Effector vector design in the Phytophthora infestans-potato pathosystem
The oomycete pathogen Phytophthora infestans is the causal agent of the devastating plant disease late blight on potato. Diverse type of transposons and many gene families are present in the genome which encodes the effector proteins involved in causing the pathogenicity. This plant pathogen is pred...
| Autor principal: | |
|---|---|
| Formato: | H2 |
| Lenguaje: | Inglés |
| Publicado: |
SLU/Dept. of Plant Biology and Forest Genetics (until 131231)
2012
|
| Materias: |
| _version_ | 1855570594300428288 |
|---|---|
| author | Tareq, Abu Sayeed |
| author_browse | Tareq, Abu Sayeed |
| author_facet | Tareq, Abu Sayeed |
| author_sort | Tareq, Abu Sayeed |
| collection | Epsilon Archive for Student Projects |
| description | The oomycete pathogen Phytophthora infestans is the causal agent of the devastating plant disease late blight on potato. Diverse type of transposons and many gene families are present in the genome which encodes the effector proteins involved in causing the pathogenicity. This plant
pathogen is predicted to secret hundreds of effector proteins inside the host plant cells to promote
infection. These proteins are sensed by the plant immune system in order to prevent pathogen growth. The effector proteins are divided into two main types, cytoplasmic effectors and apoplastic effectors based on their translocated status in the plant cell. In this study, the effectorencoding genes Avr3a, Epi1, Epi10, Inf1 and CRN8 were selected to monitor the potential in planta function of the effectors and to develop a stable transformation procedure for reporter gene constructs with effector gene promoters. The putative promoter sequences were derived
from the 5´ regions of the oomycete genes. Primers were designed to amplify the promoter regions and the amplification was confirmed by gel electrophoresis. The reporter gene GFP (encoding green fluorescent protein) was chosen for analysis of their promoter activities and to
facilitate studies on spatial and dynamic alteration of gene expression. Cloning was performed using the vector pTOR-eGFP containing a ham34 promoter and a GFP gene. The ham34 promoter was removed and the effector promoters were inserted in its place. A stable transformation procedure was examined using three vectors for the GFP-constructs and the five effector gene promoters. Transformants were obtained at similar frequencies with each combination of effector promoter and GFP; which were confirmed by gel electrophoresis.
Subsequently Agrobacterium tumefaciens (C58) mediated transformation was tried for an Avr3a promoter construct. The construct was ligated into the binary vector, but the transformation of Agrobacterium was not successful. |
| format | H2 |
| id | RepoSLU3822 |
| institution | Swedish University of Agricultural Sciences |
| language | Inglés |
| publishDate | 2012 |
| publishDateSort | 2012 |
| publisher | SLU/Dept. of Plant Biology and Forest Genetics (until 131231) |
| publisherStr | SLU/Dept. of Plant Biology and Forest Genetics (until 131231) |
| record_format | eprints |
| spelling | RepoSLU38222012-04-20T14:24:46Z Effector vector design in the Phytophthora infestans-potato pathosystem Tareq, Abu Sayeed Phytophthora infestans potato genome effectors potato pathosystem effector vector design The oomycete pathogen Phytophthora infestans is the causal agent of the devastating plant disease late blight on potato. Diverse type of transposons and many gene families are present in the genome which encodes the effector proteins involved in causing the pathogenicity. This plant pathogen is predicted to secret hundreds of effector proteins inside the host plant cells to promote infection. These proteins are sensed by the plant immune system in order to prevent pathogen growth. The effector proteins are divided into two main types, cytoplasmic effectors and apoplastic effectors based on their translocated status in the plant cell. In this study, the effectorencoding genes Avr3a, Epi1, Epi10, Inf1 and CRN8 were selected to monitor the potential in planta function of the effectors and to develop a stable transformation procedure for reporter gene constructs with effector gene promoters. The putative promoter sequences were derived from the 5´ regions of the oomycete genes. Primers were designed to amplify the promoter regions and the amplification was confirmed by gel electrophoresis. The reporter gene GFP (encoding green fluorescent protein) was chosen for analysis of their promoter activities and to facilitate studies on spatial and dynamic alteration of gene expression. Cloning was performed using the vector pTOR-eGFP containing a ham34 promoter and a GFP gene. The ham34 promoter was removed and the effector promoters were inserted in its place. A stable transformation procedure was examined using three vectors for the GFP-constructs and the five effector gene promoters. Transformants were obtained at similar frequencies with each combination of effector promoter and GFP; which were confirmed by gel electrophoresis. Subsequently Agrobacterium tumefaciens (C58) mediated transformation was tried for an Avr3a promoter construct. The construct was ligated into the binary vector, but the transformation of Agrobacterium was not successful. SLU/Dept. of Plant Biology and Forest Genetics (until 131231) 2012 H2 eng https://stud.epsilon.slu.se/3822/ |
| spellingShingle | Phytophthora infestans potato genome effectors potato pathosystem effector vector design Tareq, Abu Sayeed Effector vector design in the Phytophthora infestans-potato pathosystem |
| title | Effector vector design in the Phytophthora infestans-potato pathosystem |
| title_full | Effector vector design in the Phytophthora infestans-potato pathosystem |
| title_fullStr | Effector vector design in the Phytophthora infestans-potato pathosystem |
| title_full_unstemmed | Effector vector design in the Phytophthora infestans-potato pathosystem |
| title_short | Effector vector design in the Phytophthora infestans-potato pathosystem |
| title_sort | effector vector design in the phytophthora infestans-potato pathosystem |
| topic | Phytophthora infestans potato genome effectors potato pathosystem effector vector design |