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雨生红球藻细胞形态转换和藻落形成差异研究
程天佑1,2, 徐晓莹1,2, 张文蕾1, 张 维1, 陈 林1, 袁冠华1,2, 刘天中1
1.中国科学院 青岛生物能源与过程研究所, 中国科学院 生物燃料重点实验室;2.中国科学院大学
摘要:
基于雨生红球藻(Haematococcus pluvialis)培养过程中游动细胞和不动细胞的形态差异与转换,对比研究了两种细胞类型在离心收集与藻落形成效率上的差异。结果表明, 雨生红球藻SCCAP K-0084在BYA培养基中培养4~6 d和9~12 d 分别以绿色游动细胞与绿色不动细胞为主; 早期游动细胞离心收集需要更高的离心力, 但500~1 000 g离心5 min可以在不影响细胞存活率的情况下有效收获所有类型细胞; 相比于传统直接涂布法, 双层琼脂平板法可有效提高平板藻落形成效率, 且游动细胞藻落形成效率比不动细胞藻落形成效率更高。TAP培养基由于藻落形成效率较低并不适合于雨生红球藻平板培养与筛选, 自养型培养基虽然具有较好的藻落形成效率但是藻落形成速度较慢, 相比而言, BYA培养基藻落形成效率较高且生长速度快, 更适合用于雨生红球藻细胞的后期平板培养与筛选过程。
关键词:  雨生红球藻(Haematococcus pluvialis)  游动细胞  不动细胞  形态转换  藻落形成差异
DOI:10.11759/hykx 20161109002
分类号:
基金项目:国家自然科学基金中泰合作研究基金资助项目(51561145015)
Effect of morphological shift on phycocolony forming in Haematococcus pluvialis
CHENG Tian-you,XU Xiao-ying,ZHANG Wen-lei,ZHANG Wei,CHEN Lin,YUAN Guan-hua,LIU Tian-zhong
Abstract:
Difference in cell collection and phycocolony forming between motile and non-motile cells of Haematococcus pluvialis was studied considering the morphological shift under mixotrophic conditions. The results showed that motile and non-motile cells could be enriched separately after 4-6 days and 9-12 days, respectively, through mixotrophic cultivation in BYA. Although a greater centrifugal force was required for motile cell collection, motile and non-motile cells could both be collected by centrifugation at 500-1000 g for 5 min without obvious negative effects on the cell viability. Furthermore, the plate data indicated that the phycocolony forming rate of H. pluvialis could be significantly improved using double layer of the apical low melting point agarose in comparison with that using traditional direct coating, and the phycocolony forming rate of motile cells was significantly higher than that of non-motile cells. Besides, the phycocolony forming rate in TAP was lower than that in the other three mediums, indicating that TAP was not suitable for the screening of H. pluvialis, and the growth rate of colonies in BG11 and BBM was significantly lower than that in TAP and BYA. Above results indicated that BYA is more suitable for phycocolony formation and the screening of H. pluvialis considering the high phycocolony forming rate and rapid growth.
Key words:  Haematococcus pluvialis  motile cell  non-motile cell  morphological shift  phycocolony forming
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