title: Molecular cloning of a coiled-coil-nucleotide-binding-site-leucine-rich repeat gene from pearl millet and its expression pattern in response to the downy mildew pathogen.
abstract: Downy mildew caused by Sclerospora graminicola is a devastating disease of pearl millet. Based on candidate gene approach, a set of 22 resistance gene analogues were identified. The clone RGPM 301 (AY117410) containing a partial sequence shared 83% similarity to rice R-proteins. A full-length R-gene RGA RGPM 301 of 3552 bp with 2979 bp open reading frame encoding 992 amino acids was isolated by the degenerate primers and rapid amplification of cDNA ends polymerase chain reaction (RACE-PCR) approach. It had a molecular mass of 113.96 kDa and isoelectric point (pI) of 8.71. The sequence alignment and phylogenetic analysis grouped it to a non-TIR NBS LRR group. The quantitative real-time PCR (qRT-PCR) analysis revealed higher accumulation of the transcripts following inoculation with S. graminicola in the resistant cultivar (IP18296) compared to susceptible cultivar (7042S). Further, significant induction in the transcript levels were observed when treated with abiotic elicitor β-aminobutyric acid (BABA) and biotic elicitor Pseudomonas fluorescens. Exogenous application of phytohormones jasmonic acid or salicylic acid also up-regulated the expression levels of RGA RGPM 301. The treatment of cultivar IP18296 with mitogen-activated protein kinase (MPK) inhibitors (PD98059 and U0126) suppressed the levels of RGA RGPM 301. A 3.5 kb RGA RGPM 301 which is a non-TIR NBS-LRR protein was isolated from pearl millet and its up-regulation during downy mildew interaction was demonstrated by qRT-PCR. These studies indicate a role for this RGA in pearl millet downy mildew interaction.
authors: Department of Studies in Biotechnology, Manasagangotri, University of Mysore, Mysore, Karnataka, 570 006, India.
authors: Mariswamy
authors: M
authors: Veena
authors: Department of Studies in Biotechnology, Manasagangotri, University of Mysore, Mysore, Karnataka, 570 006, India.
authors: Prasad
authors: P
authors: Melvin
authors: Forestry and Agricultural Biotechnology Institute, University of Pretoria, Johannesburg, South Africa.
authors: Sreedhara Ashok
authors: SA
authors: Prabhu
authors: Institution of Excellence, Vijnana Bhavan, Manasagangotri, University of Mysore, Mysore, Karnataka, 570 006, India. shailasree@ioe.uni-mysore.ac.in.
authors: Sekhar
authors: S
authors: Shailasree
authors: Department of Studies in Biotechnology, Manasagangotri, University of Mysore, Mysore, Karnataka, 570 006, India.
authors: Hunthrike Shekar
authors: HS
authors: Shetty
authors: Department of Studies in Biotechnology, Manasagangotri, University of Mysore, Mysore, Karnataka, 570 006, India.
authors: Kukkundoor Ramachandra
authors: KR
authors: Kini
mesh_terms: D000595:Amino Acid Sequence; D000613:Aminobutyrates / Q000494:pharmacology; D001426:Bacterial Proteins; D001483:Base Sequence; D031741:Cenchrus / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology / Q000502:physiology; D003001:Cloning, Molecular; D003517:Cyclopentanes / Q000494:pharmacology; D060467:Disease Resistance; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D009868:Oomycetes; D054883:Oxylipins / Q000494:pharmacology; D031782:Pennisetum / Q000235:genetics* / Q000378:metabolism / Q000382:microbiology / Q000502:physiology; D010802:Phylogeny; D010935:Plant Diseases / Q000382:microbiology*; D010940:Plant Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D011551:Pseudomonas fluorescens; D020156:Salicylic Acid / Q000494:pharmacology; D016415:Sequence Alignment; D015854:Up-Regulation
chemical_list: D000613:Aminobutyrates; D001426:Bacterial Proteins; D003517:Cyclopentanes; D054883:Oxylipins; D010940:Plant Proteins; C047667:3-aminobutyric acid; C011006:jasmonic acid; D020156:Salicylic Acid
keywords: BABA; CC-NBS-LRR; Elicitors; Inhibitors; R genes; qRT-PCR