Somatic embryogenesis in the commercial papaya hybrid UENF/Caliman 01 relying on plantlet production from sexed adult hermaphrodite donor plants.

Somatic embryogenesis from explants from hermaphrodite papaya mother plants is an alternative for the production of true-to-type plants without the need for sexing. This study aimed to analyze hormonal and osmotic inducers in different somatic embryogenesis stages in the commercial hermaphrodite hybrid papaya UENF/Caliman 01. Leaf disks from in vitro shoots originated from ex vitro hermaphrodite plants were cultured in induction medium supplemented with different concentrations of 2,4-D (6, 9, 12, 15, and 18 µM) and 4-CPA (19, 22, 25, 28, and 31 µM). After 90 days, the formation of somatic embryos was verified. The 2,4-D induced the formation of light brown calli with low frequency (20%) of somatic embryogenesis. However, 4-CPA (25 µM) induced 96% of embryogenic calli, which were transferred to maturation medium (MM) and cultured for 30 days. The MM contained ABA (0.5 µM) and AC (15 g L-1) and produced 36.6 somatic embryos callus-1, mainly on cotyledonary stage. Cotyledonary embryos were transferred to germination medium supplemented with gibberellic acid (GA3) (0.0, 1.44, 2.88, and 4.32 µM), and the conversion into plantlets was enhanced with GA3 at 2.88 µM.

The effects of an osmoregulator, carbohydrates and polyol on maturation and germination of 'Golden THB' papaya somatic embryos.

This study evaluated the effect of osmoregulators and carbohydrates on the maturation and germination of somatic embryos of papaya 'Golden THB'. Cotyledon explants from papaya seedlings germinated in vitro on basal MS medium were cultured on somatic embryogenesis induction medium (IM) containing MS salts, myo-inositol, sucrose, agar and p-chlorophenoxyacetic acid. After 50 days, embryogenic calli were transferred onto maturation media (MM) for 45 additional days. For experiment 1, a MS-based medium supplemented with abscisic acid, activated charcoal and concentrations of PEG 6000 (0; 40; 50; 60 and 70 g L-1) was used, whereas for experiment 2 malt extract concentrations (0; 0.1; 0.2; 0.3 and 0.4 g L-1) were assessed. The normal cotyledonary somatic embryos produced in experiment 2 were transferred to the germination medium (GM). The GM consisted of full-strength MS medium, sucrose, agar and was supplemented with myo-inositol at varying concentrations (0; 0.275; 0.55 and 0.825 mM). The PEG concentrations tested impaired the maturation of 'Golden THB' papaya somatic embryos. The MM, supplemented with malt extract at 0.153 g L-1, promoted the greatest development of normal somatic embryos (18.28 SE calli-1), that is, two cotyledonary leaves produced 36.56 SE calli-1. The supplementation with 0.45 mM myo-inositol provided the highest germination percentage (47.42%) and conversion to emblings.

Morphophysiology, Phenotypic and Molecular Diversity of Auxin-induced Passiflora mucronata Lam. (Passifloraceae).

Genetic diversity allows identification of potential intraspecific genotypes in the genus Passiflora. The objective of this study was to examine the morphological and genetic diversity of auxin-induced Passiflora mucronata. The experiments were arranged in a complete randomized block design, with a 9 x 2 factorial arrangement (nine genotypes x presence and absence of auxin, indole-3-butyric acid (IBA)), with four replicates of 16 cuttings. The rooting and vegetative growth responses were variable. Genotype 5 was more responsive in the absence of IBA and genotypes 3, 8 and 9 were more responsive in the presence of IBA. Auxin increased rooting rate and percentage, reducing the average time of root protrusion in eight days. IBA also contributed to increase photosynthesis and dry root and shoot mass in 55.55 and 44.44% of the genotypes, respectively. The highest relative contribution to phenotypic diversity in the absence of auxin was rate (38.75%) and percentage (20.27%) of rooting, whereas in the presence of auxin was stomatal conductance (23.19%) and root dry mass (20.91%). Similarity was found for phenotypic and molecular divergence in the presence of IBA, in which genotypes 1 and 6; genotypes 5, 8 and 9; and genotype 3 were clustered in distinct groups.

Somatic embryogenesis in Carica papaya as affected by auxins and explants, and morphoanatomical-related aspects

ABSTRACT The aim of this study was to evaluate somatic embryogenesis in juvenile explants of the THB papaya cultivar. Apical shoots and cotyledonary leaves were inoculated in an induction medium composed of different concentrations of 2,4-D (6, 9, 12, 15 and 18 µM) or 4-CPA (19, 22, 25, 28 and 31 µM). The embryogenic calluses were transferred to a maturation medium for 30 days. Histological analysis were done during the induction and scanning electron microscopy after maturing. For both types of auxin, embryogenesis was achieved at higher frequencies with cotyledonary leaves incubated in induction medium than with apical shoots; except for callogenesis. The early-stage embryos (e.g., globular or heart-shape) predominated. Among the auxins, best results were observed in cotyledonary leaves induced with 4-CPA (25 µM). Histological analyses of the cotyledonary leaf-derived calluses confirmed that the somatic embryos (SEs) formed from parenchyma cells, predominantly differentiated via indirect and multicellular origin and infrequently via synchronized embryogenesis. The secondary embryogenesis was observed during induction and maturation phases in papaya THB cultivar. The combination of ABA (0.5 µM) and AC (15 g L-1) in maturation medium resulted in the highest somatic embryogenesis induction frequency (70 SEs callus-1) and the lowest percentage of early germination (4%).

Somatic embryogenesis in Carica papaya as affected by auxins and explants, and morphoanatomical-related aspects.

The aim of this study was to evaluate somatic embryogenesis in juvenile explants of the THB papaya cultivar. Apical shoots and cotyledonary leaves were inoculated in an induction medium composed of different concentrations of 2,4-D (6, 9, 12, 15 and 18 µM) or 4-CPA (19, 22, 25, 28 and 31 µM). The embryogenic calluses were transferred to a maturation medium for 30 days. Histological analysis were done during the induction and scanning electron microscopy after maturing. For both types of auxin, embryogenesis was achieved at higher frequencies with cotyledonary leaves incubated in induction medium than with apical shoots; except for callogenesis. The early-stage embryos (e.g., globular or heart-shape) predominated. Among the auxins, best results were observed in cotyledonary leaves induced with 4-CPA (25 µM). Histological analyses of the cotyledonary leaf-derived calluses confirmed that the somatic embryos (SEs) formed from parenchyma cells, predominantly differentiated via indirect and multicellular origin and infrequently via synchronized embryogenesis. The secondary embryogenesis was observed during induction and maturation phases in papaya THB cultivar. The combination of ABA (0.5 µM) and AC (15 g L-1) in maturation medium resulted in the highest somatic embryogenesis induction frequency (70 SEs callus-1) and the lowest percentage of early germination (4%).

Effects of a peripheral enamel margin on the long-term bond strength and nanoleakage of composite/dentin interfaces produced by self-adhesive and conventional resin cements.

PURPOSE: This study evaluated the effects of peripheral enamel margins on the long-term bond strength (µTBS) and nanoleakage in resin/dentin interfaces produced by self-adhesive and conventional resin cements. MATERIALS AND METHODS: Five self-adhesive [RelyX-Unicem (UN), RelyX-U100 (UC), GCem (GC), Maxcem (MC), Set (SET)] and 2 conventional resin cements [RelyX-ARC(RX), Panavia F(PF)] were used. An additional group included the use of a two-step self-etching adhesive (SE Bond) with Panavia F (PS). One hundred ninety-two molars were assigned to 8 groups according to luting material. Five-mm-thick composite disks were cemented and assigned to 3 subgroups according to water-exposure condition (n = 6): 24-h peripheral exposure (24h-PE-enamel margins), or 1 year of peripheral (1 yr-PE) or direct exposure (1 yr-DE-dentin margin). Restored teeth were sectioned into beams and tested in tension at 1 mm/min. Data were analyzed by two-way ANOVA and Tukey's test. Two additional specimens in each group were prepared for nanoleakage evaluation. Nanoleakage patterns were observed under SEM/TEM. RESULTS: Except for RX, no significant reduction in µTBS was observed between 24h-PE and 1 yr-PE. 1 yr-DE reduced µTBS for RX, PF, GC, MC, and SET. No significant reduction in µTBS was observed for PS, UC, and UN after 1 year. After 1 yr-DE, RX and PS presented the highest µTBS, and SET and MC the lowest. Nanoleakage was reduced when there was a peripheral enamel margin. SET and MC presented more silver deposition than other groups. CONCLUSION: The presence of a peripheral enamel margin reduced the degradation rate in resin/dentin interfaces for most materials. The µTBS values produced by the multi-step luting agents RX and PS were significantly higher than those observed for self-adhesive cement.

Analysis of hybrid layer thickness, resin tag length and their correlation with microtensile bond strength using a total etch adhesive to intact dentin.

The objective of this study was to evaluate the use of a two-step total etch and rinse adhesive, the correlation between the hyybrid layer thickness (HL) and bond strength (BS), and between resin tag length (RT) and bond strength in the same teeth, and also to evaluate the fracture patterns of the tested specimens. Ten human molars were used for the restorative procedure and then sectioned in two halves (mesio-distally). The materials used were Adper Single Bond 2, 3M ESPE, Ultra etch gel, Ultradent and Filtek Z250, 3M ESPE. One half were utilized to measure the HL thickness and RT length through light microscopy analysis (400x), and the other half was subject to a microtensile test to measure the BS. The fractured surfaces were analyzed by scanning electron microscopy and fracture patterns classified. The Pearson correlation test was applied (p = 0.05). The results of the analyses of each specimen then were correlated: mean HL thickness = 4.39 (0.48) microm, mean length of RT = 9.94 (1.69) microm, mean BS = 23.98 (10.24) MPa. A statistically significant correlation between HL thickness and bond strength was found (r = 0.93). The two step etch and rinse adhesive system, showed a strong correlation between HL thickness and bond strength. The most common fractures were adhesive, followed by cohesive in resin.

Evaluation "in situ" of tag formation in dental enamel submitted to microabrasion technique. Effect of two etching times.

The objective of this study was to analyze the formation of resin tags on enamel surfaces submitted or not to enamel microabrasion technique. Thirteen undergraduate dental students received removable acrylic palatal appliances on which four sections from intact premolars were fixed, measuring 4.0 mm x 4.0 mm. Two sections received the application of a microabrasive system and the remaining sections did not receive any surface treatment. The patients were instructed to wear the acrylic palatal appliances for 4 months during the day. After this time, the sections were etched with 37% phosphoric acid for 15 seconds (sections 1 and 2) and 60 seconds (sections 3 and 4), followed by adhesive system and composite resin applications. All the sections were prepared for light microscopy analysis to observe the degree of resin penetration (x400). The results were submitted to a three-way analysis of variance with a significance level of 5%. The difference between groups was verified by the Tukey test, at a significance level of 5%. The results showed that microabraded enamel required a longer period of acid etching for longer resin tags.

Evaluation "in situ" of tag formation in dental enamel submitted to microabrasion technique: Effect of two etching times

The objective of this study was to analyze the formation of resin tags on enamel surfaces submitted or not to enamel microabrasion technique. Thirteen undergraduate dental students received removable acrylic palatal appliances on which four sections from intact premolars were fixed, measuring 4.0 mm X 4.0 mm. Two sections received the application of a microabrasive system and the remaining sections did not receive any surface treatment. The patients were instructed to wear the acrylic palatal appliances for 4 months during the day. After this time, the sections were etched with 37% phosphoric acid for 15 seconds (sections 1 and 2) and 60 seconds (sections 3 and 4), followed by adhesive system and composite resin applications. All the sections were prepared for light microscopy analysis to observe the degree of resin penetration (x400). The results were submitted to a three-way analysis of variance with a significance level of 5%. The difference between groups was verified by the Tukey test, at a significance level of 5%. The results showed that microabraded enamel required a longer period of acid etching for longer resin tags.

O objetivo deste estudo foi analisar a formacao de tags resinosos em esmalte dental submetido ou nao a tecnica de microabrasao. Treze estudantes de Odontologia receberam a aplicacao de um aparelho ortodontico removivel contendo quatro seccoes de esmalte com dimensoes de 4.0 mm X 4.0 mm. Duas secoes receberam a aplicacao de um composto microabrasivo e as demais nao receberam nenhum tratamento. Os pacientes foram instruidos a usar os aparelhos ortodonticos durante o dia e por quatro meses. Apos este periodo, as seccoes foram condicionadas com acido fosforico 37% por 15 segundos (secoes 1 e 2) e por 60 segundos (seccoes 3 e 4), seguido pela aplicacao do sistema adesivo e de resina composta. Todas as seccoes foram preparadas para analise em microscopia optica comum para observar o grau de penetracao do material resinoso (x400). Os resultados foram submetidos a analise de variancia a tres criterios, ao nivel de 5%. A diferenca entre os grupos foi verificada pelo teste de Tukey, ao nivel de 5%. Os resultados mostraram que maiores tempos de condicionamento acido do esmalte dental sao exigidos para uma maior penetracao do sistema adesivo.

Microtensile bond strength of new self-adhesive luting agents and conventional multistep systems.

STATEMENT OF PROBLEM: Several self-adhesive luting agents have recently been introduced on the market. It is crucial to know the effectiveness of such luting agents prior to their clinical application. PURPOSE: The purpose of this study was to evaluate the microtensile bond strengths (microTBS) produced by different self-adhesive cements and compare them with conventional luting agents. MATERIAL AND METHODS: Six self-adhesive cements (RelyX Unicem (UN), RelyX U100 (UC), SmartCem 2 (SC), G-Cem (GC), Maxcem (MC), and SeT (SET), and 2 conventional luting agents, one that uses a 2-step etch-and-rinse adhesive (RelyX ARC (RX)), and one that uses a 1-step self-etching adhesive (Panavia F (PF)), were used in this study. An additional group included the use of a 2-step self-etching primer adhesive system (Clearfil SE Bond) prior to the application of Panavia F (PS). Fifty-four human molars were abraded to expose occlusal surfaces and were assigned to 9 groups according to the luting material (n=6). Five composite resin (Filtek Z250) discs (12 mm in diameter, 5 mm thick) were cemented on the teeth according to manufacturers' instructions. After 24 hours of water storage, restored teeth were serially sectioned into beams with a cross-sectional area of approximately 1 mm2 at the bonded interface and were tested in tension with a crosshead speed of 1 mm/min. Failure mode was determined using scanning electron microscopy. Data were statistically analyzed by 1-way ANOVA and Tukey's studentized range HSD test (alpha =.05). RESULTS: Mean bond strengths (SD) in MPa were: RX, 69.6 (16.6)A; PS, 49.2 (9.7)A; PF, 33.7 (13.9)AB; GC, 16.9 (10.3)BC; UC, 15.3 (3.4)BC; UN, 12.5 (2.4)C; MC 11.5 (6.8)CD; SC, 8.5 (4.9)CD; SET, 4.6 (0.5)D. Groups with different uppercase letters were significantly different from each other (P<.05). The predominant failure mode of the self-adhesive cements was adhesive between the resin cement and dentin. CONCLUSIONS: The bond strengths produced by the multistep luting agents were significantly higher than those observed for most self-adhesive cements.