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以红姜花(Hedychium coccineum)的花丝和花药为外植体,通过体细胞胚胎发生途径建立了植株再生体系。结果表明:外植体在MS+4 mg·L-1 2,4-D+4 mg·L-1 NAA+1 mg·L-1 6-BA+30 g·L-1蔗糖+7 g·L-1琼脂的培养基上经过120 d诱导出愈伤组织,愈伤组织在MS+1 mg·L-1 2,4-D+0.25mg·L-1 NAA+0.25 mg·L-1 6-BA+30 g·L-1蔗糖+7 g·L-1琼脂的培养基上经过继代筛选,获得浅黄色松散易碎的胚性愈伤组织。胚性愈伤组织在MS无机盐+B5维生素+100 mg·L-1谷氨酰胺+230 mg·L-1脯氨酸+100 mg·L-1麦芽提取物+0.02 mg·L-1 NAA+0.02 mg·L-1 TDZ+0.5~1.0 mg·L-1 2,4-D+45g·L-1蔗糖的培养基中通过3个月的悬浮培养,可得到均质稳定的胚性细胞悬浮系(ECS)。胚性悬浮细胞在SH无机盐+B5维生素+100 mg·L-1谷氨酰胺+230 mg·L-1脯氨酸+100 mg·L-1麦芽提取物+0.25mg·L-1 NAA+0~0.20 mg·L-1 TDZ+45 g·L-1蔗糖+7 g·L-1琼脂的体胚诱导培养基中培养10 d,可见到白色半透明体胚发生,20 d后体胚发育成熟。当TDZ浓度为0.15 mg·L-1时,体胚诱导率高达4 500个·mL-1 PCV ECS(PCV:细胞密实体积)。在SH无机盐+B5维生素+0.20 mg·L-1 IAA+0.25~1.0mg·L-1 6-BA+30 g·L-1蔗糖+7 g·L-1琼脂的体胚萌发培养基上,体胚萌发率高达100%。将萌发的体胚转移到1/2MS+1 g·L-1活性炭成苗培养基中,进一步发育成正常的再生植株,植株室外栽培成活率达90%。
The plant regeneration system was established through somatic embryogenesis using the filament and anther of Hedychium coccineum as explants. The results showed that explants were cultured in MS + 4 mg · L-1 2,4-D + 4 mg · L-1 NAA + 1 mg · L-1 6-BA + 30 g · L-1 sucrose +7 g · The callus was induced on the medium of L-1 agar for 120 days. The callus was cultured on MS + 1 mg · L-1 2,4-D + 0.25 mg · L-1 NAA + 0.25 mg · L-1 -BA + 30 g · L-1 sucrose +7 g · L-1 agar medium was subcultured to obtain a pale yellow, loose and fragile embryogenic callus. Embryogenic Callus in MS + B5 Vitamin +100 mg · L-1 Glutamine +230 mg · L-1 Proline +100 mg · L-1 Malt Extract +0.02 mg · L-1 NAA +0.02 mg · L-1 TDZ + 0.5-1.0 mg · L-1 2,4-D + 45g · L-1 sucrose by 3-month suspension culture, homogeneous and stable embryogenic cells can be obtained Suspension system (ECS). Embryogenic cells were suspended in SH salts + B5 vitamins +100 mg · L-1 glutamine +230 mg · L -1 proline +100 mg · L -1 malt extract + 0.25 mg · L -1 NAA + After being cultured for 10 days in 0-20.0 mg · L-1 TDZ + 45 g · L-1 sucrose + 7 g · L-1 agar, white translucent somatic embryogenesis was observed. After 20 d, the somatic embryos Mature. At the TDZ concentration of 0.15 mg · L -1, the somatic embryo induction rate was as high as 4 500 cells · mL -1 PCV ECS (PCV: dense cell volume). The somatic embryo germination medium of SH inorganic salts + B5 vitamins + 0.20 mg · L -1 IAA + 0.25-1.0 mg · L -1 6-BA + 30 g · L -1 sucrose +7 g · L -1 agar , Somatic embryo germination rate as high as 100%. The germinated somatic embryos were transferred to 1 / 2MS + 1 g · L-1 activated carbon seedling culture medium and further developed into normal regenerated plants, the outdoor survival rate of plants reached 90%.