Nicotiana benthamiana
Nb Experimental Protocols and Resources
New Publications:
New, efficient method for VIGS design
Reporter genes for investigating Nb-Pseudomonas interactions
Virus‐induced gene silencing database for phenomics and functional genomics in Nicotiana benthamiana
Virus-induced gene silencing in Nicotiana benthamiana
Virus-induced gene silencing – video
Virus-induced gene silencing – 1
(requires JoVE subscription)
PTI assay
(requires JoVE subscription)
First papers to use agroinfiltration
Article 1: Tang, X., R. Frederick, D. Halterman, J. Zhou and G. B. Martin (1996). Initiation of plant disease resistance by physical interaction of AvrPto with Pto kinase. Science 274: 2060–2063.
Article 2: Scofield, S. R., C. M. Tobias, J. P. Rathjen, J. H. Chang, D. T. Lavelle, R. W. Michelmore and B. J. Staskawicz (1996). Molecular basis of gene-for-gene specificity in bacterial speck disease of tomato. Science 274: 2063–2065.
Cell-to-cell movement assay
A cell-to-cell macromolecular transport assay in planta utilizing biolistic bombardment.
Agroinfiltration
Agroinfiltration of Nicotiana benthamiana leaf
Nb, Future Plans
Our current plan is to improve the assembly of the Nicotiana benthamiana genome sequence and link the sequence to the nineteen chromosomes. We are also working to improve the gene annotation.
Projects underway:
- Sequence an additional lane of a 5-kb mate-pair library at the Cornell-Weill facility.
- Use an F2 population (~96 individuals) developed by Dave Zaitlin (University of Kentucky) for a genotype-by-sequencing (GBS) approach using the Cornell Institute for Genomic Diversity facility.
- Sequence a 15-kb mate-pair library (University of Illinois-Urbana/Champagne Sequencing Facility).
- Create a new assembly of the genome sequence based on the data in the preceding text.
- Improve annotation using with RNA-seq data from various researchers.
Gregory Martin, BTI: PAMP- and Effector-triggered immunity transcriptome
Zach Lippman, Cold Spring Harbor Laboratory: meristem tissue
Dave Zaitlin, University of Kentucky: leaves, roots, flowers, developing seed capsules
Peter Moffett, University of Sherbrooke: ribosome-IP-RNA-Seq (defense response)
Data from others is welcome—please contact us!
This project is supported by BTI Innovation Funds and by National Science Foundation grant IOS-1025642.
Two Nicotiana benthamiana parents (outside; Nb-1 and Nb-DZ) were crossed to develop an F1 (middle) that was selfed to produce an F2 population for use in genotype-by-sequencing (GBS). Photo: Dave Zaitlin, U. of Kentucky.