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Diseases of Aquatic Organisms

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DAO 142:213-224 (2020)  -  DOI: https://doi.org/10.3354/dao03538

Association between ALFPm3 single nucleotide polymorphism and white spot syndrome virus resistance in black tiger shrimp Penaeus monodon

Chettupon Pooljun1, Pitchanee Jariyapong1,2, Sataporn Direkbusarakom1, Sudarat Saeng-ngern1, Takashi Sakamoto3, Suwit Wuthisuthimethavee1,*

1Center of Excellence for Shrimp, School of Agricultural Technology, Walailak University, Nakhon Si Thammarat 80160, Thailand
2School of Medicine, Walailak University, Nakhon Si Thammarat 80160, Thailand
3Department of Marine Biosciences, Tokyo University of Marine Science and Technology, 4-5-7 Konan, Minato-ku, Tokyo 108-8477, Japan
*Corresponding author:

ABSTRACT: Here single nucleotide polymorphisms (SNPs) were associated with white spot syndrome virus (WSSV) resistance in black tiger shrimp Penaeus monodon. SNPs were identified by single-strand conformation polymorphism (SSCP) screening and DNA sequencing of shrimp sampled from 3 families (100 shrimp per family) challenged with WSSV. Shrimp that died over the 14 d challenge trial were designated susceptible, with those remaining alive on Day 14 designated resistant. To compare SNPs, 10 samples from the susceptible and resistant groups, each comprising DNA pooled from 3 shrimp, were amplified by polymerase chain reaction (PCR) using primers to 12 selected genes and screened by SSCP. SNPs were only identified in the anti-lipopolysaccharide factor 3 (ALFPm3) gene product. Analysis of complete ALFPm3 gene sequences confirmed the existence of 3 SNPs (g.934C>G, g.1186A>G, and g.1898C>G) that were polymorphic between the susceptible and resistant groups. Further analyses using specific tetra-primer amplification refractory mutation system PCR primer sets associated these 3 SNPS, and particularly the g.1186A>G SNP, with WSSV resistance. This SNP thus has potential for use as a DNA marker to select for WSSV resistance in P. monodon breeding programs.


KEY WORDS: Single nucleotide polymorphism · SNP · Tetraprimer amplification refractory mutation system-PCR · Tetra-primer ARMS-PCR · WSSV resistance


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Cite this article as: Pooljun C, Jariyapong P, Direkbusarakom S, Saeng-ngern S, Sakamoto T, Wuthisuthimethavee S (2020) Association between ALFPm3 single nucleotide polymorphism and white spot syndrome virus resistance in black tiger shrimp Penaeus monodon. Dis Aquat Org 142:213-224. https://doi.org/10.3354/dao03538

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