Identification Of Anther-specific Expression Promotors In Arabidopisis And Brassica Napus

In this study, in order to obtain anther-specific promoters in Arabidopsis thaliana, the anther-specific candidate genes had been screened based on Arabidopsis thaliana transcriptome profiles.Through this method, eight genes were selected as anther-specific candidate genes. The promoter sequences were cloned according to the Arabidopsis genomic secquence. The promoter:GUS expression vectors of 8 candidate genes were constructed and transformed into the Arabidopsis. According to the histochemical staining results, PSG6 pro and PSG8 pro were seleceted for future study as pollen-specific expression promoters. Using the homology-based cloning strategy, the corresponding promoters of Brassica napus were cloned. The promoter:GUS expression vectors of the three Brassica napus promoters were constructed and transformed into the Arabidopsis. The three Brassica napus promoters were futher studied by 5,end deletion analyses. The mian results of this study were listed as follows:1. Eight genes of Arabidopsis were seleceted as anther-specific expression candidate genes, which were named PSG1, PSG2, PSG3, PSG4, PSG5, PSG6, PSG7 and PSG8. PSG is short for pollen-specific gene. Tissue-specific detection of the 8 candidate genes were performed by reverse transcription PCR(RT-PCR). It shows that PSG7 expresses in all tissues,while the other genes expresse only in the buds, open flowers or siliques.2. Cloning the promoter secquences of the eight candidate genes. The promoter:GUS expression vectors were constructed and transformed into Arabidopsis respectively. The expression patterns of the eight promoters were identified by GUS staining. The results showed that all the eight promoters were expressed only in the inflorescence, the PSG6 pro and PSG8 pro expresses relative late, with a higher specific-expression degree.3. Using the homology-based cloning strategy, one homologous gene of PSG6 and two homologous genes of PSG8 were identified in Brassica napus.The promoter secquences of the three genes were cloned and followed by the reporter gene GUS to construct expression vectors. The vectors were transformed into Arabidopsis. The GUS staining results showed that the three promoters were all anther-specific promoter.4. The three promoters of Brassica napus were futher studied by 5,end deletion analyses. Seven vectors were constructed for futher study. All these vectors were transformed into Arabidopsis. GUS staining results showed that 500 bp secquences were enough to conferring the anther-specific expression in two promoters, while 1000 bp secquences were enough for another one.5. Analyzing the common pollen-specific elements number of the eight Arabidopsis promoters and three Brassica napus promoters. It shows that all these promoters contain the LAT52 element and G10 element.