title: Molecular cloning, characterization, and expression analysis of resistance gene candidates in Kaempferia galanga L.
abstract: Majority of the plant disease resistance genes expresses cytoplasmic receptor-like proteins characterized by an N-terminal nucleotide-binding site (NBS) and a leucine-rich repeat (LRR) domain. Degenerative primers based on these conserved motifs were used to isolate NBS type sequences in Kaempferia galanga. Cloning and sequencing identified 12 Kaempferia NBS-type sequences called resistance gene candidates (RGCs) classified into four classes. The amino acid sequences of the RGCs detected the presence of conserved domains, viz., kinase-1a, kinase-2, and hydrophobic GLPL, categorizing them with the NBS-LRR class gene family. Structural and phylogenetic characterization grouped the RGCs with the non-toll interleukin receptor (non-TIR) subclasses of the NBS sequences. Reverse transcription PCR with 10 Kaempferia RGC specific primers revealed 7 out of 10 Kaempferia RGCs to be expressive. The isolation and characterization of Kaempferia RGCs has been reported for the first time in this study. This will provide a starting point towards characterization of candidate resistance genes in Kaempferia and can act as a source pool for disease resistance development in other asexually reproducing plants.
authors.affiliation: Centre of Biotechnology, Siksha O Anusandhan University, Bhubaneswar 751003, India.
authors.forename: Raj Kumar
authors.initials: RK
authors.lastname: Joshi
authors.forename: Basudeba
authors.initials: B
authors.lastname: Kar
authors.forename: Sujata
authors.initials: S
authors.lastname: Mohanty
authors.forename: Enketeswara
authors.initials: E
authors.lastname: Subudhi
authors.forename: Sanghamitra
authors.initials: S
authors.lastname: Nayak
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D003001:Cloning, Molecular; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics; D060467:Disease Resistance / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D014644:Genetic Variation; D008969:Molecular Sequence Data; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D030521:Zingiberaceae / Q000235:genetics* / Q000378:metabolism
chemical_list: D017931:DNA Primers; D018744:DNA, Plant; D010940:Plant Proteins