Browsing by Author "Caligari, Peter D. S."
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- ItemApplication of inter-simple sequence repeats relative to simple sequence repeats as a molecular marker system for indexing blueberry cultivars(2013) Garriga, Miguel; Parra, Pablo A.; Caligari, Peter D. S.; Retamales, Jorge B.; Carrasco Gálvez, Basilio Alejandro; Lobos, Gustavo A.; García Gonzales, Rolando
- ItemDevelopment and characterization of microsatellite markers in Gaultheria pumila Lf. (Ericaceae)(2018) Carrasco Gálvez, Basilio Alejandro; Garcia Gonzales, Rolando.; Pico Mendoza, José.; Quiroz, Karla.; Cáceres, Pablo.; Chong Perez, Borys.; Pino, Hugo.; Seiltgens, Marjorie.; Greck, Eglis.; Caligari, Peter D. S.Abstract Background Polymorphic microsatellite markers were developed for Gaultheria pumila (Ericaceae) to evaluate genetic diversity and population structure within its native range in Chile. This is a very important Ericaceae endemic to Chile with a large commercial potential. Its resistance to different abiotic conditions makes it a valuable target for genetic improvement. Results Ten polymorphic simple sequence repeat (SSR) loci were isolated from Gaultheria pumila using new-generation 454 FLX Titanium pyrosequencing technology. The mean number of alleles per locus ranged from 2 to 4. Observed and expected heterozygosity ranged from 0.00 to 1.0 and 0.00 to 0.64, respectively. Conclusions From 10 SSR markers developed for G. pumila, 9 markers are promising candidates for analyzing genetic variation within or between natural populations of G. pumila and other species from the same genus.
- ItemEfficient protocols for the extraction of microbial DNA from the rhizosphere of hydrophilic forests in Chile(PONTIFICIA UNIV CATOLICA CHILE, FAC AGRONOMIA INGENIERIA FORESTAL, 2012) Caceres, Pablo; Cordero, Cecilia; Gonzalez, Gloria; Quiroz, Karla; Bobadilla, Juan C.; Bravo, Carmen; Caligari, Peter D. S.; Carrasco, Basilio; Garcia Gonzales, RolandoP. Caceres, C. Cordero, G. Gonzalez, K. Quiroz, J.C. Bobadilla, C. Bravo, P.D.S. Caligari, B. Carrasco, and R. Garcia-Gonzales. 2012. Efficient protocols for the extraction of microbial DNA from the rhizosphere of hydrophilic forests in Chile. Cien. Inv. Agr. 38(3): 585-592. A lysis buffer-based protocol (Protocol BA), a modified lysis buffer-based protocol (Protocol BA Mod) and a commercial extraction kit (Protocol PS Kit) (Power Soil, Mo bio Laboratories, CA USA) were each evaluated for their ability to produce high-quality DNA with yields sufficient to allow its use in biodiversity studies. Similarly, the effect of liquid nitrogen on the process of cell disruption in all of the protocols that were studied. DNA yields ranged from 12.4 ng g(-1) of processed soil to 9620 ng g(-1) using the modified lysis buffer and commercial extraction kit, respectively. The quality of the DNA was determined by the ability of the DNA to produce efficient and reproducible polymerase chain reaction (PCR) products, using primers for universal 16S and 18S ribosomal RNA regions from bacteria and fungi, respectively. High-quality DNA was obtained to run PCRs in all protocols, but the efficiency of the method depended on the dilution of the DNA prior to performing the PCR. The three extraction methods generated PCR products with 90% efficiency. The DNA produced with the commercial kit was able to produce the highest PCR efficiency (95%) when the 10(-1) dilution was used. The method based on the use of lysis buffer produced the highest efficiency (90%) using a 10(-2) dilution. Meanwhile, the modified lysis buffer-based protocol generated the highest efficiency of PCR products using the 10(-3) dilution factor with 95% of efficiency. For the first time, reliable and efficient DNA isolation from the rhizosphere of hydrolic forest is documented, enabling a wide range of applications for this technique.
- ItemGenetic diversity and population structure of Chilean blueberry Gaultheria pumila (L.f.) DJ Middleton (Ericaceae)(2020) Pico-Mendoza, Jose; Garcia-Gonzales, Rolando; Quiroz, Karla; Pinoargote, Miryan; Rodriguez-Alvarez, Yohaily; Chong, Borys; Caceres-Ruz, Pablo; Pino, Hugo; Caligari, Peter D. S.; Carrasco, BasilioGaultheria pumila (L.f.) D.J. Middleton is a native shrub of Chile that produces edible berry fruits. This species is related to the cultivated Vaccinium species; for this reason it is currently called Chilean blueberry locally. Although G. pumila has important attributes, it has been largely ignored, and remains an unexplored genetic resource. This study investigates the genetic diversity to support the efforts to domesticate the species. Sampling was carried out in 11 sites collected from four Regions of Chile. In total, 160 individuals were collected and analyzed using a set of 10 simple sequence repeats (SSRs) markers. The average observed heterozygosity was Ho = 0.50, while the expected heterozygosity was He = 0.46. The fixation index (F-IS) showed an average of -0.07, and the proportion of differentiation among populations (F-ST) was 0.11. The average level of polymorphic loci in all populations (PPL) was 96.97%. AMOVA showed that the genetic diversity among populations was very low (Phi PT = 6%). Significant correlations were found between genetic and geographic distance. Multivariate and Bayesian analyses identified two genetic groups. These results will be very useful to support the efforts to domesticate and increase the value of this species.
- ItemGenetic structure of highland papayas (Vasconcellea pubescens (Lenn, et C. Koch) Badillo) cultivated along a geographic gradient in Chile as revealed by Inter Simple Sequence Repeats (ISSR)(SPRINGER, 2009) Carrasco, Basilio; Avila, Patricio; Perez Diaz, Jorge; Munoz, Patricio; Garcia, Rolando; Lavandero, Blas; Zurita Silva, Andres; Retamales, Jorge B.; Caligari, Peter D. S.In Chile Vasconcellea pubescens is cropped to produce canned fruit, juice, jam and processed sweets. Additionally this species produces latex with a high level of papain, an important and valuable proteolytic enzyme with industrial applications. In this investigation seven ISSR primers were used to study the level and organization of genetic diversity in 333 samples of V. pubescens. Out of the 114 bands recorded, 63 proved to be polymorphic (P = 55.3%). At the species level, the genetic diversity was rather low (h = 0.01 +/- A 6,80188E-05, Shannon's Index I = 0.16 +/- A 0,000148). The major portion of the genetic diversity was found within groups (65%). The genetic differentiation between the different groups was significant, as the AMOVA analysis suggested (I broken vertical bar(pt) = 0.35). When analysing the Northern area alone, the differentiation increased to I broken vertical bar(pt) = 0.40. When only the Southern area was analysed, I broken vertical bar(pt) decreased to 0.18, indicating greater genetic similarity among the samples. The results generated from Structure and Bayesian Analysis of Population Structure distinguished 8 genetically different groups, five of them located in the north and three in the south. The results are discussed in the light of the growers' practices.
- ItemGenomic structure of yellow lupin (Lupinus luteus): genome organization, evolution, gene family expansion, metabolites and protein synthesis(2025) Martinez-Hernandez, J. E.; Salvo-Garrido, Haroldo; Levicoy, Daniela; Caligari, Peter D. S.; Rupayán, Annally; Moyano Yugovic, Tomás Custodio; Carrasco, Makarena; Hernandez, Sebastián; Armijo-Godoy, Grace; Westermeyer, Fernando; Larama, GiovanniYellow lupin (Lupinus luteus) gives valuable high-quality protein and has good sustainability due to its ability in nitrogen fixation and exudation of organic acids, which reduces the need for chemical-based phosphate fertilization in acid soils. However, the crop needs further improvements to contribute in a major way to sustainable agriculture and food security. In this study, we present the first chromosome-level genome assembly of L. luteus. The results provide insights into its genomic organization, evolution, and functional attributes. Using integrated genomic approaches, we unveil the genetic bases governing its adaptive responses to environmental stress, delineating the intricate interplay among alkaloid biosynthesis, mechanisms of pathogen resistance, and secondary metabolite transporters. Our comparative genomic analysis of closely related species highlights recent speciation events within the Lupinus genus, exposing extensive synteny preservation alongside notable structural alterations, particularly chromosome translocations. Remarkable expansions of gene families implicated in terpene metabolism, stress responses, and conglutin proteins were identified, elucidating the genetic basis of L. luteus’ superior nutritional profile and defensive capabilities. Additionally, a diverse array of disease resistance-related (R) genes was uncovered, alongside the characterization of pivotal enzymes governing quinolizidine alkaloid biosynthesis, thus shedding light on the molecular mechanisms underlying “bitterness” in lupin seeds. This comprehensive genomic analysis serves as a valuable resource to improve this species in terms of resilience, yield, and seed protein levels to contribute to food and feed to face the worldwide challenge of sustainable agriculture and food security.
- ItemIN VITRO PROPAGATION OF CEDAR (Cedrela odorata L.) FROM JUVENILE SHOOTS(2011) Garcia-Gonzales, Rolando; Delgado, Miladys; Gonzalez, Yailin; Gonzalez, Anibal; Garriga, Miguel; Caligari, Peter D. S.; Carrasco, Basilio; Quiroz, KarlaCedrela odorata L. is one of the most important timber species currently traded in the Caribbean and Central America; however, it has been intensively exploited. In vitro techniques and clonal propagation can help to develop new plantations and assist in establishing improvement programs for this species. The aim of this study was to develop a protocol to establish in vitro conditions and to micropropagate this species from nodal explants from juvenile cuttings taken from field trees. Disinfection of node explants with 5% propiconazole CE 25 during 3 min resulted in 100% explant disinfection and 60% morphogenic response on those established explants. Shoot development was optimized by cultivating in vitro node explants in Murashige and Skoog basal medium supplemented with 2 mg L-1 6-bencilaminopurine and 3 mg L-1 naphthaleneacetic acid. This medium resulted in 100% shoot development from the in vitro node explants with a 3.93 cm mean height. Rooting was also stimulated 6 wk after individualization of the regenerated plants on the same micropropagation medium with a mean of 3.9 roots per plant. In vitro plants did not show morphologic differences when compared to ex vitro seeds.
- ItemIn vitro propagation of cedar (Cedrela odorata L.) from juvenile shoots(2011) García Gonzáles, Rolando; Delgado, Miladys; González, Yailín; González, Aníbal; Garriga, Miguel; Caligari, Peter D. S.; Carrasco Gálvez, Basilio Alejandro; Quiroz Bravo, Karla
- ItemMeristem culture and subsequent micropropagation of chilean strawberry (Fragaria chiloensis (L.) Duch.)(2017) Quiroz, Karla A.; Carrasco Gálvez, Basilio Alejandro; Berríos, Miguel.; Retamales, Jorge B.; Caligari, Peter D. S.; García-Gonzáles, Rolando.Abstract Background Vegetative propagation of Fragaria sp. is traditionally carried out using stolons. This system of propagation, in addition to being slow, can spread plant diseases, particularly serious being viral. In vitro culture of meristems and the establishment of micropropagation protocols are important tools for solving these problems. In recent years, considerable effort has been made to develop in vitro propagation of the commercial strawberry in order to produce virus-free plants of high quality. These previous results can serve as the basis for developing in vitro-based propagation technologies in the less studied species Fragaria chiloensis. Results In this context, we studied the cultivation of meristems and establishment of a micropropagation protocol for F. chiloensis. The addition of polyvinylpyrrolidone (PVP) improved the meristem regeneration efficiency of F. chiloensis accessions. Similarly, the use of 6-benzylaminopurine (BAP) in the culture media increased the average rate of multiplication to 3–6 shoots per plant. In addition, the use of 6-benzylaminopurine (BAP), had low levels (near zero) of explant losses due to oxidation. However, plant height as well as number of leaves and roots were higher in media without growth regulators, with average values of 0.5 cm, 9 leaves and 4 roots per plant. Conclusions For the first time in Chilean strawberry, meristem culture demonstrated to be an efficient tool for eliminating virus from infected plants, giving the possibility to produce disease free propagation material. Also, the addition of PVP into the basal MS medium improved the efficiency of plant recovery from isolated meristems. Farmers can now access to high quality plant material produced by biotech tools which will improve their technological practices.Abstract Background Vegetative propagation of Fragaria sp. is traditionally carried out using stolons. This system of propagation, in addition to being slow, can spread plant diseases, particularly serious being viral. In vitro culture of meristems and the establishment of micropropagation protocols are important tools for solving these problems. In recent years, considerable effort has been made to develop in vitro propagation of the commercial strawberry in order to produce virus-free plants of high quality. These previous results can serve as the basis for developing in vitro-based propagation technologies in the less studied species Fragaria chiloensis. Results In this context, we studied the cultivation of meristems and establishment of a micropropagation protocol for F. chiloensis. The addition of polyvinylpyrrolidone (PVP) improved the meristem regeneration efficiency of F. chiloensis accessions. Similarly, the use of 6-benzylaminopurine (BAP) in the culture media increased the average rate of multiplication to 3–6 shoots per plant. In addition, the use of 6-benzylaminopurine (BAP), had low levels (near zero) of explant losses due to oxidation. However, plant height as well as number of leaves and roots were higher in media without growth regulators, with average values of 0.5 cm, 9 leaves and 4 roots per plant. Conclusions For the first time in Chilean strawberry, meristem culture demonstrated to be an efficient tool for eliminating virus from infected plants, giving the possibility to produce disease free propagation material. Also, the addition of PVP into the basal MS medium improved the efficiency of plant recovery from isolated meristems. Farmers can now access to high quality plant material produced by biotech tools which will improve their technological practices.
- ItemMOLECULAR TOOLS FOR RAPID AND ACCURATE DETECTION OF BLACK TRUFFLE (Tuber melanosporum Vitt.) IN INOCULATED NURSERY PLANTS AND COMMERCIAL PLANTATIONS IN CHILE(2011) Cordero, Cecilia; Caceres, Pablo; Gonzalez, Gloria; Quiroz, Karla; Bravo, Carmen; Ramirez, Ricardo; Caligari, Peter D. S.; Carrasco, Basilio; Garcia-Gonzales, RolandoTruffle (Tuber melanosporum Vitt.) culture is an agroforestry sector in Chile of increasing interest due to the high prices that truffles fetch in the national market and the recent evidence that its commercial production is possible in Chilean climatic and soil conditions. In this study, the efficiency of three methods of DNA extraction from a mix of 5 g of soil and roots from both nursery and field plants of Quercus ilex L. mycorrhized with T. melanosporum were evaluated, and a simple and reproducible protocol was established. Detection of T. melanosporum was performed by the technique of cleaved amplified polymorphic sequence (CAPS) from amplicons generated with the primers ADL1 (5'-GTAACGATAAAGGCCATCTATAGG-3') and ADL3 (5'-CGTTTTTCCTGAACTCTTCATCAC-3'), where a restriction fragment of 160 bp specific for T. melanosporum was generated, which allows the discrimination of this species from the rest of the species belonging to the Tuber sp. genus. Direct detection of T. melanosporum in one step was also obtained by polymerase chain reaction (PCR) from total DNA isolated from mycorrhized roots and with the primers ITSML (5'-TGGCCATGTGTCAGATTTAGTA-3') and ITSLNG (5'-TGATATGCTTAAGTTCAGCGGG-3'), generating a single amplicon of 440 bp. The molecular detection of T. melanosporum by the methods presented here will allow the rapid and accurate detection of mycorrhization of trees, both under nursery and field conditions. This technology will also provide more security to farmers by controlling the quality of the mycorrhized trees they will plant and also by following the mycorrhization status of established orchards.
- ItemPlant tissue culture : current status, opportunities and challenges.(2010) García González, Rolando; Quiroz Bravo, Karla; Carrasco Galvez, Basil Alexander; Caligari, Peter D. S.