Study And Isolation Of Pollen-tissure Specific Promoter In Tritordeum

Wheat is one of the important crops in the world. Isolation of pollen/anther -specific expessing gene and promoter could not only understand gene regulation in the sexual reprodution of flowering plants,but also could generate male-sterility by gene engineering. As an approach to isolate novel cereal promoters, promoterless uidA constructs and particle bombardment were used to transform tritordeum. Then one transgenic material was got, primarily detection showed that it's a pollen-specific regulater being tagged by uidA gene. In order to confirm the inheritance of uidA gene, the pollen-tissue specific expression of uidA gene in the next genetation,and to prepare the materials for the promoter isolation, two generations of one positive material were analysed. All fity-one F1 and F2 progeny plants had uidA gene, and they did not separate. Histochemical analysis of progeny plants showed that GUS activity was only detected in pollen. Further RT -PCR analyses showed that uidA gene was not transcripted in the root and leaf, therefore there is no GUS activity in the root and leaf. All above indicated the material was pollen-specific promoter tagged by uidA gene and they were homozygotes. Our experiments confirmed this specificity could inherit steadily. TAIL-PCR was adoptted to isolate the upsteam promoter, which regulates the expression of uidA gene. The results showed that uidA gene was multicopy and some copies had undergone rearrangment/deletion. So they made the isolation of the promoter harder, it also gave some help to the isolation.