Project description:We analysed chromatin changes in Medicago (A17) between radicles from germinated tolreant to desiccation (R1P) or icompared to intolerant to desiccation (R1 and R5). Roots at 1mm were dried for 72hours and are desiccation sensitive, roots at 1mm plus incubation in PEG 8000 at -1.7Mpa for 72h at 10°C then dried for 72hours are desiccation tolerant and roots at 5mm dried for 72h are desiccation sensitive
Project description:In this study we have tried to utilize the unique aspects of the T. ruralis response to desiccation and rehydration to design a strategy to identify rehydrins that are of low abundance and perhaps completely novel to the desiccated or rehydration transcriptomes. We have constructed two Subtractive Suppression Hybridization (SSH) libraries (Diatchenko et al., 1996) that are designed to enrich for differentially expressed low-abundance transcripts contained within gametophytic cells either in the slow-dried state (mRNP sequestrated rehydrin transcripts) or cells that have been rapidly dried, rehydrated and sampled at 2h of hydration (rehydrin and recovery transcripts) when the translational change in gene expression is at its peak (Oliver 1991). To achieve this aim we constructed SSH libraries using PolyA RNA isolated from the polysomal (mRNP) fractions from the slow-dried and 2h rehydrated rapid dried gametophytes selected against PolyA RNA from hydrated control gametophytes as the source for driver cDNA. Collections of cDNA clones from each library were sequenced and used to generate a small T. ruralis SSH cDNA microarray for expression profiling of both total RNA extracts for transcript accumulation assessments and polysomal RNA extracts for transcript sequestration and recruitment assessments.
Project description:We performed Chromatine ImmunoPrecipitation of the Histone H3K27me3 mark in 1- and 5mm Medicago (A17) radicles which were desiccation sensitive (R1 and R5) and desiccation tolerant (R1P). Roots at 1mm were dried for 72hs and are desiccation sensitive, roots at 1mm plus incubation in PEG 8000 at -1.7Mpa for 72h at 10°C then dried for 72hs are desiccation tolerant and roots at 5mm dried for 72h are desication sensitive
Project description:We performed Chromatine ImmunoPrecipitation of the Histone H2AK119Ub mark in 1- and 5mm Medicago (A17) radicles which were desiccation sensitive (R1 and R5) and desiccation tolerant (R1P). Roots at 1mm were dried for 72hs and are desiccation sensitive, roots at 1mm plus incubation in PEG 8000 at -1.7Mpa for 72h at 10°C then dried for 72hs are desiccation tolerant and roots at 5mm dried for 72h are desication sensitive
