半夏愈伤组织诱导和分化再生植株研究.docVIP

半夏愈伤组织诱导和分化再生植株研究 　　摘要研究半夏外植体愈伤组织诱导、放置方式、外植体的苗龄对分化再生植株的影响,结果表明:块茎的愈伤组织诱导效果最佳,叶片次之,叶柄最差。当MS+6-BA 2mg/L+2,4-D 2mg/L激素组配时,愈伤组织分化率达100%。外植体的放置方式对分化出苗有很大影响,块茎垂直放置出芽率最高,而叶柄和叶片平放于培养基效果较好。无菌苗苗龄对分化率有较大的影响,最适无菌苗苗龄为10~25d。当6-BA为3mg/L时芽增殖倍数达4倍左右,而当6-BA 0.2mg/L+2,4-D 0.2mg/L组配时,生根效果最好。 　　关键词半夏;组织培养;分化再生;愈伤组织 　　中图分类号S567.239文献标识码A文章编号 1007-5739(2010)04-0136-02 　　 　　StudiesonCallusInductionandPlantletsRegenerationofPinelliaternata 　　HE Feng-mei1ZHU Yong-ping2 　　(1College of Horticulture and Landscape,Yunnan Agricultural University,Kunming Yunnan 650201; 2College of Agriculture and Biotechnology,Yunnan Agricultural University) 　　AbstractThe effect of callus induction,orientation and age of explants on plantlets regeneration of Pinellia ternata was studied.Results showed that tuber had the best ability in callus induction which was followed by leaf and petiole.The combination of MS+6-BA 2mg/L+2,4-D 2mg/L was optimized for differentiation of callus with differentiation frequency up to 100%.There were significant effect of explants orientation on optimum seedling differentiation in which the tuber vertically,and leaf and petiole horizontally placed on medium. Explants harvested from 10 to 25-day-old in vitro shots were significantly better than others in regeneration. The optimum hormone for bud proliferation was MS+6-BA 3mg/L and proliferation multiple of buds achieved 4 times.The combination of MS+6-BA 0.2mg/L+2,4-D 0.2mg/L was optimized for rooting. 　　Key wordsPinellia ternata;tissue culture;preparation and regeneration;callus 　　 　　半夏(Pinelliae ternata)为天南星科多年生草本植物,药用历史悠久,其地下块茎干燥炮制后入药,具有燥湿化痰、降逆止呕、消痞散结之功效[1]。近年来,研究发现半夏还有抗早孕、抗肿瘤、降血脂、护肝等功能,因而备受国内外关注[2]。半夏既能进行有性繁殖,又能进行无性繁殖。但半夏结实率低,出苗率低,种子在常温条件下贮藏,生活力较易丧失,因此,通常生产上不直接用种子繁殖[3]。在自然界中,半夏主要由生于叶柄下的珠芽繁殖。由于珠芽采集困难,故不能广泛栽培。因此,药源只能收集野生半夏,野生半夏也因大量采挖而近于枯竭,不能满足药用需求[4]。利用组培技术对半夏进行试管繁殖,不仅可提高繁殖系数,还可显著改良半夏品质[3]。因此,有必要利用半夏组织培养快速繁殖提供植物材料以满足市场需求。 　　1材料与方法 　　1.1试验材料 　　来源于云南农业科学院生物技术与种质资源研究所。 　　1.2试验方法 　　1.2.1激素对愈伤组织形成的影响。以MS培养基为基本培养基:琼脂9g/L+