Project description:Haploid embryos can be induced from cultured immature pollen following a stress treatment. In Brassica napus (B. napus), application of the histone/lysine deacetylase (HDAC/KDAC) inhibitor trichostatin A (TSA) to pollen cultures enhances the production of differentiated embryos and embryogenic callus when applied together with heat stress (Li et al., 2014). To identify genes associated with the induction of B. napus haploid embryogenesis, we employed fluorescence-activated cell sorting on microspore cultures of the LEAFY COTYLEDON1 embryo reporter line (LEC1:LEC1-GFP, Li et al., 2014) to select for embryogenic structures developing in culture, followed by RNA-sequencing. Our transcriptome analysis of the 3-day-old enriched embryo populations and in vitro cultured pollen revealed different regulatory pathways underlying the early development of embryogenic structures. Comparing FACS-sorted embryogenic transcriptomes from cultures treated with heat stress combined with either a low or a high concentration of TSA allowed us to assign transcription profiles to embryogenic structures that have a low or high potential to develop into histodifferentiated embryos.
Project description:Microspore embryogenesis is an in vitro system in which haploid microspores (precursor celsl of pollen grains) are reprogrammed towards embryogenesis by the application of a heat stress (HS) treatment. This process is of great interest in plant breeding as it enables accelerated production of double-haploid plants. The induction of in vitro microspore reprogramming comprises a complex chain of events that still remain unknown. The process can be induced at the responsive developmental stage of vacuolated microspore. In order to evaluate the transcriptional changes acompaying microspore embryogenesis induction in Brassica napus, an expression profiling through high throughput RNA sequencing, and a pipeline for bioinformatic analysis was performed. The analyses have been implemented for two samples: (1) vacuolated microspores (VM), before stress treatment, initial stage of the microspore culture, and (2) stress-treated microspores, 4 days after the application of the HS, when proembryos (PE) are formed; PE correspond to the first morphological sign of embryogenesis induction. The results obtained would provide valuable information about this complex process.
Project description:mRNA expression profiling of the embryo, endosperm (micropylar, peripheral, chalazal), and seed coat (outer, inner, chalazal, chalazal proliferating tissue) of the developing Brassica napus seed. Tissues were isolated using laser microdissection (LMD) from Brassica napus seeds at the globular, heart, and mature green stages of seed development.
Project description:High temperature stress results in yield loss and alterations to seed composition during seed filling in oilseed rape (Brassica napus). However, the mechanism underlying this heat response is poorly understood. In this study, we employed a microarray analysis with silique walls and seeds from the developing siliques (20 days after flowering) of Brassica napus that had undergone heat stress.
Project description:High temperature stress results in yield loss and alterations to seed composition during seed filling in oilseed rape (Brassica napus). However, the mechanism underlying this heat response is poorly understood. In this study, we employed a microarray analysis with silique walls and seeds from the developing siliques (20 days after flowering) of Brassica napus that had undergone heat stress. Two-condition experiment, control vs heat stress, 2 time points
Project description:To investigate the expression and methylation patterns of hybrid embryo development in Brassica napus, we performed pattern analysis on different developmental stages of parental and hybrid embryos
Project description:To investigate the expression and methylation patterns of hybrid embryo development in Brassica napus, we performed pattern analysis on different developmental stages of parental and hybrid embryos
Project description:To investigate the expression and methylation patterns of hybrid embryo development in Brassica napus, we performed pattern analysis on different developmental stages of parental and hybrid embryos
Project description:Untargeted metabolomic analyses were carried out on seed coat/endosperm and seed embryo (dry seeds) of two Brassica napus genotypes (Aviso and Major). Four biological replicates were analyzed for each sample.