Establishment of a cryopreserved biobank for the Culture Collection of Freshwater Microalgae (CCMA-UFSCar), São Paulo, Brazil

Tessarolli, Leticia Piton; Day, John Godfrey; Vieira, Armando Augusto Henriques

doi:10.1590/1676-0611-BN-2016-0299

Brasil

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article • Biota Neotrop. 17 (2) • 2017 • https://doi.org/10.1590/1676-0611-BN-2016-0299 linkcopy

Establishment of a cryopreserved biobank for the Culture Collection of Freshwater Microalgae (CCMA-UFSCar), São Paulo, Brazil

Estabelecimento de um biobanco criopreservado para a Coleção de Culturas de Microalgas de Água-Doce (CCMA-UFSCar), São Paulo, Brasil

Authorship SCIMAGO INSTITUTIONS RANKINGS

Abstract

The Culture Collection of Freshwater Microalgae (CCMA-UFSCar, Coleção de Culturas de Microalgas de Água-Doce), based at the Universidade Federal de São Carlos, plays an important role in underpinning Brazilian microalgal research, providing biological materials, substrates and training personnel for a large proportion of the past and current projects in this area. However, recent efforts to expand the culture catalogue/holdings have reached a ceiling because of the logistical practicality of maintaining actively growing cultures. In order to reduce the costs associated with the maintenance regime of cultures, efforts were initiated on the establishment of a cryopreserved biobank for long term maintenance of cultures, thus minimizing the efforts associated with handling of material, as successfully frozen cultures, in theory, could be maintained effectively indefinitely, with the advantage of the stability of cells characteristics. Initial tests were performed on 93 strains including exemplar taxa across the different taxonomic groups in the collection catalogue. The highest levels of success were achieved for the smaller taxa, such as the small green algae; while for the larger and more complex organisms limited to no success was obtained. For the strains tested, over 70% had positive good levels of post-thaw viability and regenerated phenotypically normal cultures.

Keywords: Cryopreservation; freshwater microalgae; biobank

Accession number		Strain	CPA	r ABS (%)
CCMA-UFSCar	003	Ankistrodesmus densus	DMSO 5%	81
CCMA-UFSCar	005	Messastrum gracile	DMSO 5%	89
CCMA-UFSCar	024	Monoraphidium arcuatum	DMSO 5%	95
CCMA-UFSCar	047	Selenastrum bibraianum	DMSO 5%	52
CCMA-UFSCar	048	Raphidocelis subcapitata	DMSO 5%	47
CCMA-UFSCar	063	Selenastrum bibraianum	DMSO 5%	7
CCMA-UFSCar	083	Ankistrodesmus flexuosus	DMSO 5%	73
CCMA-UFSCar	123	Kirchneriella aperta	DMSO 5%	33
CCMA-UFSCar	125	Selenastrum bibraianum	DMSO 5%	55
CCMA-UFSCar	128	Ankistrodesmus densus	DMSO 5%	100
CCMA-UFSCar	137	Chlorolobion cf. braunii	DMSO 5%	89
CCMA-UFSCar	168	Selenastrum bibraianum	DMSO 5%	76
CCMA-UFSCar	174	Kirchneriella irregularis var. spiralis	DMSO 5%	85
CCMA-UFSCar	176	Monoraphidium griffithii	DMSO 5%	100
CCMA-UFSCar	230	Kirchneriella irregularis	DMSO 5%	84
CCMA-UFSCar	234	Kirchneriella irregularis var. spiralis	DMSO 5%	100
CCMA-UFSCar	239	Ankistrodesmus densus	DMSO 5%	100
CCMA-UFSCar	241	Selenastrum bibraianum	DMSO 5%	62
CCMA-UFSCar	278	Ankistrodesmus stipitatus	DMSO 5%	95
CCMA-UFSCar	306	Monoraphidium contortum	DMSO 5%	100
CCMA-UFSCar	325	Monoraphidium pseudobraunii	DMSO 5%	61
CCMA-UFSCar	333	Monoraphidium kormakovae	DMSO 5%	100
CCMA-UFSCar	345	Kirchneriella obesa	DMSO 5%	75
CCMA-UFSCar	346	Kirchneriella pseudoaperta	DMSO 5%	63
CCMA-UFSCar	348	Kirchneriella irregularis	DMSO 5%	94
CCMA-UFSCar	349	Monoraphidium contortum	DMSO 5%	100
CCMA-UFSCar	350	Curvastrum pantanale	DMSO 5%	92
CCMA-UFSCar	353	Monoraphidium kormakovae	DMSO 5%	90
CCMA-UFSCar	423	Ankistrodesmus bernardii	DMSO 5%	82
CCMA-UFSCar	447	Kirchneriella contorta var. elegans	DMSO 5%	74
CCMA-UFSCar	455	Chlorolobion braunii	DMSO 5%	100
CCMA-UFSCar	462	Chlorolobion braunii	DMSO 5%	92
CCMA-UFSCar	470	Messastrum gracile	DMSO 5%	94
CCMA-UFSCar	476	Chlorolobion braunii	DMSO 5%	100
CCMA-UFSCar	478	Raphidocelis subcaptata	DMSO 5%	89
CCMA-UFSCar	492	Pseudokirchneriella elongata	DMSO 5%	100
CCMA-UFSCar	498	Pseudokirchneriella elongata	DMSO 5%	81
CCMA-UFSCar	516	Kirchneriella obesa	DMSO 5%	93
CCMA-UFSCar	549	Monoraphidium indicum	DMSO 5%	100
CCMA-UFSCar	604	Chlorolobion lunatum	DMSO 5%	100
CCMA-UFSCar	606	Monoraphidium irregularis	DMSO 5%	79
CCMA-UFSCar	609	Desmodesmus comunis	DMSO 5%	96
CCMA-UFSCar	611	Ankistrodesmus fusiformis	DMSO 5%	93
CCMA-UFSCar	622	Messastrum gracile	DMSO 5%	100

Accession number		Strain	CPA	r ABS (%)
CCMA-UFSCar	029	Scenedesmus bijugus	DMSO 5%	80	✓
CCMA-UFSCar	030	Desmodesmus comunis	DMSO 5%	86	✓
CCMA-UFSCar	046	Desmodesmus spinosus	DMSO 5%	75	✓
CCMA-UFSCar	060	Coelastrum sphaericum	DMSO 5%	78	✓
CCMA-UFSCar	062	Desmodesmus spinosus	DMSO 5%	88	✓
CCMA-UFSCar	088	Scenedesmus ecornis	DMSO 5%	82	✓
CCMA-UFSCar	244	Verrucodesmus verrucosus	DMSO 5%	79	✓
CCMA-UFSCar	326	Hariotina reticulata	DMSO 5%	100	✓
CCMA-UFSCar	493	Hariotina reticulata	DMSO 5%	81	✓
CCMA-UFSCar	609	Desmodesmus comunis	DMSO 5%	96	✓

Treatment	CPA	Cooling rate
T01	no	not frozen	✓
T02	no	LN	✗
T03	no	-1 ^oC min^-1	✗
T04	no	-5 ^oC min^-1	✗
T05	no	-0,5 ^oC min^-1	✗
T06	DMSO 5%	not frozen	✓
T07	DMSO 5%	LN	✗
T08	DMSO 5%	-1 ^oC min^-1	✗
T09	DMSO 5%	-5 ^oC min^-1	✗
T10	DMSO 5%	-0,5 ^oC min^-1	✗
T11	MeOH 5%	not frozen	✓
T12	MeOH 5%	LN	✗
T13	MeOH 5%	-1 ^oC min^-1	✓
T14	MeOH 5%	-5 ^oC min^-1	✓
T15	MeOH 5%	-0,5 ^oC min^-1	✓
T16	Glycerol 5%	not frozen	✓
T17	Glycerol 5%	LN	✗
T18	Glycerol 5%	-1 ^oC min^-1	✗
T19	Glycerol 5%	-5 ^oC min^-1	✗
T20	Glycerol 5%	-0,5 ^oC min^-1	✗

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[1] (The analysis was based on the growth rate calculated using the optical density of samples at two different points in time, compared with the control cultures (non-cryopreserved). The symbols and colors indicate the relative success of growth: green – good growth, yellow – medium success and red – poor growth.)