Isolation of Gametophytic Cells and Identification of Their Cell-Specific Markers in Torenia fournieri, T. concolor and Lindernia micrantha

Abstract

Summary Gametophytic cells in the female embryo sac, the egg, the central and the synergid cells have unique roles in sexual reproduction in higher plants. To investigate their cell-specific features, an efficient method of distinguishing and isolating each gametophytic cell is required. By testing the activities of several cellulase enzymes, we modified our previous cell isolation method and succeeded in isolating each cell from the Torenia and Lindernia species. We also established genetic markers that show specific expression patterns in each gametophytic cell. These markers, together with the cell isolation method, will be useful for analyzing the function of gametophytic cells.

Identification and characterization of TcCRP1, a pollen tube attractant from Torenia concolor

“…cDNA was synthesized using SuperScript w reverse transcriptase (Invitrogen, Carlsbad, CA, USA). The expression of cell-type specific marker genes in these mRNAs (Kawano et al, 2011) was confirmed to clarify that no contamination of other cell types in the pool of synergid cell mRNA and egg cell mRNA. TcCRP1 was first isolated from ovule cDNA by PCR using gene-specific degenerate primers (Supplementary Data Table S1, available online).…”

Dynamics of the cell fate specifications during female gametophyte development inArabidopsis

“…In most studies, female gametophyte cells were isolated with the enzyme solution containing cellulase. The optimized conditions of the enzyme solution were different for different plant species (Kawano et al, 2011). The cell-type specific RNA-seq, including the mutants defective in female gametophyte cell function, must be powerful tools to reveal the precise gene expression changes associated with each cell functions or specifications.…”