Project description:For expression profiling analyses of early stages of tuber induction, plants of Solanum tuberosum ssp andigena (7540) were used. This wild subspecies is strictly dependent on photoperiod for tuberisation, such that short days (SD) inductive conditions are required in order to trigger tuber induction in the stolons. Andigena plants were grown in the greenhouse under LD non-inductive conditions until a 10-leaf stage. They were subsequently transferred to inductive SD conditions (8 h light/16 h dark), and sampled at 0, 2, 4, 6 and 8 days after transfer to SDs. Tuber swelling was visible approximately 6-8 days after transfer to inductive conditions. The apical region of the stolons (2 cm) was collected one hour before the beginning of the light period.
Project description:For expression profiling analyses of early stages of tuber induction, plants of Solanum tuberosum ssp andigena (7540) were used. This wild subspecies is strictly dependent on photoperiod for tuberisation, such that short days (SD) inductive conditions are required in order to trigger tuber induction in the stolons. Andigena plants were grown in the greenhouse under LD non-inductive conditions until a 10-leaf stage. They were subsequently transferred to inductive SD conditions (8 h light/16 h dark), and sampled at 0, 2, 4, 6 and 8 days after transfer to SDs. Tuber swelling was visible approximately 6-8 days after transfer to inductive conditions. The apical region of the stolons (2 cm) was collected one hour before the beginning of the light period. All collected stolons (LD; and 2d, 4d, 6d and 8d under SD) were processed as described. Hybridization scheme with a loop design was applied to compare one sample against the rest of the samples in the induction time course.
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 Solanum tuberosum tissues (including leaves, flowers and stolon). 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 genome under study.
Project description:Light intensity and spectral quality are two important factors that affect the cryopreservation success of plant germplasm. In a previous study, we found that LEDs with 90% red and 10% blue (RB) light have a positive effect on potato (Solanum tuberosum L.) shoot tip recovery during post-cryopreservation regeneration. In the present study, transcriptome profiles of non-cryopreserved (-) and cryopreserved (+) potato shoot tips exposed to RB, cool white fluorescent tubes (CW) and full darkness (D) were compared. Results indicated that light spectrum significantly affects transcript numbers in post-cryopreserved samples resulting in 3,284 differentially expressed genes of recovering shoot tips exposed to CW, RB and D. Gene ontology (GO) enrichment analysis revealed that several stress-related terms were more abundant in CW+, whereas morphogenesis-related processes were activated under RB+. Overall, cryopreservation had an extensive effect on gene expression, as 12,017 genes were differentially expressed in non-cryopreserved and cryopreserved shoot tips. Among all cryopreserved samples, enriched GO terms included various defense and stress responses to biotic stimuli and oxidative stress, whereas suppressed responses were mainly related to organ morphogenesis. The present study emphasizes that the optimization of light spectral properties to decrease light-related stress may greatly benefit the cryopreservation efficiency of plant germplasm.