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红姜花体细胞胚胎发生及植株再生的研究 被引量:3

Somatic Embryogenesis and Plant Regeneration of Hedychium coccineum
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摘要 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%。 A reproducible protocol for somatic embryogenesis was established for the ornamental ginger Hedychium coccineum using filaments and anthers. After culture for 120 days,callus was induced on Murashige and Skoog(MS)medium supplemented with 4 mg · L-1 2,4-D,4 mg · L-1 NAA,1 mg · L-1 6-BA,30 g · L-1 sucrose and 7 g · L-1 agar,proliferated on MS medium containing 1 mg · L-1 2,4-D,0.25 mg · L-1 NAA,0.25 mg · L-1 6-BA,30 g · L-1 sucrose and 7 g · L-1 agar. Light yellow and friable embryogenic callus was obtained. Embryogenic calli were suspended in liquid medium containing MS basal salts,B5 vitamins,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 and 45 g · L-1sucrose. After 3 months culture,a homogeneous and stable embryogenic cell suspension(ECS),composed of small cell aggregates,was established. Planting of stable ECS on somatic embryo induction media containing Schenk and Hildebrandt(SH)basal salts,B5 vitamins,100 mg · L-1 glutamine,230 mg · L-1 proline,100 mg · L-1malt extract,0.25 mg · L-1 NAA,0–0.20 mg · L-1 TDZ,45 g · L-1 sucrose and 7 g · L-1 agar. White and translucent globular embryos were induced after 10 days culture,and mature somatic embryos were obtained after 20 days culture. The highest induction frequency of 4 500 somatic embryos · mL-1 PCV ECS(PCV:packed cell volume)was derived from medium with 0.15 mg · L-1 TDZ. A germination percentage of 100% were observed in germination media,which consisted of SH basal salts,B5 vitamins,0.20 mg · L-1 IAA and 0.25–1.0 mg · L-1 6-BA. Regenerated plantlets with normal shoot and root were developed after one month on half strength MS medium with 1 g · L-1 active charcoal. Well rooted plantlets were successfully acclimatized with a survival rate of 90% and grew vigorously.
作者 涂红艳 肖望 邓崇会 TU Hong-yan;XIAO Wang;DENG Chong-hui;Department of Biology,Guangdong University of Education;
出处 《园艺学报》 CAS CSCD 北大核心 2014年第10期2139-2146,共8页 Acta Horticulturae Sinica
基金 广东省自然科学基金博士启动项目(10451030301004286) 2013年广东省高等院校学科与专业建设专项资金项目(2013KJCX0137) 广州市科技计划项目(2014J4100151)
关键词 红姜花 胚性细胞悬浮系 体细胞胚胎发生 植株再生 Hedychium coccineum embryogenic cell suspensions somatic embryogenesis plant regeneration
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