北柴胡花药愈伤组织高效再生体系的建立

Studies on high efficient plant regeneration system from anther callus of Bupleurum chinense

以花粉发育处于单核期的北柴胡花药为外植体诱导出愈伤组织,愈伤组织经多次继代培养后,置于20种含不同植物激素的分化培养基上诱导分化。分别在培养21,49 d后,统计每种培养基中分化出苗数,同时观察再生植株生长状况,从中筛选出分化率高,分化速度快,且植株生长状态良好的分化培养基。结果共在19种培养基均可分化出苗,分化率在3%~60%,大部分低于20%。在MS＋KT 0.5 mgL-1＋蔗糖30 gL-1＋植物凝胶5 gL-1培养基上分化率最高,为60%,其次为MS＋ ZT 1.0 mgL-1＋蔗糖30 gL-1＋植物凝胶5 gL-1,为58%。另外,针对在分化培养基中未能生根的再生芽,进行了生根培养基筛选。结果显示再生芽在生根培养基MS＋蔗糖30 gL-1 ＋植物凝胶5 gL-1和1/2 MS＋ NAA 0.5 mgL-1＋蔗糖30 gL-1 ＋植物凝胶5 gL-1中均可生根,生根率均为100%。由此建立了北柴胡花药愈伤组织高效稳定的再生体系。

The callus of Bupleurum chinense with anthers at the stage of uninucleate was induced. After several subcultures, anther calli of B. chinense were cultured at 20 MS culture mediums with different plant hormones to differentiate into plantlets. Differentiation of callus was detected after 21 and 49 days to select the most effective medium. There were 19 culture mediums in which anther callus could differentiate into plantlets with differentiation rate range from 3% to 60%, and most less than 20%. MS＋KT 0.5 mgL-1＋sucrose 30 gL-1＋phytagel 5 gL-1 was the best differentiation medium with the differentiation rate of 60%, followed by MS＋ZT 1.0 mgL-1＋sucrose 30 gL-1＋phytagel 5 gL-1 with the differentiation rate of 58%. Then plantlets were transferred to rooting medium to obtain whole plant. All plantlets could root in the rooting medium of MS＋sucrose 30 gL-1＋phytagel 5 gL-1 and 1/2 MS＋NAA 0.5 mgL-1＋sucrose 30 gL-1＋phytagel of 5 gL-1 with the rooting rate of 100%. As a result, the high efficient and stable plant regeneration system was established from anther callus of B. chinense.keywords：Bupleurum chinense