Molecular Cloning and Characterization of PhaTF13 that May Involve in Virus-induced Salicylic Acid-related Plant Defense Response

• Main Authors: Tan-Tung Wang, 王丹彤
• Other Authors: 葉信宏
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/41173897266057443198

碩士 === 國立臺灣大學 === 植物病理與微生物學研究所 === 100 === Orchidaceae consists of at least 35000 species and is among one of the largest families within angiosperm; however, how they trigger plant defense and response to pathogens remains largely unsolved. In our previous experiments using a Cymbidium mosaic virus (CymMV) induced gene silencing system to screen 187 predicted Phalaenopsis aphrodite var. Formosa transcription factors on virus induced salicylic acids (SA) related plant defense response allowed us to identify PhaTF13 involved in the expression of P. aphrodite homolog of pathogenesis related gene 1, PhaPR1. To identify the transcription start site and stop site of PhaTF13, we used Rapid Amplification of cDNA Ends (RACE) to obtain the 5’- and 3’-end sequences. Phylogenetic analysis revealed that PhaTF13 protein contains conserved A20/AN1 zinc finger domain and is grouped together with other monocotyledons homologs. To more specifically analyze the role of PhaTF13 in SA-related plant defense, we delivered two 21-nt hairpin RNA into P. aphrodite by agroinfiltration, and the data suggested that knockdown of PhaTF13 reduced the expression of PhaPR1 and PhaNPR1. The PhaTF13 only slightly expressed in column and leaf but induced to higher levels afterz CymMV infection in all P. aphrodite parts. Expression of PhaTF13::GFP or GFP::PhaTF13 in P. aphrodite protoplasts revealed that the PhaTF13 is mainly localized in nucleolus.