| Crocus sativus L, known as saffron, is a perennial herb of Iradaceae family with highmedical value and its ecological habits, genetic diversity, the content and types of activeingredient in stigma have been widely studied, but there is little study about its influenceon different germplasm characteristics and active ingredients, at the same time ornamentalcrocus on ecological habits, landscape greening, genetic diversity, the content and types ofactive ingredient in stigma has not been paid much attention to. In this thesis, biologicalcharacteristics, genetic relationship and genetic diversity of crocus were studied bymorphology research and molecular technology. Meanwhile HPLC was applied to studythe content of crocin-1in stigma of Crocus sativus, and plant tissue culture technology andits application feasibility of rapid reproduction for the test-tube croms in Crocus sativuswere discussed. The results below were obtained mainly:1. Chosen croms of medicinal and ornamental crocus imported from different area asexperimental material. It was investigated and analysed on the growth and development,morphological characteristics, and landscape application of crocus. Also, their biologicaland ornamental characteristics were studied systematically. It was clear of its growth anddevelopment regularity of leaf and flower.2. In order to identify genetic diversity and genetic relationship of crocus, RAPDtechnique was used to analyze genetic diversity between medicinal and ornamental crocuson DNA level. Genetic diversity was detected abundantly in leaf genomic DNA andmolecular clustering results can effectively identify medicinal and ornamental crocus.3. In this study, the HPLC assay was optimized on crocin-1, the mobile phase wascomposed of a mixture of methanol and1%aqueous acetic acid （linear gradient elution）, which can quantitatively determined the content of crocin-1. HPLC was used to explorehow the producing area, storing time, baking temperature, picking time and drawn partsaffect the content of crocin-1in Crocus sativus stigma. The results showed that the bakingtemperature makes a little influence,105℃adopted in the pretreatment remarkablyimproved the content of the stigma of crocin-1. According to the analysis of HPLC spectra,the component types in ornamental crocus stigma have been confirmed preliminarily.4. Basing on the tissue culture the present research was carried out to study the in vitroculture technology of Crocus sativus and establish the sterile tissue culture system.Technical conditions about tissue culture of Crocus sativus were selected from thesterilization treatments, the varied of explants, medium composition, hormone conditionsand so on. Finally appropriate condition were obtainted, and the test-tube croms in vitroculture were got under above congdition. Auxin was an important factor affecting theinduction of callus in primary culture. Good exterior conditions and a fast-growing couldbe acheived by the following conditions: MS+6-BA5.0mg·L-1+NAA5.0mg·L-1+2,4-D0.5mg·L-1. In the stage of induction of adventitious buds and tube seedlings subculture, theappropriate culture medium was MS+6-BA5.0mg·L-1+NAA5.0mg·L-1. In the stage ofrooting culture, the appropriate culture medium for rooting was MS+NAA2.0mg·L-1, andthe occurrence of the root were thick,had good quality. These lay the foundation of thory ofthe tissue of Crocus sativus. |
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