Cristina Vieira de Almeida

Pre-procambial cells are niches for pluripotent and totipotent stem-like cells for organogenesis and somatic embryogenesis in the peach palm: a histological study

AbstractThe direct induction of adventitious buds and somatic embryos from explants is a morphogenetic process that is under the influence of exogenous plant growth regulators and its interactions with endogenous phytohormones. We performed an in vitro histological analysis in peach palm (Bactris gasipaes Kunth) shoot apexes and determined that the positioning of competent cells and their interaction with neighboring cells, under the influence of combinations of exogenously applied growth regulators (NAA/BAP and NAA/TDZ), allows the pre-procambial cells (PPCs) to act in different morphogenic pathways to establish niche competent cells. It is likely that there has been a habituation phenomenon during the regeneration and development of the microplants. This includes promoting the tillering of primary or secondary buds due to culturing in the absence of NAA/BAP or NAA/TDZ after a period in the presence of these growth regulators. Histological analyses determined that the adventitious roots were derived from the dedifferentiation of the parenchymal cells located in the basal region of the adventitious buds, with the establishment of rooting pole, due to an auxin gradient. Furthermore, histological and histochemical analyses allowed us to characterize how the PPCs provide niches for multipotent, pluripotent and totipotent stem-like cells for vascular differentiation, organogenesis and somatic embryogenesis in the peach palm. The histological and histochemical analyses also allowed us to detect the unicellular or multicellular origin of somatic embryogenesis. Therefore, our results indicate that the use of growth regulators in microplants can lead to habituation and to different morphogenic pathways leading to potential niche establishment, depending on the positioning of the competent cells and their interaction with neighboring cells. Key message Our results indicate that the use of growth regulators in microplants can lead to habituation and to different morphogenic pathways leading to potential niche establishment, depending on the positioning of the competent cells and their interaction with neighboring cells.UNLABELLED The direct induction of adventitious buds and somatic embryos from explants is a morphogenetic process that is under the influence of exogenous plant growth regulators and its interactions with endogenous phytohormones. We performed an in vitro histological analysis in peach palm (Bactris gasipaes Kunth) shoot apexes and determined that the positioning of competent cells and their interaction with neighboring cells, under the influence of combinations of exogenously applied growth regulators (NAA/BAP and NAA/TDZ), allows the pre-procambial cells (PPCs) to act in different morphogenic pathways to establish niche competent cells. It is likely that there has been a habituation phenomenon during the regeneration and development of the microplants. This includes promoting the tillering of primary or secondary buds due to culturing in the absence of NAA/BAP or NAA/TDZ after a period in the presence of these growth regulators. Histological analyses determined that the adventitious roots were derived from the dedifferentiation of the parenchymal cells located in the basal region of the adventitious buds, with the establishment of rooting pole, due to an auxin gradient. Furthermore, histological and histochemical analyses allowed us to characterize how the PPCs provide niches for multipotent, pluripotent and totipotent stem-like cells for vascular differentiation, organogenesis and somatic embryogenesis in the peach palm. The histological and histochemical analyses also allowed us to detect the unicellular or multicellular origin of somatic embryogenesis. Therefore, our results indicate that the use of growth regulators in microplants can lead to habituation and to different morphogenic pathways leading to potential niche establishment, depending on the positioning of the competent cells and their interaction with neighboring cells. KEY MESSAGE Our results indicate that the use of growth regulators in microplants can lead to habituation and to different morphogenic pathways leading to potential niche establishment, depending on the positioning of the competent cells and their interaction with neighboring cells.

Fungos endofíticos isolados de ápices caulinares de pupunheira cultivada in vivo e in vitro

Losses of micropropagated plants occur in its majority due to presence of microorganisms responsible for plant death both in culture beginning and plant establishment in field. Two years in vitro cultivated pejibaye shoot tips, showed the presence of fungi colonies after transference for new culture medium. This paper aimed at identifying fungi, by classical taxonomy and by molecular methods, present in shoot tip of healthy pejibaye plants, cultivated in the field, and to compare them with isolated ones of in vitro plants. The isolates of endophytic fungi community of the in vitro plants were Fusarium oxysporum, Neotyphodium sp. and Epicoccum nigrum; from the in vivo plants were Fusarium sp., F. proliferatum, F. oxysporum, Colletotrichum sp., Alternaria gaisen, Neotyphodium sp. and E. nigrum. The seven species of fungi were reinoculated in host plant, revealing different behaviour. Neotyphodium sp. and E. nigrum established an endophytic interaction with the host; the other fungi acted as pathogens causing decrease in seedlings development when compared to the non-inoculated plants. Species acting as endophytes present a potential use in biocontrol of pejibaye pathogens.

Somatic embryogenesis and in vitro plant regeneration from pejibaye adult plant leaf primordia

O objetivo deste trabalho foi avaliar um protocolo de regeneracao de plantas por meio de embrioes somaticos, obtidos a partir de primordios foliares de pupunheiras adultas e identificar a origem histologica dos embrioes e descrever as etapas morfogeneticas. Os explantes foram cultivados em meio MS modificado. Celulas parenquimaticas do mesofilo originaram meristemoides com BAP (7,1 µM) nos dois primeiros periodos de subcultura. A polarizacao das estruturas ocorreu com ANA (12,9 µM) e BAP (3,55 µM) no terceiro periodo de subcultura. Meristemoides se desenvolveram em embrioes somaticos, regenerando plantas normais. As celulas parenquimaticas do mesofilo apresentam elevada capacidade de resposta direta a auxina e a citocinina.

Anatomical analysis of peach palm (Bactris gasipaes) leaves cultivated in vitro, ex vitro and in vivo

The present work characterized and compared the anatomical structures of the leaves of Bactris gasipaes (Arecaceae) plants grown under different cultivation conditions (in vitro, ex vitro and in vivo) with the goal of identifying the origins of the difficulties encountered in acclimatizing micro-plants. The Quant program was used to determine leaf tissue thicknesses and areas, and histochemical tests were performed on leaf sections and analyzed using light microscopy. Stomatal and trichome densities were determined using the epidermal impression method and by scanning electronic microscopy. Our results indicated that there were no discernible alterations of the anatomical characteristics of the leaves of micro-plants cultivated under differing conditions and that the thickening of the mesophyll and the vascular fibers indicated adaptive responses to ex vitro conditions. As such, the observed difficulties in acclimatizing peach palm micro-plants to ex vitro conditions cannot be attributed to plant anatomical characteristics acquired during in vitro cultivation.

Endophytic bacterial communities associated with two explant sources of Eucalyptus benthamii Maiden & Cambage

Micropropagation has been applied in the recovery and rejuvenation of adult trees, which is achieved by various subcultures in the multiplication phase. This strategy has brought questions about the endophytic microbiota associated with these plants along its manipulation. Therefore, the aim of this study was to evaluate the composition of the endophytic bacterial communities associated with two explants sources [the canopy branches (CB) and the trunk base of the tree (TB)] under prolonged in vitro cultivation. In addition we analyzed the bacterial community dynamic along the subcultures in different micropropagation phases. Bacterial DNA was extracted from samples of mini-stumps (in vivo) from CB and TB and in micro-stumps produced by in vitro cultivations of these explants sources—both originated from one single matrix plant of Eucalyptus benthamii. In vitro establishment occurred in two dates and the evaluation of endophytic bacterial communities was made in vivo and in vitro samples (on 10th, 13th and 16th subcultures), when elongated shoots and roots were analyzed. Analysis was performed by PCR–DGGE based on the V6 region of ribosomal gene 16S rDNA. Bands profiles showed differences in communities between in vivo and in vitro samples, and also distinctions of communities assessed in the subcultures, elongated and rooted samples. Distinctions in the composition of endophytic bacterial communities were greater in CB micro-stumps. These results indicate a differential colonization of explants by endophytic bacteria, with predominance of common (ever-present) endophytes in TB samples and casual, here named opportunistic, in CB samples.

Nitrato de amônio e nitrato de potássio no desenvolvimento in vitro de embriões somáticos de pupunheiras

Pejibaye is a good alternative for the extractive exploration of species such as jucara and acai. However, when it is produced by seeds its planting is heterogeneous, which makes micropropagation a good alternative for cultivation in large scale. This study aimed to evaluate the influence of the interaction between ammonium nitrate and potassium nitrate on peach palm somatic embryos rooting in vitro cultivated, for optimization of the micropropagation protocol. The embryos were inoculated in MS medium with different concentrations of NH4NO3 and KNO3. Morphophisyologic parameters of root development were measured at 120 and 240 days of cultivation. At 120 days, at lower nitrogen concentrations, roots were stimulated to grow and major root branching occurred at low concentrations of NH4NO3 and high concentrations of KNO3. At 240 days, there was a reduction of the root growing, with more percentage of thin roots. It was concluded that up to 120 days, the microplants should be cultivated in culture medium with lower concentrations of NH4NO3 and higher concentrations of KNO3 than those recommended by MS medium, and after this period they should be cultivated at the recommended doses.

Morfologia da raiz de plantas com sementes

O estudo da Morfologia Vegetal abrange a descricao das formas e estruturas dos orgaos constituintes das plantas, oferecendo subsidios para melhor compreender as estrategias adaptativas das mais variadas especies e seus diferentes habitats.