@article{fdi:010081584,
  title = {{I}mproved cryopreservation using droplet-vitrification and histological changes associated with cryopreservation of madder ({R}ubia akane nakai)},
  author = {{Y}i, {J}. {Y}. and {S}ylvestre, {I}sabelle and {C}olin, {M}. and {S}alma, {M}. and {L}ee, {S}. {Y}. and {K}im, {H}. {H}. and {P}ark, {H}. {J}. and {E}ngelmann, {F}lorent},
  editor = {},
  language = {{ENG}},
  abstract = {{A}n efficient protocol for cryopreservation of madder hairy root cultures has been developed using droplet-vitrification. {I}n previous study, combining loading solution {C}4 (35% {PVS}3) and vitrification solution {B}5 (80% {PVS}3) was the most effective method. {I}n this study, we tried three types of vitrification solution, {B}5, {A}3 (90% {PVS}2, on ice), and {A}5 (70% {PVS}2, on ice). {C}ombining loading solution {C}4 and vitrification solution {A}5 (on ice) showed the best regeneration rate in this study. {H}istological changes of the cells within the hairy root of madder were also observed in different steps. {T}he cells from the hairy roots of the control treatment were full and intact with different size of vacuoles and obvious cell nucleus having a dark nucleolus. {A}fter the stage of preparing for cryopreservation (after preculturing, loading, followed by dehydration solution {AS} or {B}5), intercellular spaces had become distinct, and within cells, the cytoplasms had become denser and week plasmolyses had appeared. {T}he cell plasmolyses were much more apparent and we measured the degree of plasmolysis by calculating, the area of cell/the area of cytoplasm. {T}he value of plasmolysis degree was the highest in the combination of preculture, loading solution {C}4, and dehydration solution {A}5, 1.97. {B}ecause the highest regeneration rates appeared in the treatment of {AS} for 20 min, we could assume that the optimal degree of plasmolysis for cryopreservation might be around 1.97. {T}he changes in cell structure during cryopreservation might be a useful basis for the development of a proper long-term preservation method for madder germplasms.},
  keywords = {hairy root ; loading solution ; plasmolysis ; regeneration rate ; vitrification solution},
  booktitle = {},
  journal = {{K}orean {J}ournal of {H}orticultural {S}cience and {T}echnology},
  volume = {30},
  numero = {1},
  pages = {79--84},
  ISSN = {1226-8763},
  year = {2012},
  DOI = {10.7235/hort.2012.11087},
  URL = {https://www.documentation.ird.fr/hor/fdi:010081584},
}