Molecular characterization and functional analysis of four β‐1, 4‐endoglucanases from the root‐lesion nematode Pratylenchus vulnus. (10th April 2014)

• Record Type: Journal Article
• Authors: Fanelli, E.; Troccoli, A.; Picardi, E.; Pousis, C.; De Luca, F.
• Is Part Of: Plant pathology. Volume 63:Number 6(2014:Dec.)
• Page Start: 1436
• Page End: 1445
• Publication Date: 2014-04-10
• Subjects: Agricultural pests -- Periodicals; Plant diseases -- Periodicals; 632
• DOI: 10.1111/ppa.12222
• Languages: English
• ISSNs: 0032-0862

Abstract

<abstract abstract-type="main" id="ppa12222-abs-0001"> <title> <x xml:space="preserve">Abstract</x> </title> <p>This study reports on the isolation and characterization of four different endoglucanases in the root‐lesion nematode <italic>Pratylenchus vulnus</italic>. The gene structures of two of these, <italic>Pv</italic>‐<italic>eng</italic>‐<italic>1</italic> and <italic>Pv</italic>‐<italic>eng</italic>‐<italic>2</italic>, were fully determined, and <italic>Pv‐eng</italic>‐<italic>3</italic> and <italic>Pv‐eng</italic>‐<italic>5</italic> were partially sequenced. Spatial expression of <italic>Pv</italic>‐<italic>eng</italic>‐<italic>1</italic>, <italic> Pv</italic>‐<italic>eng</italic>‐<italic>2</italic> and <italic>Pv‐eng</italic>‐<italic>5</italic> was examined by <italic>in situ</italic> hybridization. <italic>Pv</italic>‐<italic>eng</italic>‐<italic>1</italic> and <italic>Pv</italic>‐<italic>eng</italic>‐<italic>2</italic> transcripts were localized in the subventral oesophageal glands, whilst the <italic>Pv‐eng</italic>‐<italic>5</italic> transcript was localized in the intestine. Real‐time RT‐PCR showed that three of the four endoglucanases had the highest transcriptional level in adult males and females, thus demonstrating that adults are also parasitic in <italic>P. vulnus</italic>. Mixed stages of <italic>P. vulnus</italic> were treated with double‐stranded RNA (dsRNA) of <italic>Pv</italic>‐<italic>eng</italic>‐<italic>1</italic> in order to study the effect of gene silencing (RNAi). Silencing <italic>Pv</italic>‐<italic>eng</italic>‐<italic>1</italic> by dsRNA targeting of the carbohydrate‐binding module (CBM) resulted in a significant reduction (88–98%) of the transcript level, suggesting that <italic>P. vulnus</italic> is susceptible to RNAi. Furthermore, silencing <italic>P. vulnus</italic> showed a reduction (54%) in nematode reproduction on carrot minidiscs over a 5 week period. These results suggest that silencing of <italic>Pv</italic>‐<italic>eng</italic>‐<italic>1</italic> may result in reduction of the ability of the nematode to locate and invade roots and, therefore, to establish and reproduce.</p> </abstract>