TaDAD2, a negative regulator of programmed cell death, is important for the interaction between wheat and the stripe rust fungus.

Defender against cell death (DAD) genes are known to function as negative regulators of cell death in animals. In plants, DAD orthologs are conserved but their role in cell death regulation is not well understood. Here, we report the characterization of the TaDAD2 gene in wheat. The predicted amino acid sequence of TaDAD2 contains typical structural features of DAD proteins, including a signal peptide, three transmembrane regions, and a subunit of oligosaccharyltransferase. Transcripts of TaDAD2 were detected in wheat leaves, culms, roots, florets, and spikelets. The expression level of TaDAD2 was reduced in the initial contact with the stripe rust fungus, subsequently induced and peaked at 18 h postinoculation (hpi), gradually reduced at 24 to 48 hpi, and restored to control level at 72 to 120 hpi. In addition, TaDAD2 exhibited positive transcriptional responses to abiotic stresses after the initial reduction at 1 hpi. Overexpression of TaDAD2 in tobacco leaves inhibited cell death. Furthermore, knocking down TaDAD2 expression by virus-induced gene silencing enhanced the susceptibility of wheat cv. Suwon11 to avirulent race CYR23 and reduced necrotic area at the infection sites. These results indicate that TaDAD2 may function as a suppressor of cell death in the early stages of wheat-stripe rust fungus interaction. However, it is dispensable for or plays an opposite role in hypersensitive response or cell death triggered by an avirulent race of stripe rust fungus at late-infection stages.

[1]  Xianming Chen,et al.  Identification of expressed genes during compatible interaction between stripe rust (Puccinia striiformis) and wheat using a cDNA library , 2009, BMC Genomics.

[2]  H. Yoshioka,et al.  Molecular mechanisms of generation for nitric oxide and reactive oxygen species, and role of the radical burst in plant immunity , 2009, Molecules and cells.

[3]  Xianming Chen,et al.  cDNA-AFLP analysis reveals differential gene expression in compatible interaction of wheat challenged with Puccinia striiformis f. sp. tritici , 2009, BMC Genomics.

[4]  Ling Zhu,et al.  Molecular cloning and responsive expression to injury stimulus of a defender against cell death 1 (DAD1) gene from bay scallops Argopecten irradians , 2008, Molecular Biology Reports.

[5]  Shiv D. Kale,et al.  Conserved C-Terminal Motifs Required for Avirulence and Suppression of Cell Death by Phytophthora sojae effector Avr1b[W] , 2008, The Plant Cell Online.

[6]  O. Mittapalli,et al.  Molecular characterization and responsive expression of a defender against apoptotic cell death homologue from the Hessian fly, Mayetiola destructor. , 2008, Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology.

[7]  Lili Huang,et al.  The development of a PCR-based method for detecting Puccinia striiformis latent infections in wheat leaves , 2008, European Journal of Plant Pathology.

[8]  Lili Huang,et al.  Histochemical studies on the accumulation of reactive oxygen species (O2− and H2O2) in the incompatible and compatible interaction of wheat—Puccinia striiformis f. sp. tritici , 2007 .

[9]  B. Rao,et al.  A homologue of the defender against the apoptotic death gene (dad1) in UV-exposed Chlamydomonas cells is downregulated with the onset of programmed cell death , 2007, Journal of Biosciences.

[10]  F. Van Breusegem,et al.  Reactive oxygen species as signals that modulate plant stress responses and programmed cell death , 2006, BioEssays : news and reviews in molecular, cellular and developmental biology.

[11]  E. Lam,et al.  Arabidopsis Bax inhibitor-1 functions as an attenuator of biotic and abiotic types of cell death. , 2006, The Plant journal : for cell and molecular biology.

[12]  M. Delledonne NO news is good news for plants. , 2005, Current opinion in plant biology.

[13]  H. Uchimiya,et al.  Mitochondrial behaviour in the early stages of ROS stress leading to cell death in Arabidopsis thaliana. , 2005, Annals of botany.

[14]  K. Shirasu,et al.  Virus-Induced Gene Silencing-Based Functional Characterization of Genes Associated with Powdery Mildew Resistance in Barley1 , 2005, Plant Physiology.

[15]  B. Gill,et al.  Development of a Virus-Induced Gene-Silencing System for Hexaploid Wheat and Its Use in Functional Analysis of the Lr21-Mediated Leaf Rust Resistance Pathway1 , 2005, Plant Physiology.

[16]  Kathryn A. O’Donnell,et al.  c-Myc-regulated microRNAs modulate E2F1 expression , 2005, Nature.

[17]  S. Mulero-Navarro,et al.  Resveratrol‐induced apoptosis in MCF‐7 human breast cancer cells involves a caspase‐independent mechanism with downregulation of Bcl‐2 and NF‐κB , 2005, International journal of cancer.

[18]  D. Yun,et al.  Mammalian Bax initiates plant cell death through organelle destruction , 2005, Plant Cell Reports.

[19]  R. Mittler,et al.  Reactive oxygen gene network of plants. , 2004, Trends in plant science.

[20]  Sang Yeol Lee,et al.  Bax-induced cell death of Arabidopsisis meditated through reactive oxygen-dependent and -independent processes , 2004, Plant Molecular Biology.

[21]  Y. Higuchi Chromosomal DNA fragmentation in apoptosis and necrosis induced by oxidative stress. , 2003, Biochemical pharmacology.

[22]  A. Heck,et al.  Homodimeric galectin-7 (p53-induced gene 1) is a negative growth regulator for human neuroblastoma cells , 2003, Oncogene.

[23]  D. Klessig,et al.  The Pathogen-Inducible Nitric Oxide Synthase (iNOS) in Plants Is a Variant of the P Protein of the Glycine Decarboxylase Complex , 2003, Cell.

[24]  Ji Hee Han,et al.  cDNA cloning of a defender against apoptotic cell death 1 (DAD1) homologue, responsive to external temperature stimulus from the spider, Araneus ventricosus , 2003 .

[25]  H. Matsumura,et al.  Overexpression of Bax inhibitor suppresses the fungal elicitor-induced cell death in rice (Oryza sativa L) cells. , 2003, The Plant journal : for cell and molecular biology.

[26]  S. Kamoun,et al.  Expression of a Phytophthora sojae necrosis-inducing protein occurs during transition from biotrophy to necrotrophy. , 2002, The Plant journal : for cell and molecular biology.

[27]  S. Holzberg,et al.  Barley stripe mosaic virus-induced gene silencing in a monocot plant. , 2002, The Plant journal : for cell and molecular biology.

[28]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[29]  F. J. Geske,et al.  The biology of apoptosis. , 2001, Human pathology.

[30]  Naohiro Kato,et al.  Programmed cell death, mitochondria and the plant hypersensitive response , 2001, Nature.

[31]  S. Sakurai,et al.  Ecdysteroid-inducible genes in the programmed cell death during insect metamorphosis. , 2001, Insect biochemistry and molecular biology.

[32]  C. Leaver,et al.  Programmed cell death in cell cultures , 2000, Plant Molecular Biology.

[33]  M. C. Heath Hypersensitive response-related death , 2000, Plant Molecular Biology.

[34]  M. Nei,et al.  Molecular Evolution and Phylogenetics , 2000 .

[35]  E. Lam,et al.  Nitric oxide and salicylic acid signaling in plant defense. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[36]  J. Brewster,et al.  Deletion of Dad1 in mice induces an apoptosis‐associated embryonic death , 2000, Genesis.

[37]  G. Bolwell Role of active oxygen species and NO in plant defence responses. , 1999, Current opinion in plant biology.

[38]  L. C. Loon,et al.  The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins , 1999 .

[39]  M. Miura,et al.  Animal cell-death suppressors Bcl-xL and Ced-9 inhibit cell death in tobacco plants , 1999, Current Biology.

[40]  S. Santa Cruz,et al.  Bax-induced cell death in tobacco is similar to the hypersensitive response. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[41]  J. Sample,et al.  Epstein-Barr Virus Regulates c-MYC, Apoptosis, and Tumorigenicity in Burkitt Lymphoma , 1999, Molecular and Cellular Biology.

[42]  J. Dangl,et al.  Dead cells do tell tales. , 1998, Current opinion in plant biology.

[43]  M. C. Heath,et al.  Changes in the cytoskeleton accompanying infection-induced nuclear movements and the hypersensitive response in plant cells invaded by rust fungi. , 1998, The Plant journal : for cell and molecular biology.

[44]  R. Dixon,et al.  A Plant Homolog of the Neutrophil NADPH Oxidase gp91phox Subunit Gene Encodes a Plasma Membrane Protein with Ca2+ Binding Motifs , 1998, Plant Cell.

[45]  T. Makishima,et al.  An Arabidopsis thaliana cDNA complementing a hamster apoptosis suppressor mutant. , 1997, The Plant journal : for cell and molecular biology.

[46]  L. C. Loon Induced resistance in plants and the role of pathogenesis-related proteins , 1997, European Journal of Plant Pathology.

[47]  N. Tapon,et al.  A programmed cell death pathway activated in carrot cells cultured at low cell density , 1997 .

[48]  A. Granell,et al.  The plant homologue of the defender against apoptotic death gene is down‐regulated during senescence of flower petals 1 , 1997, FEBS letters.

[49]  J. Rothman,et al.  dad‐1, an endogenous programmed cell death suppressor in Caenorhabditis elegans and vertebrates. , 1995, The EMBO journal.

[50]  E. Ward,et al.  A Central Role of Salicylic Acid in Plant Disease Resistance , 1994, Science.

[51]  J. Thompson,et al.  CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. , 1994, Nucleic acids research.

[52]  F. Ausubel,et al.  The A. thaliana disease resistance gene RPS2 encodes a protein containing a nucleotide-binding site and leucine-rich repeats , 1994, Cell.

[53]  S. Komiyama,et al.  Molecular cloning of a human cDNA encoding a novel protein, DAD1, whose defect causes apoptotic cell death in hamster BHK21 cells , 1993, Molecular and cellular biology.

[54]  Leslie Friedrich,et al.  Requirement of Salicylic Acid for the Induction of Systemic Acquired Resistance , 1993, Science.

[55]  R. W. Jones,et al.  Identification of barley stripe mosaic virus genes involved in viral RNA replication and systemic movement. , 1990, The EMBO journal.

[56]  J. Parlevliet Pleiotropic association of infection frequency and latent period of two barley cultivars partially resistant to barley leaf rust , 1986, Euphytica.

[57]  L. Hong,et al.  Inheritance of Yellow Rust Resistance in an Elite Wheat Germplasm Xingzi 9104 , 2006 .

[58]  Liu Hongmei,et al.  Inheritance of yellow rust resistance in an elite wheat germplasm Xingzi 9104 , 2006 .

[59]  W. Dawson,et al.  Tobamovirus Transient Expression Vectors: Tools for Plant Biology and High-Level Expression of Foreign Proteins in Plants , 1998 .

[60]  T. Makishima,et al.  dad-1, A putative programmed cell death suppressor gene in rice. , 1997, Plant & cell physiology.