Study On Chuanxiong Rhizoma Lingzhong Standardization And Its Germplasm Conservation Techology

Chuanxiong Rhizoma, the rhizome of Ligusticum Chuanxiong Hort., is a famous trueborn Chinese medicinal materials in Sichuan, China which has been cultivating and using for two thousands years. Ligusticum Chuanxiong Hort. is an asexual reproductive plant. Its overground swollen node called Lingzhong is used as reproductive material in production. Because of lacking stain selection and breeding, its genetic characterization degenerated to some extent over thousands years of cultivation. In Sichuan, Chuanxiong Rhizoma’s main production area, Lingzhong breeding is still absent of unified technology protocol and scientific organization and management. And the absence of classification criterion and testing rules for Lingzhong result in its uneven quality. Meanwhile, traditional natural conservation method is still widely used in production that can not preserve the medicinal herb for long. Traditional field conservation method consumes much manpower and material resources and may lead to the loss of germplasm, while relevant germplasm conservation technology is still lack of research.Seed and seedling of good variety and good quality are warranties to realize good agricultural practice for Chinese medicinal materials which can ensure herb’s quality in return. In view of the bottleneck in standardized production of Chuanxiong Rhizoma, the article took Lingzhong’standardization as the breakthrough point, studied the standardization of seedling of Ligusticum Chuanxiong Hort.(Lingzhong) and its germplasm conservation technology and aimed to promote Lingzhong’s standardization, to provide Lingzhong of high quality for production of Chuanxiong Rhizoma, and finally to produce Chuanxiong Rhizoma with high, stable and controllable quality as well as high yield thus to promote the continuable development of Chuanxiong Rhizoma production industry. The research results are as follows:1Lingzhong search and collectingOn the basis of early germplasm investigation around China, the article did field survey at Chuanxiong Rhizoma’s main production and breeding area including more than20villages and towns belonging to5cities and counties of Sichuan. Its distribution, evolution and migration of main production area, local cultivation technology, germplasm breeding and conservation technology, quality evaluation method and marketing situation of Lingzhong in Sichuan are basically made clear. The search showed that its production area had alternated at resent years especially after Wenchuan earthquake. Lingzhong in Sichuan are mainly gathered at Zhongxing town, Hongyan at back peak of Qingcheng mountain, Hongkou village of Dujiangyan city, Xiaoyudong town, Longmenshan town, Tongji town of Pengzhou city, Shuimo town, Sanjiangkou town of Wenchan county, Sanlang town of Pengzhou city, Bingchuan town of shifang city, whose average elevation are between900m and1500m. Among them, Lingzhong’s breeding area at Xiaoyudong town of Pengzhou city is bigger.In Chuanxiong Rhizoma’s major production area, Lingzhong is mainly bred by famers in mountainous area who are spot authorized by farmers in plain area. Most collected Lingzhong are planted at plain while few flew into the market for else need. Good and bad Lingzhong at the market are intermingled without unified criterion. Recent years, it was discovered that a few Ba Lingzhong, Dong Lingzhong, Moutain Lingzhong for own need in Chuanxiong Rhizoma’s main production area as well as the market had flown into many planting area of Chuanxiong Rhizoma in and out of Sichuan province.Lingzhong germplasm coming from34production areas and markets are colleted in the investigation which are partly preserved in early-established germplasm bank, partly provided to National Germplam Bank and partly stayed behind for project research.2Lingzhong storage and germplasm conservation technologyStudied normal temperature storage technology and low temperature storage technology to conserve Lingzhong used for produciton. Results showed that Lingzhong could be conserved for6to10days under normal temperature which will not influence its germination percentage, which provided scientific references for its short term conservation after recovered. Established refrigeration conservation method that could prolong Lingzhong’s short term conservation time to3months. This method lay a solid foundation for its further spreading and commercialization which can satisfy the need of different area’s farmers for Lingzhong’s different storage time.Growth restriction conservation method was first applied to Chuanxiong tube seedling for its middle term conservation. Taking its stem as explant and MS as solid medium, Chuanxiong tube seedling’s culture method was established. The article studied the influences on Chuangxiong tube seedlings by altering culture circumstance, adjusting medium nutrient composition, adding permeable compound and growth inhibitor. And the best growth restriction conservation method had been established. The recovering growth experiment showed that Chuanxiong germplasm could survive after six months’reservation. Variation was not found between preserved seedling and controlled seedling by morphological characteristics comparison and isoenzyme analysis. Conserved seedling had kept good heredity stability. This had been lay a good foundation for Lingzhong’s middle term conservation.Explored the practicability of preserving Lingzhong under ultralow temperature adopting embedding vitrification method. The best explant for embedding vitrification method is the stem tip. This has been established a culture method for Chuanxiong tissue culture seedling and lay a foundation for its further study.3New variety breedingChuanxiong breed colony from1300fine individuals and8000single-plants is constructed on the basis of early Chuanxiong resouce identification. Using this colony, systematic selection method is adopted to breed new Chuanxiong variety of high quality. Over6six years, after good single-plant selection, good stains selection, strain comparison test and variety(stain) muti-location trials, Ivxiong1was finally selected out and certified by the Crops Variety Appraisal Committee of Sichuan Province. Lvxiong I’s mature plant grows dark green leaves that could be kept for a long time. It showed good colony regularity, strong disease-resistance, fine yielding ability, stability and adaptability. Quality of Chuanxiong Rhizoma from the strain fits to pharmacopeia’s need. It had lain a foundation for Chuanxiong variety’s standardization and provided good provenance for Chuanxiong Rizoma’s standardized production.4Lingzhong breeding technologyOn the basis of early research, Lingzhong breeding technical specification including suitable plant area, environmental condition of producing area, field selection and soil preparation, Fuxiong’s selection and treatment, planting, field management, extermination of disease and insect pest, processing in production place and so on were deeply researched. The best methods and measures in every process of Lingzhong breeding were determined. Standard operation practicee for Lingzhong breeding is formulated on basis of it, and this provide technical support for the production of fine Lingzhong at mountain breeding area.5Quality standard study of LingzhongStudied the quality testing protocol of Lingzhong. Taking account into Chuanxiong Rhizoma breeding material’s particularity and using medicinal herb grower’ traditional experience for references, comprehensive indexes were put forward to evaluate Lingzhong’s quality which include morphological appearance,100grain weight, seedling coefficient, mixture rate, insect pest and disease. And quality testing protocol of Lingzhong was formulated which contains authenticity identification, shape index, weight determination method, seedling coefficient, impurity rate and insect pest and disease. It provide technical support for the evaluation of quality of Lingzhong.First studied classification criterion of Lingzhong by compare influences of Lingzhong of different classification on its growth, yield and quality. The result showed that classification significantly influenced its growth and the influence factor varies at its different growth period. The first and second classification apparently excel third classification at Chuanxiong’s growth and yield. Different classification showed little differences at the herbal properties, Thin-Layer Chromatography, content of main component. Through summarizing herb farmer’s long practical experience and early research achievement, and referring to the research result above as well as actual production, Lingzhong classification criterion was drawn up.On the basis of above research results, Chuanxiong Lingzhong Quality Criterion (draft) was drawn up. It provide standard criterion for Chuanxiong Lingzhong’s production, circulation, examination and application.