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Introducción: Los arrecifes de coral son ecosistemas altamente degradados, por lo que ha sido necesario implementar acciones de restauración activa para recuperar su estructura y funcionamiento. Se ha implementado la propagación clonal para obtener fragmentos pequeños (~ 10 cm) de las ramas distales de colonias donadoras de corales de la especie Acropora palmata, para posteriormente fijarlos en el sustrato arrecifal, simulando el efecto de dispersión que ocurre de manera natural en esta especie, a lo que en este trabajo se denomina ''dispersión asistida". Sin embargo, es necesario evaluar los efectos de esta técnica como son: la cantidad de fragmentos que se puede obtener de cada colonia, el periodo de recuperación de tejido de las colonias donadoras y los fragmentos sembrados. Objetivo: Evaluar el efecto de poda en las colonias donadoras estimando el porcentaje de tejido podado de colonias donadoras de A. palmata y su tasa de recuperación 30 meses después. Métodos: Se realizaron cuatro monitoreos: antes, inmediatamente después de la poda, un mes después de la siembra, y 30 meses después, en cuatro colonias de A. palmata localizadas en el Parque Nacional Costa Occidental de Isla Mujeres, Punta Cancún y Punta Nizuc en el Caribe mexicano. La modelación 3D basada en fotogrametría se realizó con el software Agisoft Metashape Pro, mientras que las métricas de área de superficie de tejido, extensión radial y apical se obtuvieron mediante el software CloudCompare. Resultados: Posterior a la colecta de fragmentos de las colonias, se observó que el material utilizado en la dispersión asistida representa menos del 12% del tejido vivo. Después de un mes, las colonias donadoras presentaban una recuperación del 5% con tejido nuevo recubriendo las áreas de corte. Las colonias donadoras perdieron, en promedio, 65% de tejido vivo tras el impacto de cuatro huracanes, y en un caso la colonia fue totalmente eliminada, pero con los fragmentos sembrados se pudo conservar el genotipo. Conclusiones: La dispersión asistida podría incrementar el tejido vivo de corales ramificados en intervalos de tiempo relativamente cortos, sin comprometer la integridad de la colonia donadora, si se poda menos del 12%.
Introduction: Coral reefs are highly degraded ecosystems, for which it has been necessary to implement active restoration actions to recover their structure and functioning. Asexual propagation has been implemented to obtain small fragments (~10 cm) from the distal branches of donor colonies of corals of the species Acropora palmata, to subsequently relocate them in the reef substrate, simulating the dispersion effect that occurs naturally in the species, which in this work is called assisted propagation. However, it is necessary to evaluate the effects of this technique, such as the number of fragments that can be obtained from each colony, the tissue recovery period of the donor colonies and fragments. Objective: To address the effect of pruning on donor colonies by estimating the percentage of live tissue removed from donor colonies of A. palmata and their recovery rate after 30-months. Methods: Four surveys were carried out: before, immediately after pruning, one month after outplanting, and 30 months after pruning on four colonies of A. palmata located in the Parque Nacional Costa Occidental de Isla Mujeres, Punta Cancún and Punta Nizuc in the Mexican Caribbean. Photogrammetry-based 3D modeling was performed using Agisoft Metashape Pro software, while tissue surface area, radial and apical growth were obtained using CloudCompare software. Results: After fragment collection, the material used in the assisted propagation represents less than 12% of the living tissue. After one month, the donor colonies showed a recovery of 5%, with new tissue covering the cut areas. The donor colonies lost on average 65 % of living tissue after four hurricanes, and in one case the colony was lost all together, but with the outplanted fragments the genotype could be preserved. Conclusions: Assisted propagation could increase living tissue of branching corals in relatively short intervals of time, without serious damage to the donor colony if less than 12 % is removed.
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Objective:Stems,petioles,stem sections with axillary and leaves of Scrophularia ningpoensis were taken as the material in vitro to screen out the suitable plantlet regeneration system and optimal exercising seedling conditions. Method:Different explants,hormones and concentrations on the induction and proliferation of cluster bud were studied by L16(45) orthogonal test. One factor analysis of variance (ANOVA) was made on the induction of adventitious buds rooted with different concentrations of hormones. At the same time,different substrates,watering cycles and transition modes were selected to optimize key technologies of exercising seedlings of S. ningpoensis. Result:Stem sections with axillary was the best explant,which was followed by stems,leaves and petioles. The suitable media for the induction of adventitious buds was MS+6-BA 0.5 mg·L-1+NAA 0.2 mg·L-1,with the induction rate of 100.0% and the proliferation multiple of 9.84.The suitable media for root induction was 1/2 MS+IBA 0.2 mg·L-1,with the rooting rate of 100.0% and the number of roots of 39.45.For matrix,they were transplanted with nutrient soil,vermiculite and perlite (5:2:1) as the media,to keep proper matching of fertility,permeability and water retention. The container seedlings can grow well,and the survival rate was more than 95% when they were watered every 2 days,the acclimatization of plantlets took 20 days indoor and 10 days in shaded greenhouses. Conclusion:The clonal propagation system of S. ningpoensis was established to provide an effective way for the efficient,rapid and steady plantlet regeneration,the breeding of high-quality seedlings and the suitable exercising seedling conditions of S. ningpoensis.
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Se presentan los procedimientos para la propagación in vitro de Perezia pinnatifida (Humb. & Bonpl.) Wedd., conocida como "valeriana". Se utilizaron las metodologías de multiplicación de brotes y embriogénesis somática indirecta. El medio de cultivo basal para todas las etapas fue Murashige y Skoog a mitad de sales, suplementado con sacarosa 2.0%, phytagel 0.3% y pH 5.67; y fue usado con o sin fitohormonas en los diferentes tratamientos. Los suplementos hormonales fueron: para la multiplicación de brotes BAP 1.0 mg.L-1 + ANA 0.01 mg.L-1 ó BAP 1.0 mg.L-1; para la inducción de callos embriogénicos ANA ó 2,4-D (1.0 mg.L-1 y 2.0 mg.L-1); y para la germinación de embriones BAP (0.5 y 1.0 mg.L-1) o BAP 0.5 mg.L-1 + ANA 0.05 mg.L-1. El mayor número de brotes se obtuvo en el medio suplementado con BAP 1.0 mg.L-1. En la embriogénesis somática, ANA a 1.0 mg.L-1 indujo mayor área de callos embriogénicos, y BAP a 0.5 mg.L-1 permitió mayor germinación de los embriones somáticos
This work inform on in vitro propagation of the "valeriana" Perezia pinnatifida (Humb. & Bonpl.) Wedd. Shoot multiplication and indirect somatic embryogenesis methodologies were performed. The basal culture medium for all stages was Murashige and Skoog, middle of salts supplemented with 2.0% sucrose, 0.3% phytagel and pH 5.67; the treatments were prepared with or without phytohormones. The hormonal supplements for the shoot multiplication were: BAP 1.0 mg.L-1 + ANA 0.01 mg.L-1, and BAP 1.0 mg.L-1; for embryogenic callus induction were: ANA or 2,4-D (1.0 mg.L-1 and 2.0 mg.L-1); and for the embryo germination were: BAP (0.5 and 1.0 mg.L-1) or BAP 0.5 mg.L-1 + ANA 0.05 mg.L-1. BAP 1.0 mg.L-1 produced the higher number buds. For somatic embryogenesis, ANA 1.0 mg.L-1 induced a greater area of embryogenic callus, and BAP 0.5 mg.L-1 allowed major germination of the somatic embryos
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Aims: Sweet potato (Ipomoea batatas) cv. “Miyazakibeni” was used as material for shoot apex culture, reverse transcription-polymerase chain reaction (RT-PCR) and clonal propagation to establish an efficient and practical virus-free seedling supply system in production of vegetatively reproductive plants. Study Design: At first, efficient plant regeneration was achieved from shoot apex culture of sweet potato. Secondly, RT-PCR method was used to detect the sweet potato feathery mottle virus (SPFMV) viral infection of tuber surface of edible sweet potato using the RNAs from the plants obtained from shoot apex culture. Finally, the virus-free plants verified by RT-PCR were propagated clonally by culture of suckers cut from stems of the virus-free plants. Place and Duration of Study: Faculty of Environmental and Horticultural Science, Minami Kyushu University, between June 2008 and December 2012. Methodology: The best efficiency for material sterilization was tested using different concentrations (0.1% - 1.5%) of sodium hypochlorite solution (SHS) and the treated times (5 min – 20 min). Theshoot apexes less than 0.3mm in size were cultured on Komamine and Nomura (1998) (KN) medium and Murashige and Skoog (1962) (MS) medium. The regenerated plants were used for RNA extraction and then, used for RT-PCR for detection of SPFMV. Based on the result of RT-PCR, the suckers cut from stems of virus-free plants were cultured and propagated clonally and routinely within a short period. Results: The combination of 0.3% of SHS and 10 and/or 20 min gave the best result (100%) of surviving rate for material sterilization. The culture of shoot apexes less than 0.3 mm in size gave plant regenerating rates of 82% and 65% on KN and MS medium, respectively. The results of RT-PCR of RNAs from plants obtained from shoot apex culture and plants of SPFMV infection showed that SPFMV virus was clearly removed by shoot apex culture conducted in this study. For clonal propagation, 80-100% of suckers cut from the stems of the virus-free plants detected grew into complete plants after 6 weeks of culture, indicating that the virus-free plants could be routinely propagated 5 times in number each time and repeatable by the short circle. The sweet potato produced in field showed no symptom called as russet crack-like symptom (RC-LS) even after cultivation two seasons. Conclusion: Overall, an efficient and practical virus-free seedling supply system was established in sweet potato by the three steps of 1) virus-free plant regeneration from shoot apex culture, 2) quick detection of SPFMV using RNA of the regenerated plants by RT-PCR, and 3) the verified virus-free plants were propagated clonally and routinely within a short period using culture of suckers cut from the stems of virus-free plants.
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<p><b>OBJECTIVE</b>To develop the reproducible in vitro propagation protocols for the medicinally important plants viz., Achyranthes aspera (A. aspera) L. and Achyranthes bidentata (A. bidentata) Blume using nodal segments as explants.</p><p><b>METHODS</b>Young shoots of A. aspera and A. bidentata were harvested and washed with running tap water and treated with 0.1% bavistin and rinsed twice with distilled water. Then the explants were surface sterilized with 0.1% (w/v) HgCl2 solutions for 1 min. After rinsing with sterile distilled water for 3-4 times, nodal segments were cut into smaller segments (1 cm) and used as the explants. The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% (w/v) agar (Hi-Media, Mumbai) and different concentration and combination of 6-benzyl amino purine (BAP), kinetin (Kin), naphthalene acetic acid (NAA) and indole acetic acid (IAA) for direct regeneration.</p><p><b>RESULTS</b>Adventitious proliferation was obtained from A. aspera and A. bidentata nodal segments inoculated on MS basal medium with 3% sucrose and augmented with BAP and Kin with varied frequency. MS medium augmented with 3.0 mg/L of BAP showed the highest percentage (93.60±0.71) of shootlets formation for A. aspera and (94.70±0.53) percentages for A. bidentata. Maximum number of shoots/explants (10.60±0.36) for A. aspera and (9.50±0.56) for A. bidentata was observed in MS medium fortified with 5.0 mg/L of BAP. For A. aspera, maximum mean length (5.50±0.34) of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A. bidentata (5.40±0.61) was observed in the very same concentration. The highest percentage, maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of IBA. Seventy percentages of plants were successfully established in polycups. Sixty eight percentages of plants were well established in the green house condition. Sixty five percentages of plants were established in the field.</p><p><b>CONCLUSIONS</b>The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A. aspera and A. bidentata. The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can be easily adopted for commercial large scale cultivation.</p>
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Achyranthes , Culture Media , Chemistry , Plant Roots , Plant Shoots , Plants, Medicinal , Survival AnalysisABSTRACT
Cecropia glaziovii is a tree with used in Brazilian popular medicine. Methods allowing the clonal propagation of this species are of great interest for superior genotype multiplication and perpetuation. For this reason, we examined the effect of different culture media and different types of explants on adventitious shoot regeneration from callus and buds of C. glaziovii. Leaves, petioles and stipules obtained from aseptically grown seedlings or from pre-sterilized plants were used to initiate cultures. Adventitious shoot regeneration was achieved when apical and axillary buds were inoculated on gelled Murashige & Skoog (MS) medium supplemented with 6-benzylaminopurine alone (BAP) (1.0, 5.0 or 10.0 mg L-1) or combined with -naphthalene acetic acid (NAA) (1.0 or 2.0 mg L-1), after 40 days of culture. Best callus production was obtained after 30 days of petioles' culture on gelled MS medium with 2,4 dichlorophenoxyacetic acid (2,4-D) (5.0 mg L-1) combined with BAP (1.0 mg L-1). Successful shoot regeneration from callus was achieved when MS medium supplemented with zeatin (ZEA) (0.1 mg L-1) alone or combined with 2,4-D (1.0 or 5.0 mg L-1) was inoculated with friable callus obtained from petioles. All shoots were rooted by inoculation on MS medium supplemented with indole-3-acetic acid (IAA) (1.0 mg L-1). Rooted plants transferred to potting soil were successfully established. All in vitro regenerated plantlets showed to be normal, without morphological variations, being also identical to the source plant. Our study has shown that C. glaziovii can be propagated by tissue culture methods, allowing large scale multiplication of superior plants for pharmacological purposes.
Cecropia glaziovii é uma planta lenhosa, popularmente usada no Brasil como medicinal. Métodos que visem a sua propagação clonal podem ser de grande utilidade na preservação de seus genótipos de elite. Foram examinados efeitos de diferentes reguladores de crescimento e explantes na formação de brotações múltiplas a partir de calos e a partir de brotos apicais. Folhas, pecíolos e estípulas obtidas de plântulas assépticas e da esterilização de sementes ou através da esterilização direta dos explantes foram utilizados para iniciar os cultivos. Brotações múltiplas foram obtidas quando brotos apicais ou axilares foram inoculados em meios compostos de sais de Murashige & Skoog (MS), suplementado com apenas 6-benzilaminopurine (BAP) (1,0, 5,0 ou 10,0 mg L-1) ou combinado com ácido -naftaleno acético (ANA) (1,0 ou 2,0 mg L-1), depois de 40 dias. A produção de calos foi obtida após 30 dias, quando pecíolos foram inoculados em meio MS acrescido de acido 2,4 diclorofenoxiacético (2,4-D) (5,0 mg L-1) combinado com BAP (1,0 mg L-1). A regeneração de brotos a partir de calos foi obtida quando meio MS acrescido de apenas zeatina (ZEA) (0,1 mg L-1) ou combinado com 2,4-D (1,0 ou 5,0 mg L-1) foi inoculado com calos friáveis obtidos de pecíolos. As plântulas foram enraizadas em MS suplementado com ácido 3-indol acético (AIA) (1.0 mg L-1). A aclimatação das plântulas enraizadas foi estabelecida pela transferência para vasos contendo substrato orgânico e vermiculita sob umidade relativa 100 por cento. As plântulas regeneradas "in vitro" apresentaram aparência normal e idênticas à planta-mãe. Nosso estudo concluiu que genótipos de elite de C. glaziovii podem ser propagados em larga escala através de métodos "in vitro", proporcionando uma fonte confiável de matéria prima para estudos farmacológicos.
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The roots of the understorey shrub Carapichea ipecacuanha (ipecac) have medicinal properties, and the uprooting of wild plants has supplied most of the world demand for this species. Although under severe population decline, C. ipecacuanha lacks legal protection. In the wild, the aerial stems of ipecac clump together to form clusters with well-defined borders. Cluster size may range from several to hundreds of aerial stems. To investigate the extent of clonality among aerial stems in ipecac clusters, we sampled 50 wild clusters (a total of 291 aerial stems) and screened them with 89 inter-simple sequence repeat (ISSR) markers. The 291 aerial stems were grouped into 42 putative clones. The clonal groups generally consisted of aerial stems from the same cluster, and there was little or no genetic differentiation among aerial stems at the cluster level. These findings suggest that strategies designed to conserve ipecac in situ should not rely upon census data, which are based on the number of aerial stems per cluster and the number of clusters per population, because such data greatly underestimate the species effective population size and genetic diversity. Our results also indicate that this species needs protection at a federal level.
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Cross prediction trials or progeny tests for family selection are commonly employed at the beginning of each breeding cycle in clonally propagated crops such as sugarcane (Saccharum spp. Hybrid). When selecting sugarcane families, the factors affecting variability between and within those families should be considered. This research examined the influence of family and intra-row plant spacing on the efficiency of the index selection procedure as a method of simultaneous improvement of a population for multiple traits at the first selection stage of the Louisiana Sugarcane Variety Development Program (LSVDP). Therefore, the main objective of this study was to develop a selection index for selecting sugarcane families within the LSVDP. Expected genetic advance values for plant weight were greater in the wide-spaced indices than in the narrow-spaced ones. Irrespective of plant spacing, selection indices revealed that an increase in efficiency was observed over direct selection for plant weight when all four plant weight contributing traits were included along with plant weight. The efficiency in selection tended to decrease when indices were based on fewer traits. Nevertheless, a few of the indices that included two traits had relative efficiencies comparable to the best indices and the majority certainly was as effective as direct selection for plant weight.
Las pruebas de progenie son comúnmente utilizadas para la selección de familias en cultivos de reproducción asexual como la caña de azúcar (Saccharum spp. Híbrido). En la selección de familias de caña es importante considerar los factores que afectan la variabilidad existente, no solo entre familias, sino también dentro de cada familia. Se examinó la influencia de familias y del espacio entre plántulas de caña de azúcar en la eficiencia del índice de selección como una metodología para mejorar simultáneamente una población por múltiples características. El trabajo se llevó a cabo en la etapa inicial del Programa de Desarrollo de Variedades de Caña de Azúcar de Louisiana, EEUU (LSVDP, por sus siglas en inglés). El objetivo principal fue desarrollar un índice para selección de familias de caña de azúcar dentro del LSVDP. El avance genético esperado para el peso de planta fue mayor en los índices obtenidos usando datos provenientes de familias donde la distancia entre individuos fue mayor. Independientemente de la distancia entre plántulas dentro de la hilera, los índices de selección donde intervinieron los cuatro componentes del carácter peso de planta, incluyendo a éste, resultaron ser más eficientes que la selección directa para ese mismo carácter. La eficiencia en la selección tendió a disminuir en la medida en que los índices contenían menos caracteres. Sin embargo, algunos de los índices que incluían solo dos caracteres mostraron eficiencias comparables a los mejores índices y la mayoría de ellos fueron más efectivos que la selección directa por peso de planta.
Ensaios de predição de cruzamentos o testes de progênie são comumente utilizados no início de cada ciclo de melhoramento genético das culturas de reprodução vegetativa como a cana-de-açúcar (Saccharum spp. Híbrido). Na seleção de famílias de cana-de-açúcar é importante considerar os fatores que afetam a variabilidade, não só entre as famílias, mais também dentro de cada família. Analisou-se a influência das famílias e do espaçamento entre plantas na eficiência do índice de seleção como uma metodologia para melhorar simultaneamente mais de uma característica em uma população. O trabalho foi feito na fase inicial do Programa de Desenvolvimento de Variedades de Cana-de-Açúcar de Louisiana, EEUU (LSVDP, por sua sigla em Inglês). Portanto, o principal objetivo foi desenvolver um índice para a seleção de famílias de cana-de-açúcar dentro do LSVDP. O ganho genético esperado para o peso da touceira foi maior nos índices obtidos a partir de dados de famílias onde a distancia entre indivíduos foi maior. Os resultados indicaram que, quando os quatro caracteres que contribuem para o peso da touceira foram incluídos, independentemente do espaçamento entre touceiras, os índices de seleção apresentaram aumento na eficiência na seleção, em comparação com a seleção direta para o peso da touceira. Á medida que os índices de seleção foram baseados em menos caracteres, a eficiência na seleção tendeu a diminuir. No entanto, alguns dos índices que incluía dois caracteres tiveran eficiências comparáveis a os melhores índices e certamente a maioria deles foram tão eficazes como a seleção direta para peso da touceira.