http://hdl.handle.net/2440/16 Fri, 24 May 2013 23:33:32 GMT 2013-05-24T23:33:32Z http://hdl.handle.net/2440/77890 Title: Transcriptomics on small samples Author: Roy, Stuart John; Conn, Simon James; Mayo, Gwenda Mary; Athman, Asmini Njeri; Gilliham, Matthew Abstract: Interrogating the cell-specific transcriptome forms an important component of understanding the role that specific cells play in assisting a plant to overcome abiotic stress. Among the challenges arising when extracting RNA from individual plant cells are: the isolation of pure cell populations; the small yield of material when isolating specific cell types, and ensuring an accurate representation of the transcriptome from each cell type after amplification of RNA. Here we describe two approaches for isolating RNA from specific cell types—single cell sampling and analysis (SiCSA) and laser capture microdissection. Isolated RNA can then be directly sampled qualitatively using reverse transcription PCR (RT-PCR) or amplified for profiling ¬multiple specific genes using quantitative RT-PCR and genome-wide transcript analyses. Sat, 31 Dec 2011 13:30:00 GMT http://hdl.handle.net/2440/77890 2011-12-31T13:30:00Z http://hdl.handle.net/2440/77889 Title: Fluorescence-activated cell sorting for analysis of cell type-specific responses to salinity stress in Arabidopsis and rice Author: Evrard, Aurelie Christiane Patricia; Bargmann, Bastiaan O. R.; Birnbaum, Kenneth; Tester, Mark Alfred; Baumann, Ute; Johnson, Alexander Arthur Theodore Sat, 31 Dec 2011 13:30:00 GMT http://hdl.handle.net/2440/77889 2011-12-31T13:30:00Z http://hdl.handle.net/2440/77846 Title: Sporophytic ovule tissues modulate the initiation and progression of apomixis in Hieracium Author: Tucker, Matthew Robert; Okada, Takashi; Johnson, Susan D.; Takaiwa, Fumio; Koltunow, Anna Maria Grazyna Abstract: Apomixis in Hieracium subgenus Pilosella initiates in ovules when sporophytic cells termed aposporous initial (AI) cells enlarge near sexual cells undergoing meiosis. AI cells displace the sexual structures and divide by mitosis to form unreduced embryo sac(s) without meiosis (apomeiosis) that initiate fertilization-independent embryo and endosperm development. In some Hieracium subgenus Pilosella species, these events are controlled by the dominant LOSS OF APOMEIOSIS (LOA) and LOSS OF PARTHENOGENESIS (LOP) loci. In H. praealtum and H. piloselloides, which both contain the same core LOA locus, the timing and frequency of AI cell formation is altered in derived mutants exhibiting abnormal funiculus growth and in transgenic plants expressing rolB which alters cellular sensitivity to auxin. The impact on apomictic and sexual reproduction was examined here when a chimeric RNAse gene was targeted to the funiculus and basal portions of the ovule, and also when polar auxin transport was inhibited during ovule development following N-1-naphthylphthalamic acid (NPA) application. Both treatments led to ovule deformity in the funiculus and distal parts of the ovule and LOA-dependent alterations in the timing, position, and frequency of AI cell formation. In the case of NPA treatment, this correlated with increased expression of DR5:GFP in the ovule, which marks the accumulation of the plant hormone auxin. Our results show that sporophytic information potentiated by funiculus growth and polar auxin transport influences ovule development, the initiation of apomixis, and the progression of embryo sac development in Hieracium. Signals associated with ovule pattern formation and auxin distribution or perception may influence the capacity of sporophytic ovule cells to respond to LOA. Sat, 31 Dec 2011 13:30:00 GMT http://hdl.handle.net/2440/77846 2011-12-31T13:30:00Z http://hdl.handle.net/2440/77823 Title: Stem and gall forming nematodes Author: Subbotin, Sergei A.; Riley, Ian Timothy Sat, 31 Dec 2011 13:30:00 GMT http://hdl.handle.net/2440/77823 2011-12-31T13:30:00Z