Identification and molecular characterization of novel anther-specific genes in Oryza sativa L. by using cDNA microarray

The complicated genetic pathway regulates the developmental programs of male reproductive organ, anther tissues. To understand these molecular mechanisms, we performed cDNA microarray analyses and in situ hybridization to monitor gene expression patterns during anther development in rice. Microarray analysis of 4,304 cDNA clones revealed that the hybridization signal of 396 cDNA clones (271 non-redundant groups) increased more than six-fold in every stage of the anthers compared with that of leaves. Cluster analysis with the expression data showed that 259 cDNA clones (156 non redundant groups) were specifically or predominantly expressed in anther tissues and were regulated by developmental stage-specific manners in the anther tissues. These co-regulated genes would be important for development of functional anther tissues. Furthermore, we selected several clones for RNA in situ hybridization analysis. From these analyses, we found several novel genes that show temporal and spatial expression patterns during anther development in addition to anther-specific genes reported so far. These results indicate that the genes identified in this experiment are controlled by different programs and are specialized in their developmental and cell types.

复杂的遗传途径调控雄性生殖器官花药组织的发育过程。为了解这些分子机制，我们进行了cDNA微阵列分析和原位杂交，以监测水稻花药发育过程中的基因表达模式。对4304个cDNA克隆的微阵列分析显示，与叶片相比，396个cDNA克隆（271个非冗余组）在花药的各个阶段的杂交信号增强了六倍以上。对表达数据进行的聚类分析表明，259个cDNA克隆（156个非冗余组）在花药组织中特异性或主要表达，并在花药组织中受发育阶段特异性方式调控。这些共同调控的基因对于功能性花药组织的发育很重要。此外，我们选择了几个克隆进行RNA原位杂交分析。通过这些分析，我们发现了除了目前已报道的花药特异性基因外，还有几个在花药发育过程中表现出时空表达模式的新基因。这些结果表明，本实验中鉴定的基因受不同程序控制，并在其发育和细胞类型上具有特异性。