Genetic Diversity On Wild Germplasm Resources And Pharmacognosy On Cultivation Of Anoectochilus Herba In Fujian Province

Objective: Anoectochilus herba is a rare Chinese herbal medicine with characteristics of Fujian and Taiwan.The wild resources have different phenotypes and cultivation has complex strains.In this study,the genetic diversity on the wild germplasm resources and the pharmacognosy on the cultivation were analyzed in order to explore the genetic characteristics of the germplasm resources and the pharmacognosy identification characteristics of the cultivated products in Fujian Province.Methods: The genetic diversity of different geographic groups in Fujian was analyzed by using SSR markers based on the Anoectochilus roxburghii transcriptome data.The traditional methods of pharmacognosy,microscopic identification and molecular pharmacognosy were used to identify some common cultivation include Da-yuan-ye,Xiao-yuan-ye,Jian-ye and Tai-wan strains.Result: 44 pairs of SSR primers were selected based on the transcriptome data of A.roxburghii,cluster analysis showed that the genetic similarity coefficient among different geographical groups varied from 0.29 to 0.90,all samples of Fujian could be divided into four major groups at 0.65,and there was genetic differentiation among different regions.The character identification results of different strains showed that the Da-yuan-ye has larger leaves,1.2-5.1cm long and 1.1-3.2cm wide,with a continuous white vein on the leaf margin;The Xiao-yuan-ye has relatively smaller leaves,0.9-3.4cm long and wide 0.7-2.7cm,with continuous or discontinuous veins,or no veins;The leaves of Jian-ye strains are long oval,1.3-4.6cm long,0.9-2.6cm wide,with dense veins and yellowish midrib area;The A.formosanus leaves are broadly oval,1.3-4.2cm long,0.9-2.9cm wide,with white veins.The microscopic identification results showed that some cells in leaves close to the upper epidermis containes red substances of A.roxburghii,while these substances are abundant in the cells of the sponge tissues of A.formosanus.The identification result of SSR markers showed that there exist a certain genetic differentiation between cultivation and the genetic differentiation ability was greater between A.formosanus and others strains.In the condition of planting over 4 months,the color of stem and leaf is deepened and the plant height is decreased;and the casparin strip is more obvious,the vessel wall thickness of xylem is increased,and the cell cavity is increased.Conclusion: This study demonstrated that there exist genetic differentiation of wild germplasm resources in different areas of Fujian Province through molecular marker technology;the differences in the characters,microscopes and molecular levels of different strains Anoectochilus herba were illustrated;and the characters and microscopical identification points of planting for 4 months were clarified,which could be used for the conservation of wild resources,genetic breeding,pharmacognosy identification,quality evaluation and control of Anoectochilus.