Project description:Spanish fir (Abies pinsapo) is a relict tree that is found in southern Spain mountains. In natural conditions two phenotypes can be observed, trees with blue needles and trees with green needles. The aim of the present work is to elucidate at transcriptomic levels the possible causes of the differences between these phenotypes.
Project description:Transcriptome analysis of small RNA was performed using pollen and embryonic callus, and vegetative tissues, needles of Picea abies to address differences in small RNA profiles between reproductive tissues and vegetative tissues in gymnosperm.
Project description:Small RNAs (21-24 nt) are pivotal regulators of gene expression that guide both transcriptional and post-transcriptional silencing mechanisms in diverse eukaryotes, including most if not all plants. MicroRNAs (miRNAs) and short interfering RNAs (siRNAs) are the two major types, both of which have a demonstrated and important role in plant development, stress responses and pathogen resistance. In this work, we used a deep sequencing approach (Sequencing-By-Synthesis, or SBS) to develop sequence resources of small RNAs from Picea abies tissues (including needles, immature cones and lateral bud meristem). The high depth of the resulting datasets enabled us to examine in detail critical small RNA features as size distribution, tissue-specific regulation and sequence conservation between different organs in this species. We also developed database resources and a dedicated website (http://smallrna.udel.edu/) with computational tools for allowing other users to identify new miRNAs or siRNAs involved in specific regulatory pathways, verify the degree of conservation of these sequences in other plant species and map small RNAs on genes or larger regions of the maize genome under study. Small RNA libraries were derived from needles, immature female cones and lateral bud meristem of Picea abies. Total RNA was isolated using the Plant RNA Purification Reagent (Invitrogen), and submitted to Illumina (Hayward, CA, http://www.illumina.com) for small RNA library construction using approaches described in (Lu et al., 2007) with minor modifications. The small RNA libraries were sequenced with the Sequencing-By-Synthesis (SBS) technology by Illumina. PERL scripts were designed to remove the adapter sequences and determine the abundance of each distinct small RNA. We thank David Neale for providing the plant material as well as Kan Nobuta and Gayathri Mahalingam for assistance with the computational methods.
Project description:Diversification of defense-related compounds is a plant defense mechanism against the diverse array of natural enemies in the course of evolution, and is known to involve various sources of tissue. Here a large-scale survey of gene expression is conducted from three different sources of tissue; needles, phloem with the attached bark, and xylem, with an emphasis on the expression of the secondary metabolites of the core phenylpropanoid pathway and derivative pathways, taken together as phenolic defense-related compounds. Five species of spruce; P. abies, P. glauca, P. jezoensis, P. mariana, and P. omorika, spanning much of their known phylogeny, are chosen for cross-species microarray hybridizations. The objectives are: first) to explore the tissue-related differences in the expression of phenolic genes and their coherent role in tree defense; second) to describe the diversification trends in the expression of phenolic gene families with respect to their position in the pathway; and third) to infer the mode of evolution underlying the expression of the selected genes.