| RRC ID |
70484
|
| 著者 |
Nozaki H, Mori F, Tanaka Y, Matsuzaki R, Yamaguchi H, Kawachi M.
|
| タイトル |
Cryopreservation of vegetative cells and zygotes of the multicellular volvocine green alga Gonium pectorale.
|
| ジャーナル |
BMC Microbiol
|
| Abstract |
Background
Colonial and multicellular volvocine green algae have been extensively studied recently in various fields of the biological sciences. However, only one species (Pandorina morum) has been cryopreserved in public culture collections.
Results
Here, we investigated conditions for cryopreservation of the multicellular volvocine alga Gonium pectorale using vegetative colonies or cells and zygotes. Rates of vegetative cell survival in a G. pectorale strain after two-step cooling and freezing in liquid nitrogen were compared between different concentrations (3% and 6%) of the cryoprotectant N,N-dimethylformamide (DMF) and two types of tubes (0.2-mL polymerase chain reaction tubes and 2-mL cryotubes) used for cryopreservation. Among the four conditions investigated, the highest rate of survival [2.7 ± 3.6% (0.54–10%) by the most probable number (MPN) method] was obtained when 2.0-mL cryotubes containing 1.0 mL of culture samples with 6% DMF were subjected to cryogenic treatment. Using these optimized cryopreservation conditions, survival rates after freezing in liquid nitrogen were examined for twelve other strains of G. pectorale and twelve strains of five other Gonium species. We obtained ≥ 0.1% MPN survival in nine of the twelve G. pectorale strains tested. However, < 0.1% MPN survival was detected in eleven of twelve strains of five other Gonium species. In total, ten cryopreserved strains of G. pectorale were newly established in the Microbial Culture Collection at the National Institute for Environmental Studies. Although the cryopreservation of zygotes of volvocine algae has not been previously reported, high rates (approximately 60%) of G. pectorale zygote germination were observed after thawing zygotes that had been cryopreserved with 5% or 10% methanol as the cryoprotectant during two-step cooling and freezing in liquid nitrogen.
Conclusions
The present study demonstrated that cryopreservation of G. pectorale is possible with 6% DMF as a cryoprotectant and 1.0-mL culture samples in 2.0-mL cryotubes subjected to two-step cooling in a programmable freezer.
|
| 巻・号 |
22(1)
|
| ページ |
103
|
| 公開日 |
2022-4-14
|
| DOI |
10.1186/s12866-022-02519-9
|
| PII |
10.1186/s12866-022-02519-9
|
| PMID |
35421922
|
| PMC |
PMC9008917
|
| MeSH |
Chlorophyta*
Cryopreservation
Nitrogen
Phylogeny
Zygote*
|
| IF |
2.989
|
| リソース情報 |
| 藻類 |
NIES-288
NIES-290
NIES-468
NIES-469
NIES-569
NIES-570
NIES-645
NIES-646
NIES-652
NIES-653
NIES-654
NIES-655
NIES-737
NIES-851
NIES-852
NIES-885
NIES-2261
NIES-2263
NIES-4121
NIES-4499
NIES-4500
NIES-4501
NIES-4502 |
