Abstract
Salt-alkali tolerance is one of the important breeding traits of Portunus trituberculatus. Identification of molecular markers linked to salt-alkali tolerance is prerequisite to develop such molecular marker-assisted breeding. In this study, Bulked Segregant Analysis (BSA) was used to screen molecular markers associated with salt-alkali tolerance trait in P. trituberculatus. Two DNA mixing pools with significant difference in salt-alkali tolerance were prepared and 94.83G of high-quality sequencing data was obtained. 855 SNPs and 1051 Indels were firstly selected as candidate markers by BSA analysis, out of which, 20 markers were further selected via △index value (close to 0 or 1) and eight of those were successfully verified. In addition, based on the located information of the markers in genome, eight candidate genes related to salt-alkali tolerance were anchored including ubiquitin-conjugating enzyme, aspartate–tRNA ligase, vesicle-trafficking protein, and so on. qPCR results showed that the expression patterns of all these genes changed significantly after salt-alkali stress, suggesting that they play certain roles in salt-alkali adaptation. Our results will provide applicable markers for molecular marker-assisted breeding and help to clarify the mechanisms of salt-alkali adaptation of P. trituberculatus.
Highlights
Portunus trituberculatus (Juan and Gaoli, 2021; Ye and Fangmin, 2021), which belongs to Crustacea, Decapoda, Portunus family, commonly known as Portunus, is an important breeding species along the coast of China (Wu, 2014)
We explored the salt-alkali tolerance molecular markers in P. trituberculatus by the Bulked Segregant Analysis (BSA) strategy
A total of 855 SNPs (Supplementary Table S1) and 1051 Indels candidate polymorphic sex sites were selected with 95% confidence level (Supplementary Table S2)
Summary
Portunus trituberculatus (Juan and Gaoli, 2021; Ye and Fangmin, 2021), which belongs to Crustacea, Decapoda, Portunus family, commonly known as Portunus, is an important breeding species along the coast of China (Wu, 2014). Due to its high salinity, high alkalinity, high pH, and complex ion composition (Li, 2020), common aquatic animals cannot survive and reproduce in this environment suitably. The use of this type of water resource is greatly hindered. Based on the location information of the salt-alkali tolerance molecular markers on genome, we tried to anchor genes related to salt-alkali adaptation in crab. Our results will provide applicable markers for molecular marker-assisted breeding and help to clarify the mechanism of salt-alkali adaptation of P. trituberculatus
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