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Title: Tight regulation of plant immune responses by combining promoter and suicide exon elements

Abstract

Effector-triggered immunity (ETI) is activated when plant disease resistance (R) proteins recognize the presence of pathogen effector proteins delivered into host cells. The ETI response generally encompasses a defensive 'hypersensitive response' (HR) that involves programmed cell death at the site of pathogen recognition. While many R protein and effector protein pairs are known to trigger HR, other components of the ETI signaling pathway remain elusive. Effector genes regulated by inducible promoters cause background HR due to leaky protein expression, preventing the generation of relevant transgenic plant lines. By employing the HyP5SM suicide exon, we have developed a strategy to tightly regulate effector proteins such that HR ischemically inducible and non-leaky. This alternative splicing-based gene regulation system was shown to successfully control Bs2/AvrBs2-dependent andRPP1/ATR1-51-dependent HR in Nicotiana benthamiana and Nicotiana tabacum, respectively. It was also used to generate viable and healthy transgenic Arabidopsis thaliana plants that inducibly initiate HR. Beyond enabling studies on the ETI pathway, our regulatory strategy is generally applicable to reduce or eliminate undesired background expression of transgene.

Authors:
 [1];  [2];  [1];  [1];  [2];  [3]
  1. Univ. of California, Berkeley, CA (United States)
  2. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  3. Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
OSTI Identifier:
1257376
Alternate Identifier(s):
OSTI ID: 1512114
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Nucleic Acids Research
Additional Journal Information:
Journal Volume: 43; Journal Issue: 14; Journal ID: ISSN 0305-1048
Publisher:
Oxford University Press
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES

Citation Formats

Gonzalez, Tania L., Liang, Yan, Nguyen, Bao N., Staskawicz, Brian J., Loqué, Dominique, and Hammond, Ming C. Tight regulation of plant immune responses by combining promoter and suicide exon elements. United States: N. p., 2015. Web. doi:10.1093/nar/gkv655.
Gonzalez, Tania L., Liang, Yan, Nguyen, Bao N., Staskawicz, Brian J., Loqué, Dominique, & Hammond, Ming C. Tight regulation of plant immune responses by combining promoter and suicide exon elements. United States. doi:10.1093/nar/gkv655.
Gonzalez, Tania L., Liang, Yan, Nguyen, Bao N., Staskawicz, Brian J., Loqué, Dominique, and Hammond, Ming C. Thu . "Tight regulation of plant immune responses by combining promoter and suicide exon elements". United States. doi:10.1093/nar/gkv655. https://www.osti.gov/servlets/purl/1257376.
@article{osti_1257376,
title = {Tight regulation of plant immune responses by combining promoter and suicide exon elements},
author = {Gonzalez, Tania L. and Liang, Yan and Nguyen, Bao N. and Staskawicz, Brian J. and Loqué, Dominique and Hammond, Ming C.},
abstractNote = {Effector-triggered immunity (ETI) is activated when plant disease resistance (R) proteins recognize the presence of pathogen effector proteins delivered into host cells. The ETI response generally encompasses a defensive 'hypersensitive response' (HR) that involves programmed cell death at the site of pathogen recognition. While many R protein and effector protein pairs are known to trigger HR, other components of the ETI signaling pathway remain elusive. Effector genes regulated by inducible promoters cause background HR due to leaky protein expression, preventing the generation of relevant transgenic plant lines. By employing the HyP5SM suicide exon, we have developed a strategy to tightly regulate effector proteins such that HR ischemically inducible and non-leaky. This alternative splicing-based gene regulation system was shown to successfully control Bs2/AvrBs2-dependent andRPP1/ATR1-51-dependent HR in Nicotiana benthamiana and Nicotiana tabacum, respectively. It was also used to generate viable and healthy transgenic Arabidopsis thaliana plants that inducibly initiate HR. Beyond enabling studies on the ETI pathway, our regulatory strategy is generally applicable to reduce or eliminate undesired background expression of transgene.},
doi = {10.1093/nar/gkv655},
journal = {Nucleic Acids Research},
number = 14,
volume = 43,
place = {United States},
year = {2015},
month = {7}
}

Journal Article:
Free Publicly Available Full Text
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Cited by: 2 works
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Figures / Tables:

Figure 1 Figure 1: Dual transcription and splicing regulation eliminates leaky hypersensitive response. (A) Nicotiana benthamiana leaf with three spots transiently co-transformed with Agrobacterium tumefaciens to introduce SuperPromoter::Bs2-$HA$ resistance gene and either (i) constitutive (35S) avrBs2-$HA$, (ii) Dex-inducible (6$xUAS$) avrBs2-$HA$ or (iii) Dex-inducible avrBs2-$HyP5SM$-$HA$. The leaf was not treated with Dex andmore » $Os$L5 is not co-expressed. (B) Scheme representing how a two input system that combines promoter and suicide exon elements to control transcription and alternative splicing, respectively, results in tighter regulation of phenotype compared to a one input system (promoter alone). See Supplementary Figure S1 for a related scheme showing the case in the presence of inputs.« less

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    Works referencing / citing this record:

    Agrobacterium-mediated transient transformation of sorghum leaves for accelerating functional genomics and genome editing studies
    journal, February 2020


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