The expression pattern of a rice proteinase inhibitor gene OsPI8-1 implies its role in plant development

A rice proteinase inhibitor (PI) gene OsPI8-1 was identified. Belonging to the potato inhibitor I family, this gene contains a 201bp coding region with no introns and encodes a deduced protein of 66 amino acids which holds a PI domain. There are two uniform gene copies, OsPI8-1a and OsPI8-1b, with direct-repeat arrangement and an interval span of 13 kb on rice chromosome 8, corresponding to the site of BAC clone P0528B09 (Accession No. AP004703). Reverse transcription polymerase chain reaction (RT-PCR) assays showed that both OsPI8-1a and OsPI8-1b can be expressed in wild-type 'Zhonghua No.11'. To investigate the physiological functions of OsPI8-1 in plant development, we analyzed the expression patterns of the reporter gene beta-glucuronidase (GUS) driven by OsPI8-1 promoter at different developmental stages and tissues. It was demonstrated that no GUS signals were detected in the roots. Despite that very high GUS expression was examined in the shoot apical meristem, no detectable GUS activity in the developmental domains of leaf primordium was observed. OsPI8-1 promoter showed an obvious wound-induced response in mature leaves. Little GUS activity was detected in young nodes and internodes at the seedling stage, but active GUS expression was observed near the nodes on mature culms. In the developing stage of the anther, GUS signal was specifically located in the middle layer and the endothecium between the epidermis and tapetum. In the germinating seed, GUS expression was gradually accumulated in the side of scutellar epithelium close to the embryo. These tissue-specific accumulations suggested that OsPI8-1 has multiple endogenous roles on developmental regulation. In this report, the inhibitor function of OsPI8-1 to proteolytic enzymes and the potential influence of their poise on plant development (such as seed germination, tapetum degeneration, programmed cell death, etc.) were discussed.