烘乾處理對葉下珠有效成分含量影響及葉下珠無菌繁殖技術建立

Abstract

葉下珠，學名Phyllanthus urinaria L.，是大戟科(Euphorbiaceae) 油柑屬( genus Phyllanthus ) 植物，為一年生草本植物。本研究以0.1% 磷酸+ 0.1%乙腈的去離子水-100%乙腈二種溶液為流動相，建立葉下珠Gallic acid、Corilagin、Rutin三種含量相對較多之指標成分HPLC分析方法。結果顯示，Gallic acid滯留時間約為4.5 min，Corilagin 11.8 - 12.1 min間，Rutin約在18.3 min。90℃水萃30分鐘為葉下珠三種指標活性成分最佳萃取方法。乾燥處理試驗結果，葉下珠以70℃熱風乾燥24小時，為最佳且快速的乾燥方法，鮮草採收後不萎凋及室內萎凋2.5%可得到較高指標活性成分量，乾燥前不需經紫外光照射。 葉下珠無菌播種的平均發芽率為46.6%。 誘導無菌初生根及胚軸發根試驗結 果，以NAA 2 mg．L-1處理較佳。MS培養基可誘導葉下珠多芽產生，可用於大量繁殖無性株；以NAA 0.5, 1.0, 2.0 mg．L-1加BA1.0、2.0或5.0 mg．L-1處理培養，可分化成癒傷組織，有助建立懸浮細胞培養系統時應用。NAA濃度和BA濃度之間，彼此存在交感作用，會相互影響葉下珠莖節培殖體分化方向，使形成根、癒傷組織或多芽，導致產生不同組織分化結果。

Shatterstone is an annual plant belongs to the family Euphorbiaceae and genus Phyllanthus, which scientific name is Phyllanthus urinaria. In this study, we use 0.1% H3PO4 + 0.1% Acetonitrile distilled H2O and 100% Acetonitrile as mobile phases to establish HPLC analysis method to analyze the content of active compounds of Phyllanthus urinaria. Results show that the elution retention time of Gallic acid is 4.5 min, Corilagin is from 11.8 min to 12.1 min, and Rutin was separated at 18.3 min. In drying experiments, 70 ℃ oven drying is the recommended way to process Phyllanthus urinaria. No withering and withering about 2.5% fresh weight have the same consequence in three active compounds content. Before drying, it need not to do UV exposure. The average germinating rate in Phyllanthus urinaria sterile seed culture is 46.6%. Primary roots and hypocotyls as explants to compare rooting rate, NAA mg．L-1 treatment has the best result. MS medium promote multipleshooing and can be use in magnifying Phyllanthus urinaria clone.On the other hand, NAA 0.5, 1.0, 2.0 mg．L-1 with BA1.0、2.0 or 5.0 mg．L-1 can induce callus, which could be used in suspension culture system. There are interactions among NAA and BA concentrations to the inductions of Phyllanthus urinaria nodal explants, leading then to rooting, callusing, or multipleshooting.