In silico identification and functional study of long non-coding RNA involved in acute hepatopancreatic necrosis disease caused by Vibrio parahaemolyticus infection in white shrimp, Litopenaeus vannamei.

Acute hepatopancreatic necrosis disease (AHPND) caused by toxin-producing Vibrio parahaemolyticus (VpAHPND) has severely affected shrimp production. Long non-coding RNA (lncRNA), a regulatory non-coding RNA, which can play important function in shrimp disease responses. This study aimed to identify and investigate the role of lncRNA involved in VpAHPND infection in Pacific white shrimp, Litopenaeus vannamei. From a total of 368,736 de novo assembled transcripts, 67,559 were identified as putative lncRNAs, and only 72 putative lncRNAs showed differential expression between VpAHPND-infected and normal shrimp. The six candidate lncRNAs were validated for their expression profiles during VpAHPND infection and tissue distribution using RT-qPCR. The role of lnc2088 in response to VpAHPND infection was investigated through RNA interference. The result indicated that the suppression of lnc2088 expression led to an increase in shrimp mortality after VpAHPND infection. To explore the set of genes involved in lnc2088 knockdown, RNA sequencing was performed. A total of 275 differentially expressed transcripts were identified in the hepatopancreas of lnc2088 knockdown shrimp. The expression profiles of five candidate metabolic and immune-related genes were validated in lnc2088 knockdown and VpAHPND-infected shrimp. The result showed that the expression of ChiNAG was significantly increased, while that of NCBP1, WIPF2, and NFKB1 was significantly downregulated in ds2088-injected shrimp. Additionally, the expression of NFKB1, NCBP1 and WIPF2 was significantly increased, whereas that of ChiNAG and CUL5 were significantly decreased after infection with VpAHPND. Our work identified putative lncRNA profiles in L. vannamei in response to VpAHPND infection and investigated the role of lncRNA in shrimp immunity.

Development of Gene-Based InDel Markers on Putative Drought Stress-Responsive Genes and Genetic Diversity of Durian (Durio zibethinus).

Insertion-deletion (InDel) markers are co-dominant, relatively abundant and practical for agarose gel genotyping. InDel polymorphism usually affects gene functions. Nucleotide sequences of durian (Durio zibethinus) are available, but InDel makers have not been well established. This study aimed to develop drought-related gene-based InDel markers for durian through bioinformatic analysis of RNA-Seq datasets. The polymorphism of the markers was verified in 24 durian genotypes local to Thailand. Bioinformatic analysis indicated 496 InDel loci having lengths more than 9 bp. To evaluate these InDel markers, 15 InDel loci were selected. Nine markers were successfully amplified a clear polymorphic band pattern on 2% agarose gel. The polymorphic information content (PIC) of these nine markers ranged from 0.1103 to 0.5808. The genetic distance between the 24 genotypes ranged from 0.222 to 0.889. The phylogeny based on the nine InDel loci distinguished the 24 genotypes and divided samples into four groups. This set of gene-based InDel markers on putative drought-responsive genes will be useful for genetic studies.

Transcriptome profiling of gonad-stimulating factors in thoracic ganglia and a potential role of Indian hedgehog gene in vitellogenesis of banana shrimp Fenneropenaeus merguiensis.

Shrimp reproduction is controlled by several factors. Central nervous tissues, especially thoracic ganglia and brain, are known sources of gonad stimulating factors (GSFs) in crustaceans, but the GSFs in shrimp have not yet been clarified. Hence, we aimed to characterize and study putative GSFs from thoracic ganglia of adult female Fenneropenaeus merguiensis. An analysis of thoracic ganglia transcriptome revealed 3224 putative GSFs of a total 77,681 unigenes. Only 376 putative GSFs were differentially expressed during ovarian developmental stages. Eight candidate GSFs were validated for their expression patterns in thoracic ganglia, including the Indian hedgehog gene. F. merguiensis Indian hedgehog (FmIHH) was then investigated for its role in vitellogenesis. The obtained full-length cDNA of FmIHH was similar to other crustacean IHHs rather than Sonic and Desert HHs. The FmIHH was dominantly expressed in thoracic ganglia, and its expression was significantly increased in the vitellogenic stages before being downregulated at the mature stage of ovarian development. Injection of the recombinant FmIHH (His-TF-IHH) protein stimulated vitellogenin expression in ovaries on day 3 and 7, and also increased the gonadosomatic index. In addition, crustacean hyperglycemic hormone expression and total sugar were significantly decreased in eyestalks and hemolymph, respectively, after injection of His-TF-IHH, while lactic acid was increased. Both total sugar and lactic acid were unchanged in ovaries of His-TF-IHH injected shrimp. These results suggested that FmIHH plays a crucial role in vitellogenesis and regulate sugar uptake during ovarian development.