaria | And 2 more authors.
Journal of Virological Methods | Year: 2016

Standard molecular methods for plant virus diagnosis require the purification of RNA or DNA extracts from a large number of samples, with sufficient concentration and quality for their use in PCR, RT-PCR, or qPCR analysis. Most methods are laborious and use either hazardous and/or costly chemicals. A previously published protocol for RNA isolation from several plant species yields high amounts of good quality RNA-DNA mixture in a simple, safe and inexpensive manner. In the present work, this method was tested to obtain RNA-DNA extracts from leaves of tomato, potato and three species of citrus, and was compared with two commercial kits. The results demonstrated that this protocol offers at least comparable nucleic acid quality, quantity and purity to those provided by commercial phenol-based or spin column systems and that are suitable to be used in PCR, RT-PCR and qPCR for virus and viroid detection. Because of its easy implementation and the use of safe and inexpensive reagents, it can be easily implemented to work in plant virus and viroid detection in different plant species. © 2016 Elsevier B.V.


Soto M.B.,Institute Investigaciones en Fruticultura Tropical | Castillo A.B.,Institute Investigaciones en Fruticultura Tropical | Rodriguez Y.A.,Institute Investigaciones en Fruticultura Tropical | Espinosa D.H.,Institute Investigaciones en Fruticultura Tropical | Rubial M.R.,Institute Investigaciones en Fruticultura Tropical
Acta Horticulturae | Year: 2015

The phytosanitary surveillance plays an important role in the early detection to prevent the establishment of harmful pests of economic and quarantine importance. The presence of fruit flies belonging to the genera Anastrepha, Bactrocera or Ceratitis (Diptera: Tephritidae) on the American continent constitute a potential risk of introduction, threatening the production of citrus and other hosts of countries in the region. In Cuba, the presence of some Anastrepha spp., as well as Bactrocera carambolae or Ceratitis capitata is not registered, thus surveillance for these quarantine pests and for other species of national interest continues. Results have been obtained from research on preferred host fruits located in the surroundings of citrus plots, revealing the presence and potential risk to citrus of A. suspensa, identified as of quarantine importance in countries such as the United States and others. During the period 2010-2012 three activities were conducted to characterize, diagnose and train staff to strengthen phytosanitary surveillance of Anastrepha fruit flies in citrus production areas in: Ceiba Citrus Enterprises (Artemisa), Victoria de Giron (Matanzas), Arimao (Cienfuegos) and Ciego de Avila citrus enterprises. To fulfill this objective, the composition of host fruit presence in production areas with respect to A. suspensa was evaluated. Host preferences of the main Anastrepha species detected and the effectiveness of traps and attractants were determined, and the monitoring system was relocated. The information obtained allowed designing the monitoring strategy in production areas. Training, one of the most important tools, was developed through seminars and workshops. These results provide inputs to the monitoring programs and the procedures that are essential in the development and implementation of measures for the phytosanitary surveillance and integrated management of fruit flies in Cuban citrus production.


Beovides Y.,Institute Investigaciones Of Viandas Tropicales Inivit | Fregene M.,Donald Danforth Plant Science Center | Gutierrez J.P.,Aereo | Milian M.D.,Institute Investigaciones Of Viandas Tropicales Inivit | And 10 more authors.
Biotechnology, Agronomy and Society and Environment | Year: 2015

A total of 36 microsatellites (SSR) markers were used to analyze the genetic diversity of 163 accessions of cultivated cassava (Manihot esculenta Crantz), 94 accessions of them from the Cuban Cassava Germplasm Collection and 69 genotypes from different countries and conserved at the International Center for Tropical Agriculture (Colombia). This study was carried out to determine genetic diversity within and between all accessions to promote their better use and conservation strategies. Thirty-four of those markers were used for the genetic diversity study based on their higher polymorphism. The Cuban cultivars showed the highest average allele number per loci with 5.8 and 100% of the loci were polymorphic, as well as those from Guatemala. The average proportion of individual heterozygocity observed (HO) was high (0.5918 ± 0.0351), while the highest HO rates were observed in groups of genotypes from Cuba (0.6016) and Tanzania (0.6459). The total heterozygocity (HT) was high (0.6538 ± 0.1770), but only 7.4% (GST = 0.0740 ± 0.0377) was due to differences between the five countries studied. Genetic differentiation coefficients (estimated by F-statistics) were low to moderate (FST > 0.04) and 17 unique alleles with low frequency were found in Cuban cultivars. The results provide the first molecular characterization of Cuban cassava genotypes and showed a wide diversity among landraces from Cuba. Application of this valuable information can be used for genetic diversity conservation and genotype identification studies for the genetic breeding program of cassava. © 2015, FAC UNIV SCIENCES AGRONOMIQUES GEMBLOUX .All Rights Reserved.


Pino J.,Institute Investigaciones para la Industria Alimentaria | Fuentes V.,Institute Investigaciones en Fruticultura Tropical | Barrios O.,Institute Investigaciones Fundamentales En Agricultura Tropical Alejandro Of Humboldt
Food Chemistry | Year: 2011

The steam volatile components of three cultivars of Cachucha mature peppers (Capsicum chinense Jacq.) were isolated by steam-distillation-continuous-extraction and analysed using GC and GC-MS. The composition of volatile compounds of the peppers differs clearly for the different cultivars. The content of volatile compounds, responsible for the flavour of Cachucha peppers, varied between 110.71 and 302.53mgkg-1. One hundred and thirty-six compounds were identified, from which hexyl isopentanoate, hexyl pentanoate, hexyl 2-methylbutanoate, 3,3-dimethylcyclohexanol, γ-himachalene and germacrene D were the major ones. © 2010 Elsevier Ltd.


Canales E.,Center for Genetic Engineering and Biotechnology | Coll Y.,University of Habana | Hernandez I.,Center for Genetic Engineering and Biotechnology | Portieles R.,Center for Genetic Engineering and Biotechnology | And 24 more authors.
PLoS ONE | Year: 2016

Huanglongbing (HLB) constitutes the most destructive disease of citrus worldwide, yet no established efficient management measures exist for it. Brassinosteroids, a family of plant steroidal compounds, are essential for plant growth, development and stress tolerance. As a possible control strategy for HLB, epibrassinolide was applied to as a foliar spray to citrus plants infected with the causal agent of HLB, 'Candidatus Liberibacter asiaticus'. The bacterial titers were reduced after treatment with epibrassinolide under both greenhouse and field conditions but were stronger in the greenhouse. Known defense genes were induced in leaves by epibrassinolide. With the SuperSAGE technology combined with next generation sequencing, induction of genes known to be associated with defense response to bacteria and hormone transduction pathways were identified. The results demonstrate that epibrassinolide may provide a useful tool for the management of HLB. This is an open access article, free of all copiright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.


PubMed | Shinshu University, Center for Genetic Engineering and Biotechnology, University of Habana, Michigan State University and 5 more.
Type: Journal Article | Journal: PloS one | Year: 2016

Huanglongbing (HLB) constitutes the most destructive disease of citrus worldwide, yet no established efficient management measures exist for it. Brassinosteroids, a family of plant steroidal compounds, are essential for plant growth, development and stress tolerance. As a possible control strategy for HLB, epibrassinolide was applied to as a foliar spray to citrus plants infected with the causal agent of HLB, Candidatus Liberibacter asiaticus. The bacterial titers were reduced after treatment with epibrassinolide under both greenhouse and field conditions but were stronger in the greenhouse. Known defense genes were induced in leaves by epibrassinolide. With the SuperSAGE technology combined with next generation sequencing, induction of genes known to be associated with defense response to bacteria and hormone transduction pathways were identified. The results demonstrate that epibrassinolide may provide a useful tool for the management of HLB.


Cabrera R.I.,Institute Investigaciones en Fruticultura Tropical | Decock C.,Catholic University of Louvain | Herrera S.,Institute Ecologia y Sistematica | Ferrer J.,Institute Investigaciones en Fruticultura Tropical | And 3 more authors.
Crop Protection | Year: 2014

Cuban citriculture has recently been affected by important pests and diseases including black citrus aphid Toxoptera citricida Kirkaldy, citrus leafminer Phyllocnistis citrella Stainton, Asiatic citrus psyllid Diaphorina citri Kuwayama, Citrus tristeza disease, and huanglongbing. This complex of 'invasive species' is thought to be contributing to the increasing decline of commercial citrus groves. During several surveys carried out in commercial citrus areas of several provinces in Cuba, symptoms of dieback, characterized by the presence of abundant dry branches and wood rot were detected on Criolla Valencia orange, Marsh grapefruit trees on sour orange, and Cleopatra mandarin rootstocks. Samples of branches and trunk wood were processed and a fungus was isolated from all samples. Morphological, physiological and molecular characterization demonstrated that the isolated agent is Fomitiporia maxonii Murrill. The incidence of affected trees was high in most surveyed groves, reaching 100% in old groves. Symptoms similar to the ones found in naturally infected trees were reproduced on trees inoculated with pure cultures of the fungus, and the fungus was recovered in culture from inoculated trees, thus fulfilling Koch's postulates. This is the first report of an association of F.maxonii with citrus wood rot in Cuba. © 2014 Elsevier Ltd.


Betancourt M.,Institute Investigaciones en Fruticultura Tropical | Sistachs V.,University of Habana | Martinez-Fuentes A.,Polytechnic University of Valencia | Mesejo C.,Polytechnic University of Valencia | And 2 more authors.
Journal of Horticultural Science and Biotechnology | Year: 2014

Grapefruit grown in tropical climates reach a suitable fruit size and juice content for fresh consumption during August - October in the northern hemisphere. However, some tropical plantations delay harvesting until November or December, with the fruit then being used for processing. In our experiments, delaying the harvest from October to December reduced the average mature fruit weight by 10% and increased abscission from 29 to 70 fruit per tree. Juice contents decreased slightly, from 44.5% (w/w) to 43.2% (w/w), while total soluble solids (TSS) contents barely changed, from 10.4 oBrix to 10.2 oBrix. Delaying the harvest date also reduced return flowering by 20% in the following Spring, and the number of fruit set by 20%. Mature fruit abscission and reduced flowering were not dependent on weather conditions.The former was spontaneous and due to senescence, while the latter was due to fruit remaining on the tree. Over a 4-year period, our results showed an average reduction of 30% in fruit yield per tree when harvest dates were delayed from October (153 kg tree-1) to December (105 kg tree-1). As juice content and TSS content values were suitable for processing in October, there was no reason to delay the harvest date.


Fernandez-Calienes Valdes A.,Institute Medicina Tropical Pedro Kouri Ipk | Cuesta Rubio O.,Technical University of Machala | Fuentes Fiallo V.,Institute Investigaciones en Fruticultura Tropical | Monzote Fidalgo L.,Institute Medicina Tropical Pedro Kouri Ipk | Mendiola Martinez J.,Institute Medicina Tropical Pedro Kouri Ipk
Revista Cubana de Medicina Tropical | Year: 2015

Introduction: malaria control mostly depends on an effective therapy. Many current antimalarials are of natural origin. Cuban flora species contain anti-Plasmodium metabolites. This study identifies Solanaceae extracts with promising antiplasmodial activity. Objective: to evaluate anti-Plasmodium berghei schizonticidal activity of 31 extracts of seven species corresponding to five genera of Solanaceae plants collected in the west of our country, without history of a similar study. Methods: 31 hydroalcoholic (30%/90% ethanol) extracts were prepared with different body parts from Brunfelsia undulata Sw., Datura stramonium L. var. tatula (L.) Torr., Physalis solanaceus (Schltdl.) Axelius, Solandra longiflora Tuss., Solanum myriacanthum Dunal, Solanum seaforthianum And. and Solanum umbellatum Mill. The extracts activity was evaluated in vitro for Plasmodium berghei and their cytotoxicity was determined for human fibroblasts MRC-5. Results: brunfelsia undulata and Solanum umbellatum extracts were inactive. The Solanum seaforthianum stems extract showed the strongest antiplasmodial activity (CI50=3.9 ?g/ml) with excellent selectivity (18.2). Conclusions: the antiplasmodial in vitro activity of extracts from five species of Solanaceae was demonstrated, without any history of this pharmacological action. An extract was identified to have a powerful schizontocidal activity against Plasmodium berghei and excellent selectivity. This result encourages us to continue the study of the plant preparation of Solanum seaforthianum. © 2015, Editorial Ciencias Medicas. All rights reserved.

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