Isolation and characterization of a mannose-binding insecticidal lectin gene from Allium sativum (garlic) and its putative role in insect resistance using bioinformatics tools.
Monocot mannose-binding lectins (MMBLs) or agglutinins are an extended superfamily of structurally and evolutionarily related proteins. They play important roles in plant defenses. Here we describe the synthesis of full-length cDNA of monocot mannose-binding insecticidal agglutinin isolated from Allium sativum, a traditional herb known to be of great applications in Africa, using reverse transcription polymerase chain reaction (RT-PCR) with specific primers designed based on the insecticidal sequence (NCBI primary accession no. AY866499.1). Sequence analysis revealed a 327bp open reading frame (ORF) encoding a putative 108-aa agglutinin precursor with a C-terminal domain. Multiple alignments of BLEC1 amino acids with those of eight other MMBLs revealed three highly conserved domains among them, indicating BLEC1 belongs to a member of the MMBL superfamily. Tertiary structure analysis showed that BLEC1 had three potential equal mannose-binding sites. Phylogenetic analysis indicated that 20 MMBLs including BLEC1 belonged to an extended superfamily. Gene ontology analyses indicate one biological process with GO ID: 0006952 representing defense response, with two secondary IDs GO: 0002217 GO: 0042829. The child terms has both negative and positive regulation some of which include GO: 0002242 defense response to parasitic plant and GO: 0002213 defense response to insect. The cloning and characterization of BLEC1 will enable us to study its potential use in plant genetic engineering in the development of insect resistance plant.
Centre for Biotechnology Research and Training, Ahmadu Bello University, Zaria, Nigeria. nbafolabi-balogun@abu.edu.ng
N B
NB
Afolabi-Balogun
H M
HM
Inuwa
M F
MF
Ishiyaku
M T
MT
Bakare-Odunoola
A J
AJ
Nok
D000595:Amino Acid Sequence; D001483:Base Sequence; D019295:Computational Biology; D060467:Disease Resistance / Q000235:genetics*; D005737:Garlic / Q000235:genetics* / Q000276:immunology; D037241:Mannose-Binding Lectins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D010935:Plant Diseases / Q000276:immunology; D037121:Plant Lectins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D037241:Mannose-Binding Lectins; D037121:Plant Lectins