Expression and Functional Roles of the Pepper Pathogen–Induced bZIP Transcription Factor CabZIP2 in Enhanced Disease Resistance to Bacterial Pathogen Infection

A pepper bZIP transcription factor gene, CabZIP2, was isolated from pepper leaves infected with a virulent strain of Xanthomonas campestris pv. vesicatoria. Transient expression analysis of the CabZIP2-GFP fusion protein in Nicotiana benthamiana revealed that the CabZIP2 protein is localized in the cytoplasm as well as the nucleus. The acidic domain in the N-terminal region of CabZIP2 that is fused to the GAL4 DNA-binding domain is required to activate the transcription of reporter genes in yeast. Transcription of CabZIP2 is induced in pepper plants inoculated with virulent or avirulent strains of X. campestris pv. vesicatoria. The CabZIP2 gene is also induced by defense-related hormones such as salicylic acid, methyl jasmonate, and ethylene. To elucidate the in vivo function of the CabZIP2 gene in plant defense, virus-induced gene silencing in pepper and overexpression in Arabidopsis were used. CabZIP2-silenced pepper plants were susceptible to infection by the virulent strain of X. campestris pv. vesicatoria, which was accompanied by reduced expression of defense-related genes such as CaBPR1 and CaAMP1. CabZIP2 overexpression in transgenic Arabidopsis plants conferred enhanced resistance to Pseudomonas syringae pv. tomato DC3000. Together, these results suggest that CabZIP2 is involved in bacterial disease resistance.

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Functional analysis of the heterotrimeric NF-Y transcription factor complex in cassava disease resistance

Annals of Botany ◽

10.1093/aob/mcz115 ◽

2019 ◽

Vol 124 (7) ◽

pp. 1185-1197 ◽

Cited By ~ 2

Author(s):

Xinyi He ◽

Guoyin Liu ◽

Bing Li ◽

Yanwei Xie ◽

Yunxie Wei ◽

...

Keyword(s):

Transcription Factor ◽

Disease Resistance ◽

Protein Interactions ◽

Transcriptional Activation ◽

Plant Disease Resistance ◽

Plant Defence ◽

Virus Induced Gene Silencing ◽

Cassava Bacterial Blight ◽

Transcription Factor Complex

Abstract Background and Aims The nuclear factor Y (NF-Y) transcription factor complex is important in plant growth, development and stress response. Information regarding this transcription factor complex is limited in cassava (Manihot esculenta). In this study, 15 MeNF-YAs, 21 MeNF-YBs and 15 MeNF-YCs were comprehensively characterized during plant defence. Methods Gene expression in MeNF-Ys was examined during interaction with the bacterial pathogen Xanthomonas axonopodis pv. manihotis (Xam). The yeast two-hybrid system was employed to investigate protein–protein interactions in the heterotrimeric NF-Y transcription factor complex. The in vivo roles of MeNF-Ys were revealed by virus-induced gene silencing (VIGS) in cassava. Key Results The regulation of MeNF-Ys in response to Xam indicated their possible roles in response to cassava bacterial blight. Protein–protein interaction assays identified the heterotrimeric NF-Y transcription factor complex (MeNF-YA1/3, MeNF-YB11/16 and MeNF-YC11/12). Moreover, the members of the heterotrimeric NF-Y transcription factor complex were located in the cell nucleus and conferred transcriptional activation activity to the CCAAT motif. Notably, the heterotrimeric NF-Y transcription factor complex positively regulated plant disease resistance to Xam, confirmed by a disease phenotype in overexpressing plants in Nicotiana benthamiana and VIGS in cassava. Consistently, the heterotrimeric NF-Y transcription factor complex positively regulated the expression of pathogenesis-related genes (MePRs). Conclusions The NF-Y transcription factor complex (MeNF-YA1/3, MeNF-YB11/16 and MeNF-YC11/12) characterized here was shown to play a role in transcriptional activation of MePR promoters, contributing to the plant defence response in cassava.

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PPI1: A Novel Pathogen-Induced Basic Region-Leucine Zipper (bZIP) Transcription Factor from Pepper

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.2002.15.6.540 ◽

2002 ◽

Vol 15 (6) ◽

pp. 540-548 ◽

Cited By ~ 30

Author(s):

Sang Jik Lee ◽

Mi Yeon Lee ◽

So Young Yi ◽

Sang Keun Oh ◽

Soon Ho Choi ◽

...

Keyword(s):

Transcription Factor ◽

Dna Binding ◽

Plant Defense ◽

Pseudomonas Syringae ◽

Xanthomonas Campestris ◽

Leucine Zipper ◽

Binding Protein ◽

Dna Binding Protein ◽

Bzip Transcription Factor ◽

Basic Region

We have isolated a full-length cDNA, PPI1 (pepper-PMMV interaction 1), encoding a novel basic region-leucine zipper (bZIP) DNA-binding protein, from expressed sequence tags differentially expressed in Capsicum chinense PI257284 infected with Pepper mild mottle virus (PMMV). PPI1 encodes a predicted protein of 170 amino acids and contains a putative DNA-binding domain that shares significant amino acid identity with ACGT-binding domains of members of the bZIP DNA-binding protein family. PPI1 was localized in the nucleus and had transcriptional activation activity in yeast. Transcripts of the PPI1 gene were preferentially induced during an incompatible interaction by inoculation with PMMV, Pseudomonas syringae pv. syringae 61, and Xanthomonas campestris pv. vesicatoria race 3. However, the PPI1 gene was not induced by abiotic stressors that activate the plant defense-signaling pathway. Our data provide the first evidence that a bZIP transcription factor is preferentially induced by pathogen attack, suggesting that PPI1 may play a specific functional role in the regulation of expression of plant defense-related genes.

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Comparative Mapping of Three Bacterial, Three Fungal, and One Virus Disease-resistance Genes in Common Bean

HortScience ◽

10.21273/hortsci.33.3.547a ◽

1998 ◽

Vol 33 (3) ◽

pp. 547a-547

Author(s):

Geunhwa Jung ◽

James Nienhuis ◽

Dermot P. Coyne ◽

H.M. Ariyarathne

Keyword(s):

Disease Resistance ◽

Common Bean ◽

Pseudomonas Syringae ◽

Resistance Genes ◽

Fungal Pathogens ◽

Xanthomonas Campestris ◽

Virus Disease ◽

Disease Resistance Genes ◽

Brown Spot ◽

Viral Pathogen

Common bacterial blight (CBB), bacterial brown spot (BBS), and halo blight (HB), incited by the bacterial pathogens Xanthomonas campestris pv. phaseoli (Smith) Dye, Pseodomonas syringae pv. syringa, and Pseudomonas syringae pv. phaseolicola, respectively are important diseases of common bean. In addition three fungal pathogens, web blight (WB) Thanatephorus cucumeris, rust Uromyces appendiculatus, and white mold (WM) Sclerotinia sclerotiorum, are also destructive diseases attacking common bean. Bean common mosaic virus is also one of most major virus disease. Resistance genes (QTLs and major genes) to three bacterial (CBB, BBS, and HB), three fungal (WB, rust, and WM), and one viral pathogen (BCMV) were previously mapped in two common bean populations (BAC 6 × HT 7719 and Belneb RR-1 × A55). The objective of this research was to use an integrated RAPD map of the two populations to compare the positions and effect of resistance QTL in common bean. Results indicate that two chromosomal regions associated with QTL for CBB resistance mapped in both populations. The same chromosomal regions associated with QTL for disease resistance to different pathogens or same pathogens were detected in the integrated population.

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Identification of Three Putative Signal Transduction Genes Involved in R Gene-Specified Disease Resistance in Arabidopsis

Genetics ◽

10.1093/genetics/152.1.401 ◽

1999 ◽

Vol 152 (1) ◽

pp. 401-412 ◽

Cited By ~ 1

Author(s):

Randall F Warren ◽

Peter M Merritt ◽

Eric Holub ◽

Roger W Innes

Keyword(s):

Disease Resistance ◽

Pseudomonas Syringae ◽

Resistance Genes ◽

Virulent Strain ◽

Disease Resistance Genes ◽

Avirulence Gene ◽

Detectable Effect ◽

Disease Resistance Gene ◽

Protein Functions ◽

Signal Transduction Genes

Abstract The RPS5 disease resistance gene of Arabidopsis mediates recognition of Pseudomonas syringae strains that possess the avirulence gene avrPphB. By screening for loss of RPS5-specified resistance, we identified five pbs (avrPphB susceptible) mutants that represent three different genes. Mutations in PBS1 completely blocked RPS5-mediated resistance, but had little to no effect on resistance specified by other disease resistance genes, suggesting that PBS1 facilitates recognition of the avrPphB protein. The pbs2 mutation dramatically reduced resistance mediated by the RPS5 and RPM1 resistance genes, but had no detectable effect on resistance mediated by RPS4 and had an intermediate effect on RPS2-mediated resistance. The pbs2 mutation also had varying effects on resistance mediated by seven different RPP (recognition of Peronospora parasitica) genes. These data indicate that the PBS2 protein functions in a pathway that is important only to a subset of disease-resistance genes. The pbs3 mutation partially suppressed all four P. syringae-resistance genes (RPS5, RPM1, RPS2, and RPS4), and it had weak-to-intermediate effects on the RPP genes. In addition, the pbs3 mutant allowed higher bacterial growth in response to a virulent strain of P. syringae, indicating that the PBS3 gene product functions in a pathway involved in restricting the spread of both virulent and avirulent pathogens. The pbs mutations are recessive and have been mapped to chromosomes I (pbs2) and V (pbs1 and pbs3).

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Overexpression of Pto Induces a Salicylate-Independent Cell Death But Inhibits Necrotic Lesions Caused by Salicylate-Deficiency in Tomato Plants

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.2002.15.7.654 ◽

2002 ◽

Vol 15 (7) ◽

pp. 654-661 ◽

Cited By ~ 16

Author(s):

Jianxiong Li ◽

Libo Shan ◽

Jian-Min Zhou ◽

Xiaoyan Tang

Keyword(s):

Cell Death ◽

Disease Resistance ◽

Pseudomonas Syringae ◽

Virulent Strain ◽

Gene Activation ◽

Cellular Damage ◽

Minor Role ◽

Disease Resistance Gene ◽

Tomato Plants ◽

Spontaneous Cell Death

Tomato plants overexpressing the disease resistance gene Pto (35S∷Pto) exhibit spontaneous cell death, accumulation of salicylic acid (SA), elevated expression of pathogenesis-related genes, and enhanced resistance to a broad range of pathogens. Because salicylate plays an important role in the cell death and defense activation in many lesion mimic mutants, we investigated the interaction of SA-mediated processes and the 35S∷Pto-mediated defense pathway by introducing the nahG transgene that encodes salicylate hydroxylase. Here, we show that SA is not required for the 35S∷Pto-activated microscopic cell death and plays a minor role in defense gene activation and general disease resistance in 35S∷Pto plants. In contrast, temperature greatly affects the spontaneous cell death and general resistance in 35S∷Pto plants, and high temperature inhibits the cell death. The NahG tomato plants develop spontaneous, unconstrained necrotic lesions on leaves. These lesions also are initiated by the inoculation of a virulent strain of Pseudomonas syringae pv. tomato. However, the NahG-dependent necrotic lesions are inhibited in the NahG/35S∷Pto plants. This inhibition is most pronounced under conditions favoring the 35S∷Pto-mediated spontaneous cell death development. These results indicate that the signaling pathways activated by Pto overexpression suppress the cellular damage that is caused by SA depletion. We also found that ethylene is dispensable for the 35S∷Pto-mediated general defense.

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The Hypersensitive Induced Reaction and Leucine-Rich Repeat Proteins Regulate Plant Cell Death Associated with Disease and Plant Immunity

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-02-10-0030 ◽

2011 ◽

Vol 24 (1) ◽

pp. 68-78 ◽

Cited By ~ 45

Author(s):

Hyong Woo Choi ◽

Young Jin Kim ◽

Byung Kook Hwang

Keyword(s):

Cell Death ◽

Xanthomonas Campestris ◽

Plant Immunity ◽

Yeast Cells ◽

Leucine Rich Repeat ◽

Virus Induced Gene Silencing ◽

Susceptible Cell ◽

Gene Transcripts ◽

Induced Reaction ◽

Pepper Plants

Pathogen-induced programmed cell death (PCD) is intimately linked with disease resistance and susceptibility. However, the molecular components regulating PCD, including hypersensitive and susceptible cell death, are largely unknown in plants. In this study, we show that pathogen-induced Capsicum annuum hypersensitive induced reaction 1 (CaHIR1) and leucine-rich repeat 1 (CaLRR1) function as distinct plant PCD regulators in pepper plants during Xanthomonas campestris pv. vesicatoria infection. Confocal microscopy and protein gel blot analyses revealed that CaLRR1 and CaHIR1 localize to the extracellular matrix and plasma membrane (PM), respectively. Bimolecular fluorescent complementation and coimmunoprecipitation assays showed that the extracellular CaLRR1 specifically binds to the PM-located CaHIR1 in pepper leaves. Overexpression of CaHIR1 triggered pathogen-independent cell death in pepper and Nicotiana benthamiana plants but not in yeast cells. Virus-induced gene silencing (VIGS) of CaLRR1 and CaHIR1 distinctly strengthened and compromised hypersensitive and susceptible cell death in pepper plants, respectively. Endogenous salicylic acid levels and pathogenesis-related gene transcripts were elevated in CaHIR1-silenced plants. VIGS of NbLRR1 and NbHIR1, the N. benthamiana orthologs of CaLRR1 and CaHIR1, regulated Bax- and avrPto-/Pto-induced PCD. Taken together, these results suggest that leucine-rich repeat and hypersensitive induced reaction proteins may act as cell-death regulators associated with plant immunity and disease.

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Environmental Control in Tea Fields to Reduce Infection by Pseudomonas syringae pv. theae

Phytopathology ◽

10.1094/phyto-99-2-0209 ◽

2009 ◽

Vol 99 (2) ◽

pp. 209-216 ◽

Cited By ~ 6

Author(s):

T. Tomihama ◽

T. Nonaka ◽

Y. Nishi ◽

K. Arai

Keyword(s):

Low Temperature ◽

Pseudomonas Syringae ◽

Xanthomonas Campestris ◽

Environmental Control ◽

Microbial Interactions ◽

Ice Formation ◽

Bacterial Disease ◽

Environmental Controls ◽

Frost Protection ◽

Tea Plants

Bacterial shoot blight (BSB) disease, caused by Pseudomonas syringae pv. theae, is a major bacterial disease of tea plants in Japan. BSB mainly occurs in the low-temperature season, and lesion formation by P. syringae pv. theae is enhanced by both low temperature and the presence of ice nucleation-active Xanthomonas campestris (INAX), which catalyzes ice formation at –2 to –4°C and is frequently co-isolated with P. syringae pv. theae from tea plants. Low temperature is thus the most important environmental factor influencing the incidence of BSB; however, the effects of low temperature on infection of the host by P. syringae pv. theae and of environmental controls in fields on the occurrence of the disease are poorly understood. In this study, we show that ice formation on tea leaves by INAX enhanced P. syringae pv. theae invasion into leaf tissue. The natural incidence of BSB in the field was closely related to early autumn frost. Frost protection in late autumn, which prevented ice formation on tea plants, significantly decreased the incidence of BSB, and frost protection combined with bactericide application held the incidence under the economic threshold level. Our data indicate that environmental control in the field based on microbial interactions in the host offers a new strategy for plant disease control.

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MAPK-directed activation of the whitefly transcription factor CREB leads to P450-mediated imidacloprid resistance

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1913603117 ◽

2020 ◽

Vol 117 (19) ◽

pp. 10246-10253 ◽

Cited By ~ 7

Author(s):

Xin Yang ◽

Shun Deng ◽

Xuegao Wei ◽

Jing Yang ◽

Qiannan Zhao ◽

...

Keyword(s):

Transcription Factor ◽

Signaling Pathway ◽

Gene Families ◽

Mapk Signaling ◽

Mitogen Activated Protein Kinase ◽

Bzip Transcription Factor ◽

Camp Response Element ◽

Response Element

The evolution of insect resistance to pesticides poses a continuing threat to agriculture and human health. While much is known about the proximate molecular and biochemical mechanisms that confer resistance, far less is known about the regulation of the specific genes/gene families involved, particularly by trans-acting factors such as signal-regulated transcription factors. Here we resolve in fine detail the trans-regulation of CYP6CM1, a cytochrome P450 that confers resistance to neonicotinoid insecticides in the whitefly Bemisia tabaci, by the mitogen-activated protein kinase (MAPK)-directed activation of the transcription factor cAMP-response element binding protein (CREB). Reporter gene assays were used to identify the putative promoter of CYP6CM1, but no consistent polymorphisms were observed in the promoter of a resistant strain of B. tabaci (imidacloprid-resistant, IMR), which overexpresses this gene, compared to a susceptible strain (imidacloprid-susceptible, IMS). Investigation of potential trans-acting factors using in vitro and in vivo assays demonstrated that the bZIP transcription factor CREB directly regulates CYP6CM1 expression by binding to a cAMP-response element (CRE)-like site in the promoter of this gene. CREB is overexpressed in the IMR strain, and inhibitor, luciferase, and RNA interference assays revealed that a signaling pathway of MAPKs mediates the activation of CREB, and thus the increased expression of CYP6CM1, by phosphorylation-mediated signal transduction. Collectively, these results provide mechanistic insights into the regulation of xenobiotic responses in insects and implicate both the MAPK-signaling pathway and a transcription factor in the development of pesticide resistance.

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C-Terminal Region of Plant Ferredoxin-Like Protein Is Required to Enhance Resistance to Bacterial Disease in Arabidopsis thaliana

Phytopathology ◽

10.1094/phyto-08-10-0220 ◽

2011 ◽

Vol 101 (6) ◽

pp. 741-749 ◽

Cited By ~ 8

Author(s):

Yi-Hsien Lin ◽

Hsiang-En Huang ◽

Yen-Ru Chen ◽

Pei-Luan Liao ◽

Ching-Lian Chen ◽

...

Keyword(s):

Disease Resistance ◽

Transgenic Plants ◽

Nadph Oxidase ◽

Pseudomonas Syringae ◽

Induced Defense ◽

Defense Responses ◽

Bacterial Disease ◽

Oxidase Gene ◽

Induced Defense Responses ◽

Resistance To Ralstonia Solanacearum

Protein phosphorylation is an important biological process associated with elicitor-induced defense responses in plants. In a previous report, we described how plant ferredoxin-like protein (PFLP) in transgenic plants enhances resistance to bacterial pathogens associated with the hypersensitive response (HR). PFLP possesses a putative casein kinase II phosphorylation (CK2P) site at the C-terminal in which phosphorylation occurs rapidly during defense response. However, the contribution of this site to the enhancement of disease resistance and the intensity of HR has not been clearly demonstrated. In this study, we generated two versions of truncated PFLP, PEC (extant CK2P site) and PDC (deleted CK2P site), and assessed their ability to trigger HR through harpin (HrpZ) derived from Pseudomonas syringae as well as their resistance to Ralstonia solanacearum. In an infiltration assay of HrpZ, PEC intensified harpin-mediated HR; however, PDC negated this effect. Transgenic plants expressing these versions indicate that nonphosphorylated PFLP loses its ability to induce HR or enhance disease resistance against R. solanacearum. Interestingly, the CK2P site of PFLP is required to induce the expression of the NADPH oxidase gene, AtrbohD, which is a reactive oxygen species producing enzyme. This was further confirmed by evaluating the HR on NADPH oxidase in mutants of Arabidopsis. As a result, we have concluded that the CK2P site is required for the phosphorylation of PFLP to enhance disease resistance.

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The Zipper Region of Epstein-Barr Virus bZIP Transcription Factor Zta Is Necessary but Not Sufficient To Direct DNA Binding

Journal of Virology ◽

10.1128/jvi.77.14.8173-8177.2003 ◽

2003 ◽

Vol 77 (14) ◽

pp. 8173-8177 ◽

Cited By ~ 15

Author(s):

Matthew R. Hicks ◽

Salama S. Al-Mehairi ◽

Alison J. Sinclair

Keyword(s):

Transcription Factor ◽

Dna Binding ◽

Epstein Barr Virus ◽

Bzip Transcription Factor ◽

Barr Virus ◽

Binding Function ◽

Sequence Elements ◽

Epstein Barr ◽

Bzip Proteins

ABSTRACT The viral bZIP transcription factor Zta (BZLF1, EB1, ZEBRA) mediates the switch between the latent and lytic cycles of Epstein-Barr virus (EBV). In part, its activity requires the formation of homodimers and interaction with specific DNA sequence elements (ZREs). Zta has an atypical zipper motif that has a lower stability than do typical bZIP proteins. Here we show that a synthetic peptide directed against the zipper can disrupt the DNA-binding function of Zta. This highlights the relevance of this region for the function of Zta and demonstrates that the zipper region is a potential target for therapeutic agents. We also unmask the relevance of a region adjacent to the zipper (CT region), which is required to direct the interaction of Zta with DNA and to transactivate ZRE-dependent promoters in vivo.

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