Ditylenchus destructor
is a migratory plant-parasitic nematode that causes huge damage to global root and tuber production annually. The main plant hosts of
D
.
destructor
contain plenty of starch, which makes the parasitic environment of
D
.
destructor
to be different from those of most other plant-parasitic nematodes. It is speculated that
D
.
destructor
may harbor some unique pathogenesis-related genes to parasitize the starch-rich hosts. Herein, we focused on the multi-copy alpha-amylase genes in
D
.
destructor
, which encode a key starch-catalyzing enzyme. Our previously published
D
.
destructor
genome showed that it has three alpha-amylase encoding genes,
Dd_02440
,
Dd_11154
, and
Dd_13225
. Comparative analysis of alpha-amylases from different species demonstrated that the other plant-parasitic nematodes, even
Ditylenchus dipsaci
in the same genus, harbor only one or no alpha-amylase gene, and the three genes from
D
.
destructor
were closely clustered in the phylogenetic tree, indicating that there was a unique expansion of the alpha-amylase gene in
D
.
destructor
. The enzymatic activity of the three alpha-amylase proteins was verified by an enzyme assay. Quantitative real-time PCR assay showed that the expression of the three alpha-amylase genes in the post-hatching stage of
D
.
destructor
was found to be significantly higher than that in eggs. In the
in situ
hybridization assay, the expression of the genes was localized to the intestine, implying the association of these genes with nematode digestion. An infection assay in sweet potato demonstrated that RNA interference of any one alpha-amylase gene had no influence on the infectivity of
D
.
destructor
. Using the multi-target dsRNA cocktail method, it was found that silencing of two of the three genes inhibited nematode infection, and the infectivity of worms treated with three dsRNA simultaneously changed the most, which decreased by 76.6%. Thus, the multi-copy alpha-amylase genes in
D
.
destructor
are compensatory and crucial for nematodes to parasitize the plant host.