Light spectra affect the in vitro shoot development of Cedrela fissilis Vell. (Meliaceae) by changing the protein profile and polyamine contents.

The light spectrum quality is an important signal for plant growth and development. We evaluated the effects of different light spectra on the in vitro shoot development of Cedrela fissilis and its proteomic and polyamine (PA) profiles. Cotyledonary and apical nodal segments were grown under different light emitting diodes (LED) and fluorescent lamps. Shoots from cotyledonary nodal segments cultured with 6-benzyladenine (BA) that were grown under WmBdR LED showed increased length and higher fresh and dry matter compared to shoots grown under fluorescent lamps. A nonredundant protein databank generated by transcriptome sequencing and the de novo assembly of C. fissilis improved, and almost doubled, the protein identification compared to a Citrus sinensis databank. A total of 616 proteins were identified, with 23 up- and 103 down-accumulated in the shoots under WmBdR LEDs compared to fluorescent lamps. Most differentially accumulated proteins in shoots grown under the WmBdR LED lamp treatment compared to the fluorescent lamp treatment are involved in responding to metabolic processes, stress, biosynthetic and cellular protein modifications, and light stimulus processes. Among the proteins, the up-accumulation of argininosuccinate synthase was associated with an increase in the free putrescine content and, consequently, with higher shoot elongation under WmBdR LED. The down-accumulation of calreticulin, heat shock proteins, plastid-lipid-associated protein, ubiquitin-conjugating enzymes, and ultraviolet-B receptor UVR8 isoform X1 could be related to the longer shoot length noted under LED treatment. This study provides important data related to the effects of the light spectrum quality on in vitro morphogenesis through the modulation of specific proteins and free putrescine biosynthesis in C. fissilis, an endangered wood species from the Brazilian Atlantic Forest of economic and ecological relevance. The nonredundant protein databank of C. fissilis is available via ProteomeXchange under identifier PXD018020.

Biomass allocation and gas exchange are affected by light conditions in endangered Cedrela salvadorensis (Meliaceae) seedlings / Asignación de biomasa e intercambio de gases son afectados por las condiciones de luz en plántulas de Cedrela salvadorensis (Meliaceae) en peligro de extinción

Abstract:The determination of favorable light habitat conditions per species and life stage is transcendental, for both ex situ and in situ conservation strategies of endangered forest tree species, and for their utilization as plantation trees. This becomes especially important when planting material is scarce. We studied the multivariate responses in biomass allocation and in gas exchange to light and to CO2 in Cedrela salvadorensis seedlings, grown under similar light conditions as those this species faces in nature. During a period of 135 days, groups of ten seedlings were put under 75, 45, 15 and 3.5 % of full sun exposure obtained with neutral shade cloth, under nursery conditions. A series of biomass allocation variables and detailed gas exchange parameters (photosynthesis response curves to light and to internal carbon concentration) were measured at the end of the growth period in plants of the four treatments. According to the principal component analyses, highest values of gas exchange response were associated with the lower values of biomass allocation traits. These changes can be associated with resource-conservative and resource-acquisitive strategies, where the C. salvadorensis seedlings acclimatize their traits for the exploration and exploitation of light, to high or to dim light environment, respectively. The multivariate analyses also showed that the plants had a high performance at 45 % of light environments. These results suggest that 45 % of light environment was the optimal light habitat of this species at the tested developing stage. Our results have important implications to choose the best natural habitat for a successful establishment of C. salvadorensis. We propose practical considerations for programs of reforestation or reintroduction where this species be involved. Rev. Biol. Trop. 64 (3): 1143-1154. Epub 2016 September 01.

ResumenLa determinación de las condiciones lumínicas favorables por especie y estado de vida es trascendental para las estrategias de conservación ex situ y in situ de especies de árboles en peligro de extinción, y su utilización como plantaciones forestales. Esto se vuelve especialmente importante cuando el material de siembra es escaso. Aquí, nosotros estudiamos las respuestas en asignación de biomasa y en intercambio de gases a luz y CO2 en plántulas de Cedrela salvadorensis crecidas bajo condiciones lumínicas similares a las que esta especie enfrenta en la naturaleza. Durante 135 días, grupos de diez plántulas fueron colocadas bajo condiciones de 75, 45, 15 y 3.5 % de exposición total al sol obtenidas por medio de sarán bajo condiciones de vivero. Una serie de variables de asignación de biomasa y parámetros de intercambio de gases (fotosíntesis según curvas de respuesta a la luz y a la concentración de carbono interno) fueron medidas al final del periodo de crecimiento en las plantas de los cuatro tratamientos. De acuerdo con los análisis de componentes principales, valores altos de respuesta de intercambio de gases están asociados con valores bajos de rasgos de asignación de biomasa. Los cambios observados se encuentran asociados con las estrategias recurso-conservativas y recurso-adquisitivas donde C. salvadorensis aclimata sus rasgos para la exploración y explotación de luz en ambientes con escasa o excesiva radiación lumínica, respectivamente. Los análisis multivariados muestran también que las plantas tienen un alto rendimiento a 45 % de luz ambiental. Estos resultados sugieren que el 45 % de ambiente lumínico es el hábitat lumínico óptimo de esta especie en el estado de desarrollo estudiado. Nuestros resultados tienen importantes implicaciones para escoger el mejor hábitat natural para un exitoso establecimiento de C. salvadorensis. Por esto, proponemos consideraciones prácticas para programas de reforestación y reintroducción donde esta especie estaría involucrada.

Biomass allocation and gas exchange are affected by light conditions in endangered Cedrela salvadorensis (Meliaceae) seedlings.

The determination of favorable light habitat conditions per species and life stage is transcendental, for both ex situ and in situ conservation strategies of endangered forest tree species, and for their utilization as plantation trees. This becomes especially important when planting material is scarce. We studied the multivariate responses in biomass allocation and in gas exchange to light and to CO2 in Cedrela salvadorensis seedlings, grown under similar light conditions as those this species faces in nature. During a period of 135 days, groups of ten seedlings were put under 75, 45, 15 and 3.5 % of full sun exposure obtained with neutral shade cloth, under nursery conditions. A series of biomass allocation variables and detailed gas exchange parameters (photosynthesis response curves to light and to internal carbon concentration) were measured at the end of the growth period in plants of the four treatments. According to the principal component analyses, highest values of gas exchange response were associated with the lower values of biomass allocation traits. These changes can be associated with resource-conservative and resource-acquisitive strategies, where the C. salvadorensis seedlings acclimatize their traits for the exploration and exploitation of light, to high or to dim light environment, respectively. The multivariate analyses also showed that the plants had a high performance at 45 % of light environment. These results suggest that 45 % of light environment was the optimal light habitat of this species at the tested developing stage. Our results have important implications to choose the best natural habitat for a successful establishment of C. salvadorensis. We propose practical considerations for programs of reforestation or reintroduction where this species be involved.

Re-induction of desiccation tolerance after germination of Cedrela fissilis Vell. seeds.

This work aimed to characterize the re-induction of desiccation tolerance (DT) in germinated seeds, using polyethylene glycol (PEG 8000). Cell changes were investigated through cytological assays (cell viability and transmission electronic microscopy) as well as DNA integrity during loss and re-establishment of DT. The loss of DT was characterized by drying germinated seeds with different radicle lengths (1, 2, 3, 4 and 5 mm) in silica gel, decreasing the moisture content to ten percentage points intervals, followed by pre-humidification (100% RH / 24 h) and rehydration. To re-induce DT, germinated seeds were treated for 72 h with PEG (-2.04 MPa) and PEG (-2.04 MPa) + ABA (100 µM) before dehydration. Germinated seeds did not tolerate desiccation to 10% moisture content, irrespectively of the radicle length. However, when incubated in PEG, those with 1 and 2 mm long radicle attained 71% and 29% survival, respectively. The PEG+ABA treatment was efficient to re-establish DT in seeds with 1 mm long radicles (100% survival). The ultrastructural assays of the cells of germinated seeds with 2 and 5 mm length confirmed the obtained physiological results. Germinated seeds of C. fissilis constitute a useful tool for desiccation tolerance investigations.

Water use in four model tropical plant associations established in the lowlands of Costa Rica

We examined soil water use patterns of four model plant associations established in the North Caribbean lowlands of Costa Rica by comparing the stable hydrogen isotope composition, δD, in xylem sap and in soil water at different depths, under rainy and dry conditions. Four 5-year-old model plant associations composed of 2 tree species (Hyeronima alchorneoides and Cedrela odorata) having different architecture and phenology were studied. Average tree height was 8.9 and 7.6 m, respectively. Each tree species was grown in monoculture and in polyculture with 2 perennial monocotyledons (Euterpe oleracea and Heliconia imbricata). Maximum rooting depth at the time of δD determination was ~ 2 m for almost all species. Most roots of all species were concentrated in the upper soil layers. Stomatal conductance to water vapor (gS) was higher in the deciduous C. odorata than in the evergreen H. alchorneoides; within each species, gS did not differ when the trees were grown in mono or in polyculture. During the rainy season, gradients in soil water δD were not observed. Average rainy season xylem sap δD did not differ among members of the plant combinations tested (-30 ‰), and was more similar to δD values of shallow soil water. Under dry conditions, volumetric soil water content declined from 50 to ~ 35%, and modest gradients in soil water δD were observed. xylem sap δD obtained during dry conditions was significantly lower than rainy season values. xylem sap δD of plants growing in the four associations varied between -9 and -22‰, indicating that shallow water was predominantly absorbed during the dry period too. Differences in xylem sap δD of trees and monocots were also detected, but no significant patterns emerged. The results suggest that: a) the plant associations examined extracted water predominantly from shallow soil layers (<1 m), b) the natural isotopic variation in soil and plant water at the study site was low, and c) the plant mixes obtain water from more than a single soil layer simultaneously. Temporal factors were important in determining the competition and complementary relations observed among the trees and the perennial monocots. Under the prevailing environmental conditions, water use in these plant associations was determined largely by species-specific attributes such as biomass allocation to fine roots, phenology, and canopy architecture, and to a lesser extent by water limitations. Rev. Biol. Trop. 56 (4): 1947-1957. Epub 2008 December 12.

Examinamos los patrones de uso de agua del suelo de cuatro asociaciones vegetales establecidas en el Caribe norte de Costa Rica, comparando la composición isotópica del hidrógeno, δD, en la savia del xilema y en el agua del suelo en condiciones lluviosas y secas. Estudiamos cuatro asociaciones de cinco años de edad compuestas por dos árboles (Hyeronima alchorneoides y Cedrela odorata) con diferente arquitectura y fenología, cultivados en mono y policultivo con dos monocotiledóneas perennes (Euterpe oleracea y Heliconia imbricata). Las excavaciones mostraron que la profundidad máxima de las raíces fue de 2 m para casi todas las especies, y que la mayor densidad de raíces se encontraba en la superficie del suelo. La conductividad estomática (gS) fue mayor en el árbol caducifolio (C. odorata) que en el perennifolio (H. alchorneoides); dentro de cada especie, gS no difirió cuando los árboles fueron cultivados en mono o en policultivo. Los resultados sugieren que: a) las asociaciones examinadas extrajeron agua predominantemente de las capas superficiales del suelo (<1 m), b) la variación natural en el acceso al agua del suelo por parte de las especies, y en las propiedades del suelo, fue baja, y c) las combinaciones de plantas obtuvieron agua de varias capas del perfil del suelo simultáneamente. Los factores relacionados con el tiempo fueron importantes en la determinación de las relaciones de competencia y complementariedad observadas entre los árboles y las monocotiledóneas perennes. En las condiciones ambientales prevalecientes, el uso del agua por parte de estas asociaciones de plantas fue determinado más por atributos, como la asignación de biomasa a las raíces finas, la fenología, y las propiedades del dosel, que por limitaciones en la disponibilidad de agua.

Water use in four model tropical plant associations established in the lowlands of Costa Rica.

We examined soil water use patterns of four model plant associations established in the North Caribbean lowlands of Costa Rica by comparing the stable hydrogen isotope composition, deltaD, in xylem sap and in soil water at different depths, under rainy and dry conditions. Four 5-year-old model plant associations composed of 2 tree species (Hyeronima alchorneoides and Cedrela odorata) having different architecture and phenology were studied. Average tree height was 8.9 and 7.6 m, respectively. Each tree species was grown in monoculture and in polyculture with 2 perennial monocotyledons (Euterpe oleracea and Heliconia imbricata). Maximum rooting depth at the time of 6D determination was approximately 2 m for almost all species. Most roots of all species were concentrated in the upper soil layers. Stomatal conductance to water vapor (gS) was higher in the deciduous C. odorata than in the evergreen H. alchorneoides; within each species, g, did not differ when the trees were grown in mono or in polyculture. During the rainy season, gradients in soil water 6D were not observed. Average rainy season xylem sap deltaD did not differ among members of the plant combinations tested (-30% per thousand), and was more similar to deltaD values of shallow soil water. Under dry conditions, volumetric soil water content declined from 50 to approximately 35%, and modest gradients in soil water deltaD were observed. Xylem sap deltaD obtained during dry conditions was significantly lower than rainy season values. Xylem sap deltaD of plants growing in the four associations varied between -9 and -22% per hundred, indicating that shallow water was predominantly absorbed during the dry period too. Differences in xylem sap deltaD of trees and monocots were also detected, but no significant patterns emerged. The results suggest that: (a) the plant associations examined extracted water predominantly from shallow soil layers (<1 m), (b) the natural isotopic variation in soil and plant water at the study site was low, and (c) the plant mixes obtain water from more than a single soil layer simultaneously. Temporal factors were important in determining the competition and complementary relations observed among the trees and the perennial monocots. Under the prevailing environmental conditions, water use in these plant associations was determined largely by species-specific attributes such as biomass allocation to fine roots, phenology, and canopy architecture, and to a lesser extent by water limitations.

The tropical cedar tree (Cedrela fissilis Vell., Meliaceae) homolog of the Arabidopsis LEAFY gene is expressed in reproductive tissues and can complement Arabidopsis leafy mutants.

A homolog of FLORICAULA/LEAFY, CfLFY (for Cedrela fissilis LFY), was isolated from tropical cedar. The main stages of the reproductive development in C. fissilis were documented by scanning electron microscopy and the expression patterns of CfLFY were studied during the differentiation of the floral meristems. Furthermore, the biological role of the CfLFY gene was assessed using transgenic Arabidopsis plants. CfLFY showed a high degree of similarity to other plant homologs of FLO/LFY. Southern analysis showed that CfLFY is a single-copy gene in the tropical cedar genome. Northern blot analysis and in situ hybridization results showed that CfLFY was expressed in the reproductive buds during the transition from vegetative to reproductive growth, as well as in floral meristems and floral organs but was excluded from the vegetative apex and leaves. Transgenic Arabidopsis lfy26 mutant lines expressing the CfLFY coding region, under the control of the LFY promoter, showed restored wild-type phenotype. Taken together, our results suggest that CfLFY is a FLO/LFY homolog probably involved in the control of tropical cedar reproductive development.

Chloroplast DNA phylogeography reveals colonization history of a Neotropical tree, Cedrela odorata L., in Mesoamerica.

Spanish Cedar (Cedrela odorata L.) is a globally important timber species which has been severely exploited in Mesoamerica for over 200 years. Using polymerase chain reaction-restriction fragment length polymorphisms, its chloroplast (cp) DNA phylogeography was studied in Mesoamerica with samples from 29 populations in six countries. Five haplotypes were characterized, phylogenetically grouped into three lineages (Northern, Central and Southern). Spatial analysis of ordered genetic distance confirmed deviation from a pattern of isolation by distance. The geographically proximate Northern and Central cpDNA lineages were genetically the most differentiated, with the Southern lineage appearing between them on a minimum spanning tree. However, populations possessing Southern lineage haplotypes occupy distinct moist habitats, in contrast to populations possessing Northern and Central lineage haplotypes which occupy drier and more seasonal habitats. Given the known colonization of the proto-Mesoamerican peninsula by South American flora and fauna prior to the formation of the Isthmus of Panama, it seems most likely that the observed population structure in C. odorata results from repeated colonization of Mesoamerica from South American source populations. Such a model would imply an ancient, pre-Isthmian colonization of a dry-adapted type (possessing the Northern lineage or a prototype thereof), with a secondary colonization via the land bridge. Following this, a more recent (possibly post-Pleistocene) expansion of moist-adapted types possessing the Southern lineage from the south fits the known vegetation history of the region.