Project description:Successful pollination brings together the mature pollen grain and stigma papilla to initiate an intricate series of molecular processes meant to eventually enable sperm cell delivery for fertilization and reproduction. At maturity, the pollen and stigma cells have acquired proteomes comprising the primary molecular effectors required upon their meeting. In Brassica species, knowledge of the roles and global composition of these proteomes is largely lacking. To address this gap, gel-free shotgun proteomics was performed on the mature pollen and stigma of Brassica carinata, a representative of the Brassica family and its many crop species (e.g. B. napus, B. oleracea, B. rapa), which holds considerable potential as a bio-industrial crop. 5608 and 7703 B. carinata mature pollen and stigma proteins were identified, respectively. The pollen and stigma proteomes were found to reflect not only their many common functional and developmental objectives, but also important differences underlying their cellular specialization. Isobaric tag for relative and absolute quantification (iTRAQ) was exploited in the first analysis of a developing Brassicaceae stigma, and uncovered 251 B. carinata proteins that were differentially abundant during stigma maturation, providing insight into proteins involved in the initial phases of pollination.
Project description:The purpose of the present study is to determine the effect of Phosphorus deficiency on gene expression level using microarray analysis to identify genes responsible for root hair development. Phosphorus deficiency induced the formation of root hairs to explore a greater soil volume but molecular mechanisms were unknown. Therefore, microarray experiments were performed using root tips of Brassica carinata cultivars Bale and Bacho, respectively differing in root hair length during Phosphorus deficiency. Experimental design was carried out in nutrient solution in a climate chamber with controlled environmental conditions (20°C, 16h day/8h night cycle, 70% relative humidity) in a randomized design. 25 root tips from 10 day old seedlings grown without Phosphorus of 1cm length were harvested and immediately frozen in liquid nitrogen. Gene expression analyses were performed
Project description:The purpose of the present study is to determine the effect of Phosphorus deficiency on gene expression level using microarray analysis to identify genes responsible for root hair development. Phosphorus deficiency induced the formation of root hairs to explore a greater soil volume but molecular mechanisms were unknown. Therefore, microarray experiments were performed using root tips of Brassica carinata cultivars Bale and Bacho, respectively differing in root hair length during Phosphorus deficiency. Experimental design was carried out in nutrient solution in a climate chamber with controlled environmental conditions (20°C, 16h day/8h night cycle, 70% relative humidity) in a randomized design. 25 root tips from 10 day old seedlings grown without Phosphorus of 1cm length were harvested and immediately frozen in liquid nitrogen. Gene expression analyses were performed Results from xy microarrays are summarized in this study. The samples originate from roots of cultivars Bale and Bacho grown in Phosphorus deficient conditions. Microarrays were hybridized with Cy3 and Cy5 labeled cDNA from Bale and Bacho both during Phosphorus deficiency using a dye swap approach
Project description:Purpose:Comparative cellular and transcriptome analyses was applied to characterize gene expression during male gametophytic development in Brassica carinata. Methods: floral buds (contain two developmental progress,1.1-1.6 mm and 1.8-6.5 long floral buds) were collected, then Separated male organs were kept in liquid nitrogen immediately until use. Total RNA was extracted using the TRIzol reagent (Invitrogen, Waltham, MA, USA). DNase (Promega, USA) was used to remove potential DNA contamination. For the quantitative real-time polymerase chain reaction (qRT-PCR) analysis Results: In this study, Up-regulated expression of DNA methylation probably affected pollen abortion in synthetic allohaploid B. carinata,and Down-regulated expression of cytokinin may affect pollen division and growth in synthetic allohaploid B. carinata Conclusions: Genes were shown male-preferred implies the dynamic changes of DNA methylation during the development of male gametes. The DEGs, related to CK signaling pathway and BR synthesis pathway were highly enriched in developmental male gametes, suggesting that CK played pivotal roles in male gamete development.