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.

Cryptomeria japonica (Thunb. ex L.f.) D. Don is a commercial tree native to Japan and is one of the most important forest species in that country and the Azores (Portugal). Because of the quality of C. japonica timber, several genetic improvement programs have been performed. Recently, some studies focusing on C. japonica somatic embryogenesis have been carried out. However, in this species, this process uses immature seeds as initial explants, and for this reason, it is not possible to achieve the maximum genetic gain (100% genetic of the donor plant). Although some studies have been made applying organogenesis to this species, the success of the process in adult trees is low. For this reason, our main goal was to optimize the micropropagation method by using trees older than 30 years as a source of plant material. In this sense, in a first experiment, we studied the effect of different types of initial explants and three basal culture media on shoot induction; then, two sucrose concentrations and two light treatments (LEDs versus fluorescent lights) were tested for the improvement of rooting. In a second experiment, the effects of different plant growth regulators (6-benzylaminopurine, meta-topolin, and thidiazuron) on shoot induction and the subsequent phases of the organogenesis process were analyzed. The cultures produced the highest number of shoots when QL medium (Quoirin and Lepoivre, 1977) and long basal explants (>1.5 cm) were used; the shoots obtained produced a higher number of roots when they were grown under red LED lights. Moreover, root induction was significantly higher in shoots previously induced with meta-topolin.

Genetic improvement programs for conifer forest species face the challenge of propagating elite individuals with superior characteristics in the present landscape of climate change; the problem is focused on the fact that when these individuals have shown the desirable traits, they have changed phase and therefore have lost the ability to be propagated by traditional methods. Based on our previous works on Pinus spp. regeneration of adult trees through organogenesis and trying to improve the protocol in Pinus radiata, our objective was to analyze the influence of collection dates and different 6-benzyladenine (BA) concentrations in the first phase of shoot induction, as well as the effect of different light types on the success of root induction. Moreover, we were interested in studying the effect of the abovementioned physico-chemical factors on the amino acid and carbohydrate content in the shoots developed in vitro. Reinvigorated shoots were obtained in both BA concentrations (22 or 44 μM), although the highest BA concentration showed the best results in terms of shoot induction (explants forming shoots (46%) and number of shoots per explant (1.95 ± 0.52)) when using initial explants collected in the first week of February. The percentage of explants forming shoots (EFS) was genotype-dependent. Explants from genotype A induced with the highest BA concentration showed the highest EFS (91%). With respect to the light treatment applied, significant differences in root induction (20%) and in the number of roots per explant (4.62 ± 0.65) were observed in shoots cultured under white FL. Finally, significant differences in different phases of the rooting process were detected in the amounts of fructose, glucose and sucrose and in the content of threonine and tyrosine.

Teak has become one of the most widely planted species in tropical regions of the world, given its high price for its timber in international markets. This has motivated the development of tree improvement programs in the Latin American region and in the tropical world in general. The latest advances have achieved clonal forestry at an operational scale. Recently, important efforts are being made to advance towards the next breeding generation, since knowledge about floral biology and pollen management have become important issues. A breeding program is being developed through the Tree Improvement Cooperative GENFORES—a vinculation model between the academy and forestry companies that was initiated in Costa Rica and now involves six Latin American countries. In order to advance into the next breeding generations, building capacities in topics such as pollen banks requires pollen quality, thus enabling the exchange of pollen among cooperative members. Pollen fertility studies are of considerable value in breeding programs, in order to determine pollen viability and germination in collections of genotypes, before going into mating operational activities. In this study, we optimized pollen quality analysis protocols in terms of the viability and germination of fresh teak pollen. Results of this research show that 90% viability and 28% germination can be achieved in fresh pollen grains, both inside and outside the anther, previously dehydrated in silica gel for 2 to 4 h (40% and 33% humidity, respectively). Brewbacker and Kwack (BK) medium at 10% of its salts + 10% sucrose and at a pH of 7 must be used as the germination medium. It is possible to evaluate teak pollen quality using the parameters defined in this study, which will in turn allow pollen management and purification, providing an opportunity for carrying out controlled crosses at an operational scale as part of teak breeding programs.

Introduction: A protocol for the in vitro culture of the anxiolytic medicinal plant Souroubea sympetala (Marcgraviaceae) was developed, representing one of the first in vitro cultures for the family. This species was previously very difficult to cultivate from seed or cuttings. Methods: Methods included (1) the improvement of seed germination by axenic culture (2) development of regenerative cultures in vitro, then cultivation under greenhouse and finally field conditions and (3) creation of cell suspensions. Phytochemical analysis was undertaken by liquid chromatography coupled to mass spectrometry (HPLC-MS). Results: The percentage of seed germination was improved from 2% to 59% in axenic culture and the full development of the seedling with its apical shoot and root took twenty-four days. The best seedling development was obtained in Gamborg B5 culture medium. Most friable callus formation, (66.7%) was obtained in the Murashige and Skoog medium supplemented with naphthalene acetic acid (1 mg · L–1) and kinetin (0.5 mg · L–1) from which viable cell cultures were developed. Analysis identified 4 main triterpenes with both in vitro plants and greenhouse grown plants derived from them. The triterpenes were betulinic acid, ursolic acid, alpha-amyrin and beta-amyrin. The betulinic acid found in greenhouse plants was comparable to wild plants. The cell suspension cultures had much lower levels of betulinic acid than plants and are not at present a viable source of this anxiolytic triterpene. Discussion: The improvement in seed germination of this recalcitrant tropical species was highly successful. The subsequent in vitro propagation and progression of plants through greenhouse and field conditions to provide mature plants with active principle concentrations comparable to wild plants was promising. Friable callus was achieved but phytochemical analysis showed that the level of betulinic acid in callus was much lower than that found in mature plant tissue. Conclusion: The method provides healthy plants for cultivation of this new medicinal plant and consequently harvesting of wild plants is not required.

La caoba es la especie forestal nativa económicamente más importante en Latinoamérica, se encuentra en el Apéndice II de CITES, debido a que registra un elevado índice de explotación, producto de la alta extracción sufrida a lo largo de décadas. El objetivo de esta investigación fue desarrollar un protocolo para el establecimiento in vitro de 2 clones (33 y 80), pertenecientes a un programa de mejoramiento genético forestal. Se lograron establecer segmentos nodales de los clones 83 y 33 en condiciones in vitro, al emplear hipoclorito de calcio al 15 % con un tiempo de exposición de 20 minutos. El mejor medio de cultivo para el establecimiento in vitro de los nudos fue el MS complementado con 3 % de sacarosa, 0.016 g/L de agrimicin y benlate, obteniéndose un 55 % de nudos establecidos a los 16 días de cultivo. En ambos clones se evaluó el efecto de 5 concentraciones de 6-Bencilaminopurina (6-BAP) (0.0, 0.44, 0.88, 1.33, 1.77 y 2.21 μM L-1) sobre la brotación. Se obtuvo en promedio un brote por explante en los 5 tratamientos evaluados. No se observaron diferencias estadísticas significativas.