Reasearch On The Quality Standard And Genetic Diversity Of Hovenia Acerba Lindl

Objective（1）To determine the quality of H.acerba seeds from 25 habitats in China,establish methods to detect the index components of H.acerba seeds,provide technical support to evaluate the quality of H.acerba seeds.（2）To investigate the relationship of apparent traits on the quality of H.acerba seeds,verify the quality evaluation method of the index components.（3）Establishing comprehensive evaluation method for chemical composition of H.acerba seeds.（4）Elucidating the genetic diversity of H.acerba Lindl from different habitat in China.Methods（1）Establish a method to evaluate the index components of dihydromyricetin and total flavonoids of H.acerba seeds by using high performance liquid chromatography（HPLC）and sodium nitrite colorimetry.（2）The H.acerba seeds materials were sorted according to different colors and 1000-grain weights,determine the chemical composition of H.acerba seeds with different appearance traits.Verify index evaluation methods by using traditional Chinese medicine classification standards.（3）The HPLC fingerprint of H.acerba seeds was established,and verified by methodology.（4）Study the genetic diversity of Chinese H.acerba seeds with SRAP molecular marker.Results（1）Clear differences in the quality of H.acerba seeds from different habitatThe method for the determination of dihydromyricetin and total flavonoids was established.By geographical clustering of different habitat,the average content of dihydromyricetin in the southwestern habitats of North China Plain was the highest,being 7.97 mg.g^-1.The average total flavonoid content in the Qinling Mountains was the highest,which was 18.75 mg.g^-1.The variation of dihydromyricetin and total flavonoid content in different habitats was the most stable in the Qinling Mountains.The variation of dihydromyricetin content was only 18.92%,and the variation of total flavonoid content was only 9.71%.（2）Completion of the correlation between appearance traits and quality of medicinal materialsCorrelation between red-black ratio and 1000-seed weight with quality of H.acerba seedsAmong the statistics of the proportions of red and black in the 25 habitats,the red-black ratio of Xinyang in Henan was the highest.H.acerba seeds of Panzhihua Sichuan Province and Ya’an Sichuan Province were all yellow.As the 1000-seed weight in 25 habitats of H.acerba seeds,the 1000-seed weight of H.acerba seeds in Xinyang,Henan Province had the largest.Panzhihua,Sichuan Province and Ya’an,Sichuan Province had the lowest 1000-seed weigh.The red-black ratio and 1000-seed weight combined with the contents of dihydromyricetin and total flavonoids from 25habitats of H.acerba seeds were analyzed by Pearson correlation.The correlation coefficients of dihydromyricetin with red-black ratio and 1000-seed weight were0.733 and 0.560,respectively,P<0.01.It was shown that there was a significant correlation between appearance characteristics such as color and 1000-seed weight and dihydromyricetin content.Correlation of color grading and quality of H.acerba seedsThe dihydromyricetin content in different colors of different batches of H.acerba seeds was analyzed,rufous>gray>yellow>black,and the difference between different colors was significant.Based on the above results,the color of H.acerba seeds was graded,and examine the Pearson correlation coefficient of color and index components.It was found that the correlation coefficient was greater than 0.95,and the P value was less than 0.01.Explain that the color grading has a significant correlation with quality.（3）Established HPLC fingerprint of H.acerba seedsFingerprints of H.acerba seeds were constructed using HPLC.The nine common peaks in the HPLC fingerprint of H.acerba seeds were calibrated,three were identified,and one characteristic peak was found.It could well separate the main compounds of H.acerba seeds,and it has better discrimination for H.acerba seeds in different habitats.（4）Elucidated genetic diversity of H.acerba Lindl.The seed germination of H.acerba Lindl was carried out to obtain the genomic DNA of each plant.A method for the study of H.acerba Lindl SRAP molecular markers was established,The ratio of 27 pairs of primers selected was 91.90%,and the PIC was greater than 0.64,description of genetic diversity of H.acerba Lindl.Genetic similarity cluster analysis showed that the 16 habitats were clustered into five categories,and the southwestern region and the western part of South China which included Guilin Guangxi Province,Hechi Guangxi Province,Zunyi Guizhou Province,Qujing Yunnan Province,Yongzhou Hunan Province,and Chenzhou Hunan Province were clustered into a category I.Conclusion（1）The contents of dihydromyricetin and total flavonoids in H.acerba seeds were the highest in the southwestern part of North China Plain,Henan Province and Xinyang,Hubei Province.In this collection of samples,the quality of H.acerba seeds in southwestern part of North China Plain was superior.The variation of dihydromyricetin and total flavonoid content in the Qinling area is relatively low,indicating that the quality of H.acerba seeds in the Qinling producing area is stable,and the average content of total flavonoids is the highest.The average content of dihydromyricetin is second only to the southwestern part of the North China Plain.（2）Through the study of the appearance of H.acerba seeds,it was concluded that the bigger the red-black ratio,the larger the 1000-seed weight,the better the quality of medicinal herbs.The quality of H.acerba seeds was graded as rufous,gray,yellow,and black.The color have significant correlation to the index components of dihydromyricetin and total flavonoids in H.acerba seeds,and At the same time,it showed that the evaluation of the index components was consistent with the traditional medication habits and the traditional medicine grading standards.（3）Establishing the HPLC fingerprint of H.acerba seeds,provided a more comprehensive basis for the quality control of H.acerba seeds.（4）No relationship was found between the germination rate and the quality of H.acerba seeds.A method for studying the genetic diversity of H.acerba seeds has been established initially.The experimental results showed that the genetic composition of H.acerba seeds was affected by geographic isolation,but there was also genetic exchange among populations,which provided a reference for future breeding and resource utilization.