Cryopreservation of plant germplasm in Argentina

This review describes the current status of development of methods for cryopreservation (at -196ºC) of plants germplasm in Argentina. Arachis pintoi, a forage legume, has been maintained as seeds using vitrification method. Additionally, apical meristems, shoot tips, and somatic embryos have been cryopreserved using encapsulation-dehydration. Zygotic embryos, encapsulated and dehydrated, have permitted the cryopreservation of seven species of the genus Ilex. Various explants (apical meristems, uninodal segments, buds and somatic embryos) of Melia azedarach have been cryopreserved using the encapsulation-dehydration method. Protocols based in encapsulation-dehydration have also been developed for shoot tips of Citrus sinensis, seeds and protocorms of Oncidum bifolium and anthers of Oryza sativa. Vitrification protocols have been developed forcryopreservation of shoot tips of Solanum tuberosum and seeds of Toona ciliata.Fil: Mroginski, Luis Amado. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Rey de Badaró, Hebe Yolanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentin

Regeneration of plants from apical meristem tips and nodal segments of Arachis pintoi

The in vitro regeneration potential of shoot apical tips (2 to 3 mm in length), meristems (0.3 to 0.5 mm in length), and nodal segments (4 to 7 mm long with an axillary bud) of diploid (2n = 2x = 20) and triploid (2n = 3x = 30) cytotypes of Arachis pintoi was evaluated. Explants were cultured on MS medium supplemented with different concentrations and combinations of naphthaleneacetic acid (NAA) and benzyladenine (BA). In one experiment the effect of gibberellic acid was tested. The cultures were done in liquid and solid media. Plant regeneration can be readily achieved from all explants in one step of 30 d culture on MS + 0.01 mg/L each of NAA and BA or two steps consisting of 1) shoots regeneration through culture of explants on MS + 0.01 mg/L each of NAA and BA, and 2) induction of rooting in regenerated shoots by reculture on MS + 0.01 mg/L NAA. The plantlets were successfully transferred to pots in a greenhouse.Fil: Rey de Badaró, Hebe Yolanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Mroginski, Luis Amado. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentin

Organogenesis and plant regeneration of Arachis villosa Benth. (Leguminosae) through leaf culture

With the aim of developing an efficient plant regeneration protocol, leaflet explants of three accessions of Arachis villosa Benth. (S2866, S2867 and L97) were cultured on basic Murashige and Skoog medium supplemented with different combinations of plant growth regulators: α-naphthalenacetic acid, indole-3-butyric acid, 6-benzylaminopurine, kinetin and thidiazuron. The accession L97 was the only one able to differentiate buds through indirect organogenesis. The most suitable combination for bud regeneration was the basic medium added with 13.62 μM thidiazuron and 4.44 μM 6-benzylaminopurine. These results show the important role of the genotype in morphogenetic responses and the organogenetic effect of thidiazuron in Arachis villosa accession L97. A thidiazuron lacking media (only 0.54 μM α-naphthalenacetic acid, 13.95 μM kinetin and 13.32 μM 6-benzylaminopurine were added) promoted the elongation of the regenerated buds. Adventitious rooting was achieved 90 days after the isolated shoots were transferred to a rooting medium containing 0.54 μM α-naphthalenacetic acid.Fil: Fontana, María Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Mroginski, Luis Amado. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Rey de Badaró, Hebe Yolanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentin

Enhanced seed germination of Ilex dumosa R. (Aquifoliaceae) through in vitro culture of cut pyrenes

An in vitro culture protocol was developed that increased the germination percentage and decreased the lag time to germination for Ilex dumosa R. pyrenes as a tool for replacing the laborious task of embryo rescue technique. This method involves transversely cutting surface-sterilized pyrenes with a scalpel blade, then placing the micropylar one-third end with the rudimentary embryo (0.25 mm long) on solidified (agar 0.65%) quarter-strength salts and vitamins of Murashige and Skoog, 1962 medium with 3% sucrose, and incubating in a growth room at 27 ± 2 8C with a 14-h photoperiod (116 mmolm?2 s?1). Most of the cut pyrenes (greater than 50%) germinated within the first month after inoculation and achieved maximum germination (70%) in 2 months compared with whole pyrenes, which began to germinate 3 months after sowing and required more than 8 months for maximum germination (37%). Moreover, the germination percentage of cut pyrenes was significantly higher than the germination of isolated embryos (34%). Thus, the cut pyrenes culture is a simpler and more effective technique than embryo rescue. Easily, on average, a trained operator is able to culture ~1000 cut pyrenes per day instead of ~100 isolated embryos.Fil: Dolce, Natalia Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; ArgentinaFil: Mroginski, Luis Amado. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; ArgentinaFil: Rey, Hebe Yolanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Nordeste. Instituto de Botánica del Nordeste (i); Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentin

Plant regeneration of tea (Camellia sinensis) by in vitro culture of meristems, axillary buds and uninodal segments

Tres tipos de explantes de dos clones ( C H 1 4 I N TA y C H 3 1 8 I N TA ) d e t é (Camellia sinensis (L.) O. Kuntze) fueron evaluados para su regeneración in vitro, bajo la influencia de dos citocininas (BAP y CIN) y una giberelina (AG3). Previa desinfección, con etanol 70% (1 minuto) e hipoclorito de sodio 1,5% (20 minutos) y tres enjuagues con agua destilada estéril, los explantes fueron aislados y cultivados en los distintos medios de cultivo. Las mejores respuestas en formación de vástagos se registraron con los segmentos uninodales de ambos clones cultivados en el medio ½ MS + 1 mg/L de BAP o con el cultivo de yemas axilares del clon CH 14 INTA en el medio ½ MS + 1 mg/L de BAP o del clon CH 318 INTA en el medio ½ MS + 1 mg/L BAP + 1 mg/L AG3. Los mejores resultados con el empleo de meristemas caulinares se obtuvieron en el medio ½ MS + 1 mg/L de CIN y 1 mg/L de AG3. Los vástagos obtenidos fueron enraizados mediante su cultivo en ¼ MS + 6 mg/L de IBA.Plants of two clones (CH 14 INTA and CH 318 INTA) of tea (Camellia sinensis (L.) O. Kuntze) were regenerated by in vitro culture of three types of explants disinfected by immersion in 70% ethanol (1 min) and 1.5% sodium hypochlorite (20 min). The best medium for shoot regeneration from uninodal segments, for both clones, as well as for axillary buds of CH 14 INTA clone was ½ MS + 1 mg/L BAP. While the best medium for axillary buds of CH 318 clone was ½ MS + 1 mg/L BAP + 1 mg/L AG3. For meristems culture, the best medium, for both clones was ½ MS + 1 mg/L KIN + 1 mg/L AG3. Rooting of regenerated shoots were achieved by culture them on ¼ MS + 6 mg/L IBA.Fil: Molina, Sandra P.. Instituto Nacional de Tecnología Agropecuaria (Argentina). Estación Experimental Agropecuaria Cerro AzulFil: Pérez, María Laura. Universidad Nacional del Nordeste. Facultad de Ciencias AgrariasFil: Rey, Hebe Y.. Universidad Nacional del Nordeste. Facultad de Ciencias AgrariasFil: Mroginski, Luis A.. Universidad Nacional del Nordeste. Facultad de Ciencias Agraria

A cryopreservation protocol for immature zygotic embryos of species of Ilex (Aquifoliaceae)

Tropical Ilex species have recalcitrant seeds. This work describes experiments demonstrating the feasibility of long-term conservation of Ilex brasiliensis, I. brevicuspis, I. dumosa, I. intergerrima, I. paraguariensis, I. pseudoboxus, I. taubertiana, and I. theezans through cryopreservation of zygotic rudimentary embryos at the heart developmental stage. The embryos were aseptically removed from the seeds and precultured (7 days) in the dark, at 27± 2ºC on solidified (0.8% agar) 1/4MS medium, [consisting of quarterstrength salts and vitamins of Murashige and Skoog (1962) medium] with 3% sucrose and 0.1 mg/l Zeatin. The embryos were then encapsulated in 3% calcium alginate beads and pretreated at 24 h intervals in liquid medium supplemented with progressively increasing sucrose concentrations (0.5, 0.75 and 1 M). Beads were dehydrated for 5 h with silicagel to 25% water content (fresh weight basis) and then placed in sterile 5 ml cryovials. Then the beads were either plunged rapidly in liquid nitrogen were they were kept for 1 h (rapid cooling) or cooled at 1ºC min-1 to -30ºC. Then the beads were immersed in liquid nitrogen for 1 h (slow cooling). The beads were rewarmed by immersion of the cryovials for 1 min in a water bath thermostated at 30ºC. Finally, beads were transferred onto culture medium (1/4MS, 3% sucrose, 0.1 mg/l zeatin, solidified with 0.8% agar) and incubated in a growth room at 27 ± 2ºC under a 14 h light (116 μmol. m-2.s-1)/ 10 h dark photoperiod. Maximum recovery percentages between 15 and 83% (depending on de the species and the treatment) were obtained with the cryopreserved embryosFil: Mroginski, Luis Amado. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Sansberro, Pedro Alfonso. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Scocchi, Adriana M.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Luna, Claudia Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Rey de Badaró, Hebe Yolanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentin

Regeneración de plantas de té (Camellia sinensis) por cultivo invitro de meristemas, yemas axilares y segmentos uninodales *

Plants of two clones (CH 14 INTA and CH 318 INTA) of tea (Camellia sinensis (L.) O. Kuntze) were regenerated by in vitro culture of three types of explants disinfected by immersion in 70% ethanol (1 min) and 1.5% sodium hypochlorite (20 min). The best medium for shoot regeneration from uninodal segments, for both clones, as well as for axillary buds of CH 14 INTA clone was ½ MS + 1 mg/L BAP. While the best medium for axillary buds of CH 318 clone was ½ MS + 1 mg/L BAP + 1 mg/L AG3. For meristems culture, the best medium, for both clones was ½ MS + 1 mg/L KIN + 1 mg/L AG3. Rooting of regenerated shoots were achieved by culture them on ¼ MS + 6 mg/L IBA.  Tres tipos de explantes de dos clones ( C H 1 4 I N TA y C H 3 1 8 I N TA ) d e t é (Camellia sinensis (L.) O. Kuntze) fueron evaluados para su regeneración in vitro, bajo la influencia de dos citocininas (BAP y CIN) y una giberelina (AG3). Previa desinfección, con etanol 70% (1 minuto) e hipoclorito de sodio 1,5% (20 minutos) y tres enjuagues con agua destilada estéril, los explantes fueron aislados y cultivados en los distintos medios de cultivo. Las mejores respuestas en formación de vástagos se registraron con los segmentos uninodales de ambos clones cultivados en el medio ½ MS + 1 mg/L de BAP o con el cultivo de yemas axilares del clon CH 14 INTA en el medio ½ MS + 1 mg/L de BAP o del clon CH 318 INTA en el medio ½ MS + 1 mg/L BAP + 1 mg/L AG3. Los mejores resultados con el empleo de meristemas caulinares se obtuvieron en el medio ½ MS + 1 mg/L de CIN y 1 mg/L de AG3. Los vástagos obtenidos fueron enraizados mediante su cultivo en ¼ MS + 6 mg/L de IBA

Técnicas de cultivo in vitro y almacenamiento a baja temperatura para la conservación de explantes de té

Los resultados de este trabajo permiten concluir que es posible la conservación a corto/mediano plazo de explantes de té mediante el uso de la baja temperatura. La sobrevivencia, formación de vástagos y longitud de vástagos son influenciados por el tiempo de exposición a las bajas temperaturas. Es posible la conservación de plantas de té utilizando segmentos uninodales a bajas temperaturas, por un período máximo de 5 meses sin subcultivo, momento a partir del cual la sobrevivencia y calidad de los explantes declinan rápidamente. Los segmentos uninodales y yemas axilares son los explantes que permiten una mejor conservación y generación de vástagos.Fil: Molina, Sandra Patricia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Cerro Azul; ArgentinaFil: Pérez, María Laura. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; ArgentinaFil: Rey, Hebe Yolanda. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; ArgentinaFil: Mroginski, Luis Amado. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentin

A cryopreservation protocol for immature zygotic embryos of species of Ilex (Aquifoliaceae)

ABSTRACT: Tropical Ilex species have recalcitrant seeds. This work describes experiments demonstrating the feasibility of long-term conservation of Ilex brasiliensis, I. brevicuspis, I. dumosa, I. intergerrima, I. paraguariensis, I. pseudoboxus, I. taubertiana, and I. theezans through cryopreservation of zygotic rudimentary embryos at the heart developmental stage. The embryos were aseptically removed from the seeds and precultured (7 days) in the dark, at 27± 2ºC on solidified (0.8% agar) 1/4MS medium, [consisting of quarterstrength salts and vitamins o

Measuring the health-related Sustainable Development Goals in 188 countries : a baseline analysis from the Global Burden of Disease Study 2015

Background In September, 2015, the UN General Assembly established the Sustainable Development Goals (SDGs). The SDGs specify 17 universal goals, 169 targets, and 230 indicators leading up to 2030. We provide an analysis of 33 health-related SDG indicators based on the Global Burden of Diseases, Injuries, and Risk Factors Study 2015 (GBD 2015). Methods We applied statistical methods to systematically compiled data to estimate the performance of 33 health-related SDG indicators for 188 countries from 1990 to 2015. We rescaled each indicator on a scale from 0 (worst observed value between 1990 and 2015) to 100 (best observed). Indices representing all 33 health-related SDG indicators (health-related SDG index), health-related SDG indicators included in the Millennium Development Goals (MDG index), and health-related indicators not included in the MDGs (non-MDG index) were computed as the geometric mean of the rescaled indicators by SDG target. We used spline regressions to examine the relations between the Socio-demographic Index (SDI, a summary measure based on average income per person, educational attainment, and total fertility rate) and each of the health-related SDG indicators and indices. Findings In 2015, the median health-related SDG index was 59.3 (95% uncertainty interval 56.8-61.8) and varied widely by country, ranging from 85.5 (84.2-86.5) in Iceland to 20.4 (15.4-24.9) in Central African Republic. SDI was a good predictor of the health-related SDG index (r(2) = 0.88) and the MDG index (r(2) = 0.2), whereas the non-MDG index had a weaker relation with SDI (r(2) = 0.79). Between 2000 and 2015, the health-related SDG index improved by a median of 7.9 (IQR 5.0-10.4), and gains on the MDG index (a median change of 10.0 [6.7-13.1]) exceeded that of the non-MDG index (a median change of 5.5 [2.1-8.9]). Since 2000, pronounced progress occurred for indicators such as met need with modern contraception, under-5 mortality, and neonatal mortality, as well as the indicator for universal health coverage tracer interventions. Moderate improvements were found for indicators such as HIV and tuberculosis incidence, minimal changes for hepatitis B incidence took place, and childhood overweight considerably worsened. Interpretation GBD provides an independent, comparable avenue for monitoring progress towards the health-related SDGs. Our analysis not only highlights the importance of income, education, and fertility as drivers of health improvement but also emphasises that investments in these areas alone will not be sufficient. Although considerable progress on the health-related MDG indicators has been made, these gains will need to be sustained and, in many cases, accelerated to achieve the ambitious SDG targets. The minimal improvement in or worsening of health-related indicators beyond the MDGs highlight the need for additional resources to effectively address the expanded scope of the health-related SDGs.Peer reviewe