Silencing of prophenoloxidase (proPO) gene in freshwater prawn, Macrobrachium rosenbergii, makes them susceptible to white spot syndrome virus (WSSV)
Silencing of prophenoloxidase (proPO) gene in freshwater prawn, Macrobrachium rosenbergii, makes them susceptible to white spot syndrome virus (WSSV)
Prophenoloxidase (proPO) is very important to protect the invertebrates from microbial infections. Our previous studies revealed that proPO was up-regulated in WSSV-injected Macrobrachium rosenbergii and is responsible for protecting M. rosenbergii from WSSV. In order to prove this mechanism, an attempt was made in the present study to silence the proPO gene in freshwater prawn by injection of dsRNA-proPO followed by WSSV challenge. Two partial fragments of proPO with the size of 251 and 331 bp were used to synthesize dsRNA using LITMUS38i vector and E. coli. The bacterially synthesized dsRNA-proPO was used to silence proPO gene to determine its involvement in developing resistance in prawn against WSSV. In proPO gene-silenced prawn, 100% mortality was observed after WSSV challenge whereas no mortality was observed in prawn injected with WSSV alone. The WSSV infection in gene-silenced prawn was confirmed by PCR, and its propagation was quantified by ELISA and real-time PCR at different time intervals. Real-time PCR assay revealed a significant reduction in the expression of proPO gene in WSSV-challenged proPO-silenced prawn when compared to normal prawn. Level of proPO was reduced significantly in the haemolymph of proPO-silenced prawn when compared to prawn injected with PBS.
Macrobrachium rosenbergii, gene silencing, prophenoloxidase, resistance, white spot syndrome virus
573-584
Thamizhvanan, S.
120cdeaa-0213-47ac-bfc5-321aae723ea9
Nafeez Ahmed, A.
a4a2ffdb-0a47-493f-b593-e97a04a5d89e
Vinoth Kumar, D.
0161f93a-8183-4bf0-a2a5-d05b8b87b103
Vimal, S.
7e4fa71a-fa89-4f08-ac09-0c09b2a1909f
Majeed, S. Abdul
7f2312e8-8bd7-41ee-a7b9-e37d2d89c41d
Taju, G.
3b3be448-2eba-4eda-9254-cec64db2f8d6
Hauton, Chris
7706f6ba-4497-42b2-8c6d-00df81676331
Hameed, A.S. Sahul
8af1f9f5-408e-4ef6-80be-8c446ffc6634
Thamizhvanan, S.
120cdeaa-0213-47ac-bfc5-321aae723ea9
Nafeez Ahmed, A.
a4a2ffdb-0a47-493f-b593-e97a04a5d89e
Vinoth Kumar, D.
0161f93a-8183-4bf0-a2a5-d05b8b87b103
Vimal, S.
7e4fa71a-fa89-4f08-ac09-0c09b2a1909f
Majeed, S. Abdul
7f2312e8-8bd7-41ee-a7b9-e37d2d89c41d
Taju, G.
3b3be448-2eba-4eda-9254-cec64db2f8d6
Hauton, Chris
7706f6ba-4497-42b2-8c6d-00df81676331
Hameed, A.S. Sahul
8af1f9f5-408e-4ef6-80be-8c446ffc6634
Thamizhvanan, S., Nafeez Ahmed, A., Vinoth Kumar, D., Vimal, S., Majeed, S. Abdul, Taju, G., Hauton, Chris and Hameed, A.S. Sahul
(2020)
Silencing of prophenoloxidase (proPO) gene in freshwater prawn, Macrobrachium rosenbergii, makes them susceptible to white spot syndrome virus (WSSV).
Journal of Fish Diseases, 44 (5), .
(doi:10.1111/jfd.13297).
Abstract
Prophenoloxidase (proPO) is very important to protect the invertebrates from microbial infections. Our previous studies revealed that proPO was up-regulated in WSSV-injected Macrobrachium rosenbergii and is responsible for protecting M. rosenbergii from WSSV. In order to prove this mechanism, an attempt was made in the present study to silence the proPO gene in freshwater prawn by injection of dsRNA-proPO followed by WSSV challenge. Two partial fragments of proPO with the size of 251 and 331 bp were used to synthesize dsRNA using LITMUS38i vector and E. coli. The bacterially synthesized dsRNA-proPO was used to silence proPO gene to determine its involvement in developing resistance in prawn against WSSV. In proPO gene-silenced prawn, 100% mortality was observed after WSSV challenge whereas no mortality was observed in prawn injected with WSSV alone. The WSSV infection in gene-silenced prawn was confirmed by PCR, and its propagation was quantified by ELISA and real-time PCR at different time intervals. Real-time PCR assay revealed a significant reduction in the expression of proPO gene in WSSV-challenged proPO-silenced prawn when compared to normal prawn. Level of proPO was reduced significantly in the haemolymph of proPO-silenced prawn when compared to prawn injected with PBS.
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Accepted/In Press date: 20 October 2020
e-pub ahead of print date: 10 November 2020
Keywords:
Macrobrachium rosenbergii, gene silencing, prophenoloxidase, resistance, white spot syndrome virus
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Local EPrints ID: 485445
URI: http://eprints.soton.ac.uk/id/eprint/485445
ISSN: 0140-7775
PURE UUID: 55d32695-c61a-48c3-81eb-00dcb9a153e0
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Date deposited: 06 Dec 2023 17:48
Last modified: 17 Mar 2024 02:43
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Author:
S. Thamizhvanan
Author:
A. Nafeez Ahmed
Author:
D. Vinoth Kumar
Author:
S. Vimal
Author:
S. Abdul Majeed
Author:
G. Taju
Author:
A.S. Sahul Hameed
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