Study On Variety Selection Of Magnolia Officinalis And Its Identification Of Provenance Of RAPD

Chinese medicinal materials Magnolia officinalis mainly originated from Magnolia officinalis and M.officinalis var biloba. In history, according to origination of Magnolia officinalis (produced in Hubei and Sichuan, and M.officinalis var biloba (produced in Zhejiang and Fujian).Authenticity is one of the important criterions for quality evaluation of varieties, Identification techniques is also the base of solving the problem about rights and interests protection of new varieties of seeds (seedlings) as well as its economic disputes. Selecting fine varieties, establishing evaluation system with good and comprehensive traits for Magnolia officinalis, collecting elite germplasm resources, improving high-yield cultivation technique is the prerequisite to develop chinese medicinal materials.In this paper plus tree with high comprehensive evaluation from high-quality provenance area and high effective ingredient plus good traits were selected on the base of studying investigation and morphological index from different provenances of Fujian Province Magnolia officinalis. In addition different provenance of the country Magnolia officinalis for RAPD molecular markers and genetic diversity were analyzed.The major results were as follows:1. Magnolia phenol extraction was experimentally studied through orthogonal design L9(34). The result showed that ethanol concentration is a key factor of extraction, followed by ultrasonic time. The optimal method for extraction was: ethanol concentration 90%, water-curing treatment time 50min, water-curing treatment temperature 60℃, ultrasonic time for 20min.2. The investigation showed that the distribution of Magnolia officinalis was generally in dispersed state, mainly in the west and north of Fujian Province. 147 plus trees primarily selected in Magnolia officinalis from 24 town and citieswere depleted analysis. The result indicated that the content of Magnolia phenol varied greatly with different provenances, with the minimum at 0.0907 mg.g-1 but the maximum at 5.4759 mg.g-1.The impact of basic climatic factors existed in sampling land on the content of Magnolia phenol was analyzed. The result showed that annual temperature and frostless season were the greatest impact factors.3. Seasonal changes have an impact on the content of secondary metabolites during the period of plant growth. The content of phenol in shoot cortex of Magnolia was highest in January. Afterward with the passage of time, the content would decrease at first then increased but in downward trend on September. In the Magnolia leaves honokiol content is always higher than that magnolol content,the trend of which is more obvious in tender leaves.Finally the total phenol content distribution from tender leaves to old leaves was in Increasing trend until it reached the maximum before the deciduous period.4.There are some differences of Magnolia content among the different parts of Magnolia but the differences would be minimal among the same organs. Polyphenol content in different organs ranked in the order of root bark ,dry hide ,branch bark, tender bud , seed, fruit, stem wood and needle.5. With the increase of Magnolia trees age, magnolol and honokiol content trend in both branch bark and stem bark were shown as increasing - decreasing– increasing. while in both middle-aged and young growth plantations, the phenol content would increased with their ages increasing. The value reached the maximum approximately at their ages of 25 but afterward it would be shown as downtrend. However, in hundred years of aged trees, the active component content would show increasing trend in the last stage. The phenol content of Magnolia leaves decreased with age increasing, which reached higher level in their young growth tree leaves.6. The variation of phenol contents between Magnolia individuals in the same population with the diameter of the similar size were quite large and the difference of phenol contents in various parts between Magnolia individuals were significantly.When excellent magnolia provenance was selected, there exited very significant or significant positive correlation with tree height, crown, bark thick, and branches rather than straight degrees, round degree and health degree. Meanwhile, the positive correlation between phenol content of branch bark and traits of trees was very significant or significant.7. Four main components were extracted, according to the principle in which characteristic root is equal to or greater than 1. Then the principal components were comprehensive evaluated from 24 different provenance. The evaluation result by ranking order was as follow: top front group was Zherong,Youxi, Licheng, Fuzhou, which is followed by the Wuping,Yongtai, Shouning,Qingliu.Shaxian,Yongan,Pucheng, campus of Fujian Agricultural and Forestry University,Pingnan, Mingxi,Ninghua, Zhangping, and at least was Taining,Nanping,Shunchang,Jianyang,Datian, Jianou, Fuan, Guangze.8. Correlation between various traits in plus tree selection was analyzed. The results were shown as follow: there exited very significant positive correlation between DBH and crown width, tree height and phenol content, very significant positive correlation between tree height and crown width, clear bole height, and branches, very significant positive correlation between crown width and clear bole height, significant positive correlation between stem straightness and bark thickness, rather than other indexes.9. Six principal components extracted by principal component analysis of cumulative contribution rate of greater than or equal to 85% were comprehensively scored from 54 plus tree. The result was shown as follow: the higher scores of plus tree family was the number 11,6,131,146,16,followed by 123,33,49,114, 2. Cluster analysis results were divided into two categories Provenance, the first category was the number 2,14,99,107,113,8,141,114,37,75,44,117,135,109,16,33,49,123, 6,11,131,146, the second category was the number 13,102,111,43,116,145,50,70,93,136,76,90,132,147,38,115,119,45,101,20,110,142,125,21,46,120, 138,129,97,94,29,112.10. DNA fingerprinting of 89 copies of Magnolia resources was constructed based on the data matrix obtained out of 96 reproducible RAPD bands of Magnolia resources amplified by six primers. The results indicated that 11 combinations were respectively S60 and S69, S60 and S91, S60 and S1412, S60 and S330, S60 and S1422, S69 and S91, S69 and S1412, S69 and S1422, S91 and S1412, S1412 and S1422, S1412 and S330.In each combination , data matrix formed by amplification can be relatively easy for multiple authentication to distinguish from the 89 resources.11.Source of 89 samples from the whole nation was analyzed. The analysis result can be concluded to three categories. The first category was Jiangyou,Fujian in which genetic structure possessed particularity while the second category included Jiangyou Sichuan,Daoxian Hunan,Mingxi Fujian, Shexian Anhui and part of Provenance individual of Jianyang Fujian. In addition, the third category can be divided into two sub-categories.In the first sub-category, main source region included Enshi Hubei, Jingning Zhejiang, Ziyuan Guangxi, Quanzhou Guangxi, Lushan Jiangxi, Haozhou Anhui, Hefei Anhui, Yaan Sichuan, Changan Shaanxi, Shexian Anhui,Jiangyou Sichuan and part of the Provenance individual of Yongzhou Hunan and the the second sub-category included Zhangjiajie Hunan, Jiangjin district of Chongqing, Shunchang Fujian, Pucheng Fujian, Nanping Fujian, Zherong Fujian, Yongtai Fujian, Forest Park of Fuzhou, Liancheng Fujian, Guangze Fujian,Taining Fujian, Shouning Fujian,Daoxian Hunan and part of the Provenance individualof Fujian Jianyang.