Determination Of Effective Components And SRAP Analysis Of Anoectochilus Roxburghii In Different Geographical Provenances

Anoectochilus roxburghii is the valuable Chinese medicinal materials in our country folk, and is an ornamental plant as well. In this study, we optimized the technique of extracting flavonoids and polysaccharides from Anoectochilus roxburghii by three methods which were ultrasonic extraction, microwave extraction and ultrasonic-microwave cooperative extraction. Then using the best method and technique to detect flavonoids and polysaccharides content in10geographical provenances of Anoectochilus roxburghii, and analyzed the genetic diversity of these10provenances by SRAP. The results of the study are as follows:1. The optimization of extracting flavonoids from Anoectochilus roxburghii in different methods. The best technique of ultrasonic extraction method:the ratio of liquid to solid was10,30min extraction time, the extraction times was5,83.77%of ethanol concentration and ultrasonic was opened throughout the extraction process. The best technique of microwave extraction method:the ratio of liquid to solid was18.38,23.71min extraction time, the extraction times was5,100%of ethanol concentration and the power of microwave was100W. The best technique of ultrasonic-microwave cooperative extraction method:the ratio of liquid to solid was10,24.88min extraction time, the extraction times was5,96.17%of ethanol concentration, the power of microwave was500W and ultrasonic was opened throughout the extraction process.2. The optimization of extracting polysaccharides from Anoectochilus roxburghii in different methods. The best technique of ultrasonic extraction method:the ratio of liquid to solid was10,30min extraction time, the extraction times was4.47and ultrasonic was opened throughout the extraction process. The best technique of microwave extraction method:the ratio of liquid to solid was10,30min extraction time, the extraction times was5and the power of microwave was100W. The best technique of ultrasonic-microwave cooperative extraction method:the ratio of liquid to solid was10,30min extraction time, the extraction times was5, the power of microwave was100W and ultrasonic was opened throughout the extraction process.3. The comparison of the extraction efficiency between the different methods. The yield of flavonoids in Anoectochilus roxburghii used ultrasonic-microwave cooperative extraction was1.85and1.25times as high as ultrasonic and microwave extraction. The yield of polysaccharides in Anoectochilus roxburghii used ultrasonic-microwave cooperative extraction was1.65and1.1times as high as ultrasonic and microwave extraction. Both flavonoids and polysaccharides had the best yield by using ultrasonic-microwave cooperative extraction method.4. The flavonoids and polysaccharides content in the different geographical provenances of Anoectochilus roxburghii. The flavonoids contents in10provenances of Anoectochilus roxburghii were from1.7438%to2.5075%, with the lowest content in Wuping and highest in Qingliu provenances. The polysaccharides contents in10provenances of Anoectochilus roxburghii were from16.9333%to25.6373%, with the lowest content in Taiwan and highest in Guangxi provenances.5. The SRAP marker analysis of10geographical provenances of Anoectochilus roxburghii. The result showed that a total of93bands based on17pairs of primer were detected, of which68were polymorphic (73.1%), and per primer pair amplified5.5bands and4polymorphic bands in average. The genetic similarity coefficients of proven ances were from0.5161to1.0000.10Anoectochilus roxburghii geographical provenances could be divided into two main groups, group Ⅰ was Taiwan provenances and group Ⅱ was the rest of9provenances. Group Ⅱ could be divided into two sub groups. Group A consisted of Guangxi and Nanjing provenances and the rest of7provenances made up of group B. Group B could be divided two small groups, too. Group a was Wuping provenances, and group b consisted of Yongan, Ning hua, Qingliu, Mingxi and Jiangle Anoectochilus roxburghii.