Comparative morphological, physiological, biochemical and genomic studies reveal novel genes of Dunaliella bioculata and D. quartolecta in response to salt stress.

Background: Salinity is an essential abiotic stress in plants. Dunaliella is a genus of high-salt-tolerant microalgae. The present study aimed to compare the characterizations of D. bioculata and D. quartolecta at different levels and investigate novel genes response to salt stress.

Methods And Results: High chlorophyll contents were detected in D. bioculata on the 35 d of salt stress, while high lipid and carotenoid contents were detected in D. quartolecta via morphological and biochemical analyses. Physiological analysis showed that D. quartolecta cells had a smaller increase in osmotic potential, a smaller decrease in the Na+/K+ ratio and photochemical efficiency (Fv/Fm), and a lower relative conductivity than D. bioculata cells. The genomic lengths of D. quartolecta and D. bioculata were 396,013,629 bp (scaffold N50 = 1954 bp) and 427,667,563 bp (scaffold N50 = 3093 bp) via high-throughput sequencing and de novo assembly, respectively. Altogether, 25,751 and 26,620 genes were predicted in their genomes by annotation analysis with various biodatabases. The D. bioculata genome showed more segmental duplication events via collinearity analysis. More single nucleotide polymorphisms and insertion-deletion variants were detected in the D. bioculata genome. Both algae, which showed a close phylogenetic relationship, may undergo positive selection via bioinformatics analysis. A total of 382 and 85 novel genes were screened in D. bioculata and D. quartolecta, with 138 and 51 enriched KEGG pathways, respectively. Unlike the novel genes adh1, hprA and serA, the relative expression of livF and phbB in D. bioculata was markedly downregulated as salinity increased, as determined by qPCR analysis. The relative expression of leuB, asd, pstC and proA in D. quartolecta was markedly upregulated with the same salinity increase.

Conclusion: Dunaliella quartolecta is more halophilic than D. bioculata, with more effective responses to high salt stress based on the multiphase comparative data.