Separated Transcriptomes of Male Gametophyte and Tapetum in Rice: Validity of a Laser Microdissection (LM) Microarray

Abstract

In flowering plants, the male gametophyte, the pollen, develops in the anther. Complex patterns of gene expression in both the gametophytic and sporophytic tissues of the anther regulate this process. The gene expression profiles of the microspore/pollen and the sporophytic tapetum are of particular interest. In this study, a microarray technique combined with laser microdissection (44K LM-microarray) was developed and used to characterize separately the transcriptomes of the microspore/pollen and tapetum in rice. Expression profiles of 11 known tapetum specific-genes were consistent with previous reports. Based on their spatial and temporal expression patterns, 140 genes which had been previously defined as anther specific were further classified as male gametophyte specific (71 genes, 51%), tapetum-specific (seven genes, 5%) or expressed in both male gametophyte and tapetum (62 genes, 44%). These results indicate that the 44K LM-microarray is a reliable tool to analyze the gene expression profiles of two important cell types in the anther, the microspore/pollen and tapetum.

A novel epistatic interaction at two loci causing hybrid male sterility in an inter-subspecific cross of rice (Oryza sativa L.)

“…Recent pollen transcriptome studies have reported that 40% of about 1000 pollen-expressed mRNA were detected specifically in Arabidopsis pollen (Honys and Twell, 2004). In rice, 71 genes have been reported to express specifically in male gametophyte (Suwabe et al, 2008). If loss-offunction mutations occur at two related pollen specific genes in each parental species, these genes act as complementary genes causing hybrid male sterility.…”

Morphological and gene expression analysis under cool temperature conditions in rice anther development

“…Fluorescent probe labeling, hybridization, and scanning were performed according to the manufacturer's instructions (Agilent Technologies), with slight modification (Hobo et al, 2008;Suwabe et al, 2008). Biological replications with independently isolated samples and Cyanine3 (Cy3)/ Cyanine5 (Cy5) dye-swap were taken four times to confirm an experimental reproducibility of the analysis.…”

Induction of Male-Sterility by Controlling of Gibberellin Biosynthesis in Rice (Oryza sativa)

“…Use of an appropriate tissue specific promoter is a critical factor to control gene expression at specific stage and organ-specific manners. In rice, many kinds of pollen/anther/tapetumspecific genes with their promoters have been reported using traditional approaches and recently have been identified via transcriptome study through microarray analysis (Endo et al 2004;Suwabe et al 2008;Tang et al 2010). Most of the genes that are expressed at early stage of microspore development are usually tapetum-specific since their cells usually play roles in leading the initial development of microspore.…”