Identification of RAPD‐SCAR marker linked to white spot syndrome virus resistance in populations of giant black tiger shrimp, Penaeus monodon Fabricius

Abstract

White spot disease (WSD) caused by white spot syndrome virus (WSSV) creates severe epizootics in shrimp aquaculture industry worldwide. Despite several efforts, no such permanent remedy was yet developed. Selective breeding using DNA markers would be a cost-effective strategy for long-term solution of this problem. In the present investigation, out of 30 random primers, only one primer produced a statistically significant (P < 0.01) randomly amplified polymorphic DNA (RAPD) marker of 502 bp, which provided a good discrimination between disease resistant and disease susceptible populations of Penaeus monodon from three geographical locations along the East coast of India. Because RAPD markers are dominant, a sequence characterized amplified region (SCAR) marker was developed by cloning and sequencing of 502 bp RAPD fragment, which generates a single 457 bp DNA fragment after PCR amplification only in the disease resistant shrimps. Challenge experiment was also conducted to validate this 457 bp SCAR marker, and the results suggested that the WSSV loads were 2.25 × 10(3) fold higher in disease susceptible than that in disease resistant shrimps using real-time PCR. Therefore, this 457 bp DNA SCAR marker will be very valuable towards the development of disease-free shrimp aquaculture industry.

Study on the Distribution of Disease-Resistant Shrimp Identified by DNA Markers in Respect to WSSV Infection in Different Seasons Along the Entire East Coast of India Aiming to Prevent White Spot Disease inPenaeus monodon

“…Previously, a prevalence study has been performed among the different important coasts along the east coast of India namely Digha, West Bengal; Chilika, Orissa; Visakhapatnam, Andhra Pradesh; and Chennai, Tamil Nadu. Similar observation with the current study was found except for Digha and Chennai (Dutta et al., ). This disparity in the result may be due to, in the previous study, shrimp samples were collected throughout the year, not in a particular season.…”

“…Additionally, a 457 bp DNA marker developed by RAPD‐SCAR method was also found in P. monodon , which is mostly present in the disease‐resistant shrimp. WSSV challenge experiment to prove its significant association with disease resistance showed 2.25 × 10 3 fold lower WSSV propagation occurred in disease‐resistant shrimp populations (Dutta et al., ).…”

Development of RAPD-SCAR markers for different Ganoderma species authentication by improved RAPD amplification and molecular cloning

“…A combination of SCAR and RAPD has significantly improved the stability and authenticity of the technique (Dnyaneshwar et al, 2006;Su et al, 2007;Li et al, 2010;Kumla et al, 2012;Rajesh et al, 2013;Fu et al, 2013;Noormohammadi et al, 2013;Mei et al, 2014a,b). Previous studies successfully developed RAPD markers for different species of animals, plants, and microbes (Lee et al, 2013;Yang et al, 2013Yang et al, , 2014Dutta et al, 2014). However, no SCAR marker for the identification of different Ganoderma species has been reported.…”

Genomics and genetic breeding in aquatic animals: progress and prospects

“…Norwegian scientists initiated the study of resistance-related MHC genes in Atlantic salmon [95] . Rodriguez-Ramilo et al found an anti-ciliate related SSR marker, Sma-USC256, in turbot [96] and Duttaet et al found a RAPD-SCAR marker related to white spot disease (WSD) resistance in Penaeus monodon, and subsequently identified another 71bp SSR marker associated with WSD [97,98] . In rainbow trout, Campbell et al isolated SNP markers related to bacterial cold water disease and infectious hematopoietic necrosis virus [99] .…”