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The papain-like cysteine proteases (PLCPs), the most important group of cysteine proteases, have been reported to participate in the regulation of growth, senescence, and abiotic stresses in plants. However, the functions of PLCPs and their roles in stress response in microalgae was rarely reported. The responses to different abiotic stresses in  Haematococcus pluvialis  were often observed, including growth regulation and astaxanthin accumulation. In this study, the cDNA of  HpXBCP3  containing 1515 bp open reading frame (ORF) was firstly cloned from  H. pluvialis  by RT-PCR. The analysis of protein domains and molecular evolution showed that  HpXBCP3  was closely related to  AtXBCP3  from  Arabidopsis . The expression pattern analysis revealed that it significantly responds to NaCl stress in  H  . pluvialis . Subsequently, transformants expressing  HpXBCP3  in  Chlamydomonas reinhardtii  were obtained and subjected to transcriptomic analysis. Results showed that  HpXBCP3  might affect the cell cycle regulation and DNA replication in transgenic  Chlamydomonas , resulting in abnormal growth of transformants. Moreover, the expression of  HpXBCP3  might increase the sensitivity to NaCl stress by regulating ubiquitin and the expression of WD40 proteins in microalgae. Furthermore, the expression of  HpXBCP3  might improve chlorophyll content by up-regulating the expression of NADH-dependent glutamate synthases in  C. reinhardtii . This study indicated for the first time that  HpXBCP3  was involved in the regulation of cell growth, salt stress response, and chlorophyll synthesis in microalgae. Results in this study might enrich the understanding of PLCPs in microalgae and provide a novel perspective for studying the mechanism of environmental stress responses in  H. pluvialis .

international journal of molecular sciences

The Papain-like Cysteine Protease HpXBCP3 from Haematococcus pluvialis Involved in the Regulation of Growth, Salt Stress Tolerance and Chlorophyll Synthesis in Microalgae