Micropropagation of Calophyllum brasiliense Cambess. (Clusiaceae) is a way to overcome difficulties in achieving large-scale plant production, given the recalcitrant nature of the seeds, irregular fructification and absence of natural vegetative propagation of the species. Cultures were established using nodal segments 2 cm in length, obtained from 1-2 year old seedlings, maintained in a greenhouse. Mercury chloride and Plant Preservative Mixture™ were used in the surface sterilizing stage, better results being achieved with Plant Preservative Mixture™ incorporation in culture medium, at any concentration. Polyvinylpyrrolidone, activated charcoal, cysteine, ascorbic acid or citric acid were added to the culture medium to avoid oxidation. After 30 days of culture, polyvinylpirrolidone and ascorbic acid gave better results, eliminating oxidation in most explants. For shoot multiplication, benzylaminopurine was used in concentrations of 4.4 and 8.8 µM in Woody Plant Medium, resulting in an average of 4.43 and 4.68 shoots per explant, respectively, after 90 days. Indole-3-butyric acid and α-naphthalene acetic acid were used to induce root formation, reaching a maximum rooting rate of 24% with 20µM α-naphthalene acetic acid. For acclimatization. the rooted plants were transferred to Plantmax  substrate and cultured in a greenhouse, reaching 79% of survival after 30 days and 60% after one year.

Plinia peruviana is a species that is native to Brazil and is important due to the taste and medicinal properties of its fruits. Young leaves and split mature seeds were used as explants to initiate somatic embryogenesis to obtain a large number of plants in a short period of time. Leaf discs were cultured in MS medium containing various concentrations of 2,4-D (2,4-dichlorophenoxyacetic acid) or picloram (4-amino-3,5,6-trichloro-2-pyridinecarboxylic acid). In the case of the mature seeds, various concentrations of glutamine, 2,4-D and a combination of auxin and BAP (6-benzylaminopurine) were tested for somatic embryogenesis induction. For somatic embryo maturation, several concentrations of PEG 6000 (polyethylene glycol; up to 90 g L-1) were tested. After 60 days of culture using leaf discs, callus formation occurred in all treatments, with the highest averages obtained with 10 μM 2,4-D. However, these calluses did not form somatic embryos. For the cultured seeds, the best treatment was the MS medium with 1,000 mg L-1 glutamine and 10 μM 2,4-D without BAP. The supplementation of 60 g L-1 PEG 6000 was sufficient to promote the maturation of the somatic embryos. Histological analyses of the calluses that were formed from leaf discs showed nonembryogenic characteristics. In contrast, the calluses that originated from mature seeds had small and round cells with little vacuolation, which are characteristics of embryogenic structures.

ResumoMarcadores RAPD (Random amplified polymorphic DNA) são simples, eficientes, baratos e não requerem conhecimento prévio do genoma do organismo estudado, o que é particularmente importante para espé-cies nativas. O objetivo deste estudo é identificar marcadores moleculares que possam ser usados para caracterizar genótipos responsivos às auxinas usadas para induzir o enraizamento in vitro de explantes de guanandi (Calophyllum brasiliense). Amostras de DNA foram extraídas de folhas de plantas mantidas em casa de vegetação e plantas micropropagadas após a fase de enraizamento. Quatro primers foram usados e 26 loci encontrados, dois quais 21 são polimórficos (87.5%), com um número de alelos observado de 1.87 e efetivo de 1.39. A diversidade genética de Nei foi 0.24, valor similar ao encontrado em populações naturais. A substituição de nitrogênio líquido por armazenamento a -80°C por meia hora, com a adição de polivinilpirrolidona durante a maceração, foi efetiva na prevenção da degradação das amostras. Os resultados das análises sugerem uma interação entre o genótipo e a resposta à auxina usada durante a cultura in vitro, bem como a possibilidade de um grupo de plantas particularmente responsivo. Novas técnicas são necessárias para determinar a presença de um marcador que identifique genótipos mais suscetíveis ao enraizamento in vitro. Palavras-chave:Guanandi, cultura in vitro, marcador molecular, polivinilpirrolidona Abstract RAPD (Random amplified polymorphic DNA) markers are simple, efficient, inexpensive and do not require prior knowledge of the genome of the organism studied, which is especially important for native species. The objective of this study was to characterize molecular markers that may be used to identify genotypes responsive to the auxin used to induce the in vitro rooting of shoots of guanandi (Calophyllum brasiliense). DNA samples were extracted from leaves of greenhouse and micropropagated plants after the rooting phase. Four primers were used and 26 loci found, of which 21 were polymorphic (87.5%) with an average number of alleles observed of 1.87 and an effective number of 1.39. Genetic diversity using Nei's measure was 0.24, similar to the values of natural populations. The replacement of liquid nitrogen by storage at -80°C for half an hour, with the addition of polyvinylpyrrolidone during maceration, was effective in preventing the degradation of the samples. The results of the analyses suggest an interaction between the genotype and the response to the auxin used during in vitro culture, as well as the possibility of one particular group of responsive arrays. New techniques are needed to determine the presence of a marker that identifies some genotypes more susceptible to in vitro rooting.

Calophyllum brasiliense is a tree species with limited natural reproduction. In vitro germination may be an alternative for obtaining high-quality seedlings. Seeds were maintained in water before surface disinfestation and compared with control seeds (i.e. not immersed), without differences between treatments. HgCl2 used during surface-disinfestation reduced contamination rates of cultures. Fungal contamination was reduced with fungicide added to culture medium (23 to 6.4%), although bacterial contamination increased (24 to 36%). In another experiment, seeds were immersed in plant preservative mixture (PPM™) prior to surface disinfestation. By combining immersion for 48 h and 2 mL L-1 in culture medium, contamination was only 6%. Seeds immersion in GA3 prior to surface disinfestation reduced root formation as concentration increased. Germination rate and GSI were reduced, respectively, from 72% and 0.129 (24 h) to 60% and 0.092 (48 h) according to exposure time to GA3. After 90 days in multiplication medium containing benzylaminopurine, average number of shoots per nodal segment was 3.4. In conclusion, in vitro germination of C. brasiliense seeds is feasible in sucrose-free WPM medium and reaches a high contamination-free rate (up to 93.3%).