Researches On Biological Trait, The Physical-Chemical Composition And Molecular Identification Technology Of Germplasm Resources Of Fritillaria

Fritillaria thunbergii miq, alias like shell and pearl, belonging to lily family(Liliaceae), which is one of perennial herbs. The Fritillaria collected from Zhangshui town of Yinzhou district at Ningbo city by using the conventional survey method was investigated and analyzed. Bulbs of Broad leaf, Multi-seed, Three-seed, Narrow leaf,new Meiyuan and high Three-seed varieties of Fritillaria thunbergii were used to investigate nutrition composition(including water content, soluble sugar, protein,amino acid and mineral content) and alkaloid content of different varieties via conventional physiological-biochemical technology and automatic amino acid analyzer, HPLC-ELSD and ICP-AES determination. Finally, the application value of DNA barcode ITS1 and ITS2 technology in germplasm resources evaluation of Fritillaria thunbergii was preliminarily discussed. The major results were as follows:(1)The order of growth status of Fritillaria species was as follows from best to worst: High three seed, three seed, narrow leaves, New Meiyuan and width leaves.The total alkaloid content of mature Fritillaria bulbs of different varieties was as follows: new Meiyuan, Narrow leaves, Multi-seed, Three-seed and Broad leaves were0.2421%, 0.2088%, 0.1859%, 0.1811% and 0.1607%, respectively. Offspring traits of? radiation population of Fritillaria thunbergii appeared segregating and its performance was the increase of main stalk's height and numbers.(2)The protein content of different varieties of Fritillaria was up to the highest in pre-blossom period, and then was decreased continuously. However, soluble sugar content of different varieties of Fritillaria was increasing during the whole growth stage. The protein content existed the peak at the dormant ending period, while total soluble sugars content was the highest at the dormant beginning period. It was also found that the change of alkaloids content was not significant during dormant period.It was also found that there existed big difference of physical and chemical ingredients among different parts of the bulb, in which the protein, total soluble sugars and alkaloids were mainly concentrated in the outer bulb. The results showedthat protein, soluble sugar and alkaloid content of Fritillaria bulbs was not significantly linear relationship with its size.(3) DNA barcoding is a new biometric identification system, which can automatic identification of differential species within a standard, with sufficient variability, and relatively short and easily amplified DNA fragments of biological species quickly. Firstly, genomic DNA of 79 samples from 17 kinds of Fritillaria were extracted by kit method and its concentration and purity were detected; then,ITS1, ITS2 primers were used for PCR amplification and PCR products were sequenced; finally the sequences of DNA were compared and analyzed in this study.The results showed that the sequence differences among local Fritillaria species were small and the resolution among different sample was low. Therefore, ITS1 and ITS2 sequence was not suitable for DNA barcode of Fritillaric thunbergii.