De novo transcriptome assembly and comparative transcriptomic analysis provide molecular insights into low temperature stress response of Canarium album.

A de novo transcriptome analysis was performed in C. album, a temperature sensitive fruit tree in China, after treatment with varied temperatures. A total number of 168,385 transcripts were assembled, comprising of 109,439 unigenes, of which 70,530 were successfully annotated. Compared with control check group (CK), which was treated under 25 °C, the chilling stress (4 °C) treated group (CT), showed about 2810 up-regulated and 2567 down-regulated genes. Whereas, group treated under freezing (- 3 °C) stress (FT) showed an up-regulation and a down-regulation of 1748 and 1459 genes, respectively. GO classification analysis revealed that DEGs related to metabolic processes, single-organism metabolic process, and catalytic activity are significantly enriched in both CT and FT conditions. KEGG pathway enrichment analysis for both CT and FT treatments showed an enrichment of genes encoding or related to glycine/serine and threonine metabolism, alpha-linolenic acid metabolism, carotenoid biosynthesis, photosynthesis-antenna proteins, and circadian rhythm. However, genes related to photosynthesis, carbon fixation in photosynthetic organisms, glutathione metabolism, pyruvate metabolism, nicotinate and nicotinamide metabolism were specifically enriched in CT condition. Nevertheless, FT treatment induced genes related to plant-pathogen interaction, linoleic acid metabolism, plant hormone signal transduction and pentose phosphate pathway. Many of the genes involved in plant hormone signal transduction showed significantly different expression in both FT and CT conditions. However, the change was more evident in FT. Here we present the first of the reports for a de novo transcriptomic analysis in C. album, suggesting that the plant shows differential responses in chilling and freezing temperatures, where the hormone signaling and transduction contribute greatly to FT responses. Our study thus paves way for future research regarding functions of these potentially identified genes.

Pollination ecology and reproductive biology of Canarium strictum Roxb. from evergreen forests of Central Western Ghats, India.

Pollination and reproductive biology of a dioecious tree Canarium strictum Roxb. (Burseraceae) was extensively studied within the Agumbe forest range of Western Ghats, Karnataka to identify primary pollen vectors and to enumerate interrelationship with the pollinators. The study also investigated phenology, floral biology, pollen production, pollen viability, stigma receptivity and nectar production. Trees produced functionally unisexual flowers with white petals, organized densely on inflorescences. Staminate flowers produced high percentage of viable pollen and relatively abundant nectar (15.75 µl) as a reward to the pollinators, while pistillate flowers produced only nectar (12 µl). Successful fruit set with wind pollination was facilitated by synchronization of flowering male and female trees, long term receptivity of stigma in female flowers and extended lifespan of flowers. The highest mean percent of fruit set with hand cross-pollination (µ = 91.06) suggests the influence of local male tree density, as well as, frequency and abundance of pollinator community on fruit set by open pollination.

Experimental evidence for an intraspecific Janzen-Connell effect mediated by soil biota.

The negative effect of soil pathogens on seedling survival varies considerably among conspecific individuals, but the underlying mechanisms are largely unknown. For variation between heterospecifics, a common explanation is the Janzen-Connell effect: negative density dependence in survival due to specialized pathogens aggregating on common hosts. We test whether an intraspecific Janzen-Connell effect exists, i.e., whether the survival chances of one population's seedlings surrounded by a different conspecific population increase with genetic difference, spatial distance, and trait dissimilarity between them. In a shade-house experiment, we grew seedlings of five populations of each of two subtropical tree species (Castanopsis fissa and Canarium album) for which we measured genetic distance using intersimple sequence repeat (ISSR) analysis and eight common traits/characters, and we treated them with soil material or soil biota filtrate collected from different populations. We found that the relative survival rate increased with increasing dissimilarity measured by spatial distance, genetic distance, and trait differences between the seedling and the population around which the soil was collected. This effect disappeared after soil sterilization. Our results provide evidence that genetic variation, trait similarity, and spatial distance can explain intraspecific variation in plant-soil biotic interactions and suggest that limiting similarity also occurs at the intraspecific level.

Size-dependent changes in leaf and wood chemical traits in two Caribbean rainforest trees.

Tree functional traits and their link to patterns of growth and demography are central to informing trait-based analyses of forest communities, and mechanistic models of forest dynamics. However, few data are available on how functional traits in trees vary through ontogeny, particularly in tropical species; and less is known about how patterns of size-dependent changes in traits may differ across species of contrasting life-history strategies. Here we describe size-dependent variation in seven leaf functional traits and four wood chemical traits, in two Dominican rainforest tree species (Dacryodes excelsa Vahl. and Miconia mirabilis (Aubl.) L.O. Williams), ranging from small saplings to the largest canopy trees. With one exception, all traits showed pronounced variation with tree size (diameter at breast height, DBH). Leaf mass per area (LMA), thickness and tissue density increased monotonically with DBH in both species. Leaf area, leaf nitrogen (N) and carbon (C) : nitrogen (N) ratios also varied significantly with DBH; however, these patterns were unimodal, with peak trait values preceding the DBH at reproductive onset in both species. Size-dependent changes in leaf structural traits (LMA and leaf thickness) were generally similar in both species, while traits associated with leaf-level investment in C gain (leaf area, leaf C : N ratio) showed contrasting ontogenetic trends between species. Wood starch concentration varied with DBH in both species, also showing unimodal patterns with peaks preceding size at reproductive onset. Wood C concentration increased linearly with DBH in both species, though significantly only in M. mirabilis. Size-dependent patterns in wood chemical traits were similar between both species. Our data demonstrate pronounced variation in functional traits through tree ontogeny, probably due to a combination of environmental factors and shifts in resource allocation. Such ontogenetic variation is comparable in magnitude with interspecific variation, and so should be accounted for in trait-based studies of forest dynamics, structure and function.

Insect herbivores, chemical innovation, and the evolution of habit specialization in Amazonian trees.

Herbivores are often implicated in the generation of the extraordinarily diverse tropical flora. One hypothesis linking enemies to plant diversification posits that the evolution of novel defenses allows plants to escape their enemies and expand their ranges. When range expansion involves entering a new habitat type, this could accelerate defense evolution if habitats contain different assemblages of herbivores and/or divergent resource availabilities that affect plant defense allocation. We evaluated this hypothesis by investigating two sister habitat specialist ecotypes of Protium subserratum (Burseraceae), a common Amazonian tree that occurs in white-sand and terra firme forests. We collected insect herbivores feeding on the plants, assessed whether growth differences between habitats were genetically based using a reciprocal transplant experiment, and sampled multiple populations of both lineages for defense chemistry. Protium subserratum plants were attacked mainly by chrysomelid beetles and cicadellid hemipterans. Assemblages of insect herbivores were dissimilar between populations of ecotypes from different habitats, as well as from the same habitat 100 km distant. Populations from terra firme habitats grew significantly faster than white-sand populations; they were taller, produced more leaf area, and had more chlorophyll. White-sand populations expressed more dry mass of secondary compounds and accumulated more flavone glycosides and oxidized terpenes, whereas terra firme populations produced a coumaroylquinic acid that was absent from white-sand populations. We interpret these results as strong evidence that herbivores and resource availability select for divergent types and amounts of defense investment in white-sand and terra firme lineages of Protium subserratum, which may contribute to habitat-mediated speciation in these trees.

The contribution of edaphic heterogeneity to the evolution and diversity of Burseraceae trees in the western Amazon.

Environmental heterogeneity in the tropics is thought to lead to specialization in plants and thereby contribute to the diversity of the tropical flora. We examine this idea with data on the habitat specificity of 35 western Amazonian species from the genera Protium, Crepidospermum, and Tetragastris in the monophyletic tribe Protieae (Burseraceae) mapped on a molecular-based phylogeny. We surveyed three edaphic habitats that occur throughout terra firme Amazonia: white-sand, clay, and terrace soils in eight forests across more than 2000 km in the western Amazon. Twenty-six of the 35 species were found to be associated with only one of three soil types, and no species was associated with all three habitats; this pattern of edaphic specialization was consistent across the entire region. Habitat association mapped onto the phylogenetic tree shows association with terrace soils to be the probable ancestral state in the group, with subsequent speciation events onto clay and white-sand soils. The repeated gain of clay association within the clade likely coincides with the emergence of large areas of clay soils in the Miocene deposited during the Andean uplift. Character optimizations revealed that soil association was not phylogenetically clustered for white-sand and clay specialists, suggesting repeated independent evolution of soil specificity is common within the Protieae. This phylogenetic analysis also showed that multiple cases of putative sister taxa with parapatric distributions differ in their edaphic associations, suggesting that edaphic heterogeneity was an important driver of speciation in the Protieae in the Amazon basin.