培养方式对川贝母胚性愈伤组织生长及活性物质积累的影响OA

The aim of this study was to screen an optimal culture method for the growth and bioactive metabolite accumulation of Fritillaria cirrhosa D.Don embryogenic callus,thereby providing a reference for the rapid proliferation and efficient enrichment of its secondary metabolites within a short period.Three culture methods,namely"solid-solid","solid-liquid"and"solid-liquid-solid",were adopted to culture the embryogenic callus of F.cirrhosa.After 40 days of culture,structural differentiation,growth yield,in vitro antioxidant capacity,and bioactive metabolite accumulation were compared between the treatments.Significant differences were observed among the three culture methods for the embryogenic callus of F.cirrhosa,with the"solid-liquid-solid"mode showing the best overall performance.(1)Structural differentiation:The callus cultured by"solid-liquid-solid"mode showed uniform morphology,compact texture,and bright color.The cell nucleus was regular in shape and evenly distributed.The procambial bundle structure was well-defined,and the cells exhibited typical embryogenic characteristics.(2)Proliferation coefficient:The proliferation multiple of the callus under this system was 1.45,which was 2.84 and 1.93 times that of the"solid-solid"and"solid-liquid"cultures,respectively.(3)In vitro antioxidant capacity:The callus from the"solid-liquid-solid"culture exhibited strong DPPH radical scavenging activity(47.55％)and a high FRAP value(720.95 μmol·L-1),both of which were significantly superior to those of the"solid-solid"culture(40.41％,482.86 μmol·L-1)and the"solid-liquid"culture(44.15％,694.68 μmol·L-1).(4)Bioactive metabolite accumulation:The embryogenic callus in this group exhibited the highest levels of soluble sugar(56.15 mg·g-1),total flavonoids(2.06 mg·g-1),total phenolic(1.44 mg·g-1),imperialine(0.25 mg·g-1).These values were 1.07,1.15,1.66,and 1.39 times as high as those of the"solid-solid"culture,and 1.23,1.11,1.13,and 1.19 times as high as those of the"solid-liquid"culture,respectively.This study provides a theoretical basis and technical support for the large-scale production of secondary metabolites and the establishment of a plant regeneration system for F.cirrhosa.