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Identification and molecular characterization of a β-1,4-endoglucanase gene (Rr-eng-1) from Rotylenchulus reniformis
- Abstract::
- β-1,4-endoglucanses, a.k.a. cellulases, are parasitism genes that facilitate root penetration and migration by plant-parasitic nematodes. Rotylenchulus reniformis is a sedentary semi-endoparasite for which little molecular data has been collected. In this report, we describe the isolation and characterization of a predicted glycosyl hydrolase family 5 cellulase from R. reniformis that we have named Rr-eng-1. The Rr-eng-1 cDNA was 1,341 bp long and was comprised of a 19 bp 5′-untranslated region (UTR), a 1,245 bp open reading frame (ORF), and an 80 bp 3′-UTR. The Rr-eng-1 genomic sequence was 2,325 bp. Alignment of the cDNA and genomic sequences revealed seven introns and eight exons for Rr-eng-1. BLASTN analysis showed the Rr-eng-1 cDNA was most homologous to the Hg-eng-6 mRNA from Heterodera glycines. Southern blot analysis indicated that at least three Rr-eng-1-like sequences were present in the R. reniformis genome. Translation of the Rr-eng-1 ORF yielded a 414 amino acid peptide (Rr-ENG-1) having an N-terminal signal sequence for secretion. No cellulose binding module (CBM) was detected in Rr-ENG-1; however, a putative CBM linker sequence N-terminal to the catalytic domain was present. Rr-ENG-1 was most homologous to Hg-ENG-6 but also shared a number of intron splice positions with Mi-ENG-2. Quantitative RT-PCR indicated that Rr-eng-1 was highly expressed in the J2 and adult vermiform life-stages with a sharp decline in expression detected in sedentary females.
- Author(s):
-
Wubben, Martin J. , Ganji, Satish , Callahan, Franklin E.
- Subject(s):
-
Heterodera glycines , Rotylenchulus reniformis , Southern blotting , active sites , cellulose , complementary DNA , endo-1,4-beta-glucanase , exons , females , gene expression regulation , genes , introns , messenger RNA , open reading frames , secretion , signal peptide
- Description:
- Includes references
- Source:
- Journal of nematology 2010 Dec., v. 42, no. 4
- Language:
- English
- Year:
- 2010
- Collection:
-
Journal Articles, USDA Authors, Peer-Reviewed
- File:
-
Download [PDF]
- Rights:
- Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted.
