Study On Regeneration System Construction And Total Phenol Contents Change In Callus Of Magnolia Officinalis Subsp. Biloba (Rehd.Et Wils.)Law

Magnolia officinalis subsp. biloba (Rehd.et Wils.) Law. is original woody plant in taxonomy and important traditional medicine in China which belonged to Magnoliaceae. The barks of root, branch and stem in M. officinalis subsp. biloba (Rehd.et Wils.) Law. are medicinal, used for sterilization, thoracic-abdominal distension and phlegm asthma, and occupy an important place in the clinical application. Be from Hunan, Hubei, Jiangxi and widely spread in the country, M. officinalis subsp. biloba (Rehd.et Wils.) Law. is an unique economic species in China, medicinal, timber used and ornamental. As a secondary protective plant in China, the industry of M. officinalis subsp. biloba (Rehd.et Wils.) Law. has broad prospects and great potentions. The study provided productive methods for rapid propagation in qualitive provences through in virto regeneration system construction of M. officinalis subsp. biloba (Rehd.et Wils.) Law.; by callus induction of different explants cultured under different conditions and detected the change of total phenol contents, to provide references for development of cell culture being extracted total phenols in future. The main conclusions were as follows:1. Before removing testa artificially, rubbing repeatedly, immersing in warm water and preliminary disinfecting by0.2%KMnO4. To adopt75%alcohol in15s and0.1%HgCl2in8min on sterile desk, the seeds were injured at least and the pollution rate reached22.6%;2. The optimum initial culture medium was:3/4B5+6-BA0.5mg·L-1+NAA0.1mg·L-1+sucrose20g·L-1+agar6.5g·L-1, PH:5.8-6.0,temperature:24±1℃, illumination time:12h·d-1, illumination intensity:1500-20001ux. The stem with leaves was more suitable for multiple buds proliferation and the optimum culture medium was: MS+6-BA2.5mg·L-1+NAA0.5mg·L-1with the induced rate reaching93.18%and multiplication coefficient reaching6.25; the optimum rooting medium was:1/2MS+NAA1.Omg·L-1, sucrose25g·L-1and the rooting rate reached81.5%.3. Hypocotyl was the most suitable explants for callus induction, being the best state in the medium of B5+6-BA2.0mg·L-1+2,4-D1.5mg·L-1+sucrose25g·L-1+agar6.5g·L-1, and the induced rate reached over90%; the optimum culture medium for callus proliferation was:B5+6-BA2.0mg·L-1+2,4-D1.5mg·L-1+NAA0.5mg·L-1+sugar25g·L-1+agar6.0g·L-1. 4. The total phenol contents of callus varied greatly from0.01%to0.25%in different provenances; the same as different organs in the same provenance. The contents of young stem induced callus was the highest and hypocotyl had the lowest contents; different browning level also had effect and the influence degree was:serious browning> mild browning> not browning; the addition of two precursor compound (L-phenylalanine and DL-β-phenylalanine) could increase total phenol contents effectively in callus, of which DL-β-phenylalanine was better and it can increase the contents from0.06%to0.16%, more than1.69%-7.27%as control.