Sesbania Mosaic Virus (SeMV) Infectious Clone: Possible Mechanism of 3 ` and 5 ` End Repair and Role of Polyprotein Processing in Viral Replication

Sesbania mosaic virus (SeMV) is a positive stranded RNA virus belonging to the genus Sobemovirus. Construction of an infectious clone is an essential step for deciphering the virus gene functions in vivo. Using Agrobacterium based transient expression system we show that SeMV icDNA is infectious on Sesbania grandiflora and Cyamopsis tetragonoloba plants. The efficiency of icDNA infection was found to be significantly high on Cyamopsis plants when compared to that on Sesbania grandiflora. The coat protein could be detected within 6 days post infiltration in the infiltrated leaves. Different species of viral RNA (double stranded and single stranded genomic and subgenomic RNA) could be detected upon northern analysis, suggesting that complete replication had taken place. Based on the analysis of the sequences at the genomic termini of progeny RNA from SeMV icDNA infiltrated leaves and those of its 3' and 5' terminal deletion mutants, we propose a possible mechanism for 3' and 5' end repair in vivo. Mutation of the cleavage sites in the polyproteins encoded by ORF 2 resulted in complete loss of infection by the icDNA, suggesting the importance of correct polyprotein processing at all the four cleavage sites for viral replication. Complementation analysis suggested that ORF 2 gene products can act in trans. However, the trans acting ability of ORF 2 gene products was abolished upon deletion of the N-terminal hydrophobic domain of polyprotein 2a and 2ab, suggesting that these products necessarily function at the replication site, where they are anchored to membranes.

Interaction of Sesbania mosaic virus (SeMV) RNA-dependent RNA polymerase (RdRp) with the p10 domain of polyprotein 2a and its implications in SeMV replication

Identification of viral encoded proteins that interact with RNA-dependent RNA polymerase (RdRp) is an important step towards unraveling the mechanism of replication. Sesbania mosaic virus (SeMV) RdRp was shown to interact strongly with p10 domain of polyprotein 2a and moderately with the protease domain. Mutational analysis suggested that the C-terminal disordered domain of RdRp is involved in the interaction with p10. Coexpression of full length RdRp and p10 resulted in formation of RdRp-p10 complex which showed significantly higher polymerase activity than RdRp alone. Interestingly, C Delta 43 RdRp also showed a similar increase in activity. Thus, p10 acts as a positive regulator of RdRp by interacting with the C-terminal disordered domain of RdRp. (C) 2014 The Authors. Published by Elsevier B.V.

Intracellular delivery of antibodies by chimeric Sesbania mosaic virus (SeMV) virus like particles

The therapeutic potential of antibodies has not been fully exploited as they fail to cross cell membrane. In this article, we have tested the possibility of using plant virus based nanoparticles for intracellular delivery of antibodies. For this purpose, Sesbania mosaic virus coat protein (CP) was genetically engineered with the B domain of Staphylococcus aureus protein A (SpA) at the beta H-beta I loop, to generate SeMV loop B (SLB), which self-assembled to virus like particles (VLPs) with 43 times higher affinity towards antibodies. CP and SLB could internalize into various types of mammalian cells and SLB could efficiently deliver three different monoclonal antibodies-D6F10 (targeting abrin), anti-a-tubulin (targeting intracellular tubulin) and Herclon (against HER2 receptor) inside the cells. Such a mode of delivery was much more effective than antibodies alone treatment. These results highlight the potential of SLB as a universal nanocarrier for intracellular delivery of antibodies.

Structural studies on chimeric Sesbania mosaic virus coat protein: Revisiting SeMV assembly

The capsid protein (CP) of Sesbania mosaic virus (SeMV, a T=3 plant virus) consists of a disordered N-terminal R-domain and an ordered S-domain. Removal of the R-domain results in the formation of T=1 particles. In the current study, the R-domain was replaced with unrelated polypeptides of similar lengths: the B-domain of Staphylococcus aureus SpA, and SeMV encoded polypeptides P8 and P10. The chimeric proteins contained T=3 or larger virus-like particles (VLPs) and could not be crystallized. The presence of metal ions during purification resulted in a large number of heterogeneous nucleoprotein complexes. N Delta 65-B (R domain replaced with B domain) could also be purified in a dimeric form. Its crystal structure revealed T=1 particles devoid of metal ions and the B-domain was disordered. However, the B-domain was functional in N Delta 65-B VLPs, suggesting possible biotechnological applications. These studies illustrate the importance of N-terminal residues, metal ions and robustness of the assembly process. (C) 2015 Elsevier Inc. All rights reserved.

Status and management of the sea cucumber fishery of La Grande Terre, New Caledonia

From October 2006 to May 2008, The WorldFish Center coordinated a ZoNéCo project to provide support to the Southern and Northern Provinces for decisions about how best to manage the sea cucumber fishery around La Grande Terre. We collected data during underwater population surveys, questionnaire-based interviews with fishers and processors, and landing catch surveys. A core aim was to furnish the Provinces with ‘ballpark’ estimates of the abundance and density of commercially important sea cucumbers on 50 lagoon and barrier reefs. Analysis and synthesis of the ecological and sociological data provide the basis for informed recommendations for fisheries management. Counts of trochus and giant clams on the reefs allow us to also describe the general status of those resources. We propose 13 recommendations for management actions and fishery regulations and advocate an adaptive management approach. This multidisciplinary study should serve as a useful template for assessing other fisheries, and we provide a series of generic ‘lessons learnt’ to aid future programmes. (PDF has 140 pages.)

Value chain analysis for sea cucumber in the Philippines

This study examined the sea cucumber industry in the Philippines through the value chain lens. The intent was to identify effective pathways for the successful introduction of sandfish culture as livelihood support for coastal communities. Value chain analysis is a high-resolution analytical tool that enables industry examination at a detailed level. Previous industry assessments have provided a general picture of the sea cucumber industry in the country. The present study builds on the earlier work and supplies additional details for a better understanding of the industry's status and problems, especially their implications for the Australian Center for International Agricultural Research (ACIAR) funded sandfish project "Culture of sandfish (Holothuria scabra) in Asia- Pacific" (FIS/2003/059). (PDF contains 54 pages)

A study of tagging methods for the sea cucumber Cucumaria frondosa in the waters off Maine

The sea cucumber fishery in waters off Maine is developing and has recently experienced great increases in landings, corresponding to expanding export markets. Between 1994 and 1996, reported landings ranged from one to three million pounds (Fig. 1). In 1999, reported landings were over eight million pounds and rose to over nine million in 2000 (Feindel1). Like other developing fisheries, we have little information about the biology and ecology of the sea cucumber off Maine, limited data on the fishery, and little knowledge about the key life history processes that characterize its population dynamics. Therefore, we have a limited understanding of the current status of the resource and the impacts the fishery may have on the stock.

Hatchery and culture technology for the sea cucumber Holothuria scabra Jaeger, in India

The seed of the sea cucumber Holothuria scabra jaeger is being produced at the Central Marine Fisheries Research Institute in India. This article describes the techniques being used in the production of seed and the experiments being carried out for the rearing of juveniles. Trials to grow juveniles in hatcheries on prawn farms have shown spectabular results that are both cost efficient and environmentally friendly.

A Review of Recent Developments in the World Sea Cucumber Fisheries

Sea cucumbers (Holothuridae and Stichopodidae) have been harvested commercially for at least 1,000 years. The world fisheries for sea cucumbers, however, are not well documented and in general are poorly managed. Depending upon the species exploited, there are two processing procedures for the sea cucumber product. Some species are eaten raw, while most commercial species are processed into a dry product called beche-de-mer or trepang. This dry product is exported to a central market such as Hong Kong and then re-exported to the consumers. In this review, recent statistics on the world sea cucumber fisheries, collected from different services, are detailed for each major fishing area. Case studies for each fishing area are also presented. Recent major changes in the Indo-Pacific fishery include the participation of new producer countries, the shift in the species being exploited, and an increase in the Chinese market. The expansion of the largely monospecific temperate North Pacific fisheries is also described. Statistics from Hong Kong, Singapore, Taiwan, and the Food and Agriculture Organization provide valuable information on the producer and importer countries. Particular attention is paid to the reciprocal trade of beche-de-mer between Hong Kong and Singapore. An evaluation of the world sea cucumber landings and beche-de-mer production is presented. Recent developments include an expansion of the Hong Kong market due to increased demand by China, the importance of Indonesia as a major world producer, and an increase in the fisheries of Tropical Pacific nations. This increase is best documented for New Caledonia and Fiji. Ways to improve the access and the reliability of the statistics for the sea cucumber fishery are discussed, as is the potential for management of artisanal fisheries.