Studies On The Germplasm Resources Evaluation And Utilization On Medical Lily In Hunan

Lily is a kind of perennial corm herb assembling medicinal,edible and ornamental value. The studies on morphological characteristics, tissue cultures and DNA fingerprints, genetic breeding and many other aspects of Lily flowers have been very thorough and the food value of edible Lily has been known, but the study on medicinal Lily is more shallow, especially its germplasm research is still blank. This thesis has carried out systematic studies on biological characteristics, water-soluble polysaccharide, total phospholipids and sapogenin and so on in Lily,and analyzed the HPLC fingerprint from extracted sapogenin, the genetic diversity with RAPD analysis and pharmacological action in order to comprehensively evaluate the germplasm resources about medical Lily in Hunan, provide the scientific basis for better protection and utilization of Lily resources, provide an experiment basis for formulation of manufacturing quality management about Hunan medical Lily.The main results are as follows:1. In the long-term natural evolution, hybridization, mutation, introduction and domestication process,it formed abundant Lily resources in Hunan Province. Lily resources in Hunan Province are divided into three kinds,including Lilium brownii F.E.Brown var.viridulum Baker,L.lancifolium Thunb. and L. pumilum DC. as well as wild species Cardiocrinum giganteum (Wall.) Makino by surveying, collecting, dollating and classifying the resources from Long-shan County, Long-hui County,Heng-shan County and so on. By observating and evaluating the biological characteristics ( growth period traits, morphological characteristics and so on) of 14 Lily samples introduced a fine variety from different places,the cultivation of suitable varieties have Lily and Dan vol in Hunan Province.2. 27 medical Lily samples from 8 different areas in Hunan Province were collected, and analyzed its main components such as water-soluble polysaccharide, total phospholipids and total sapogenin. Every component contents among samples were different. Total phospholipids and total sapogenin contents of Dan vol from Xiang-xi autonomous prefecture are the highest, with 4.69 mg/10g and 3.70 mg/10g; Water-soluble polysaccharide content of Lily from Shao-yang County is the highest=with 6.41%. So we think that Dan vol resources from Xiang-xi autonomous prefecture are suitable as high quality germplasm resources of medical Lily in Hunan Province. Lily resources from Shao-yang County are higher edible value, and more suitable as a food development. The content change ranges of three kinds in components are different,the total content changes on sapogenin are the biggest and the water-soluble carbohydrates the smallest. These resulted showed that total sapogenin as an index is better than water-soluble carbohydrate on breeding Lily varieties.3. Hunan has a good periodic biological conditions,Lily germplasms are widely distributed in the territory of Hunan Province. Long-shan and Long-hui County have a long history planting Lily. After a long-term natural selection and artificial selection, the farmer varieties (or types) of Lily are formed separately. Their ingredients varied. Through examining and analyzing water-soluble polysaccharide,total phospholipids, total sapogenin, water content, ash content and extracted content in Lily of 5 different species in Hunan province, the level of various ingredients mixed. All component contents came to significance levels, but didn't reach a extremely significance level(0.01 < P < 0.05). Total phospholipids and total sapogenin contents of Dan vol 'jiantou' were the tallest and had 4.97 mg/10g and 3.51 mg/10g; Water-soluble polysaccharide content of 'LongYa' Lily was the highest and had 7.39%. Dan vol 'jiantou' germplasm was better than other varieties as medicine; But LongYa Lily germplasm was more excellent as edible. Though comparative analysis of the relevance between active ingredients and water content, we found that the water content of medicinal herbs was positively correlated with one of total phospholipids, total sapogenin content, but with water-soluble polysaccharide content was negatively correlated. Recommending in the formulation of quality standards Lily process, containing amount of water with no more than 13.69%, ash content with no more than 3.86% , cold water extract content with no lower than 18.24% and the ethanol extract content with no lower than 11.06% are proper according to 95% allowing zone.4.With diosgenin for evaluating index sign, we have built up HPLC fingerprint of Lily material. 15 samples of Lily and L. lancifolium Thunb have demarcated 14 or 18 mutual fingerprint peaks, the similarities of their peak area were all more than 90%. In demarcated the sapogenin extracts of 26 samples, 14 mutual fingerprint peaks were gained and the similarities of peak area were more than 90%. The diagram obtained were divided into 2 groups: L. brownii F.E.Brown var.viridulum Baker and L. lancifolium Thunb.. Diosgenin contents existed greater difference in each sample, Most of L. lancifolium Thunb. contents were higher than L. brownii F.E.Brown var.viridulum Baker and peaks were also more complicated which explained its effective components complicated. 6# L. lancifolium Thunb. fumed with sulfur from LongShan existed greater differences compared with the same kind other samples, its similarity was 0.877, but diosgenin content was the highest,reaching to 40.5μg/g. Through evaluating diosgenin contents and fingerprints of the peaks, we think vol Dan has better medicinal value for development.5. By using scale leaves of Lily as material firstly,genetic diversities and genetic relationship of 16 different ecological sources of Lily were analyzed by RAPD molecular maker technology. 35 effective primers were selected from 120 random primers, and a total of 769 DNA bands were gained. of which 767 were polymorphic, accounting for 99.7%. Cluster analysis showed that 16 different ecological sources of Lily have been divided into two groups in the threshold 0.9407, they were two different plants from Liliaceae, one was from Lilium, the other was from Cardiocrinum. And Liliaceae were divided into three different species in the threshold 0.5790: L. brownii F.E.Brown var.viridulum Baker, L. lancifolium Thunb. and L. pumilum DC. The above results were consistent with the taxonomy. Cluster results and genetic relationship suggested that genetic diversities are higher and genetic relationship farther between different Lily. And germplasm and geography distance present definitely related trend.6.Though comparing and analyzing pharmacology action about relieving cough, dispelling phlegm and soothing the nerves among L. brownii F.E.Brown var.viridulum Baker, L. lancifolium Thunb., Platycodongrandiflorum(Jacq. ) A. DC. and Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H.F.Chow. , The resulted suggested that both of L. lancifolium Thunb. and L. brownii F.E.Brown var.viridulum Baker could relieve a cough, dispel the phlegm and soothe the nerves obviously, but the ratio of L. lancifolium Thunb. relieving a cough is 58.04%, L. brownii F.E.Brown var.viridulum Baker was 18.57% ; The discharging phenolsulfonphthalein content with L. lancifolium Thunb. was 6.93μg/mL,and L. brownii F.E.Brown var.viridulum Baker 5.85μg/mL; The sleep extension time caused by sodium pentobarbital was 36.4 min and 34.8 min with L. lancifolium Thunb. or with L. brownii F.E.Brown var.viridulum Baker respectively. The pharmacological effects of Vol Dan are better than Lily. From the point of view of resource utilization as medicinal plants, L. lancifolium Thunb. was more suitable to use as a medicine and have a higher on developing and using value.The research revealed the diversities of Hunan Lily germplasms from the biological characteristics, chemical composition, molecular level and so on, accumulated data for preservation of Lily germplasm resources in Hunan province and well-bred breeding selection, and have great significance for protection and utilization of Hunan Lily varieties. According to the results by analysis, the author suggested that L. lancifolium Thunb. is more suitable to carry out reasonable development and utilization in which Hunan medicinal germplasm Lily's.