Long-Read Sequencing Reveals the Repertoire of Long-Chain Polyunsaturated Fatty Acid Biosynthetic Genes in the Purple Land Crab, Gecarcoidea lalandii (H. Milne Edwards, 1837)

Ting, Seng Yeat; Lau, Nyok-Sean; Sam, Ka-Kei; Quah, Evan S. H.; Ahmad, Amirrudin; Mat-Isa, Mohd-Noor; Shu‐Chien, Alexander Chong

doi:10.3389/fmars.2021.713928

Cited by 4 publications

(6 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While PUFA elongases have been successfully functionally characterized in other crustacean lineages such as decapods, branchiopods and amphipods (Mah et al, 2019; Ribes‐Navarro et al, 2021; Sun et al, 2020; Ting et al, 2020), the occurrence and functionality of methyl‐end and front‐end desaturases in these lineages remains highly questionable (Chen et al, 2017; Kabeya et al, 2021; Lin et al, 2017; Monroig & Kabeya, 2018; Nielsen et al, 2019; Ting et al, 2021; Wu et al, 2018; Yang et al, 2013). In our phylogenetic study, we detected a putative front‐end desaturase in a transcriptome of the decapod Eurypanopeus depressus clustering together with the T. californicus Δ5 front‐end desaturase “ Fed5 ” with high bootstrap support (86%).…”

Section: Discussionmentioning

confidence: 99%

Functional characterization reveals a diverse array of metazoan fatty acid biosynthesis genes

et al. 2022

View full text Add to dashboard Cite

Long‐chain (≥C20) polyunsaturated fatty acids (LC‐PUFAs) are physiologically important fatty acids for most animals, including humans. Although most LC‐PUFA production occurs in aquatic primary producers such as microalgae, recent research indicates the ability of certain groups of (mainly marine) invertebrates for endogenous LC‐PUFA biosynthesis and/or bioconversion from dietary precursors. The genetic pathways for and mechanisms behind LC‐PUFA biosynthesis remain unknown in many invertebrates to date, especially in non‐model species. However, the numerous genomic and transcriptomic resources currently available can contribute to our knowledge of the LC‐PUFA biosynthetic capabilities of metazoans. Within our previously generated transcriptome of the benthic harpacticoid copepod Platychelipus littoralis, we detected expression of one methyl‐end desaturase, one front‐end desaturase, and seven elongases, key enzymes responsible for LC‐PUFA biosynthesis. To demonstrate their functionality, we characterized eight of them using heterologous expression in yeast. The P. littoralis methyl‐end desaturase has Δ15/17/19 desaturation activity, enabling biosynthesis of α‐linolenic acid, eicosapentaenoic acid and docosahexaenoic acid (DHA) from 18:2 n−6, 20:4 n−6 and 22:5 n−6, respectively. Its front‐end desaturase has Δ4 desaturation activity from 22:5 n−3 to DHA, implying that P. littoralis has multiple pathways to produce this physiologically important fatty acid. All studied P. littoralis elongases possess varying degrees of elongation activity for saturated and unsaturated fatty acids, producing aliphatic hydrocarbon chains with lengths of up to 30 carbons. Our investigation revealed a functionally diverse range of fatty acid biosynthesis genes in copepods, which highlights the need to scrutinize the role that primary consumers could perform in providing essential nutrients to upper trophic levels.

show abstract

Section: Discussionmentioning

confidence: 99%

Functional characterization reveals a diverse array of metazoan fatty acid biosynthesis genes

et al. 2022

View full text Add to dashboard Cite

show abstract

“…To date, studies on crustacean Elovl have unveiled orthologues of Elovl1, Elovl4, Elovl6, and Elovl7 (Kabeya et al, 2021;Lee et al, 2020;Mah et al, 2019;Ribes-Navarro et al, 2021;Sun et al, 2020;Ting et al, 2020). Interestingly, while aquatic brachyurans contain all the four elovl mentioned above, terrestrial brachyuran crabs such as G. lalandii appear to possess only two elovl-like genes in their genomes, namely Elovl1 and Elovl6 (Ting et al, 2021).…”

Section: Discussionmentioning

confidence: 99%

“…One µg of DNase-treated RNA was reverse transcribed into cDNA using SuperScript IV kit (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). Full-length elovl1 and elovl6 nucleotide sequences were identified and acquired from a transcriptome database (GIKV00000000) (Ting et al, 2021). The open reading frames (ORF) of these putative G. lalandii elovl were amplified by PCR (MyTaq™ Redmix, Bioline, Massachusetts, USA), using hepatopancreas cDNA as template and gene specific primers containing restriction enzyme sites for subsequent cloning into the yeast expression vector pYES2 (Thermo Fisher Scientific, Waltham, MA, USA) (Supplementary Table 1).…”

Section: Isolation and Cloning Of Elovl Full-length Cdnamentioning

confidence: 99%

“…Our previous work in the mud crab characterised the capacity of Elovl1/7 and Elovl6 in PUFA elongation (Mah et al, 2019;Ting et al, 2020). A transcriptomic approach to identify putative LC-PUFA biosynthesis genes in G. lalandii also identified the presence of these two orthologs (Ting et al, 2021). Specifically, we isolated and functionally characterised two elovl from G. lalandii with putative roles in LC-PUFA biosynthesis and determined their mRNA transcript abundance in different tissues and the fatty acid composition of selected tissues.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Long-chain polyunsaturated fatty acid biosynthesis in a land-crab with advanced terrestrial adaptations: Molecular cloning and functional characterization of two fatty acyl elongases

Ting

Lau²,

Sam³

et al. 2022

Comparative Biochemistry and Physiology Part B: Biochemistry an

View full text Add to dashboard Cite

“…The use of RNA-seq data has become a cost-effective and useful approach for functional genomics research in species with or without a reference genome. De novo transcriptome assembly has been applied in a terrestrial crustacean species to mine for Fads or Elovl orthologs(Ting et al 2021). Through the P. micromegethes transcriptome assembly, a single Fads2 and four Elovl sequences were putatively identi ed.…”

mentioning

confidence: 99%

A Complete Inventory of Long-Chain Polyunsaturated Fatty Acids Biosynthesis Pathway Enzymes in the Miniaturized Cyprinid Paedocypris Micromegethes

Sam

Lau

Kuah

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

The capacity for long-chain polyunsaturated fatty acid (LC-PUFA) biosynthesis activity in a species depends on the enzymatic activities of fatty acyl desaturase (Fads) and elongation of very long-chain fatty acid (Elovl). The miniaturized fish Paedocypris micromegethes is a developmentally truncated cyprinid living in highly acidic water conditions in tropical peat swamps. The capacity for LC-PUFA biosynthesis in this species, which has a reduced genome size, is unknown. A high-quality de novo transcriptome assembly enabled the identification of a putative Fads2 and four Elovl. The Fads2 was verified as a P. micromegethes Fads2 ortholog with in vitro Δ5 and Δ6 activities. The Elovl sequences were established as an Elovl5, Elovl2, and two Elovl4 paralogs, namely Elovl4a and Elovl4b. These Elovl enzymes, mainly Elovl5 and Elovl2, fulfill the necessary C18, C20, and C22 PUFA elongation steps for LC-PUFA biosynthesis. Collectively, these results validate the presence of a complete repertoire of LC-PUFA biosynthesis enzymes in a tropical miniatured freshwater fish.

show abstract

Long-Read Sequencing Reveals the Repertoire of Long-Chain Polyunsaturated Fatty Acid Biosynthetic Genes in the Purple Land Crab, Gecarcoidea lalandii (H. Milne Edwards, 1837)

Cited by 4 publications

References 38 publications

Functional characterization reveals a diverse array of metazoan fatty acid biosynthesis genes

Functional characterization reveals a diverse array of metazoan fatty acid biosynthesis genes

Long-chain polyunsaturated fatty acid biosynthesis in a land-crab with advanced terrestrial adaptations: Molecular cloning and functional characterization of two fatty acyl elongases

A Complete Inventory of Long-Chain Polyunsaturated Fatty Acids Biosynthesis Pathway Enzymes in the Miniaturized Cyprinid Paedocypris Micromegethes

Contact Info

Product

Resources

About