In Vitro Propagation of Threatened Agave Species Using a Two-Stage System.

Agaves are plants with multiple possibilities of use and are naturally tolerant to low water availability conditions and high temperatures. This makes them species of great interest in the context of the necessary substitution of crops due to climate change. Unfortunately, the overexploitation of wild specimens has endangered many species of the genus that have not been domesticated or cultivated intensively. In vitro mass culture and propagation techniques have emerged as a very efficient option to produce agave plants that can be used without damage to the natural populations. A protocol is presented here for the in vitro micropropagation of agaves in a two-stage process. In the first step, clusters of slightly differentiated shoots are generated from stem segments cultivated on a semisolid medium added with cytokinin. In a second step, these shoot clusters are cultured in temporary immersion bioreactors where they grow and complete their differentiation, and then the shoots are rooted and transferred to soil. This protocol has been successfully applied to several threatened species of the Agave genus.

Delayed senescence and crop performance under stress: always a functional couple?

Exposure to abiotic stresses accelerates leaf senescence in most crop plant species, thereby reducing photosynthesis and other assimilatory processes. In some cases, genotypes with delayed leaf senescence (i.e. 'stay-green') show stress resistance, particularly in cases of water deficit, and this has led to the proposal that senescence delay improves crop performance under some abiotic stresses. In this review, we summarize the evidence for increased resistance to abiotic stress, mostly water deficit, in genotypes with delayed senescence, and specifically focus on the physiological mechanisms and agronomic conditions under which the stay-green trait may ameliorate grain yield under stress.

Comparative transcriptome analysis of high- and low-embryogenic Hevea brasiliensis genotypes reveals involvement of phytohormones in somatic embryogenesis.

BACKGROUND: Rubber plant (Hevea brasiliensis) is one of the major sources of latex. Somatic embryogenesis (SE) is a promising alterative to its propagation by grafting and seed. Phytohormones have been shown to influence SE in different plant species. However, limited knowledge is available on the role of phytohormones in SE in Hevea. The anther cultures of two Hevea genotypes (Yunyan 73477-YT and Reken 628-RT) with contrasting SE rate were established and four stages i.e., anthers (h), anther induced callus (y), callus differentiation state (f), and somatic embryos (p) were studied. UPLC-ESI-MS/MS and transcriptome analyses were used to study phytohormone accumulation and related expression changes in biosynthesis and signaling genes. RESULTS: YT showed higher callus induction rate than RT. Of the two genotypes, only YT exhibited successful SE. Auxins, cytokinins (CKs), abscisic acid (ABA), jasmonic acid (JA), salicylic acid (SA), gibberellins (GAs), and ethylene (ETH) were detected in the two genotypes. Indole-3-acetic acid (IAA), CKs, ABA, and ETH had notable differences in the studied stages of the two genotypes. The differentially expressed genes identified in treatment comparisons were majorly enriched in MAPK and phytohormone signaling, biosynthesis of secondary metabolites, and metabolic pathways. The expression changes in IAA, CK, ABA, and ETH biosynthesis and signaling genes confirmed the differential accumulation of respective phytohormones in the two genotypes. CONCLUSION: These results suggest potential roles of phytohormones in SE in Hevea.

The Cytokinins BAP and 2-iP Modulate Different Molecular Mechanisms on Shoot Proliferation and Root Development in Lemongrass (Cymbopogon citratus).

The known activities of cytokinins (CKs) are promoting shoot multiplication, root growth inhibition, and delaying senescence. 6-Benzylaminopurine (BAP) has been the most effective CK to induce shoot proliferation in cereal and grasses. Previously, we reported that in lemongrass (Cymbopogon citratus) micropropagation, BAP 10 µM induces high shoot proliferation, while the natural CK 6-(γ,γ-Dimethylallylamino)purine (2-iP) 10 µM shows less pronounced effects and developed rooting. To understand the molecular mechanisms involved, we perform a protein-protein interaction (PPI) network based on the genes of Brachypodium distachyon involved in shoot proliferation/repression, cell cycle, stem cell maintenance, auxin response factors, and CK signaling to analyze the molecular mechanisms in BAP versus 2-iP plants. A different pattern of gene expression was observed between BAP- versus 2-iP-treated plants. In shoots derived from BAP, we found upregulated genes that have already been demonstrated to be involved in de novo shoot proliferation development in several plant species; CK receptors (AHK3, ARR1), stem cell maintenance (STM, REV and CLV3), cell cycle regulation (CDKA-CYCD3 complex), as well as the auxin response factor (ARF5) and CK metabolism (CKX1). In contrast, in the 2-iP culture medium, there was an upregulation of genes involved in shoot repression (BRC1, MAX3), ARR4, a type A-response regulator (RR), and auxin metabolism (SHY2).

Multiplication, morphogenesis and foliar abscision of Schinopsis brasiliensis Engl. in vitro / Multiplicación, morfogénesis y abscisión foliar de Schinopsis brasiliensis Engl. in vitro

The objective was to evaluate plant growth regulators and ethylene inhibitors on the development and leaf abscission of Schinopsis brasiliensis Engl. Zeatin (ZEA) was evaluated in concentrations combined with concentrations of indolacetic acid (IAA), naphthalene acetic acid (NAA) and indolbutyric acid (IBA). ZEA and 6-benzylamino purine (BAP) were evaluated in concentrations plus a control. Ethylene inhibitors, silver nitrate and cobalt chloride were evaluated in four concentrations. The addition of 0.2 µL-1 of NAA to 0.4 µL-1 of ZEA promotes a greater number of baraúna sprouts. At concentrations of 5 and 10 µM, cobalt chloride is more efficient than silver nitrate for reducing leaf abscission in baraúna. IAA is the most suitable auxin to be associated with ZEA for higher shoot length and number of buds. Silver nitrate from a concentration of 20 µM completely avoids leaf abscission whilecobalt chloride has a maximum reduction in abscission at a concentration of 40 µM.

El objetivo fue evaluar reguladores de crecimiento e inhibidores de etileno sobre el desarrollo y abscisión foliar en Schinopsis brasiliensis Engl. La zeatina (ZEA) se evaluó en concentraciones combinadas con concentraciones de ácido indolacético (IAA), ácido naftaleno acético (NAA) y ácido indolbutírico (IBA). Se evaluaron ZEA y 6-bencilamino purina (BAP) en concentraciones más un control. Se evaluaron inhibidores de etileno, nitrato de plata y cloruro de cobalto, en cuatro concentraciones. La adición de 0.2 µL-1 de NAA a 0.4 µL-1 de ZEA promueve un mayor número de brotes de baraúna. A concentraciones de 5 y 10 µM, el cloruro de cobalto es más eficaz que el nitrato de plata para reducir la abscisión de las hojas en baraúna. IAA es la auxina más adecuada para asociar con ZEA para una mayor longitud de brotes y número de brotes. El nitrato de plata a partir de una concentración de 20 µM evita completamente la abscisión de las hojas, mientras que el cloruro de cobalto tiene una reducción máxima en la abscisión a una concentración de 40 µM.

Testing Explant Sources, Culture Media, and Light Conditions for the Improvement of Organogenesis in Pinus ponderosa (P. Lawson and C. Lawson).

Pinus. ponderosa (P. Lawson and C. Lawson) is a commercial tree and one of the most important forest species in North America. Ponderosa pine suffers hardship when going through vegetative propagation and, in some cases, 15-30 years are needed to achieve full reproductive capacity. Based on previous works on P. ponderosa regeneration through in vitro organogenesis and trying to improve the published protocols, our objective was to analyze the influence of different types of explants, basal culture media, cytokinins, auxins, and light treatments on the success of shoot multiplication and rooting phases. Whole zygotic embryos and 44 µΜ 6-benzyladenine showed the best results in terms of explants survival. For shoot organogenesis, whole zygotic embryos and half LP (LP medium, Quoirin and Lepoivre, 1977, modified by Aitken-Christie et al., 1988) macronutrients were selected. A significant positive interaction between whole zygotic embryos and half LP macronutrients was found for the percentage of explants forming shoots. Regarding the light treatments applied, a significantly higher percentage of shoots elongated enough to be rooted was detected in shoots growing under blue LED at a light intensity of 61.09 µmol m-2 s-1. However, the acclimatization percentage was higher in shoots previously cultivated under fluorescent light at a light intensity of 61.71 µmol m-2 s-1. Anatomical studies using light microscopy and scanning electron microscopy showed the light treatments promoted differences in anatomical aspects in in vitro shoots; needles of plantlets exposed to red and blue LEDs revealed less stomata compared with needles from plantlets exposed to fluorescent light.

The source, level, and balance of nitrogen during the somatic embryogenesis process drive cellular differentiation.

Since the discovery of somatic embryogenesis (SE), it has been evident that nitrogen (N) metabolism is essential during morphogenesis and cell differentiation. Usually, N is supplied to cultures in vitro in three forms, ammonium (NH4+), nitrate (NO3-), and amino N from amino acids (AAs). Although most plants prefer NO3- to NH4+, NH4+ is the primary form route to be assimilated. The balance of NO3- and NH4+ determines if the morphological differentiation process will produce embryos. That the N reduction of NO3- is needed for both embryo initiation and maturation is well-established in several models, such as carrot, tobacco, and rose. It is clear that N is indispensable for SE, but the mechanism that triggers the signal for embryo formation remains unknown. Here, we discuss recent studies that suggest an optimal endogenous concentration of auxin and cytokinin is closely related to N supply to plant tissue. From a molecular and biochemical perspective, we explain N's role in embryo formation, hypothesizing possible mechanisms that allow cellular differentiation by changing the nitrogen source.

Auxin-Cytokinin Cross Talk in Somatic Embryogenesis of Coffea canephora.

Cytokinins (CK) are plant growth regulators involved in multiple physiological processes in plants. One less studied aspect is CK homeostasis (HM). The primary genes related to HM are involved in biosynthesis (IPT), degradation (CKX), and signaling (ARR). This paper demonstrates the effect of auxin (Aux) and CK and their cross talk in a Coffea canephora embryogenic system. The transcriptome and RT-qPCR suggest that Aux in pre-treatment represses biosynthesis, degradation, and signal CK genes. However, in the induction, there is an increase of genes implicated in the CK perception/signal, indicating perhaps, as in other species, Aux is repressing CK, and CK are inducing per se genes involved in its HM. This is reflected in the endogenous concentration of CK; pharmacology experiments helped study the effect of each plant growth regulator in our SE system. We conclude that the Aux-CK balance is crucial to directing somatic embryogenesis in C. canephora.

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

Micropropagación de Calibanus hookeri (lem.) trel (1911). Una especie amenazada / Micropropagation of Calibanus hookeri (lem.) trel (1911). A threatened species

RESUMEN La especie Calibanus hookerii perteneciente a la familia Asparagaceae, está registrada en la NOM-059-SEMARNAT-2010 catalogada como planta amenazada. Sus poblaciones naturales se han visto reducidas de manera importante debido a una explotación excesiva y destrucción de su hábitat, por lo que se requiere de métodos de propagación eficaz que aseguren su conservación. La propagación in vitro es una alternativa viable para especies vegetales amenazadas. En la presente investigación se reporta el protocolo para la micropropagación de Calibanus hookerii mediante la germinación de semilla sin testa inoculada en medio MS complementado con 2.5, 5.0 y 7.0 mg L-1 de 6 benciladenina (BA), cinetina (K), 2-isopentil-adenina (2iP) y tidiazuron (TDZ). Las variables a medir fueron porcentaje de germinación, número y longitud de brotes producidos por semilla. El tratamiento más eficiente fue de 5.0 mg L-1 de BA produciéndose un promedio de 26 brotes por semilla; el tratamiento menos eficaz fue con 2.5 mg L-1 K en el cual solamente se obtuvieron dos brotes por semilla. De las tres concentraciones de 2iP solamente en la concentración de 7 mg. L-1 mostró resultados produciendo 6 brotes por semilla. En lo que respecta a la longitud del brote ningún tratamiento superó al testigo (8.07cm). La eficiencia la germinación in vitro fue de 56-97%.

ABSTRACT The species Calibanus hookerii belonging to the family Aspagaceae is registered in the NOM-059-SEMARNAT-2010 cataloged in danger of extinction and therefore is necessary of propagation methods that assure its conservation and its propagation. In vitro propagation is a viable alternative for endangered plant species. The present investigation reports the protocol for the micropropagation of Calibanus hookerii. This was achieved by seed germination without test in MS medium supplemented with 2.5, 5.0 and 7.0 mg L-1 of 6-benzyladenine (BA), kinetin (K), 2-isopentyl-adenine (2iP) and tidiazuron (TDZ). The variables to be measured were percentage of germination, number and length of shoots produced by seed. The most efficient treatment was 5 mg L-1 of BA producing an average of 26 shoots per seed, the worst treatment was with 2.5 K only produced 2 shoots per seed, of the four cytokinins used 2ip treatment in only one study of the Performed showed results (7 mg L-1) producing 6 shoots per seed. Regarding the length of the shoot, no treatment exceeded the control (8.07cm). Finally, in vitro germination was high (56-97%) in all treatments.

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

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.(AU)

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.(AU)

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.

Micropropagation of Agave Species.

The genus Agave originates from the American continent and grows in arid and semiarid places, being México the center of origin. Many species of the genus are a source of diverse products for human needs, such as food, medicines, fibers, and beverages, and a good source of biomass for the production of biofuels, among many others. These plants are gaining importance as climate change becomes more evident as heat is reaching temperatures above 40 °C worldwide and rains are scarce. Many species of the genus grow in places where other plant species do not survive under severe field conditions, due to their CAM pathway for fixing CO2 where gas exchange occurs at night when stomata are open, allowing them to avoid excess loss of water. Most of the important species and varieties are usually propagated by offshoots that develop from rhizomes around the mother plant and by bulbils that develop up in the inflorescence, which are produced by the plant mostly when there is a failure in the production of seeds.Areas for commercial plantations are growing worldwide and therefore in the need of big amounts of healthy and good quality plantlets. Although many Agave species produce seeds, it takes longer for the plants to reach appropriate maturity and size for diverse purposes. Micropropagation techniques for the genus Agave offer the opportunity to produce relatively high amounts of plants year around in relatively small spaces in a laboratory. Here, a protocol for micropropagation that has proven good for several Agave species (including species from both subgenera) is presented in detail with two different kinds of explants to initiate the process: rescued zygotic embryos and small offshoots that grow around a mother plant.

Induction of Somatic Embryogenesis in Jatropha curcas.

Jatropha curcas has been a promising crop for biofuel production for the last decade. However, the lack of resistant materials to diseases and improved quality of the oil produced by the seeds has restricted the use of this promising crop. The genetic modifications in the fatty acid pathway, as well as the introduction of resistance to different diseases, would change the fate of Jatropha. To achieve these goals, we need to have a very efficient regeneration system. Here, we report a very useful protocol to induce somatic embryogenesis from leaves of Jatropha using cytokinin as the only growth regulator.

Detection of cytokinins and auxin in plant tissues using histochemistry and immunocytochemistry.

We report a new method for histochemical localization of cytokinins (CKs) in plant tissues based on bromophenol blue/silver nitrate staining. The method was validated by immunohistochemistry using anti-trans-zeatin riboside antibody. Indole-3-acetic acid (auxin, IAA) was localized by anti-IAA antibody in plant tissues as a proof for IAA histolocalization. We used root sections, because they are major sites of CKs synthesis, and insect galls of Piptadenia gonoacantha that accumulate IAA. Immunostaining confirmed the presence of zeatin and sites of accumulation of IAA indicated by histochemistry. The colors developed by histochemical reactions in free-hand sections of plant tissues were similar to those obtained by thin layer chromatography (TLC), which reinforced the reactive sites of zeatin. The histochemical method for detecting CKs is useful for galls and roots, whereas IAA detection is more efficient for gall tissues. Therefore, galls constitute a useful model for validating histochemical techniques due to their rapid cell cycles and relatively high accumulation of plant hormones.

The AP2/ERF Transcription Factor DRNL Modulates Gynoecium Development and Affects Its Response to Cytokinin.

The gynoecium is the female reproductive system in flowering plants. It is a complex structure formed by different tissues, some that are essential for reproduction and others that facilitate the fertilization process and nurture and protect the developing seeds. The coordinated development of these different tissues during the formation of the gynoecium is important for reproductive success. Both hormones and genetic regulators guide the development of the different tissues. Auxin and cytokinin in particular have been found to play important roles in this process. On the other hand, the AP2/ERF2 transcription factor BOL/DRNL/ESR2/SOB is expressed at very early stages of aerial organ formation and has been proposed to be a marker for organ founder cells. In this work, we found that this gene is also expressed at later stages during gynoecium development, particularly at the lateral regions (the region related to the valves of the ovary). The loss of DRNL function affects gynoecium development. Some of the mutant phenotypes present similarities to those observed in plants treated with exogenous cytokinins, and AHP6 has been previously proposed to be a target of DRNL. Therefore, we explored the response of drnl-2 developing gynoecia to cytokinins, and found that the loss of DRNL function affects the response of the gynoecium to exogenously applied cytokinins in a developmental-stage-dependent manner. In summary, this gene participates during gynoecium development, possibly through the dynamic modulation of cytokinin homeostasis and response.

Plant immunity: unravelling the complexity of plant responses to biotic stresses.

Background: Plants are constantly exposed to evolving pathogens and pests, with crop losses representing a considerable threat to global food security. As pathogen evolution can overcome disease resistance that is conferred by individual plant resistance genes, an enhanced understanding of the plant immune system is necessary for the long-term development of effective disease management strategies. Current research is rapidly advancing our understanding of the plant innate immune system, with this multidisciplinary subject area reflected in the content of the 18 papers in this Special Issue. Scope: Advances in specific areas of plant innate immunity are highlighted in this issue, with focus on molecular interactions occurring between plant hosts and viruses, bacteria, phytoplasmas, oomycetes, fungi, nematodes and insect pests. We provide a focus on research across multiple areas related to pathogen sensing and plant immune response. Topics covered are categorized as follows: binding proteins in plant immunity; cytokinin phytohormones in plant growth and immunity; plant-virus interactions; plant-phytoplasma interactions; plant-fungus interactions; plant-nematode interactions; plant immunity in Citrus; plant peptides and volatiles; and assimilate dynamics in source/sink metabolism. Conclusions: Although knowledge of the plant immune system remains incomplete, the considerable ongoing scientific progress into pathogen sensing and plant immune response mechanisms suggests far reaching implications for the development of durable disease resistance against pathogens and pests.

Competitive-IgY- Enzyme Linked Immuno Sorbent Assay (CIgY-ELISA) to detect the cytokinins in Gerbera jamesonii plantlets

A competitive hyper-immune yolk Immunoglobulin Y - Enzyme Linked Immune Sorbent Assay (CIgY-ELISA), was developed as an alternative method to detect zeatin and 2ip in plantlets of gerbera. The endogenous level of hormones in the plantlets in vitro of gerbera with one or six weeks after replication was determined with competitive IgY-ELISA set to detect between 1 and 100 pmoles of plant hormone for each 1.0 g tissue. The plantlets of six weeks presented sprouts and root, while the plantlets of one week presented only sprouts. The CIgY-ELISA was set with high independent variables values of sensitivity/specificity of 96/89 percent for zeatin and 94/78 percent for 2ip, with high values of reproducibility (up to 90 percent) for both the cytokinins. Zeatin content varied from 2.2 to 2.8 pmoles.g-1 and from 2.7 to 3.3 pmoles.g-1 on the plantlet with one and six weeks, respectively. The 2ip content did not vary and was detected near the detection limit in all the assays. It was concluded that the observed capabilities of CIgY-ELISA were putative and the competitive assay was a highly robust and stable method, which could be used for the studies on plant physiology for endogenous cytokinins.

Regeneração in vitro via organogênese direta de Bauhinia cheilantha / In vitro regeneration of the Bauhinia cheilantha via organogenesis

Bauhinia cheilantha, conhecida como "pata-de-vaca", possui grande relevância econômica e etnofarmacológica no semiárido brasileiro. Nas suas sementes constatou-se dormência, o que dificulta a obtenção de plantas uniformes e em curto período de tempo, diante disso, o presente trabalho objetivou estabelecer um protocolo de micropropagação para a espécie. Os segmentos cotiledonar e nodal de plântulas emergidas in vitro foram inoculados em meio de cultura WPM, suplementado com diferentes concentrações de 6-benzilaminopurina (BAP), thidiazuron (TDZ) ou cinetina (KIN) para induzir a regeneração de brotos adventícios. Na fase de enraizamento foram testadas diferentes concentrações de ácido indol-3-butírico (AIB) e de carvão ativado. Nos segmentos nodais houve maior capacidade organogênica do que no segmento cotiledonar. O maior número de brotos (4,3 e 2,1) foi obtido com 1,0mg L-1 de TDZ e 1,5mg L-1 de BAP, respectivamente. No entanto, na presença de TDZ foram observadas as menores brotações. A presença de 2,0mg L-1 de AIB com carvão ativado (CA) promoveu a maior porcentagem de enraizamento (cerca de 60,0 por cento) e maior número de raízes adventícias (2,5). As brotações enraizadas foram transferidas para casa de vegetação e aclimatizadas com sucesso.

Bauhinia cheilantha, known as "pata-de-vaca", is of great economic and ethnopharmacological importance in the semiarid Brazilian. Seeds are dormant which makes it difficult to obtain uniform plants in a short time before this, our study aimed to establish a micropropagation protocol for the species. Cotyledonary and nodal segments of seedlings grown in vitro were inoculated on Woody Plant Medium (WPM) supplemented with various concentrations of 6-benzylaminopurine (BAP), thidiazuron (TDZ) or kinetin (KN) to induce adventitious shoot regeneration. In the rooting phase, different concentrations of indole-3-butyric acid (IBA) and the activated charcoal were tested. The nodal segments showed organogenic capacity greater than the cotyledonary segment. The highest number of shoots (4.3 and 2.1) was obtained at 1.0mg L-1 TDZ and 1.5mg L-1 BAP, respectively. However, in the presence of TDZ was observed the smaller shoots.The use of 2.0mg L-1 IBA within activated charcoal (AC) promoted the highest percentage of rooting (about 60.0 percent) and number of adventitious roots (2.5). The rooted shoots were transferred to greenhouse and successfully acclimatized.

Regeneração in vitro via organogênese direta de Bauhinia cheilantha

Bauhinia cheilantha, known as "pata-de-vaca", is of great economic and ethnopharmacological importance in the semiarid Brazilian. Seeds are dormant which makes it difficult to obtain uniform plants in a short time before this, our study aimed to establish a micropropagation protocol for the species. Cotyledonary and nodal segments of seedlings grown in vitro were inoculated on Woody Plant Medium (WPM) supplemented with various concentrations of 6-benzylaminopurine (BAP), thidiazuron (TDZ) or kinetin (KN) to induce adventitious shoot regeneration. In the rooting phase, different concentrations of indole-3-butyric acid (IBA) and the activated charcoal were tested. The nodal segments showed organogenic capacity greater than the cotyledonary segment. The highest number of shoots (4.3 and 2.1) was obtained at 1.0mg L-1 TDZ and 1.5mg L-1 BAP, respectively. However, in the presence of TDZ was observed the smaller shoots.The use of 2.0mg L-1 IBA within activated charcoal (AC) promoted the highest percentage of rooting (about 60.0%) and number of adventitious roots (2.5). The rooted shoots were transferred to greenhouse and successfully acclimatized.

Bauhinia cheilantha, conhecida como "pata-de-vaca", possui grande relevância econômica e etnofarmacológica no semiárido brasileiro. Nas suas sementes constatou-se dormência, o que dificulta a obtenção de plantas uniformes e em curto período de tempo, diante disso, o presente trabalho objetivou estabelecer um protocolo de micropropagação para a espécie. Os segmentos cotiledonar e nodal de plântulas emergidas in vitro foram inoculados em meio de cultura WPM, suplementado com diferentes concentrações de 6-benzilaminopurina (BAP), thidiazuron (TDZ) ou cinetina (KIN) para induzir a regeneração de brotos adventícios. Na fase de enraizamento foram testadas diferentes concentrações de ácido indol-3-butírico (AIB) e de carvão ativado. Nos segmentos nodais houve maior capacidade organogênica do que no segmento cotiledonar. O maior número de brotos (4,3 e 2,1) foi obtido com 1,0mg L-1 de TDZ e 1,5mg L-1 de BAP, respectivamente. No entanto, na presença de TDZ foram observadas as menores brotações. A presença de 2,0mg L-1 de AIB com carvão ativado (CA) promoveu a maior porcentagem de enraizamento (cerca de 60,0%) e maior número de raízes adventícias (2,5). As brotações enraizadas foram transferidas para casa de vegetação e aclimatizadas com sucesso.