Phytophthora infestans Transformants Deficient in inf1 mRNA and INF1 Protein Production

<p>Scans of blots used in Figure 2 of van West et al. Mol Cell (1999)</p

Deletion analysis of Phytophthora infestans CRN8

<p>Data files for Figure 2 of "The Irish Potato Famine Pathogen Phytophthora infestans Translocates the CRN8 Kinase into Host Plant Cells" Mireille van Damme, Tolga O. Bozkurt, Cahid Cakir, Sebastian Schornack, Jan Sklenar, Alexandra M. E. Jones, Sophien Kamoun, PLoS Pathogens (2012)</p

Integrated model of plant-microbe interactions

<p>Adapted from:</p> <p>Win, J., Chaparro-Garcia, A., Belhaj, K., Saunders, D.G.O., Yoshida, K., Dong, S., Schornack, S., Zipfel, C., Robatzek, S., Hogenhout, S.A., and Kamoun, S. 2012. Effector Biology of Plant-associated Organisms: Concepts and Perspectives. Cold Spring Harbor Symposium on Quantitative Biology, 77: in press.</p> <p>Dodds, P.N., and Rathjen, J.P. 2010. Plant immunity: towards an integrated view of plant–pathogen interactions. Nature Reviews Genetics 11, 539-548</p> <p> </p

Transcriptome assembly of RNA-Seq data from field samples collected during wheat blast epidemic in Bangladesh 2016

In 2016, wheat fields in Bangladesh were infected by wheat blast fungus (<i>Magnaporthe oryzae</i>) for the first time. The fungus spread quickly thoroughout the country and threatened other wheat growing areas in neighbouring countries. Islam <i>et al </i>(BMC Biology 2016 14:84<strong>, </strong>DOI<strong>: </strong>10.1186/s12915-016-0309-7) collected infected wheat samples, extracted RNA, and obtained short reads by Illumina RNA-Seq technology. Here, we report assemby of these short reads using Trinity v 2.0.6.<br

Agrobacterium tumefaciens does not transform guard cells in Nicotiana benthamiana

<p>The bacterium Agrobacterium tumefaciens is widely used to genetically transform leaf cells of Nicotiana benthamiana using the agroinfiltration method (http://www.youtube.com/watch?v=GHc7PU_jG2M). Interestingly, A. tumefaciens does not transform guard cells, whereas it transforms efficiently pavement cells. The high chlorophyll content makes chloroplasts autofluorescent. In the case of live-cell imaging experiments with leaf cells transiently expressing fluorescent markers targeted to chloroplasts, it is hard to differenciate the fluorescent marker signal from chlorophyll autofluorescence. Here, I show that guard cells can be used to differentiate those. A GFP targeted to the chloroplast stroma was transiently expressed in N. benthamiana leaf cells, and epidermis cells were observed by live-cell imaging with a laser-scanning confocal microscope two days after the agro-infiltration. A 488 nm wavelength was used to illuminate the tissues, and GFP and chlorophyll signals were collected between 505-525 nm and 680-700 nm, respectively. The asterisks mark the guard cells, the cross marks a non-transformed pavement cell. The white rectangle on the merged image delimitates the close-ups presented in the lower panel. Note the absence of GFP signal in chloroplasts from the guard cells and from the non-transformed pavement cell.</p

pHIS-ATS: A Protein Expression Vector Modified from pFLAG-ATS for Secreted Expression of HIS-tagged Fusion Proteins

This paper describes a protein expression vector that allows secreted expression of HIS-tagged recombinant proteins in <i>E. coli.</i

Populus trichocarpa Coproporphyrinogen III oxidase (PtCPO) localizes in sub-chloroplastic structures

<p>The open reading frame (ORF) of PtCPO (see sequence below) was amplified by PCR from the hybrid poplar Beaupré (P. trichocarpa x P. deltoides) leaf cDNAs. The ORF was fused with the N-terminus of a green fluorescent protein (GFP) into the pICH86988 in order to create an in-frame PtCPO-GFP protein fusion, under the control of a 35S-Ω promoter. The final vector was inserted into Agrobacterium tumefaciens (GV3101 strain). The PtCPO-GFP protein fusion was transiently expressed in N. benthamiana leaf cells using A. tumefaciens leaf infiltration. Two days after infiltration, pavement cells were observed by live-cell imaging with a laser-scanning confocal microscope. Chlorophyll and GFP were excited at 488 nm, and their fluorescent signals were collected between 505-525 nm and 680-700 nm, respectively. Images present a single optical section of 0.8 µm. White arrowheads indicate discrete areas (diameter: 0.1 to 0.3 µm) within the chloroplasts. Scale bar: 5 µm. >PtCPO (98.7 % amino acid identity with Potri.011G023900.1, Phytozome portal http://www.phytozome.net/genePage.php?search=1&detail=1&crown&method=0&searchText=transcriptid%3A27003187/)</p> <p> </p

Golden-Gate compatible Magnaporthe oryzae Agrobacterium transformation vectors

The Golden Gate cloning system uses standardised parts to facilitate the assembly of multiple transcriptional units, to ensure that future work with these genes can be carried out with ease (Patron et al., 2015 New Phytologist, v. 208, p. 13-19).<br><div><br></div><div>We have developed the Golden Gate compatible vector pBHt2G-RFP (Addgene #107162) from the pCAMBIA-derived (Mullins et al., 2001) pBHt2G vector (Khang et al, 2010). The vector was domesticated through removal of BsaI cloning sites. An RFP-marker was inserted, which is expressed in E. coli, allowing for red-white selection of transformants. The marker is lost during the Golden Gate reaction, as it is replaced by the inserted transcriptional units.</div><div><br></div><div>Vector, sequence information and plasmid maps are available from Addgene https://www.addgene.org/107162//</div

Golden-Gate compatible Magnaporthe oryzae protoplast transformation vectors

<div>The Golden Gate cloning system uses standardised parts to facilitate the assembly of multiple transcriptional units, to ensure that future work with these genes can be carried out with ease (Patron et al., 2015 New Phytologist, v. 208, p. 13-19).</div><div><br></div><div>Three fungal transformation vectors have been adapted from the pCB1532 vector series (Sweigard et al., 1997. Fungal Genetics Newsletter 44: 52-53). Vector pCB1532B-RFP Addgene #101854 encodes bialaphos/basta/L-phosphinothricin resistance, pCB1532H-RFP #101855 hygromycin resistance and pCB1532S-RFP #101856 sulfonylurea/chlorimuron ethyl resistance. </div><div><br></div><div>Vectors were domesticated through removal of BsaI cloning sites. An RFP-marker was inserted. The RFP is expressed in E. coli, allowing for red-white selection of transformants. The marker is lost during the Golden Gate reaction, as it is replaced by the inserted transcriptional units. Vectors, sequence information and plasmid maps are available from Addgene https://www.addgene.org/plasmids/articles/28191792/ </div><div><br></div

Transcriptome sequencing of rice leaves with blast symptoms collected from rice fields of the Philippines in 2017 and release of raw sequence data on OpenRiceBlast website for open access

<div>Infected rice samples were collected by Bo Zhou and team, and sent to the Sainsbury Laboratory for RNA extraction and sequencing. Library preparation and RNA-Seq sequencing runs were performed by Genewiz using Illumina HiSeq-2500 machines to produce paired-end reads with ~300 bp average insert size. Here we report the release of these data to general public with open access on OpenRiceBlast website.</div><div><br></div