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International audience; A procedure was developed for in vitro propagation of Rosa genotypes along with an efficient de novo shoot organogenesis (DNSO) method. We tested, on one genotype (hybrid of Rosa wichurana), the effects of MS basal medium complemented with two growth regulators to achieve either shoot elongation or shoot multiplication of plants. These media were complemented with carbohydrate concentrations from different sources. Then, the impacts of various carbohydrates (fructose, glucose, maltose, sorbitol, sucrose) on the growth and development of several rose genotypes during donor plant subculturing were studied on SMM. The results showed high variability in growth and development between genotypes. Contrary to other members of the Rosaceae family, no correlation was found between the shoot size and number when the amount of sorbitol was increased.Murashige and Skoog medium supplemented with 3.0 mg L-1 BAP and containing fructose or glucose at 30 g L-1 was chosen to induce leaf explants for the DNSO experiments. MS basal medium complemented with TDZ/IBA at three ratios and the same range of carbohydrate sources were tested for DNSO. Significant genotypic variations with regard to the percentage of regeneration was demonstrated with six genotypes. For two genotypes, a hybrid of Rosa wichurana and Rosa 'White Pet', we defined the conditions required to obtain 100% DNSO. For Rosa chinensis 'Old blush' and the rootstock genotype Rosa 'Natal Briar', we obtained 74 and 87.5% DNSO and only 56.67% and 37.5% for Rosa GUY SAVOY (R) ('Delstrimen') and Rosa Felicite et Perpetue' respectively. This adventitious shoot regeneration method may be used for large-scale shoot propagation and genetic engineering studies in Rosa.Keywords

Our experiments have confirmed the considerable disposition of leaf explants of Actinidia chinensis Planch. for induction and intensive proliferation of callus cultures, as well as, a possibility to regulate morhogenesis in in vitro conditions. Under specific culture conditions the morphogenic potential of callus cells of Actinidia chinensis was manifested both in organogenesis and somatic embryogenesis. Organogenesis was represented by induction of adventitious buds and regeneration shoots on the modified MS culture medium (Murashige and Skoog 1962) with BAP in combination with GA3 (each 1.0 mg. l-1). Rooting of shoots was successful on modified MS medium containing IBA (0.5-1.0 mg. l-1). Histological studies of callus tissues revealed their structural heterogeneity. Morphogenic processes in the callus were characterized by the appearance of meristematic zones and vascular elements. The formation of apical meristem, leaf primordia and finally shoot development proved de novo regeneration in callus culture. The obtained results demonstrate a possibility of plant regeneration through indirect organogenesis, which can be used for propagation of Actinidia chinensis Planch.

Adventitious regeneration is a key step in the application of genetic engineering to the breeding programs of plants. In this work a method for adventitious shoot regeneration from leaves of micropropagated guava shoots has been developed and some of main factors to affect the shoot regeneration like, concentration of plant growth regulators (citoquinine), physiological state of explants and the wound are studied. Leaves from guava in vitro cultured of variety Cuban Red Dwarf 18-40 was used like explant in all experiments. The best re-sult was reached with MS basal medium supplemented with 0.75 mg/L of 6-benzylaminopurine. Was de-monstrated that for the leaves with more multiplication subculture number and taken from lower part of shoot (older); the morfogenetic potential falls significantly (p60%) and the largest number of shoot per regenerating leaf (>3) were obtained with several wounds carried out in f orm of jabs to the central nerve of leaves. This regeneration protocol constitutes an important tool that can be applied for future studies of genetic transformation in this species. Key words: Tissue culture, leaf explants, growth regulators, organogenesis, guava

The paper presents evaluation of different types and concentrations of plant growth regulators on induction of adventitious organogenesis from the leaf of blackberry cultivar Cacanska Bestrna. This genotype was introduced and maintained in tissue culture on Murashige and Skoog medium (MS) (1962), mineral salts and organic complex with in mg l-1: BAP 1.0, IBA 0.1, GA3 0.1. Young, expanded leaves taken from upper third of in vitro proliferating shoots were used as initial explants. The leaves were cut three times across the midrib and placed with adaxial side touching the regeneration medium. In this experiment 20 types of regeneration media have been evaluated with different types and concentrations of cytokinins and auxins: BAP (2.0 mg l-1) or TDZ (1.0 mg l-1) alone, or each in combination with different concentrations of IBA, NAA and 2,4-D (0.1, 1.0 and 2.0 mg l-1). In cv Cacanska Bestrna regeneration of adventitious shoots was observed on three different types of media with BAP combined with IBA (0.1 and 1.0 mg l-1) and NAA (0.1 mg l-1). TDZ alone or in combination with IBA (0.1, 1.0 and 2.0 mg l-1) and NAA (0.1 mg l-1) has been more effective at induction of regeneration than BAP. The highest percentage of regeneration was obtained precisely on medium supplemented with TDZ alone (41.66%). .

La regeneración de brotes adventicios es una etapa clave para la aplicación de las técnicas de ingeniería gené­tica. El presente trabajo tuvo como objetivo el desarrollo de un procedimiento para la regeneración de brotes por organogénesis a partir de hojas de microesquejes o brotes de guayaba (Psidium guajava L.) cultivados in vi­tro. Para ello se estudiaron algunos de los principales factores que afectan la regeneración de brotes, tales como la concentración de reguladores del crecimiento (citoquinina), el estado fisiológico del explante y la he­rida. En todos los experimentos se utilizaron como explantes hojas de vitroplantas de guayaba de la variedad Enana Roja Cubana EEA18-40 de diferentes subcultivos. El mejor resultado se alcanzó con una concentración de 0,75 mg/L de 6-bencilaminopurina como suplemento hormonal del medio MS. Se comprobó que para las hojas provenientes de microesquejes con mayor número de subcultivo y tomadas de la parte inferior de los brotes, el potencial morfogenético disminuye significativamente (p60%) y el mayor número de brotes por explante promedio (>3) se alcanzó al realizar varias heridas en forma de punteaduras en el nervio central de las hojas. Este protocolo de regeneración constituye una importante herramienta que puede ser empleada para futuros estudios de transformación genética en esta especie. Palabras clave: cultivo de tejidos, explantes de hoja, reguladores de crecimiento, organogénesis, guayaba.Adventitious regeneration is a key step in the application of genetic engineering to the breeding programs of plants. In this work a method for adventitious shoot regeneration from leaves of micropropagated guava shoots has been developed and some of main factors to affect the shoot regeneration like, concentration of plant growth regulators (citoquinine), physiological state of explants and the wound are studied. Leaves from guava in vitro cultured of variety Cuban Red Dwarf 18-40 was used like explant in all experiments. The best re-sult was reached with MS basal medium supplemented with 0.75 mg/L of 6-benzylaminopurine. Was de-monstrated that for the leaves with more multiplication subculture number and taken from lower part of shoot (older); the morfogenetic potential falls significantly (p60%) and the largest number of shoot per regenerating leaf (>3) were obtained with several wounds carried out in f orm of jabs to the central nerve of leaves. This regeneration protocol constitutes an important tool that can be applied for future studies of genetic transformation in this species. Key words: Tissue culture, leaf explants, growth regulators, organogenesis, guava

Adventitious regeneration is a key step in the application of genetic engineering to the breeding programs of plants. In this work a method for adventitious shoot regeneration from leaves of micropropagated guava shoots has been developed and some of main factors to affect the shoot regeneration like, concentration of plant growth regulators (citoquinine), physiological state of explants and the wound are studied. Leaves from guava in vitro cultured of variety Cuban Red Dwarf 18-40 was used like explant in all experiments. The best re-sult was reached with MS basal medium supplemented with 0.75 mg/L of 6-benzylaminopurine. Was de-monstrated that for the leaves with more multiplication subculture number and taken from lower part of shoot (older); the morfogenetic potential falls significantly (p60%) and the largest number of shoot per regenerating leaf (>3) were obtained with several wounds carried out in f orm of jabs to the central nerve of leaves. This regeneration protocol constitutes an important tool that can be applied for future studies of genetic transformation in this species. Key words: Tissue culture, leaf explants, growth regulators, organogenesis, guavaLa regeneración de brotes adventicios es una etapa clave para la aplicación de las técnicas de ingeniería gené­tica. El presente trabajo tuvo como objetivo el desarrollo de un procedimiento para la regeneración de brotes por organogénesis a partir de hojas de microesquejes o brotes de guayaba (Psidium guajava L.) cultivados in vi­tro. Para ello se estudiaron algunos de los principales factores que afectan la regeneración de brotes, tales como la concentración de reguladores del crecimiento (citoquinina), el estado fisiológico del explante y la he­rida. En todos los experimentos se utilizaron como explantes hojas de vitroplantas de guayaba de la variedad Enana Roja Cubana EEA18-40 de diferentes subcultivos. El mejor resultado se alcanzó con una concentración de 0,75 mg/L de 6-bencilaminopurina como suplemento hormonal del medio MS. Se comprobó que para las hojas provenientes de microesquejes con mayor número de subcultivo y tomadas de la parte inferior de los brotes, el potencial morfogenético disminuye significativamente (p60%) y el mayor número de brotes por explante promedio (>3) se alcanzó al realizar varias heridas en forma de punteaduras en el nervio central de las hojas. Este protocolo de regeneración constituye una importante herramienta que puede ser empleada para futuros estudios de transformación genética en esta especie. Palabras clave: cultivo de tejidos, explantes de hoja, reguladores de crecimiento, organogénesis, guayaba.

International audience; Shoot organogenesis from mature leaf tissues of two scented Pelargonium capitatum cultivars, 'Attar of Roses' and 'Atomic Snowflake', grown in the greenhouse, were optimized in the presence of thidiazuron (TDZ). The protocol involved preculture of leaf sections on basal Murashige and Skoog (MS) medium supplemented with 10 lM TDZ, 4.4 lM of 6-benzyladenine (BA) and 5.4 lM a-naphtaleneacetic acid (NAA) for a period of 2 weeks and followed by subculture of explants to a fresh medium containing 4.4 lM BA and 5.4 lM NAA. Frequency of regeneration reached approximately 93% for both cultivars, with the induction of more than 100 shoots per explant. Regenerated plantlets were rooted on half-strength MS medium supplemented with 4.4 mM sucrose and 8.6 lM of Indole-3-acetic acid (IAA). All regenerated shoots from both cultivars developed roots when transferred to organic soil mix, acclimatized, and successfully transferred to greenhouse conditions. When regenerated shoots were transferred to hydroponic conditions, frequency of survival was 76.2 and 61.9% for 'Attar of Roses' and 'Atomic Snowflake', respectively.

La regeneración de brotes adventicios es una etapa clave para la aplicación de las técnicas de ingeniería gené­tica. El presente trabajo tuvo como objetivo el desarrollo de un procedimiento para la regeneración de brotes por organogénesis a partir de hojas de microesquejes o brotes de guayaba (Psidium guajava L.) cultivados in vi­tro. Para ello se estudiaron algunos de los principales factores que afectan la regeneración de brotes, tales como la concentración de reguladores del crecimiento (citoquinina), el estado fisiológico del explante y la he­rida. En todos los experimentos se utilizaron como explantes hojas de vitroplantas de guayaba de la variedad Enana Roja Cubana EEA18-40 de diferentes subcultivos. El mejor resultado se alcanzó con una concentración de 0,75 mg/L de 6-bencilaminopurina como suplemento hormonal del medio MS. Se comprobó que para las hojas provenientes de microesquejes con mayor número de subcultivo y tomadas de la parte inferior de los brotes, el potencial morfogenético disminuye significativamente (p60%) y el mayor número de brotes por explante promedio ( and gt;3) se alcanzó al realizar varias heridas en forma de punteaduras en el nervio central de las hojas. Este protocolo de regeneración constituye una importante herramienta que puede ser empleada para futuros estudios de transformación genética en esta especie. Palabras clave: cultivo de tejidos, explantes de hoja, reguladores de crecimiento, organogénesis, guayaba. Adventitious regeneration is a key step in the application of genetic engineering to the breeding programs of plants. In this work a method for adventitious shoot regeneration from leaves of micropropagated guava shoots has been developed and some of main factors to affect the shoot regeneration like, concentration of plant growth regulators (citoquinine), physiological state of explants and the wound are studied. Leaves from guava in vitro cultured of variety Cuban Red Dwarf 18-40 was used like explant in all experiments. The best re-sult was reached with MS basal medium supplemented with 0.75 mg/L of 6-benzylaminopurine. Was de-monstrated that for the leaves with more multiplication subculture number and taken from lower part of shoot (older); the morfogenetic potential falls significantly (p60%) and the largest number of shoot per regenerating leaf ( and gt;3) were obtained with several wounds carried out in f orm of jabs to the central nerve of leaves. This regeneration protocol constitutes an important tool that can be applied for future studies of genetic transformation in this species. Key words: Tissue culture, leaf explants, growth regulators, organogenesis, guava