Methods to monitor post-translational gene silencing activity in plant tissues/cell types relevant for pathogen entry, propagation or replication

Patent metadata

Field	Value
Publication number	US-10550400-B2
Application number	US-201515536191-A
Country	US
Kind code	B2
Filing date	Dec 15, 2015
Priority date	Dec 15, 2014
Publication date	Feb 4, 2020
Grant date	Feb 4, 2020

Abstract

The invention relates to transgenic plants comprising an inverted-repeat construct which triggers post-transcriptional gene silencing of an endogenous visual reporter gene driven by a tissue-specific promoter wherein said tissue is relevant for pathogen entry, propagation or replication and their uses for screening natural or synthetic molecules, microorganisms or extracts from micro- or macro-organisms for their potential ability to inhibit pathogen entry, propagation or replication in plants by enhancing PTGS or for characterizing the mode of action of natural or synthetic molecules that are known to enhance plant disease resistance through an ill-defined mode of action.

First claim

The invention claimed is:

1. A method for characterizing the mode of action of natural or synthetic molecules that are known to enhance plant disease resistance comprising the following steps:
(a) providing several types of visual PTGS reporter plants transgenic for an inverted-repeat construct which triggers post-transcriptional gene silencing of an endogenous visual reporter gene, each type driven by a different tissue-specific promoter wherein said tissue is relevant for pathogen entry and/or propagation,
(b) applying said molecules on said plants, and
(c) characterizing the mode of action of said molecules by visually observing plant changes translating an increase or decrease of said post-transcriptional gene silencing activity of said endogenous visual reporter gene in one or several of said types of visual PTGS reporter plants.

2. The method of claim 1 , for characterizing the mode of action of said molecules that have a tissue-specific effect or an effect on different tissues that are relevant for pathogen entry, propagation or replication, further comprising an additional step:
(d) comparing the visual effect between said types of visual PTGS reporter plants
(i) wherein a tissue-specific effect means that said molecules are relevant for pathogen entry, propagation or replication only by said tissue or
(ii) wherein an effect on different tissues means that said molecules are relevant for pathogen entry, propagation or replication by different tissues.

3. The method of claim 1 , wherein said endogenous visual reporter gene is SUL cDNA of SEQ ID NO:1.

4. The method of claim 1 , wherein said relevant tissue for pathogen entry and/or propagation is selected from the group consisting of hydathodes, guard cells, xylem parenchyma and cambium cells, xylem parenchyma cells, cells at the base of trichomes, mesophyll cells and epidermal cells.

5. The method of claim 1 , wherein said tissue-specific promoter comprises AAP6 promoter of SEQ ID NO: 4, IRX3 promoter of SEQ ID NO: 5, GC1 promoter of SEQ ID NO: 6, HYD promoter of SEQ ID NO: 7, AHA3 promoter of SEQ ID NO: 8, MYC1 promoter of SEQ ID NO: 9, MYB60-promoter of SEQ ID NO: 10, CAB3 promoter of SEQ ID NO: 11, WOX4 promoter of SEQ ID NO: 12, and BDG promoter of SEQ ID NO: 37.

Assignees

• Paris Sciences Et Lettres—Quartier Latin
• Inst Nat Sante Rech Med
• Centre Nat Rech Scient

Inventors

• Navarro Lionel
• Thiebeauld Odon

Classifications

• C12N15/8212

  Colour markers, e.g. beta-glucoronidase [GUS], green fluorescent protein [GFP], carotenoid · CPC title

• C12N15/8279

  for biotic stress resistance, pathogen resistance, disease resistance · CPC title

• C12Q2600/13

  Plant traits · CPC title

• C12N15/8241

  Phenotypically and genetically modified plants via recombinant DNA technology · CPC title

• C12N15/8209 Primary

  Selection, visualisation of transformants, reporter constructs, e.g. antibiotic resistance markers · CPC title