Isolation and Characterization of a Microsporocyte-Specific Gene, OsMSP, in Rice

Abstract

A spikelet-specific cDNA clone, OsMSP (Oryza sativa meiosis serine protease), was isolated from a rice cDNA library constructed by suppression subtraction hybridization (SSH). Its expression level was significantly reduced in the thermo-sensitive genic male-sterile rice line at sterile temperature (30/26°C, day/night) compared with that at fertile temperature (23/21°C, day/night) conditions. In contrast, no significant differences were observed in the normal fertile line under the above two sets of temperature conditions. The cDNA is 2,867 bp in length with an open reading frame (ORF) encoding 837 amino acid residues. Northern blots revealed that OsMSP transcripts were specifically present in spikelets at meiosis, and reached the highest level at early microspore stage. mRNA in situ hybridization confirmed that OsMSP was a rice microsporocyte-specific gene and its expression was limited to early microspore developmental stage. BLAST and phylogenetic analyses revealed that OsMSP shared high similarities with subtilisin-like serine protease genes from other organisms at both nucleotide and amino acid levels.

SSH Analysis of Endosperm Transcripts and Characterization of Heat Stress Regulated Expressed Sequence Tags in Bread Wheat

“…This technique identifies the abundant DEGs as well as rare transcripts in order to facilitate the identification of novel genes (Boominathan et al, 2004 ; Clement et al, 2008 ; Liu et al, 2008 ). SSH has been widely used in the past to identify differential expression of genes in maize ( Zea mays L.; Zhang et al, 2004 ; Nguyen et al, 2009 ), cucumber ( Cucumis sativus ; Terefe and Tatlioglu, 2005 ), mustard ( Brassica napus ; Wu et al, 2007 ), wheat ( T. aestivum ; Li et al, 2008 ), Mulberry (Gulyani and Khurana, 2011 ) and rice ( Oryza sativa ; Gorantla et al, 2007 ; Jiang et al, 2009 ). SSH is preferred in cases where genome sequence information is not available.…”

Identification and expression analysis of chloroplast ribonucleoproteins (cpRNPs) in Arabidopsis and rice

“…As compared with the other methods for analyzing altered gene expression, such as mRNA differential display (Liang and Pardee, 1992), serial analysis of gene expression (Velculescu et al, 1997), and cDNA microarray (Chu et al, 1998); suppressive subtractive hybridization (SSH) (Diatchenko et al, 1996) is a productive and efficient approach for identifying and characterizing both known and unknown genes involved in complex developmental processes and stress responses (Fernandez et al, 2007;Namasivayam and Hanke, 2006;Tsuwamoto et al, 2007). In recent years, SSH method is widely employed as a method of differential expression of genes in corn (Zhang et al, 2004), rape (Wu et al, 2007), wheat (Li et al, 2008;Zhang et al, 2005), rice (Jiang et al, 2009), and cucumber (Terefe and Tatlioglu, 2005). The present study aimed at identifying heat responsive genes at anthesis stage in thermo tolerant wheat cv Raj3765 (Rane et al, 2007) by construction and sequence analysis of a subtractive cDNA library.…”