Sour orange bud regeneration and in vitro plant development related to culture medium composition and explant type

In order to evaluate the formation of adventitious buds and in vitro regeneration of sour orange plants (Citrus aurantium L.) two organogenesis-inducing experiments were conducted. In the first experiment, the induction and in vitro regeneration of adventitious buds were tested on epicotyl and internodal segments under the influence of BAP or KIN associated with NAA. The second experiment evaluated the in vitro regeneration of sour orange plants related to different explant types (epicotyls segments, internodal segments of in vitro germinated plantlets and internodal segments of greenhouse cultivated plants). Data collected on both experiments included the percentage of responsive explants (explants that formed buds), and the number of buds per explant. The addition of BAP showed the best organogenic response. In vitro germinated epicotyl segments and internodal segments are recommended as explants for sour orange in vitro organogenesis. Rooting of regenerated shoots was achieved without the need of auxin in the medium.

Residual effect of growth regulators in etiolation and regeneration of in vitro pineapple plants

This work aimed to evaluate the influence of naphthaleneacetic acid (NAA) and gibberellic acid (GA3) plant regulators in in vitro etiolation and subsequent regeneration of the PE x SC-60 pineapple hybrid. Nodal segments of in vitro plants with approximately 5-7 cm height were incubated in basic MS culture medium supplemented with 0.0; 0.5 and 1.0 mg L-1 of naphthaleneacetic acid (NAA) in combination with gibberellic acid (GA3) in concentrations of 0.0; 0.5 and 1.0 mg L-1, and maintained at 27 ºC under dark condition. Evaluations were carried out at 90 and 180 days after incubation period. The best results for length of etiolated stems were obtained with 1.0 mg L-1 of NAA. In the experiment followed by the regeneration, stems with 3 cm from the etiolation treatment, were cultivated in proliferation medium and the number of regenerated plants per treatment was evaluated at 60 days of cultivation. The treatment that promoted the best etiolation of plants also promoted the worst regeneration rates, demonstrating the residual effect of the auxin used in the previous step in the regeneration of plants of the pineapple hybrid evaluated.

Poisoning and regeneration of Pt-Pd/WO3-ZrO2 short paraffin isomerization catalysts

WO3-ZrO2 catalysts promoted with Pt and Pd were tested as paraffin isomerization catalysts using n-hexane as model compound. Sulfur and amine poisoning and regeneration tests were used to assess the impact of the addition of Pt and Pd on the deactivation resistance and regenerability. Pt and PtPd catalysts were the most active for n-hexane isomerization. The low activity of the Pd catalyst was attributed to poor Pd metal properties when supported over WO3-ZrO2 and to a decrease of the number of BrQnsted acid sites. PtPd was the only catalyst capable of full regeneration after S poisoning. Amine poisoning completely supressed the isomerization activity and the original activity could only be restored by calcination and reduction.

Regeneration of five commercially-valuable tree species after experimental logging in an Amazonian forest

This study investigated the regeneration variation of five commercially valuable tree species in relation to different intensities of felling in fourteen 4-ha plots in an area under experimental forest management. This experiment was carried out in a typical Amazonian tropical forest sample on "terra-firme," in Manaus (AM). Plots were logged 7 and 8 years (1987 and 1988), or 3 years (1993) before the study. All trees with height greater than 2 m, and diameter at breast height (DBH) smaller than 10 cm were measured. Only Aniba hostmanniana, Ocotea aciphylla, Licaria pachycarpa, Eschweilera coriacea and Goupia glabra were sufficiently common for individual analyses. These species have high timber values in the local market. Eight years after logging, the species responded differently to logging intensities. The numbers of individuals of Goupia glabra and Aniba hostmanniana were positively related to the intensity of logging, while Ocotea aciphylla, Licaria pachycarpa, and Eschweilera coriacea showed no statistically significant response. In the most recently (1993) logged areas, Goupia glabra and Aniba hostmanniana had higher numbers of individuals than the control plots.

Natural regeneration in abandoned fields following intensive agricultural land use in an Atlantic Forest Island, Brazil

The time required to regrowth a forest in degraded areas depends on how the forest is removed and on the type of land use following removal. Natural regeneration was studied in abandoned old fields after intensive agricultural land use in areas originally covered by Brazilian Atlantic Forests of the Anchieta Island, Brazil in order to understand how plant communities reassemble following human disturbances as well as to determine suitable strategies of forest restoration. The fields were classified into three vegetation types according to the dominant plant species in: 1) Miconia albicans (Sw.) Triana (Melastomataceae) fields, 2) Dicranopteris flexuosa (Schrader) Underw. (Gleicheniaceae) thickets, and 3) Gleichenella pectinata (Willd.) Ching. (Gleicheniaceae) thickets. Both composition and structure of natural regeneration were compared among the three dominant vegetation types by establishing randomly three plots of 1 x 3 m in five sites of the island. A gradient in composition and abundance of species in natural regeneration could be observed along vegetation types from Dicranopteris fern thickets to Miconia fields. The gradient did not accurately follow the pattern of spatial distribution of the three dominant vegetation types in the island regarding their proximity of the remnant forests. A complex association of biotic and abiotic factors seems to be affecting the seedling recruitment and establishment in the study plots. The lowest plant regeneration found in Dicranopteris and Gleichenella thickets suggests that the ferns inhibit the recruitment of woody and herbaceous species. Otherwise, we could not distinguish different patterns of tree regeneration among the three vegetation types. Our results showed that forest recovery following severe anthropogenic disturbances is not direct, predictable or even achievable on its own. Appropriated actions and methods such as fern removal, planting ground covers, and enrichment planting with tree species were suggested in order to restore the natural forest regeneration process in the abandoned old fields.

POPULATION STRUCTURE AND SPATIAL DISTRIBUTION OF Bauhinia cheilantha (Bong.) Steud. IN TWO FRAGMENTS AT DIFFERENT REGENERATION STAGES IN THE CAATINGA, IN SERGIPE, BRAZIL

ABSTRACTThis study aimed to analyze the vertical and diameter structure and the spatial distribution pattern of Bauhinia cheilantha in two Caatinga fragments in Sergipe, Brazil, at different regeneration stages. Thirty plots were demarcated in area I (Canindé de São Francisco and Poço Redondo), which has vegetation regeneration, and 25 plots in area II (Porto da Folha) with preserved vegetation, both having 400 m2. All B. cheilanthaindividuals had their height and circumference (circumference at breast height > 6 cm) measured. Possible differences in height and diameter at breast height were tested in the two populations by using Student’s T-test. The distribution pattern of species was calculated through Payandeh’s index. We sampled 154 B. cheilantha individuals, equivalent to 33.3% of the plots in area I and in 1,027 individuals in area II, totaling 100% frequency. Height and the diameter of the two populations were statistically different, where AI achieved all values lower than AII. The spatial distribution pattern of B. cheilantha found in both areas was aggregate, with values of 11.85 and 9.00, respectively. Thus, it became clear that the population in AII is at a more advanced successional status than AI, due to its longer conservation time.

CLONAL PROPAGATION OF NEEM (Azadirachta indica A. Juss.) VIA DIRECT AND INDIRECT IN VITRO REGENERATION

ABSTRACTThe study was conducted with shoot tip explants of neem (Azadirachta indica A. Juss) to identify a viable regenerative process. Shoot tips were obtained from neem embryos cultured alternatingly in DKW medium supplemented with BAP and medium without hormones. Initial shoot development was influenced by cotyledon presence. Basal callus, excised from in vitro stem base, also presented organogenic potential. In some cases, plant lines, obtained from each seed, presented different characteristics. The most common characteristic observed in vitro was callus formation at the stem base. However, the rarest characteristics were stem callus formation and leaf senescence. The regenerated shoot tips were further subculture and rooted on a medium supplemented with IBA so that complete plants could be obtained. The rooted plants were transplanted to a greenhouse and successfully acclimatized. No significant differences in in vivo development were observed between neem plants from callus and from shoot tip propagation.

NATURAL REGENERATION STRATUM AS AN INDICATOR OF RESTORATION IN AREA OF ENVIRONMENTAL COMPENSATION FOR MINING LIMESTONE, MUNICIPALITY OF BARROSO, MG, BRAZIL

ABSTRACT This study was conducted in a forest under restoration process, which belongs to the company Holcim Brasil S/A, in the municipality of Barroso, state of Minas Gerais (21º00'to 22º00'S and 43º00' to 44º00'W), where 40 plots (2 x 2 m) were set, spaced at 10 m, forming eight strata parallel to the watercourse present in the area. Floristic composition and natural regeneration stratum were characterized, and the formed strata allowed evaluating whether the riparian vegetation and watercourse influence on the local regeneration. It was found 162 individuals of 13 families, 18 genera and 22 species, and 10,125 individuals/ha were estimated. Successional classes from pioneer and early secondary and zoochory dispersion syndrome prevailed among species and individuals. The watercourse and riparian vegetation did not exercise significant influence (p> 0.05) on the number of species and regenerating individuals among the different strata of the forest. The diversity index of Shannon-Wiener (H') and equability of Pielou (J') were 2.691 and 0.870, respectively. The species Psidium guajava and Myrtaceae families presented the highest VI (value of importance). Natural regeneration analysis showed the low floristic diversity in the area, suggesting that corrective management actions should be adopted.

PHYTOSOCIOLOGY AND STRUCTURAL CHARACTERIZATION OF WOODY REGENERATION FROM A REFORESTATION WITH NATIVE SPECIES IN SOUTHEASTERN BRAZIL

ABSTRACT In Brazil, specifically in São Paulo State, there are guidelines based on the high diversity of tropical forests that instructs the restoration projects in the state (current SMA 32/2014). The main goal of this study was verify the importance and effectiveness of the high diversity of arboreal species originated from a reforestation, and its influence in a woody regenerating composition. We developed a phytosociologic study in a woody regenerating stratum of a nine year old reforestation at a Private Reserve of Natural Heritage (RPPN), in Mogi-Guaçu, São Paulo State. All specimens with height > 30 cm and Diameter at Breast Height (DBH) < 5 cm were evaluated. The woody regenerating diversity was smaller than the overstory diversity and the species composition was similar to the overstory. The Simpson index (1-D) was 0.85, Shannon index (H') was 2.46 and the Pielou index (J') was 0.60. The zoochoric dispersion syndrome was major among the species. Our results suggest that the use of high diversity of native seedlings in a reforestation leads to high diversity of species in woody regeneration stratum, after one decade of planting.

Liver regeneration with l-glutamine supplemented diet: experimental study in rats

OBJECTIVE: To assess liver regeneration in rats after 60% hepatectomy with and without supplementation of L-glutamine through liver weight changes, laboratory parameters and histological study.METHODS: 36 male rats were divided into two groups: glutamine group and control group. Each group was subdivided into three subgroups, with death in 24h, 72h and seven days. The glutamine group received water and standard diet supplemented with L-glutamine, and the control recieved 0.9% saline. In all subgroups analysis of liver regeneration was made by the Kwon formula, study of liver function (AST, ALT, GGT, total bilirubin, indirect and indirect bilirubin and albumin) and analysis of cell mitosis by hematoxylin-eosin.RESULTS: In both groups there was liver regeneration by weight gain. Gamma-GT increased significantly in the control group (p < 0.05); albumin increased in the glutamine group. The other indicators of liver function showed no significant differences. Histological analysis at 72h showed a higher number of mitoses in the glutamine group, with no differences in other subgroups.CONCLUSION: Diet supplementation with L glutamine is beneficial for liver regeneration.

Sirolimus influence on hepatectomy-induced liver regeneration in rats

OBJECTIVE: To evaluate the influence of sirolimus on liver regeneration triggered by resection of 70% of the liver of adult rats.METHODS: we used 40 Wistar rats randomly divided into two groups (study and control), each group was divided into two equal subgroups according to the day of death (24 hours and seven days). Sirolimus was administered at a dose of 1mg/kg in the study group and the control group was given 1 ml of saline. The solutions were administered daily since three days before hepatectomy till the rats death to removal of the regenerated liver, conducted in 24 hours or 7 days after hepatectomy. Liver regeneration was measured by the KWON formula, by thenumber of mitotic figures (hematoxylin-eosin staining) and by the immunohistochemical markers PCNA and Ki-67.RESULTS: there was a statistically significant difference between the 24h and the 7d groups. When comparing the study and control groups in the same period, there was a statistically significant variation only for Ki-67, in which there were increased numbers of hepatocytes in cell multiplication in the 7d study group compared with the 7d control group (p = 0.04).CONCLUSION: there was no negative influence of sirolimus in liver regeneration and there was a positive partial effect at immunohistochemistry with Ki-67.

Evaluation of liver regeneration diet supplemented with omega-3 fatty acids: experimental study in rats

Objective: to evaluate liver regeneration in rats after partial hepatectomy of 60% with and without action diet supplemented with fatty acids through the study of the regenerated liver weight, laboratory parameters of liver function and histological study. Methods: thirty-six Wistar rats, males, adults were used, weighing between 195 and 330 g assigned to control and groups. The supplementation group received the diet by gavage and were killed after 24h, 72h and seven days. Evaluation of regeneration occurred through analysis of weight gain liver, serum aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltranspeptidase, and mitosis of the liver stained with H&E. Results: the diet supplemented group showed no statistical difference (p>0.05) on the evolution of weights. Administration of fatty acids post-hepatectomy had significant reduction in gamma glutamyltransferase levels and may reflect liver regeneration. Referring to mitotic index, it did not differ between period of times among the groups. Conclusion: supplementation with fatty acids in rats undergoing 60% hepatic resection showed no significant interference related to liver regeneration.

Pharmacological immunomodulation enhances peripheral nerve regeneration

To assess the effect of N-Acetylmuramyl-L-Alanyl-D-Isoglutamine MDP topically administrated on the regenerating peripheral neurons, twelve male C57BL/6J adult mice were equally distributed into three groups. Four mice underwent unilateral sciatic nerve transection and polyethylene tubulization, with a 4mm gap between the proximal and distal nerve stumps and were implanted with collagen + PBS (COL). Other four animals underwent the same surgical procedure but received collagen + MDP (COL/MDP) inside the prosthesis. Four animals were not operated and served as control group (NOR). After 4 weeks, the regenerated nerve cables were processed for total myelinated axon counting and myelinated fiber diameter measurement. The L5 dorsal root ganglion (DRG) was also removed and sectioned for sensory neurons counting and measurement. The results revealed significant difference (p<0.05) in axonal counting among the groups NOR (4,355±32), COL (1,869±289) and COL/MDP (2,430±223). There was a significant reduction in the axonal diameter in the operated groups (COL=3.38µm±1.16 and COL/MDP=3.54µm±1.16) compared to NOR (6.19µm±2.45). No difference was found in the number of DRG neurons between the experimental groups (COL=564±51; COL/MDP=514±56), which presented fewer sensory neurons compared to NOR (1,097±142). Data obtained indicate that locally applied MDP stimulates peripheral nerve regeneration in mice.

The potential of natural regeneration of rocky outcrop vegetation on rupestrian field soils in "Serra do Cipó", Brazil

An area's innate potential to regenerate represents a crucial factor for its conservation and management. The seed rain and seed bank are important agents in the regeneration process. Seed banks are particularly important in communities where there is a high proportion of obligate seeders. Rocky outcrops are habitats where most part of the plant species depends on their seeds to reproduce and maintain viable populations. Therefore, seed banks ought to be important in this vegetation physiognomy. We test the hypotheses that the seed bank of the rocky outcrops found in the rupestrian fields of "Serra do Cipó", Brazil, is richer in species and denser than those formed on different vegetation physiognomies neighboring the outcrops. We then compared species abundance, species richness and composition in the rocky outcrops' seed banks with those of sandy and peaty bogs, forests, gallery forests, and "cerrados". Furthermore, we report on the natural regeneration potential of these soils by assessing a greenhouse study on seedling emergence. Soil samples were collected from 0 to 5 and 5 to 10 cm of depth. Rocky outcrops had the poorest in species and less dense seed bank and showed segregation in species composition. Emergence was greater in the most superficial layer. However, soils on rocky outcrops showed the greatest proportion of endemic threatened species in their seed banks, demonstrating their importance for biodiversity conservation of the "Serra do Cipó" rupestrian fields.

Callus induction and plant regeneration by Brazilian triticale and wheat genotypes

In order to determine the in vitro behavior of Brazilian triticale, 16 triticale genotypes, and three wheat genotypes used as checks, were sown in June 1994. The explants used were immature embryos. In addition to the genotype tests, two culture media for callus induction were also evaluated, i.e., MS (Murashige and Skoog, Physiol. Plant. 15: 473-497, 1962) medium containing 2.0 mg 2,4D/l, and MS medium containing 4.0 mg 2,4D/l. The plant regeneration protocol used was the one employed at the Laboratório de Cultura de Tecidos, Departamento de Plantas de Lavoura, Universidade Federal do Rio Grande do Sul, for wheat. Differences in plant regeneration were observed both among triticale and wheat genotypes, with triticale usually showing better regeneration than wheat. No differences were observed between the callus induction media.