莱茵衣藻蓝光受体植物类型隐花色素CRY突变体的表型鉴定

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

以莱茵衣藻(Chlamydomonas reinhardtii)野生株(CC5325)和植物类型CRY突变株(crcry)为研究对象,探究其在正常光照CK(Control)与蓝光BL(Blue light)培养条件下的表型差异。PCR检测表明,crcry突变株在编码区插入含有巴龙霉素抗性基因AphVIII的表达盒。以巴龙霉素为筛选条件的平板和液体培养体系进一步验证,crcry突变株中插入了AphVIII抗性基因并成功表达。表型鉴定表明,在CK培养条件下,野生株CC5325和突变株crcry在生长、色素、光合及油脂合成等方面没有显著差异;但是在BL条件下,突变株crcry的生长受到明显抑制,藻液颜色变黄;单位细胞叶绿素a、叶绿素b及总色素含量显著下降,相反,总类胡萝卜素含量明显增高;光合系统受到严重抑制,以及总脂含量显著降低。植物类型cry参与莱茵衣藻蓝光响应过程。

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.