Ferredoxin from sweet pepper (Capsicum annuum L.) intensifying harpinpss-mediated hypersensitive response shows an enhanced production of active oxygen species (AOS)

Badri Venkata Dayakar, Hao Jan Lin, Cheng-Hsien Chen, Mang Jye Ger, Bor Heng Lee, Chia Hwei Pai, David T. Chow, Hsiang En Huang, Shaw Yhi Hwang, Mei Chu Chung, Teng Yung Feng

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

The hypersensitive response (HR) is a form of cell death associated with plant resistance to pathogen infection. Harpinpss, an elicitor from the bacterium Pseudomonas syringae pv. syringae, induces a HR in non-host plants. Previously, we reported an amphipathic protein from sweet pepper interfering with harpinpss-mediated HR. In this report, we isolated and characterized a cDNA clone encoded that amphipathic protein from sweet pepper. This protein is designated as PFLP (plant ferredoxin-like protein) by virtue of its high homology with plant ferredoxin protein containing an N-terminal signal peptide responsible for chloroplast targeting and a putative 2Fe-2S domain responsible for redox activity. Recombinant PFLP obtained from Escherichia coli was able to significantly increase active oxygen species (AOS) generation when mixed with harpinpss in tobacco suspension cells. It also showed enhanced HR when co-infiltrated with harpinpss in tobacco leaves. We used a transgenic tobacco suspension cells system that constitutively expresses the Pflp gene driven by the CaMV 35S promoter to study the function of PFLP in enhancing harpinpss-mediated hypersensitive cell death in vivo. In response to harpinpss, suspension cells derived from Pflp transgenic tobacco showed a significant increase both in the generation of AOS and in cell death as compared to the wild type. AOS inhibitors diphenylene iodonium chloride (DPI) and lanthanum chlorate (LaCl3) were used to study the involvement of AOS in harpinpss-induced cell death. Our results demonstrate enhanced generation of AOS is necessary to cause enhanced hypersensitive cell death in Pflp transgenic tobacco cells and it is plasma membrane-bound NADPH-oxidase-dependent. Sub-cellular localization studies showed that PFLP is present in the cytoplasm and chloroplast of Pflp transgenic tobacco cells, but only in the chloroplast, not in the cytoplasm, of wild-type tobacco cells. It is possible that PFLP can change the redox state of the cell upon harpinpss inoculation to increase AOS generation and hypersensitive cell death. Overall, this study will provide a new insight in the functional properties of ferredoxin in hypersensitive cell death.

Original languageEnglish
Pages (from-to)913-924
Number of pages12
JournalPlant Molecular Biology
Volume51
Issue number6
DOIs
Publication statusPublished - Apr 2003
Externally publishedYes

Fingerprint

Ferredoxins
Capsicum
ferredoxins
hypersensitive response
sweet peppers
Capsicum annuum
Tobacco
Cell death
Reactive Oxygen Species
Cell Death
cell death
tobacco
Chloroplasts
Proteins
proteins
Suspensions
genetically modified organisms
cell suspension culture
chloroplasts
Oxidation-Reduction

Keywords

  • Active oxygen species
  • Ferredoxin
  • Hypersensitive cell death
  • PFLP

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Ferredoxin from sweet pepper (Capsicum annuum L.) intensifying harpinpss-mediated hypersensitive response shows an enhanced production of active oxygen species (AOS). / Venkata Dayakar, Badri; Lin, Hao Jan; Chen, Cheng-Hsien; Ger, Mang Jye; Lee, Bor Heng; Pai, Chia Hwei; Chow, David T.; Huang, Hsiang En; Hwang, Shaw Yhi; Chung, Mei Chu; Feng, Teng Yung.

In: Plant Molecular Biology, Vol. 51, No. 6, 04.2003, p. 913-924.

Research output: Contribution to journalArticle

Venkata Dayakar, Badri ; Lin, Hao Jan ; Chen, Cheng-Hsien ; Ger, Mang Jye ; Lee, Bor Heng ; Pai, Chia Hwei ; Chow, David T. ; Huang, Hsiang En ; Hwang, Shaw Yhi ; Chung, Mei Chu ; Feng, Teng Yung. / Ferredoxin from sweet pepper (Capsicum annuum L.) intensifying harpinpss-mediated hypersensitive response shows an enhanced production of active oxygen species (AOS). In: Plant Molecular Biology. 2003 ; Vol. 51, No. 6. pp. 913-924.
@article{5dc5c5b22dfc478195902dd03430a78e,
title = "Ferredoxin from sweet pepper (Capsicum annuum L.) intensifying harpinpss-mediated hypersensitive response shows an enhanced production of active oxygen species (AOS)",
abstract = "The hypersensitive response (HR) is a form of cell death associated with plant resistance to pathogen infection. Harpinpss, an elicitor from the bacterium Pseudomonas syringae pv. syringae, induces a HR in non-host plants. Previously, we reported an amphipathic protein from sweet pepper interfering with harpinpss-mediated HR. In this report, we isolated and characterized a cDNA clone encoded that amphipathic protein from sweet pepper. This protein is designated as PFLP (plant ferredoxin-like protein) by virtue of its high homology with plant ferredoxin protein containing an N-terminal signal peptide responsible for chloroplast targeting and a putative 2Fe-2S domain responsible for redox activity. Recombinant PFLP obtained from Escherichia coli was able to significantly increase active oxygen species (AOS) generation when mixed with harpinpss in tobacco suspension cells. It also showed enhanced HR when co-infiltrated with harpinpss in tobacco leaves. We used a transgenic tobacco suspension cells system that constitutively expresses the Pflp gene driven by the CaMV 35S promoter to study the function of PFLP in enhancing harpinpss-mediated hypersensitive cell death in vivo. In response to harpinpss, suspension cells derived from Pflp transgenic tobacco showed a significant increase both in the generation of AOS and in cell death as compared to the wild type. AOS inhibitors diphenylene iodonium chloride (DPI) and lanthanum chlorate (LaCl3) were used to study the involvement of AOS in harpinpss-induced cell death. Our results demonstrate enhanced generation of AOS is necessary to cause enhanced hypersensitive cell death in Pflp transgenic tobacco cells and it is plasma membrane-bound NADPH-oxidase-dependent. Sub-cellular localization studies showed that PFLP is present in the cytoplasm and chloroplast of Pflp transgenic tobacco cells, but only in the chloroplast, not in the cytoplasm, of wild-type tobacco cells. It is possible that PFLP can change the redox state of the cell upon harpinpss inoculation to increase AOS generation and hypersensitive cell death. Overall, this study will provide a new insight in the functional properties of ferredoxin in hypersensitive cell death.",
keywords = "Active oxygen species, Ferredoxin, Hypersensitive cell death, PFLP",
author = "{Venkata Dayakar}, Badri and Lin, {Hao Jan} and Cheng-Hsien Chen and Ger, {Mang Jye} and Lee, {Bor Heng} and Pai, {Chia Hwei} and Chow, {David T.} and Huang, {Hsiang En} and Hwang, {Shaw Yhi} and Chung, {Mei Chu} and Feng, {Teng Yung}",
year = "2003",
month = "4",
doi = "10.1023/A:1023061303755",
language = "English",
volume = "51",
pages = "913--924",
journal = "Plant Molecular Biology",
issn = "0167-4412",
publisher = "Springer Netherlands",
number = "6",

}

TY - JOUR

T1 - Ferredoxin from sweet pepper (Capsicum annuum L.) intensifying harpinpss-mediated hypersensitive response shows an enhanced production of active oxygen species (AOS)

AU - Venkata Dayakar, Badri

AU - Lin, Hao Jan

AU - Chen, Cheng-Hsien

AU - Ger, Mang Jye

AU - Lee, Bor Heng

AU - Pai, Chia Hwei

AU - Chow, David T.

AU - Huang, Hsiang En

AU - Hwang, Shaw Yhi

AU - Chung, Mei Chu

AU - Feng, Teng Yung

PY - 2003/4

Y1 - 2003/4

N2 - The hypersensitive response (HR) is a form of cell death associated with plant resistance to pathogen infection. Harpinpss, an elicitor from the bacterium Pseudomonas syringae pv. syringae, induces a HR in non-host plants. Previously, we reported an amphipathic protein from sweet pepper interfering with harpinpss-mediated HR. In this report, we isolated and characterized a cDNA clone encoded that amphipathic protein from sweet pepper. This protein is designated as PFLP (plant ferredoxin-like protein) by virtue of its high homology with plant ferredoxin protein containing an N-terminal signal peptide responsible for chloroplast targeting and a putative 2Fe-2S domain responsible for redox activity. Recombinant PFLP obtained from Escherichia coli was able to significantly increase active oxygen species (AOS) generation when mixed with harpinpss in tobacco suspension cells. It also showed enhanced HR when co-infiltrated with harpinpss in tobacco leaves. We used a transgenic tobacco suspension cells system that constitutively expresses the Pflp gene driven by the CaMV 35S promoter to study the function of PFLP in enhancing harpinpss-mediated hypersensitive cell death in vivo. In response to harpinpss, suspension cells derived from Pflp transgenic tobacco showed a significant increase both in the generation of AOS and in cell death as compared to the wild type. AOS inhibitors diphenylene iodonium chloride (DPI) and lanthanum chlorate (LaCl3) were used to study the involvement of AOS in harpinpss-induced cell death. Our results demonstrate enhanced generation of AOS is necessary to cause enhanced hypersensitive cell death in Pflp transgenic tobacco cells and it is plasma membrane-bound NADPH-oxidase-dependent. Sub-cellular localization studies showed that PFLP is present in the cytoplasm and chloroplast of Pflp transgenic tobacco cells, but only in the chloroplast, not in the cytoplasm, of wild-type tobacco cells. It is possible that PFLP can change the redox state of the cell upon harpinpss inoculation to increase AOS generation and hypersensitive cell death. Overall, this study will provide a new insight in the functional properties of ferredoxin in hypersensitive cell death.

AB - The hypersensitive response (HR) is a form of cell death associated with plant resistance to pathogen infection. Harpinpss, an elicitor from the bacterium Pseudomonas syringae pv. syringae, induces a HR in non-host plants. Previously, we reported an amphipathic protein from sweet pepper interfering with harpinpss-mediated HR. In this report, we isolated and characterized a cDNA clone encoded that amphipathic protein from sweet pepper. This protein is designated as PFLP (plant ferredoxin-like protein) by virtue of its high homology with plant ferredoxin protein containing an N-terminal signal peptide responsible for chloroplast targeting and a putative 2Fe-2S domain responsible for redox activity. Recombinant PFLP obtained from Escherichia coli was able to significantly increase active oxygen species (AOS) generation when mixed with harpinpss in tobacco suspension cells. It also showed enhanced HR when co-infiltrated with harpinpss in tobacco leaves. We used a transgenic tobacco suspension cells system that constitutively expresses the Pflp gene driven by the CaMV 35S promoter to study the function of PFLP in enhancing harpinpss-mediated hypersensitive cell death in vivo. In response to harpinpss, suspension cells derived from Pflp transgenic tobacco showed a significant increase both in the generation of AOS and in cell death as compared to the wild type. AOS inhibitors diphenylene iodonium chloride (DPI) and lanthanum chlorate (LaCl3) were used to study the involvement of AOS in harpinpss-induced cell death. Our results demonstrate enhanced generation of AOS is necessary to cause enhanced hypersensitive cell death in Pflp transgenic tobacco cells and it is plasma membrane-bound NADPH-oxidase-dependent. Sub-cellular localization studies showed that PFLP is present in the cytoplasm and chloroplast of Pflp transgenic tobacco cells, but only in the chloroplast, not in the cytoplasm, of wild-type tobacco cells. It is possible that PFLP can change the redox state of the cell upon harpinpss inoculation to increase AOS generation and hypersensitive cell death. Overall, this study will provide a new insight in the functional properties of ferredoxin in hypersensitive cell death.

KW - Active oxygen species

KW - Ferredoxin

KW - Hypersensitive cell death

KW - PFLP

UR - http://www.scopus.com/inward/record.url?scp=0037720026&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037720026&partnerID=8YFLogxK

U2 - 10.1023/A:1023061303755

DO - 10.1023/A:1023061303755

M3 - Article

C2 - 12777051

AN - SCOPUS:0037720026

VL - 51

SP - 913

EP - 924

JO - Plant Molecular Biology

JF - Plant Molecular Biology

SN - 0167-4412

IS - 6

ER -