Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (2): 243-253.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0611

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Phenotypic Characterization of Blue Photoreceptor Plant Type Cryptochrome CRY Mutant in Chlamydomonas reinhardtii

LI Wang-ning1(), ZHANG Hao-jie1, LI Ya-nan1, LIANG Meng-jing1, JI Chun-li1, Zhang Chun-hui1, LI Run-zhi1, CUI Yu-lin2, QIN Song2, CUI Hong-li1()   

  1. 1. College of Agriculture, Shanxi Agricultural University, Institute of Molecular Agriculture and Bioenergy, Taigu 030801
    2. Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Key Laboratory of Coastal Biology and Biological Resources Utilization, Yantai 264003
  • Received:2022-05-18 Online:2023-02-26 Published:2023-03-07

Abstract:

The phenotypic differences of Chlamydomonas reinhardtii wild strain(CC5325)and CRY mutant strain(crcry)under normal CK(Control)and Blue BL(Blue light)cultures were investigated. The PCR verification showed that the crcry mutant was inserted with paromomycin resistance gene AphVIII expression box in the coding region. AphVIII resistance gene was inserted into crcry mutant and successfully expressed in plate and liquid culture system with paromomycin as screening condition. Phenotypic identification showed that there were no significant differences in growth, pigment, photosynthesis and lipid synthesis between wild strain CC5325 and mutant strain crcry under CK culture condition. However, under BL condition, the growth of mutant crcry was significantly inhibited, and the color of algae liquid turned yellow. Chlorophyll a, chlorophyll b and total pigment contents of unit cells decreased significantly, while total carotenoid contents increased significantly. The photosynthetic system was severely suppressed and the total lipid content significantly reduced. In conclusion, plant cry is involved in the blue light response of C. reinhardtii.

Key words: Chlamydomonas reinhardtii, cryptochrome, blue light, mutant, phenotypic characterization