Plant regeneration from rice anthers cryopreserved by an encapsulation/dehydration technique.

MARÍA A. MARASSI; ADRIANA SCOCCHI; ANA MARÍA GONZALEZ

IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY. PLANT

Año: 2006 p. 31 - 36

1054-5476

Anther-derived rice (Oryza sativa L. ssp. japonica variety Yerua P.A.) plants were obtained after cryopreservation by an encapsulation/dehydration technique. Immature anthers, excised from spikelets pretreated at 88C for 8 d, were encapsulated in calcium alginate beads. The beads were cultured on N6 medium with 11.5 mM naphthaleneacetic acid (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. encapsulated in calcium alginate beads. The beads were cultured on N6 medium with 11.5 mM naphthaleneacetic acid (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. encapsulation/dehydration technique. Immature anthers, excised from spikelets pretreated at 88C for 8 d, were encapsulated in calcium alginate beads. The beads were cultured on N6 medium with 11.5 mM naphthaleneacetic acid (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. encapsulated in calcium alginate beads. The beads were cultured on N6 medium with 11.5 mM naphthaleneacetic acid (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. Oryza sativa L. ssp. japonica variety Yerua P.A.) plants were obtained after cryopreservation by an encapsulation/dehydration technique. Immature anthers, excised from spikelets pretreated at 88C for 8 d, were encapsulated in calcium alginate beads. The beads were cultured on N6 medium with 11.5 mM naphthaleneacetic acid (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. encapsulated in calcium alginate beads. The beads were cultured on N6 medium with 11.5 mM naphthaleneacetic acid (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. 8C for 8 d, were encapsulated in calcium alginate beads. The beads were cultured on N6 medium with 11.5 mM naphthaleneacetic acid (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. mM naphthaleneacetic acid (NAA) and 2.3mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. mM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for 3 d in liquid medium, desiccated on silica gel, slowly cooled to 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. 2308C, immersed in liquid nitrogen (LN), thawed, and recultured. The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers), which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process.