Tissue Culture Based Conservation Strategies for Litsea cubeba (Lours.) Pers: A Medicinally Importance Plant

Litsea cubeba (Lours.) Pers. belongs to the family Lauraceae, which occurs mainly in the tropical and sub-tropical regions worldwide. The plant gained importance for its essential oils, sesquiterpenoids, flavonoids, lignans volatile oils, and numerous secondary metabolites. The Essential oil extracted from its bark, stem, leaves can be used commercially for the preparation of medicines, insecticides, perfumes, flavors, and colognes. The secondary metabolites extracted from L. cubeba show potential pharmacological activities, viz., antipyretic, analgesic, antidiarrheal and anti-tumor, antimicrobial, anti-inflammatory, antioxidant, anti-HIV, hepatoprotective, antidiabetic, and hypothermic activities. In north east plants and parts sold in the local market and used by local people for various ailments and culinary purposes. Overexploitation of the plants took place due to its essential oils and medicinal value; therefore, conservation strategies are needed. Here, we are summarizing the medicinal uses of Litsea sp. and the conservation strategies for the Lauraceae family plants using various tissue culture approaches.

Enhancing Gerbera Micropropagation Efficiency and Genetic Fidelity through Cytokinin and Auxin Combination Strategies

The present investigation was carried out to assess the impact of 6-benzyladenine (BA) in conjunction with Indole-3-butyric acid (IBA) on gerbera explant establishment under micropropagation. Additionally, the effects of BA, either individually or in combination with Kinetin (KIN), on shoot proliferation in two Gerbera cultivars, namely Kormoran and Dolores was experimented too. Throughout the experiment, various morphological changes were documented occurring during these micropropagation phases and also monitored potential genetic alterations using SSR markers. The studies revealed that the combination of BA and IBA yielded exceptional results, achieving a 100% success rate in explant regeneration within the shortest time frame. Notably, when BA and IBA were applied at lower concentrations, the number of shoots generated was reduced. However, the most substantial proliferation of shoots was observed when the growth medium contained 4 mg of BA and 0.5 mg of IBA per litre. Furthermore, our investigation into genetic fidelity using SSR analysis revealed no detectable polymorphism between the mother plant and the micropropagated plantlets in both the Gerbera cultivars, affirming the reliability of the micropropagation method in preserving genetic consistency.

In-vitro Regeneration of Bamboo (Bambus balcooa): A Review

Bamboos are adaptable, fragrant, perennial, and non-wood forest plants that are extremely significant from an ecological, sociological, and economic standpoint. Bamboo may be propagated using a variety of methods, including rhizome and culm cuttings, clump division, and seed propagation, however these traditional methods have significant limitations when it comes to large- or mass-scale multiplication. These are typically inadequate and ineffective for mass scale dissemination, leaving micropropagation as the sole practical approach. The requirement for bamboo material for cultivation is so high that large-scale multiplying will inevitably necessitate micropropagation. High hopes have been placed in the ability of micropropagation for the mass propagation of bamboo, and a great deal of study has gone into the creation of procedures for large-scale, quick propagation. These include clonal fidelity, somatic embryogenesis, In vitro blooming, macro-proliferation, field efficiency as well as the optimisation and development of in vitro culture procedures. For large-scale micropropagation, which is urgently needed, this paper rapidly presents the most current knowledge on tissue culture mediated biotechnological interventions done in bamboo.

Introducción a cultivo de vitroplantas de "Peperina de las Lomas" (Hedeoma multiflora Benth) como contribución para su conservación ex situ / Field establishment of vitroplants of "Peperina de las Lomas" (Hedeoma multiflora Benth) as a contribution to its ex situ conservation

Hedeoma multiflora Benth. (Lamiaceae) is an aromatic-medicinal species native to Argentina, Uruguay and southern Brazil that is in a state of vulnerability due to overexploitation. It is used in the preparation of flavored yerba mate and in popular medicine, mainly in abdominal conditions. The objective of this work was to adjust the micropropagation technique, study the field behavior of vitroplants, compare the seeds generated and close their cultivation cycle. Different concentrations of growth regulators were evaluated on Murashige-Skoog medium. The implantation was successful. There are no differences between the evaluated plants. It was possible to efficiently close the complete cycle in vitro, with 100% survival, flowering and production of viable seeds. This methodology will serve for its introduction to the field, subsequent domestication, reintroduction into its natural environment and mitigate the process of degradation of the populations.

Hedeoma multiflora Benth. (Lamiaceae) es una especie aromatico-medicinal nativa de Argentina, Uruguay y sur de Brasil que se encuentra en estado de vulnerabilidad debido a la sobreexplotación. Es utilizada en la elaboración de yerba mate saborizada y en medicina popular, principalmente en afecciones abdominales. El objetivo de este trabajo fue ajustar la técnica de micropropagación, estudiar el comportamiento a campo de vitroplantas, comparar las semillas generadas y cerrar su ciclo de cultivo. Se evaluaron diferentes concentraciones de reguladores de crecimiento sobre medio de cultivo Murashige-Skoog. La implantación se realizó exitosamente, sin deterioro ni muerte de las plantas. No existen diferencias entre las plantas evaluadas. Se logró cerrar eficientemente el ciclo completo in vitro, con un 100% de supervivencia, floración y producción de semillas viables. Esta metodología servirá para su introducción a campo, posterior domesticación, reintroducción en su ambiente natural y mitigar el proceso de degradación de las poblaciones.

IN VITRO GERMINATION AND VEGETATIVE PROPAGATION THROUGH BUD DEVELOPMENT OF SACHA INCHI (Plukenetia volubilis) / Germinación y propagación vegetativa in vitro a través del desarrollo de yemas de Sacha Inchi (Plukenetia volubilis)

ABSTRACT This study describes the in vitro seed germination and micropropagation of Plukenetia volubilis (sacha inchi), an oilseed crop rich in omega-3 fatty acids, with health benefits and several industrial applications. Seed germination was evaluated in different culture media (MS and 1/2 MS), seed coat presence/absence and culture temperature (18 °C and 28 °C). Micropropagation was performed using axillary bud development (ABD) on nodal segments from in vitro seedlings. KIN, BAP and 2-ip were evaluated for ABD, and the effect of modified MS in 453 mg L-1 CaCl2 and 351.62 mg L-1 MgSO4 on ABD and shoot survival was assessed to improve the process. Finally, six treatments were evaluated to optimize ABD and shoot leaf formation. Seed germination of 91.6 % was achieved in MS at 28 °C when the seed coat was removed. ABD was obtained in 45 % and 40 % with 0.4 mg L-1 KIN and 0.6 mg L-1 2-ip, respectively, with the least CAL. The modification in 453 mg L-1 CaCl2 then allowed 76 % ABD and 82 % explant survival. ABD response was optimized to 95 % and 2.45 leaves with MS medium + CaCl2 modification + 10 % coconut water + 0.4 mg L-1 KIN. The same results were obtained by replacing the latter with 0.6 mg L-1 2-ip. Rooting was achieved in MS without PGR, and acclimatization was successful. The results indicate that plant production via germination and vegetative propagation is effective for commercial purposes.

RESUMEN Este estudio describe la germinación in vitro y micropropagación de Plukenetia volubilis, un cultivo oleaginoso rico en omega-3 benéfico para la salud y con múltiples aplicaciones industriales. Se evaluó en la germinación diferentes medios de cultivo (MS y 1/2 MS), presencia-ausencia de testa y temperatura de cultivo (18 ° C y 28 ° C). La micropropagación se realizó vía yemas axilares (ABD) de plántulas in vitro. Se evaluó el efecto de KIN, BAP y 2-ip sobre ABD, seguidamente, para mejorar el proceso se evaluó el efecto de MS modificado en 453 mg L-1 CaCl2 y 351.62 mg L-1 MgSO4 sobre ABD y supervivencia del brote. Finalmente, se evaluaron seis tratamientos para optimizar ABD y la formación de hojas. Se logró una germinación 91,6 % en MS a 28 °C cuando se retiró la testa. Se obtuvo 45 % y 40 % de ABD con 0,4 mg L-1 KIN y 0,6 mg L-1 2-ip respectivamente, ambos con la menor CAL. Posteriormente, la modificación de CaCl2 permitió 76 % ABD y 82 % de supervivencia. Se optimizó ABD al 95 % con 2,45 hojas por brote con el medio: MS + modificación de CaCl2 + 10 % de agua de coco + 0.4 mg L-1 KIN, los mismos resultados se obtuvieron cambiando este último con 0,6 mg L-1 2-ip, se logró enraizamiento en MS sin PGR, y la aclimatización fue exitosa. Los resultados indican que la producción de plantas vía germinación y propagación vegetativa es efectiva con fines comerciales.

In vitro establishment and multiplication of Echynochloa polystachya

The objective of this work was to carry out the in vitro establishment of Echynochloa polystachya aiming at obtaining a micropropagation protocol for works involving the selection of superior genotypes and the cultivation of the species. E. polystachya stems were collected in the municipality of Manaus-AM. Explants were inoculated in test tubes containing Murashige and Skoog (MS) medium. Thirty days after in vitro establishment, the rate of sprouting and contamination were evaluated. Experiments were also carried out to assess the effects of sucrose and 6-benzylaminopurine (BAP) concentrations on the tillering rate of explants. It was found that during the successive subcultures there was a decrease in internodes and the consequent loss of vigor. There were responses in the multiplication rate at concentrations starting from 45 g L-1 sucrose. In addition, BAP and sucrose interfered the development and in vitro multiplication. Sucrose in conjunction with BAP was harmful and shortened internodes. The physiological state of the explants for the species under study was intrinsically linked to the concentrations of sucrose used for the culture medium and the concentrations of BAP. However, the sucrose and BAP concentrations suggested for in vitro cultivation of E. Polystachya must be adjusted during successive subcultures. Absence of contamination in the in vitro establishment occurred at concentrations 15, 30 and 60 g L-1 sucrose. The combination of 1.5 mg L-1 BAP and 30 g L-1 sucrose promoted greater induction of sprouts. In addition, the in vitro rooting of E. polystachya was 45%.

Quantitative standardization of pharmacologically active components from antiplasmodial plant Solanum nudum Dunal (wild and in vitro) / Estandarización cuantitativa de componentes farmacológicamente activos de la planta antiplasmodial Solanum nudum Dunal (silvestre e in vitro)

Solanum nudum Dunal (Solanaceae) is most commonly known andused by the population of the colombian Pacific coast as an antimalarial treatment. This article study into optimization and quantitative analysis of compounds steroidal over time of development of this species when grown in vitro and wild. A new steroidal compound named SN6 was elucidated by NMR and a new method of quantification of seven steroidal compounds (Diosgenone DONA and six steroids SNs) using HPLC-DAD-MS in extracts of cultures in vitroand wild was investigated. Biology activity of extracts was found to a range of antiplasmodial activity in FCB2 and NF-54 with inhibitory concentration (IC50) between (17.04 -100µg/mL) and cytotoxicity in U-937 of CC50 (7.18 -104.7µg/mL). This method creates the basis for the detection of seven sterols antiplasmodial present in extracts from S. nudum plant as a quality parameter in the control and expression of phytochemicals.

Solanum nudum Dunal (Solanaceae) es el más conocido y utilizado por la población de la costa del Pacífico colombiano como tratamiento antipalúdico. Este artículo estudia la optimización y el análisis cuantitativo de compuestos esteroides a lo largo del tiempo de desarrollo de esta especie cuando se cultiva in vitro y en forma silvestre. Un nuevo compuesto esteroideo llamado SN6 fue dilucidado por RMN y se investigó un nuevo método de cuantificación de siete compuestos esteroides (Diosgenone DONA y seis esteroides SN) usando HPLC-DAD-MS en extractos de cultivos in vitro y silvestres. La actividad biológica de los extractos se encontró en un rango de actividad antiplasmodial en FCB2 y NF-54 con concentración inhibitoria (IC50) entre (17.04 -100 µg/mL) y citotoxicidad en U-937 de CC50 (7.18 -104.7 µg/mL). Este método crea la base para la detección de siete esteroles antiplasmodiales presentes en extractos de planta de S. nudum como parámetro de calidad en el control y expresión de fitoquímicos.

Vegetative propagation of hops (Humulus lupulus L. ): Historical approach and perspectives / Propagação vegetativa do lúpulo (Humulus lupulus L. ): Abordagem histórica e perspectivas

Hop (Humulus lupulus L.) female inflorescences are important raw materials used to produce beers, cosmetics, and medicines. Vegetative propagation is the preferred way of obtaining seedlings for commercial cultivations as female plants produce more lupulin than male plants, a component of commercial interest. It can be carried out by macropropagation (stem cuttings or rhizomes) or micropropagation. This review aimed to systematize different techniques of hop vegetative propagation, with no time frame, from searches in the main academic research bases: Capes Journal Portal, Scielo, Scopus, Web of Science, Science Direct, Google Scholar, and ResearchGate. Most studies are related to micropropagation, mainly addressing different plant regulators and concentrations, as well as types of explants and culture media, strategies to produce virus-free plants, artificial lighting, and cryopreservation. Experiments with stem cuttings are more common regarding macropropagation, but factors such as size and origin of cuttings, rooting period, and the response of different cultivars need to be better evaluated. Cultivation by cuttings allows the production of clones of female plants and micropropagation the production of virus-free clones in a short time and less physical space. Currently, micropropagation has been widely applied to cryopreservation.(AU)

As inflorescências femininas do lúpulo (Humulus lupulus L.) são matérias-primas importantes utilizadas na produção de cervejas, cosméticos e medicamentos. Como as plantas femininas produzem mais lupulina que as masculinas, componente de interesse comercial, a propagação vegetativa é a forma preferencial de obtenção de mudas para os cultivos comerciais. Esta pode ser realizada por macropropagação (estaquia caulinar ou rizomas) ou micropropagação. O objetivo desta revisão foi sistematizar as diferentes técnicas de propagação vegetativa do lúpulo, sem recorte temporal, a partir de buscas nas principais bases de pesquisa acadêmica: Portal de Periódicos Capes, Scielo, Scopus, Web of Science, Science Direct, Google Acadêmico e Research Gate. A maioria dos trabalhos são relacionados à micropropagação, abordando principalmente diferentes reguladores vegetais e concentrações, além de tipos de explantes e meios de cultura, estratégias para produzir plantas livres de vírus, iluminação artificial e criopreservação. Quanto à macropropagação, experimentos com estaquia caulinar são mais comuns, porém fatores precisam ser melhor avaliados tais como tamanho e origem das estacas, período de enraizamento e resposta de diferentes cultivares. O cultivo por estacas permite a produção de clones de plantas femininas e a micropropagação a produção de clones isentos de vírus, em pouco tempo e em menor espaço físico. Atualmente, a micropropagação tem sido muito aplicada à criopreservação.(AU)

Development of the prickly pear cactus Opuntia stricta (Haw.) Haw. (Cactaceae) in vitro in response to the replacement of potassium nitrate for a commercial kno 3 fertilizer / Desenvolvimento da palma Opuntia stricta (Haw.) Haw. (Cactaceae) in vitro em resposta à substituição de nitrato de potássio (P.A.) por fertilizante comercial KNO 3

ABSTRACT: In micropropagation, potassium nitrate (KNO3), an ACS reagent grade chemical, used in the preparation of growing mediums is expensive and its procurement depends on bureaucratic procedures, as it is controlled by the Brazilian Army. This research to assessed the effect of replacing the ACS KNO3 for a commercially available fertilizer (KNO3- based) on the micropropagation of the prickly pear cactus (Opuntia stricta (Haw.) Haw. cv. Elephant Ear. Treatments used six different fertilizer concentrations (0, 0.5, 1, 1.5, 2 and 2.5 g L-1) and a control consisting of 1.9 g L-1 KNO3, as shown in the MS salts. The survival, size and number of sprouts and the value of fresh biomass were evaluated. After seedling acclimation, we assessed the survival, number of sprouts, length, and number of roots, racket formation, average fresh biomass mass, macronutrient absorption and morphological changes of the seedlings. Explants inoculated with fertilizers at concentrations of 0.0; 2.0 and 2.5 g L-¹ did not grow. The response of explants at concentrations of 0.5 and 1.5 g L-1 of the fertilizer were the same as those developed in a KNO3 medium, and at a concentration of 1.0 g L-1, in all variables, the means were higher than those of the control medium. Therefore, it showed the feasibility of using fertilizers in the in vitro cultivation of the prickly pear cactus, which may remove bureaucratic barriers and reduce product costs by 99.12%.

RESUMO: Na micropropagação, o nitrato de potássio (KNO3), reagente puro para análise (P.A.), utilizado no preparo dos meios de cultura, possui custo elevado e a sua aquisição depende de trâmites burocráticos, por se tratar de substância controlada pelo Exército Brasileiro. O objetivo deste trabalho foi avaliar o efeito da substituição do KNO3 P.A. por fertilizante comercial (com fonte de KNO3), encontrado livremente no comércio, na micropropagação de palma (Opuntia stricta (Haw.) Haw. cv Orelha de Elefante. Os tratamentos foram de seis concentrações do fertilizante (0; 0,5; 1; 1,5; 2 e 2,5 g L-1) e um controle constituído de 1,9 g L-1 de reagente KNO3, conforme mostrado nos sais MS. Avaliou-se a sobrevivência, tamanho e número de brotações do explante, e o valor da biomassa fresca. Após a aclimatização das mudas avaliou-se a sobrevivência, número de brotações, comprimento da parte aérea, número de raízes, formação da raquete, massa média da biomassa fresca, absorção de macronutrientes e alterações morfológicas das mudas. Os explantes inoculados em meio com fertilizantes nas concentrações de 0,0; 2,0 e 2,5 g L-¹ não se desenvolveram. A resposta dos explantes nas concentrações de 0,5 e 1,5 g L-1 do fertilizante foram iguais aos desenvolvidos em meio contendo KNO3, e na concentração de 1,0 g L-1, em todas as variáveis, as médias foram superiores em relação as do controle. Dessa forma, constatou-se a viabilidade do uso do fertilizante no cultivo in vitro da palma, o que propiciou a eliminação dos entraves burocráticos e redução no custo de 99,12% na compra do produto.

Organogenesis on apical buds in common fig (Ficus carica) var. Black Jack

BACKGROUND Plant tissue culture involves the use of explants obtained from plants to induce organogenesis with the help of plant growth regulators (PGRs). Micropropagation techniques provide a faster and economical solution to the limitations associated with traditional methods of plant cultivation. The present study focuses on the multiple shoot induction and proliferation of Ficus carica var. Black Jack. Factors that influence the growth of in vitro multiple shoots on the apical buds, which include growth media and PGRs, were investigated in this study. Different concentrations of cytokinins like 6-benzylaminopurine (BAP), Thidiazuron (TDZ), and Kinetin (Kin) were used on woody plant medium (WPM) for the optimization of media for multiple shoot induction and proliferation. RESULTS Apical buds of Ficus carica var. Black Jack growing in WPM supplemented with BAP produced the healthiest plantlets, with the highest number of multiple shoots. The most efficient medium composition which produced the highest number of multiple shoots (37.8) per growing explant was WPM supplemented with 20 mM BAP. Proliferated multiple shoots were efficiently rooted using WPM + 20 mM BAP + 8 mM indole-3-acetic acid (IAA). This optimized medium composition significantly enhanced the production of multiple, disease-free plantlets using single apical bud explants of Ficus carica var. Black Jack. CONCLUSIONS In the present study the observations indicate that WPM supplemented with 20 mM BAP is the best-suited medium for organogenesis and multiple shoot culture of Ficus carica var. Black Jack, and this technique can be potentially applied for commercialization of the plant

In vitro Propagation to Conserve the Local Endemic and Endangered Medicinal Plant Helianthemum germanicopolitanum Bornm

Abstract In this study, in vitro propagation and acclimatization of Helianthemum germanicopolitanum Bornm. plant, a local endemic in Çankırı Province (Turkey) with arid and semi-arid lands, and an endangered species taking part among medicinal and aromatic plants were accomplished, which is under-researched. In this study, three basal media [a) Murashige and Skoog b) Gamborg's B5, and c) Nitsch & Nitsch], two gelling agents (agar 7 g/L, and gelrite 2.1 g/L), eight cytokinins and eight auxin doses of plant growth regulators [a) 6-benzyladenin, b) Kinetin-(0, 0.5, 1, and 2 mg/L), c) Indole-3-butyric acid, d) α-napthaleneacetic acid-(0, 0.25, 0.5, and 1 mg/L)] prepared in 64 different combinations with 30 g/L sucrose was added to the basal media and adjusted to pH 5.7 for in vitro propagation of H. germanicopolitanum. During in vitro propagation of the plant, external and internal infections were frequently encountered and this was solved by the developed protocol. The best shoot growth (1.141 cm) and shoot length (0.572 cm) were obtained in the Gamborg's B5 medium in combination with Kinetin (0.5 mg/L)+Indole-3-butyric acid (0.5 mg/L)+gelrite. The maximum number of shoots (19.50) and the best multiplication rate (94%) were obtained in the media containing benzyladenin (1 mg/L)+Indole-3-butyric acid (0.5 mg/L) plant growth regulator in Murashige and Skoog medium solidified with agar. At the rooting stage, the maximum number of roots (30) was reached in the Murashige and Skoog medium containing gelrite and the best rooting rate (92%) with agar. A hundred plants representing the best shoot and root growth were taken to acclimatization stage, and 32 of these plants adapted to external conditions.

Strategies for vegetative propagation and viral cleaning of a miniature ornamental pineapple hybrid

This study assessed and compared different methods for vegetative propagation of a miniature ornamental pineapple hybrid (ORN-MUT), seeking to determine the best method for production of plantlets, as well as for removal of the PMWaV viral complex from plants cultured in vitro, for production of healthy parent plants. Pineapple wilt is a disease that can cause large economic and is caused by a viral complex called Pineapple mealybug wilt-associated virus (PMWaV). For this, four propagation methods were evaluated (conventional, stem sectioning, micropropagation and etiolation of nodal segments). The time necessary for each method and the number of plants formed were assessed. Stem tips (0.5 mm) were cultured and indexed for three PMWaV types. Conventional propagation produced 17 plantlets per plant in 566 days, stem sectioning produced 2.3 plantlets per stem in 591 days, while the conventional micropropagation technique produced 1,284 plants after four subcultures in 778 days. Stems etiolated for 60 days showed peak production in the second subculture, with 1,224 plants. This method required 883 days to obtain plants with ideal size for transplantation to the field. In turn, stems etiolated for 120 days produced 935 plants at the end of four subcultures, with peak output in the third subculture, in which the plants could be cultivated in the field after 943 days. Conventional micropropagation and etiolation for 60 days were the best methods for production of plantlets of the ORN-MUT hybrid. The results of this work showed that the cultivation of shoot tips is an efficient strategy to remove the PMWaV complex and obtain healthy mother plants and can be a useful tool for other varieties of pineapple.

In vitro multiplication of wild Manihot species with different naphthaleneacetic acid and benzylaminopurine concentrations

In vitro multiplication is an important tissue culture technique that is capable of efficiently producing seedlings at any scale. It is a propagation method based on the aseptic culture of small propagules in a suitable culture medium to enable plant regeneration. Multiplication experiments conducted in vitro to set protocols adapted to wild Manihot species have used modified mineral salts and MS vitamins as basic culture medium. Here, 25 treatments based on combinations of the regulators benzylaminopurine (BAP) and naphthaleneacetic acid (NAA) at 0, 0.025, 0.05, 0.075, and 0.1 mg L-1 were used for in vitro multiplication of three genotypes of wild Manihot species (M. violaceae Pohl Müll. Arg., M. pseudoglaziovii Pax & Hoff., and M. flabellifolia Pohl). Plant height and the number of 1 cm minicuttings, number of roots, shoots, green leaves and senescent leaves were recorded 120 days after explant inoculation. M. violaceae Pohl. Müll. Arg. and M. flabellifolia Pohl. presented favorable results with 0.05 and 0.025 mg L-1 NAA, respectively. Culture medium lacking NAA and BAP favored the in vitro growth of M. pseudoglaziovii Pax & Hoff.

Improving micropropagation of some grape cultivars via boron, calcium and phosphate

BACKGROUND: An efficient regeneration protocol is a priority for the successful application of plant biotechnology. Grape nodal explants were used to develop a micropropagation protocol for Thompson Seedless and Taify cvs. Explants were cultured on MS medium supplemented with Kinetin or benzylaminopurine (BA) and indolebutyric acid (IBA). RESULTS: For both cultivars, axillary buds were grown, only, on a medium enriched with kinetin, moreover, shoot tip necrosis and callus formation were observed on Thompson Seedless cv. cultures grown on a medium with BA. Supplementing the growth medium with 100 mM (boron) B and 2.5 mM (calcium) Ca successfully help overcome these phenomena. The highest regenerated shoot numbers (14 and 6.2 explant 1 ) for Taify and Thompson Seedless cvs., respectively, were on media supplemented with 13.2 mM BA + 4.9 mM IBA and BA 13.2 mM + 5.8 mM IBA, respectively. Moreover, these media supported the developing shoots to have the heaviest dry weights (1.46 and 0.72 mg explant 1 ) for Taify and Thompson Seedless cvs., respectively. Thompson Seedless cv. regenerated shoot numbers and their dry weights were significantly increased by increasing the MS medium PO4 concentration. However, these two parameters were significantly decreased for Taify cv. Developing shoots were elongated and rooted on MS medium enriched with 4.9 mM, IBA 100 mM B and 2.5 mM Ca. Plantlets were acclimatized and successfully transferred to the greenhouse conditions. CONCLUSIONS: A novel promising protocol for Thomson Seedless and Taify cvs. micropropagation using single nodes has been developed.

Micropropagación de Prosopis pallida (Humb &Bonpl. Ex Willd.) Kunth a partir de yemas apicales / Micropropagation of Prosopis pallida (Humb &Bonpl. Ex Willd.) Kunth from shoot tips

RESUMEN Prosopis pallida, conocido como algarrobo, es una especie emblemática de los bosques secos del norte del Perú. Es de gran importancia económica por su uso en la producción de leña y carbón, así como en la producción de algarrobina proveniente de sus frutos. Actualmente las actividades humanas han deforestado grandes poblaciones de algarrobo en el bosque seco, por lo que es muy importante una propagación masiva para planes de reforestación a gran escala en esos ecosistemas, con la finalidad de conservar la especie y también las características genéticas de individuos élite. El objetivo de la investigación fue establecer un protocolo para la propagación in vitro de algarrobo. Previo a la siembra in vitro, se evaluaron tres tratamientos pregerminativos para poder acelerar la germinación de las semillas con la finalidad de obtener mayor material de propagación. Luego se evaluó el efecto del medio de plantas leñosas con la adición de cuatro concentraciones (0; 0,5; 1,0 y 1,5 mg/L) de citoquininas (BAP y ZEA) sobre la propagación in vitro de Prosopis pallida, habiendo realizado tres ensayos debido a la poca efectividad de brotación de las yemas apicales, resultando mejor la ausencia de citoquininas en yemas apicales con sus dos cotiledones, usando tapas de algodón en los tubos de ensayo. En esta etapa se evaluó el número de nudos, altura de plántula y número de brotes. Para el enraizamiento se ensayó con tres concentraciones (0; 0,5 y 1,0 mg/L) de auxinas (NAA, IBA y IAA), y se evaluó el porcentaje de enraizamiento, longitud de la raíz y número de raíces; obteniéndose mejores resultados con 0,5 mg/L IBA. En la fase de aclimatación se evaluó el porcentaje de aclimatación en dos tipos de sustratos, obteniéndose mejores resultados con sustrato comercial de turba y vermiculita.

ABSTRACT Prosopis pallida, known as algarrobo, is an emblematic species of the dry forests of northern Peru. It is of great economic importance for its use in the production of firewood and coal, as well as in the production of carob from its fruits. Currently human activities have deforested large populations of algarrobo in the dry forest, so it is very important a massive propagation for large-scale reforestation plans in these ecosystems, in order to conserve the species and also the genetic characteristics of elite individuals. The objective of the research was to establish a protocol for the in vitro propagation of algarrobo. Before in vitro propagation, It was evaluated the effect of three seeds treatments to accelerate seeds germination in order to get more propagation material. Then, the effect of woody plants medium with the addition four concentrations (0, 0.5, 1.0 and 1.5 mg /L) of cytokinins (BAP and ZEA) on the in vitro propagation of Prosopis was evaluated and it has performed three trials for the ineffective budding of apical buds, resulting in a better absence of cytokinins in apical buds with their two cotyledons, and using cotton lids in the test tubes. In this stage the number of nodes, seedling height and number of shoots was evaluated. For rooting, it was tested with three concentrations (0, 0.5 and 1.0 mg /L) of auxins (NAA, IBA and IAA), and the percentage of rooting, root length and amount of roots was evaluated, obtaining better results with 0.5 mg / L IBA. In the acclimation phase, the acclimation percentage was evaluated using two substrates, and the best results were commercial substrate of peat and vermiculite.

Conservation of Vriesea flammea L. B. Sm., an endemic Brazilian bromeliad: effects of nutrients and carbon source on plant development / Conservação de Vriesea flammea L. B. Sm., uma bromélia Brasileira endêmica: efeitos dos nutrientes e fonte de carbono sobre o desenvolvimento das plantas

Abstract Bromeliaceae is restricted to the Neotropical region and has a high degree of endemism, which contributes to increased biodiversity because of the diverse morphological characteristics of individuals. In order to develop an in vitro conservation technology to obtain plants for reintroduction, seeds of Vriesea flammea L.B.Sm. were collected, sterilized and germinated in culture medium. The plants obtained were cultured for 180 days in MS medium with different concentrations of mineral nutrients (25 and 50% of nitrogenous salts and macronutrients), and different concentrations of sucrose (20, 30, 40, 50 and 60 g L-1), and then acclimatized for 150 days on commercial substrate. When seeds were sterilized directly, only 4% of them were contaminated, whereas sterilization of capsules resulted in 43.6% contaminated seeds. Germination rates above 80% were recorded. Low concentrations of nitrogenous salts and macronutrients produced greater than 76% survival and promoted greater in vitro plant development than the complete MS medium. The development of the aerial system, root system, fresh mass and photosynthetic pigments were positively related to sucrose concentration in vitro. The highest sucrose concentration also indirectly promoted greater development of the aerial system and fresh mass of acclimatized plants. We established conditions for in vitro cultivation and acclimatization for efficient propagation of V. flammea with a view towards conservation of the species or reestablishment of natural populations.

Resumo Bromeliaceae é restrita à região neotropical, com alto grau de endemismo, que contribui para o aumento da biodiversidade, devido às características morfológicas dos indivíduos. Objetivando desenvolver uma tecnologia in vitro de conservação para obtenção de plantas com vistas à reintrodução, sementes de Vriesea flammea L.B.Sm. foram coletadas, esterilizadas, germinadas em meio de cultura e as plantas obtidas foram cultivadas por 180 dias em meio MS com diferentes concentrações de nutrientes minerais (25 e 50% dos sais nitrogenados e dos macronutrientes), bem como em diferentes concentrações de sacarose (20, 30, 40, 50 e 60 g L-1). Após, as plantas foram aclimatizadas por 150 dias em substrato comercial. Quando da esterilização das sementes, apenas 4% destas contaminaram. Por sua vez, a esterilização de cápsulas resultou em 43,6% de sementes contaminadas. Taxas de germinação superiores a 80% foram registradas. Baixas concentrações dos sais nitrogenados e de macronutrientes proporcionaram sobrevivência superior a 76% e promoveram maior desenvolvimento das plantas in vitro do que o meio MS completo. O desenvolvimento do sistema aéreo, radicular, a massa fresca e pigmentos fotossintéticos apresentaram relação positiva com a concentração de sacarose in vitro. A maior concentração de sacarose também propiciou indiretamente maior desenvolvimento do sistema aéreo e massa fresca das plantas aclimatizadas. Estabelecemos as condições de cultivo in vitro e aclimatização para a eficiente propagação de V. flammea com vistas à conservação da espécie ou reestabelecimento das populações naturais.

Clonal bamboo production based on in vitro culture / Produção de plantas clonais de bambu a partir do cultivo in vitro

Bamboo species are an alternative for the composition of forest plantations. However, their potential has not been explored due to the hard time in producing large-scale clonal plants. Thus, the aim this work was to evaluate the in vitro establishment, bud multiplication and ex vitro rooting of Bambusa vulgaris. The first experiment tested different systemic and contact fungicide solutions, based on exposure time, during the establishment phase. Established explants were subjected to evaluation of residual fungicide effect on subcultures during the multiplication and elongation phases. The second experiment evaluated the influence of activated carbon on ex vitro survival and on adventitious rooting. Explant immersion in liquid culture medium added with 1.0 mL of fungicide for 120 hours has favored the in vitro establishment and reduced fungal contamination. On the other hand, it favored the shoot emission of shoots per explant during the multiplication phase. Both rooting induction culture medium and mini-incubator system use were effective in enabling adventitious root formation. The presence of activated carbon in the rooting induction culture medium resulted in a higher clonal plant survival rate.

As espécies de bambus são uma alternativa para a composição de plantios florestais. Entretanto, esse potencial não tem sido explorado devido à dificuldade de produção de mudas clonais em larga escala. Assim, objetivo deste trabalho foi avaliar o estabelecimento in vitro, a multiplicação e o enraizamento ex vitro de Bambusa vulgaris. No primeiro experimento foram testadas diferentes soluções de fungicida sistêmico e de contato em relação ao tempo de exposição durante a fase de estabelecimento. Os explantes estabelecidos foram avaliados quanto ao efeito residual do fungicida durante subcultivos nas fases de multiplicação e alongamento. No segundo experimento, foi avaliada a influência do carvão ativado sobre a sobrevivência e enraizamento ex vitro. Durante o estabelecimento in vitro, a imersão de explantes em meio de cultura líquido contendo alíquota de 1,0 mL de fungicida durante 120 horas favoreceu o estabelecimento e reduziu a contaminação fúngica, enquanto na fase de multiplicação, houve o favorecimento da emissão de brotos por explante. O meio de cultura de indução ao enraizamento e uso de sistema de mini-estufim foram efetivos para a formação de raízes adventícias e a presença de carvão ativado resultou em uma maior sobrevivência das mudas clonais.

Micropropagation of Stevia rebaudiana plants / Micropropagação de plantas de Stevia rebaudiana

ABSTRACT: The aim of the study was to develop optimum composition of plant growth regulators in media for the propagation and rooting of shoots of stevia (Stevia rebaudiana Bertoni) in in vitro cultures. Single-node shoot fragments obtained from plants propagated on MS medium were placed onto media supplemented with: BAP, 2iP and KIN at concentrations: 0.5, 1, 2 and 5 mg∙dm-3, whereas at the rooting stage with addition of: IAA, IBA and NAA at concentrations 1, 2, 4 and 8 mg∙dm-3. The highest number of shoots and leaves was reported for plants propagated on MS medium enriched with 0.5 mg∙dm-3 BAP. The greatest number of the longest roots was developed by stevia on the MS medium enriched with 1 mg∙dm-3 IAA.

RESUMO: O objetivo do estudo foi desenvolver uma composição ótima de reguladores de crescimento em meios para a propagação e enraizamento de brotos de estévia (Stevia rebaudiana Bertoni) em culturas in vitro. Fragmentos de parte aérea obtidos de plantas propagadas em meio MS foram colocados em meio suplementado com: BAP, 2iP e KIN nas concentrações: 0.5, 1, 2 e 5 mg∙dm-3, enquanto no estádio de enraizamento com adição de: IAA, IBA e ANA nas concentrações 1, 2, 4 e 8 mg∙dm-3.O maior número de brotações e folhas foi encontrado para plantas propagadas em meio MS enriquecido com 0.5 mg∙dm-3 de BAP. O maior número de raízes mais longas foi desenvolvido por estévia no meio MS enriquecido com 1 mg∙dm-3 de IAA.

Germinación y multiplicación in vitro de Matricaria recutita L: los fenoles totales determinan su germinación / Germination and in vitro multiplication of Matricaria recutita L.: total phenols determine their germination

RESUMEN La manzanilla (Matricaria recutita L., Chamomilla recutita L. y Matricaria chamomilla L.), es conocida por su alto contenido de compuestos fenólicos que le confieren propiedades antiinflamatorias, antisépticas y antimutagénicas. En este estudio se evaluó el porcentaje de fenoles totales y la germinación en cinco periodos de almacenamiento de semillas de M. recutita (5, 31, 75, 96 y 128 días). Además, se evaluó el efecto de citoquininas (6-Bencil Amino Purina, BAP y Kinetina) y auxinas (α-Ácido Naftalen Acético, ANA) en la brotación in vitro de esta especie. Se evidenció que la concentración total de fenoles disminuyó de 13.8% a 1.9% en los cinco periodos de almacenamiento evaluados y que los porcentajes de germinación aumentaron de 2.2% a los cinco días a 8,9% a los 128 días de almacenamiento, mostrándose evidencia de una correlación de -0.989 entre la germinación y el contenido de fenoles totales. Los mejores resultados para inducir brotación (5 brotes/explante) fueron obtenidos en el medio de cultivo MS con citoquininas.

ABSTRACT Chamomile (Matricaria recutita L., Chamomilla recutita L., and Matricaria chamomilla L.) is known for its high content of phenolic compounds that confer anti-inflammatory, antiseptic and antimutagenic properties. This study evaluated the percentage of total phenols and germination in five storage periods of M. recutita seeds (5, 31, 75, 96 and 128 days). In addition, the effect of cytoki-nins (6-Benzyl Amino Purine, BAP and Kinetin) and auxins (α-Naphthalene Acetic Acid, ANA) on in vitro sprouting of this species was evaluated. It was evidenced that the total concentration of phenols decreased from 13.8% to 1.9% in the five storage periods evaluated and that the germination percentages increased from 2.2% at five days to 8.9% at 128 days of storage, showing evidence of a correlation of -0.989 between the germination and the content of total phenols. The best results to induce sprouting (5 shoots/ explant) were obtained in the MS culture medium with cytokinins.

Effect of Rhustox on Micropropagation of Scoparia dulcis L. through Leaf Explants Culture

Aim: To investigate the effect of Rhus toxicodendron (30CH) along with different compositions of phytohormones (Auxin and Cytokinin) on the basis of growth and multiplication of explants under optimum temperature under in-vitro conditions. Study Design: To establish and design the standard protocol for the in-vitro propagation through leaf explant of Scoparia dulcis under stress of phytohormones and homeopathic medicine Rhus toxicodendron (30CH). Place and Duration of Study: The plant materials were procured from the Herbal Botanical Garden Patna Science College, Department of Botany, Patna University, Patna, Bihar. The experimental part was carried out in Plant Tissue Culture Laboratory, between December 2017 to August 2018 in Department of Botany P.U. Patna. Methodlogy: The sterilized leaf explants were inoculated into MS media fortified with different phytohormones (Auxin and Cytokinin) and Rhus tox(30CH) under aseptic environmental conditions for the growth and development of callus, embryoids etc. Result: The explants in MS medium supplemented with auxins phytohormones and Rhus tox(30CH) exhibited that IAA (0.10 to 2.0 mg/l) and BAP (0.10 to 2.5 mg/l) induces green and compact calli. Whereas at 0.30mg/l of IAA and 0.50 mg/l BAP induced brown friable calli. 2,4-D (1.5 mg/l) and Kinetin (1.5-6.5mg/l) concentrations induced brown and friable calli. Rhus tox(30CH) (100 µl/100 ml) enhances proliferation with 2,4-D and Kinetin (1.5/1.5 mg/l.). Conclusion: After 42 days of culture initiation and establishment the callus was 520.0±1.12 mg in the mixture of 2,4-D and Kinetin (1.5 mg/l) in Rhus tox free medium. Whereas weight of callus were found to be 1092±0.74 mg after 42 days in the same medium of 2, 4-D and Kinetin (1.5/5.5 mg/l) supplemented with Rhus tox (100 µl/100 ml). Hence, the investigation proponded that the Rhus tox (CH30) has increased the rate of callus development and plantlet regeneration.