Project description:Current understanding of floral developmental genetics comes primarily from the core-eudicot model Arabidopsis thaliana. Here we explore the floral transcriptome of the basal angiosperm, Nuphar advena (water lily), for insights into the ancestral developmental program of flowers. Several thousand Nuphar genes with significantly up-regulated floral expression are identified, including homologs of the well-known ABCE floral regulators. However, strong similarities in the expression profiles of different organ categories contradict the organ-specific spatial expression domains predicted by the ABCE model. The broadly overlapping transcriptional programs observed among floral organs in Nuphar are shared with the magnoliid Persea americana (avocado), supporting the inference that this is the ancestral condition in angiosperms. Consequently, the predominantly organ-specific transcriptional programs that characterize Arabidopsis flowers (and perhaps other eudicots) are derived. The transcriptional landscapes in Arabidopsis correlate with a shift toward morphologically distinct floral organs, including differentiated sepals and petals, and a perianth distinct from stamens and carpels. In contrast to most eudicots, perianth organs are weakly differentiated in Nuphar and Persea, with staminodial intermediates between stamens and perianth in Nuphar, and between stamens and carpels in Persea. Our findings suggest that genetic regulation of more spatially discrete transcriptional programs underlies the evolution of floral morphology.
Project description:Current understanding of floral developmental genetics comes primarily from the core-eudicot model Arabidopsis thaliana. Here we explore the floral transcriptome of the basal angiosperm, Nuphar advena (water lily), for insights into the ancestral developmental program of flowers. Several thousand Nuphar genes with significantly up-regulated floral expression are identified, including homologs of the well-known ABCE floral regulators. However, strong similarities in the expression profiles of different organ categories contradict the organ-specific spatial expression domains predicted by the ABCE model. The broadly overlapping transcriptional programs observed among floral organs in Nuphar are shared with the magnoliid Persea americana (avocado), supporting the inference that this is the ancestral condition in angiosperms. Consequently, the predominantly organ-specific transcriptional programs that characterize Arabidopsis flowers (and perhaps other eudicots) are derived. The transcriptional landscapes in Arabidopsis correlate with a shift toward morphologically distinct floral organs, including differentiated sepals and petals, and a perianth distinct from stamens and carpels. In contrast to most eudicots, perianth organs are weakly differentiated in Nuphar and Persea, with staminodial intermediates between stamens and perianth in Nuphar, and between stamens and carpels in Persea. Our findings suggest that genetic regulation of more spatially discrete transcriptional programs underlies the evolution of floral morphology. Custom microarrays targeting 6,220 unique Nuphar floral transcripts were used to measure expression levels in eight tissues using an interwoven double-loop design for 16 arrays.
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 Nuphar advena tissues (including leaves, flowers and roots). 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.
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 Nuphar advena tissues (including leaves, flowers and roots). 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 leaves, flowers and fruit of Nuphar advena. Total RNA was isolated using the Plant RNA Purification Reagent (Invitrogen) for leaves and the Guanidinium-free for flowers and roots, 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 Claude dePamphilis for providing the plant material as well as Kan Nobuta and Gayathri Mahalingam for assistance with the computational methods.