The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
S Higuchi - One of the best experts on this subject based on the ideXlab platform.
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cryopreservation of Apical Meristems of white clover trifolium repens l by vitrification
Plant Science, 1991Co-Authors: Toshihiko Yamada, A Sakai, T Matsumura, S HiguchiAbstract:Abstract Apical Meristems of white clover ( Trifolium repens L.) were successfully cryopreserved by vitrification. Excised Apical Meristems from in vitro plantlets were precultured on agar-B5 medium containing 1.2 M sorbitol at 4°C for 2 days. Meristems following preculture were transferred to a 1.8 ml plastic cryotube and then a highly concentrated cryoprotective solution (designated PVS2) was added at 25°C or at 0°C. The Meristems treated with PVS2 at 25°C for 5 min or at 0°C for 15 min were directly plunged into liquid nitrogen. After rapid warming in water at 25°C, the Meristems were expelled into B5 medium containing 1.2 M sucrose. Meristems on filter paper over agar-B5 medium resumed growth within 3 days and directly developed shoots. The average rate of plant regeneration was about 80% in Meristems treated with PVS2 at 25°C for 5 min or at 0°C for 15 min. This procedure for cryopreservation is very simple and appears promising as a routine method for cryopreserving Apical Meristems.
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cryopreservation of Apical Meristems of white clover trifolium repens l
Plant Science, 1991Co-Authors: Toshihiko Yamada, T Matsumura, A Sakai, S HiguchiAbstract:Abstract Apical Meristems of white clover (Trifolium repens L.) were tested for their ability to survive immersion in liquid nitrogen (LN). High rates of survival and plant regeneration were obtained when shoot Meristems were precultured on agar-B5 medium containing 5% (v/v) dimethylsulfoxide (DMSO) and 5% (w/v) glucose at 4°C for 2 days and then frozen to −40°C at 0.3°C/min with 10% DMSO and 10% glucose as cryoprotectants. After rapid thawing in 40°C water, the regrowth of LN stored Meristems was achieved by transfer without washing onto filter paper discs over agar-solidified B5 medium. This post-thaw handling significantly improved the survival rates. Length of storage time did not significantly decrease the survival rate. The survival rates of Meristems frozen in LN under optimal conditions exceeded 80% in all cultivars tested. Survival rates were not correlated with the field cold hardiness of the cultivars tested. The cryogenic protocol reported here appears promising as a routine method for preserving genetic resources of white clover.
A Sakai - One of the best experts on this subject based on the ideXlab platform.
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Cryopreservation of in vitro-grown Apical Meristems of hybrid statice by three different procedures
Scientia Horticulturae, 1998Co-Authors: T Matsumoto, A Sakai, Chiaki Takahashi, Yoji NakoAbstract:Abstract In vitro-grown Apical Meristems of hybrid statice (Limonium cv. Blue Symphonet) were cryopreserved by three cryogenic procedures; (1) vitrification with encapsulation, (2) vitrification without encapsulation, and (3) a revised encapsulation/dehydration technique. When dehydration tolerance was well developed by preconditioning and cryogenic procedures were well optimized, these three procedures produced nearly the same levels of growth recovery (70–75%). These results support our theory that the acquisition of dehydration tolerance is sufficient for specimens to survive to cryopreservation.
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cryopreservation of in vitro grown Apical Meristems of lily by vitrification
Plant Cell Tissue and Organ Culture, 1995Co-Authors: T Matsumoto, A Sakai, K. YamadaAbstract:Apical Meristems from adventitious buds induced by culturing of bulb-scale segments of Japanese Pink Lily (Lilium japonicum Thunb.) were successfully cryopreserved by a vitrification. The excised Apical Meristems were precultured on a solidified Murashige & Skoog medium, containing 0.3 M sucrose, for 1 day at 25°C and then loaded in a mixture of 2 M glycerol plus 0.4 M sucrose for 20 min at 25°C. Cryoprotected Meristems were then sufficiently dehydrated with a highly concentrated vitrification solution (designated PVS2) at 25°C for 20 min or at 0°C for 110 min prior to a plunge into liquid nitrogen. After rapid warming in a water bath at 40°C, the Meristems were placed in 1.8 ml of 1.2 M sucrose for 20 min and then, placed on filter papers over gellan gum-solidified MS medium. The revived Meristems resumed growth within 5 days and directly produced shoots. The rate of shoot formation was approximately 80% after 4 weeks. When bulb-scale segments with adventitious buds were cold-hardened at 0°C for more than 7 days before the procedure, the rates of shoot formation were significantly increased. This vitrification method was successfully applied to five other lily cultivars. Thus, this vitrification procedure for cryopreservation appears promising as a routine method for cryopreserving Meristems of lily.
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cryopreservation of in vitro grown Apical Meristems of wasabi wasabia japonica by vitrification and subsequent high plant regeneration
Plant Cell Reports, 1994Co-Authors: T Matsumoto, A Sakai, K. YamadaAbstract:In vitro-grown Apical Meristems of wasabi (Wasabia japonica Matsumura) were successfully cryopreserved by vitrification. Excised Apical Meristems precultured on solidified M S medium containing 0.3M sucrose at 20°C for 1 day were loaded with a mixture of 2M glycerol and 0.4M sucrose for 20 min at 25°C. Cryoprotected Meristems were then sufficiently dehydrated with a highly concentrated vitrification solution (designated PVS2) for 10 min at 25°C prior to a plunge into liquid nitrogen. After rapid warming, the Meristems were expelled into 2 ml of 1.2M sucrose for 20 min and then plated on solidified culture medium. Successfully vitrified and warmed Meristems remained green after plating, resumed growth within 3 days, and directly developed shoots within two weeks. The average rate of normal shoot formation amounted to about 80 to 90% in the cryopreserved Meristems. This method was successfully applied to three other cultivars of wasabi. This vitrification procedure promises to become a routine method for cryopreserving Meristems of wasabi.
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cryopreservation of Apical Meristems of white clover trifolium repens l by vitrification
Plant Science, 1991Co-Authors: Toshihiko Yamada, A Sakai, T Matsumura, S HiguchiAbstract:Abstract Apical Meristems of white clover ( Trifolium repens L.) were successfully cryopreserved by vitrification. Excised Apical Meristems from in vitro plantlets were precultured on agar-B5 medium containing 1.2 M sorbitol at 4°C for 2 days. Meristems following preculture were transferred to a 1.8 ml plastic cryotube and then a highly concentrated cryoprotective solution (designated PVS2) was added at 25°C or at 0°C. The Meristems treated with PVS2 at 25°C for 5 min or at 0°C for 15 min were directly plunged into liquid nitrogen. After rapid warming in water at 25°C, the Meristems were expelled into B5 medium containing 1.2 M sucrose. Meristems on filter paper over agar-B5 medium resumed growth within 3 days and directly developed shoots. The average rate of plant regeneration was about 80% in Meristems treated with PVS2 at 25°C for 5 min or at 0°C for 15 min. This procedure for cryopreservation is very simple and appears promising as a routine method for cryopreserving Apical Meristems.
Toshihiko Yamada - One of the best experts on this subject based on the ideXlab platform.
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cryopreservation of Apical Meristems of white clover trifolium repens l by vitrification
Plant Science, 1991Co-Authors: Toshihiko Yamada, A Sakai, T Matsumura, S HiguchiAbstract:Abstract Apical Meristems of white clover ( Trifolium repens L.) were successfully cryopreserved by vitrification. Excised Apical Meristems from in vitro plantlets were precultured on agar-B5 medium containing 1.2 M sorbitol at 4°C for 2 days. Meristems following preculture were transferred to a 1.8 ml plastic cryotube and then a highly concentrated cryoprotective solution (designated PVS2) was added at 25°C or at 0°C. The Meristems treated with PVS2 at 25°C for 5 min or at 0°C for 15 min were directly plunged into liquid nitrogen. After rapid warming in water at 25°C, the Meristems were expelled into B5 medium containing 1.2 M sucrose. Meristems on filter paper over agar-B5 medium resumed growth within 3 days and directly developed shoots. The average rate of plant regeneration was about 80% in Meristems treated with PVS2 at 25°C for 5 min or at 0°C for 15 min. This procedure for cryopreservation is very simple and appears promising as a routine method for cryopreserving Apical Meristems.
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cryopreservation of Apical Meristems of white clover trifolium repens l
Plant Science, 1991Co-Authors: Toshihiko Yamada, T Matsumura, A Sakai, S HiguchiAbstract:Abstract Apical Meristems of white clover (Trifolium repens L.) were tested for their ability to survive immersion in liquid nitrogen (LN). High rates of survival and plant regeneration were obtained when shoot Meristems were precultured on agar-B5 medium containing 5% (v/v) dimethylsulfoxide (DMSO) and 5% (w/v) glucose at 4°C for 2 days and then frozen to −40°C at 0.3°C/min with 10% DMSO and 10% glucose as cryoprotectants. After rapid thawing in 40°C water, the regrowth of LN stored Meristems was achieved by transfer without washing onto filter paper discs over agar-solidified B5 medium. This post-thaw handling significantly improved the survival rates. Length of storage time did not significantly decrease the survival rate. The survival rates of Meristems frozen in LN under optimal conditions exceeded 80% in all cultivars tested. Survival rates were not correlated with the field cold hardiness of the cultivars tested. The cryogenic protocol reported here appears promising as a routine method for preserving genetic resources of white clover.
Ryo Akashi - One of the best experts on this subject based on the ideXlab platform.
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multiple shoot formation somatic embryogenesis and plant regeneration from seed derived shoot Apical Meristems in ruzigrass brachiaria ruziziensis
Grassland Science, 2009Co-Authors: Genki Ishigaki, Takahiro Gondo, Kazuhiro Suenaga, Ryo AkashiAbstract:We have established a plant regeneration system via multiple shoot formation, or somatic embryogenesis from seed-derived shoot Apical Meristems of ruzigrass (Brachiaria ruziziensis). Shoot apices as initial explants were isolated from aseptically germinated seedlings, and cultured in vitro. The most effective treatments for multiple shoots and embryogenic calli formation were 0.5 mg L−1 2,4-dichlorophnoxyacetic acid (2,4-D) plus 2.0 mg L−1 6-benzylaminopurine (BAP) and 4.0 mg L−1 2,4-D plus 0.2 mg L−1 BAP, respectively. Plant regeneration from multiple shoot-clumps was achieved by culturing on solid MS medium containing 1.0 mg L−1 BAP or 2.0 mg L−1 kinetin plus 2.0 mg L−1 gibberellic acid. Somatic embryos regenerated plantlets on a MS medium without plant hormones. Regenerated shoots were germinated precociously on MS medium containing 0.01 or 0.1 mg L−1 naphthalene acetic acid and 2.0 mg L−1 BAP. All regenerants were successfully grown up to maturity. The plant regeneration via multiple-shoot clumps and embryogenic calli formation can be used as an alternative regenerable target tissue for genetic transformation using particle bombardment of ruzigrass.
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somatic embryogenesis and multiple shoot formation from seed derived shoot Apical Meristems of rhodesgrass chloris gayana kunth
Grassland Science, 2007Co-Authors: Takahiro Gondo, Kazutomi Yamakawa, Shinichi Tsuruta, Jun Matsumoto, Masumi Ebina, Ryo AkashiAbstract:High-frequencies of somatic embryogenesis and multiple-shoot formation were achieved with seed-derived shoot Apical Meristems of rhodesgrass (Chloris gayana Kunth). Shoot apices as initial explants were isolated from aseptically germinated seedlings, and cultured in vitro. Embryogenic calli and the multiple shoots were induced and maintained on MS basal medium with various combinations of 2,4-dichlorophnoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP), the two most effective treatments being 2.0 mg L⁻¹ 2,4-D and 0.1 mg L⁻¹ 2,4-D + 2.0 mg L⁻¹ BAP. These embryogenic calli and multiple-shoot formations could be used as alternative regenerable target tissues for genetic transformation using particle bombardment of rhodesgrass.
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somatic embryogenesis and multiple shoot formation from seed derived shoot Apical Meristems of rhodesgrass chloris gayana kunth
Grassland Science, 2007Co-Authors: Takahiro Gondo, Kazutomi Yamakawa, Shinichi Tsuruta, Jun Matsumoto, Masumi Ebina, Ryo AkashiAbstract:High-frequencies of somatic embryogenesis and multiple-shoot formation were achieved with seed-derived shoot Apical Meristems of rhodesgrass (Chloris gayana Kunth). Shoot apices as initial explants were isolated from aseptically germinated seedlings, and cultured in vitro. Embryogenic calli and the multiple shoots were induced and maintained on MS basal medium with various combinations of 2,4-dichlorophnoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP), the two most effective treatments being 2.0 mg L⁻¹ 2,4-D and 0.1 mg L⁻¹ 2,4-D + 2.0 mg L⁻¹ BAP. These embryogenic calli and multiple-shoot formations could be used as alternative regenerable target tissues for genetic transformation using particle bombardment of rhodesgrass.
K. Yamada - One of the best experts on this subject based on the ideXlab platform.
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cryopreservation of in vitro grown Apical Meristems of lily by vitrification
Plant Cell Tissue and Organ Culture, 1995Co-Authors: T Matsumoto, A Sakai, K. YamadaAbstract:Apical Meristems from adventitious buds induced by culturing of bulb-scale segments of Japanese Pink Lily (Lilium japonicum Thunb.) were successfully cryopreserved by a vitrification. The excised Apical Meristems were precultured on a solidified Murashige & Skoog medium, containing 0.3 M sucrose, for 1 day at 25°C and then loaded in a mixture of 2 M glycerol plus 0.4 M sucrose for 20 min at 25°C. Cryoprotected Meristems were then sufficiently dehydrated with a highly concentrated vitrification solution (designated PVS2) at 25°C for 20 min or at 0°C for 110 min prior to a plunge into liquid nitrogen. After rapid warming in a water bath at 40°C, the Meristems were placed in 1.8 ml of 1.2 M sucrose for 20 min and then, placed on filter papers over gellan gum-solidified MS medium. The revived Meristems resumed growth within 5 days and directly produced shoots. The rate of shoot formation was approximately 80% after 4 weeks. When bulb-scale segments with adventitious buds were cold-hardened at 0°C for more than 7 days before the procedure, the rates of shoot formation were significantly increased. This vitrification method was successfully applied to five other lily cultivars. Thus, this vitrification procedure for cryopreservation appears promising as a routine method for cryopreserving Meristems of lily.
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cryopreservation of in vitro grown Apical Meristems of wasabi wasabia japonica by vitrification and subsequent high plant regeneration
Plant Cell Reports, 1994Co-Authors: T Matsumoto, A Sakai, K. YamadaAbstract:In vitro-grown Apical Meristems of wasabi (Wasabia japonica Matsumura) were successfully cryopreserved by vitrification. Excised Apical Meristems precultured on solidified M S medium containing 0.3M sucrose at 20°C for 1 day were loaded with a mixture of 2M glycerol and 0.4M sucrose for 20 min at 25°C. Cryoprotected Meristems were then sufficiently dehydrated with a highly concentrated vitrification solution (designated PVS2) for 10 min at 25°C prior to a plunge into liquid nitrogen. After rapid warming, the Meristems were expelled into 2 ml of 1.2M sucrose for 20 min and then plated on solidified culture medium. Successfully vitrified and warmed Meristems remained green after plating, resumed growth within 3 days, and directly developed shoots within two weeks. The average rate of normal shoot formation amounted to about 80 to 90% in the cryopreserved Meristems. This method was successfully applied to three other cultivars of wasabi. This vitrification procedure promises to become a routine method for cryopreserving Meristems of wasabi.