Studies On Morphological Characteristics, Spectral Analysis And ITS Sequence Of Main Medicinal Plants Of Valerians L. In China

To clarify main medicinal plants of Valeriana L. (MMPV) in China and to classify and identify them, the study based on morphology, chemical constituents spectroscopy and molecular genetics, and investigated 5 MMPVs and V. officinalis L. including bencaoological study, plant resources, cluster analysis of morphous characters, microstructural features, ultraviolet spectrums and infrared spectrums, and rDNA ITS sequence. The primary results are as follow:(1) Bencaoolgical researches showed that only V.jatamansi Jones was described in the ancient books on Chinese medicine(in Ming dynasty). There are 9 species of Valeriana been recorded as herbs in research articles in China today, and 5 are widespread for concentrated learning reports and large production. They are V. pseudofficinalis C. Y. Cheng et H. B. Chen, V.officinalis L. var. latifolia Miq., V. amurensis Smir. ex Komarov, V. jatamansi Jones, V. hardwickii Wall..Otherwise, V. officinalis L.was cultivated in China. The former 3 species and V.officinalis L.were used as Radix Valerianae; The later two used as other herbs. There still exists much pending questions in taxonomy of V.officinalis L., V. pseudofficinalis C. Y. Cheng et H. B. Chen and V. officinalis L. var. latifolia Miq.(2) Based on investigations of 3 typical wild habitats of Valeriana(Haituo mountain in Beijing, E'mei mountain in Sichuan and Enshi in Hubei) , the growing habits of V. pseudofficinalis C. Y. Cheng et H. B. Chen , V. jatamansi Jones , V. officinalis L. var. latifolia Miq. and V. hardwickii Wall. were recorded. Because there are rich resources of V.officinalis L. var. latifolia Miq. and selenium ore in Enshi, atomic fluorescence spectrometry was used to measure the trace selenium of Valeriana officinalis L. var. latifolia Miq. and its growing soil in 3 regions of Enshi. The results showed that selenium content of V. officinalis L. var. latifolia Miq. in Enshi reachs 0.2 mg/kg, and was conspicuously higher than in other regions, and could be a effective index to appraise it as a famous-region drug in Enshi. Meanwhile, the local advancements of cultivation were investigated.(3) Based on the specimen reviews, characters of the botanic origins of 6 MMPV were described. Through cluster analysis, it is proved that V.officinalis L.and V. pseudofficinalis C. Y. Cheng et H. B. Chen are not the same species, and V.officinalis L. var. latifolia Miq. is variety of V. pseudofficinalis C. Y. Cheng et H. B. Chen. Because of a large numbers of interim species, it can not deny that V. officinalis L. do not grows in China. The main differences of 3 species are the number of leaf lobes and the shapes of lobes. By observation of microstructural features of the fruits, the sizes, appearances, clothing hair, the base whether bias or concave, colour spots and dots in surfaces can identify the above-mentioned 6 species. By using of electron microscope to observe the microstructural features of the pollen grains, the sizes, appearances, pit canals and the shape of thorn sculpture can identify the above-mentioned 6 species. They also define that V.officinalis L.var. latifolia Miq. is similar to V. pseudofficinalis C. Y. Cheng et H. B. Chen, and both are different to V.officinalis L..(4) 20 samples of above-mentioned 6 species were studied by the ultraviolet spectrums of extraction of H2O, EtOH, CHCl3 and ligarine for the first time. The ultraviolet spectrums of extraction of polar solvent embody the differences of species, and can be used for identity; And absorptances of the nonpolarity reflect the qualities of the herbs in some degrees.Through cluster analysis of the peak positions and values, the samples were clearly classfied into 3 groups by their similar characteristics: valeriana radix, jatamans valeriana rhizome, hardwickii valeriana herb. That is same with medicinal facts. The groups of spectral lines can identify different species. Powders of above-mentioned 20 samples were directly compressed and determined by Fourier transformation intermediate infrared spectrum for the first time . Different species of Valeriana with respective spectral characteristics; Spectral characteristics between intraspecies reflect the component constancy and habitat difference. Infrared spectrum is a high-performance and volant identified method for MMPV.(5) The methods to isolate and to purify the total DNA from herbs of Valeriana and the conditions of the PCR for ITS sequence were optimized for the first time; At the same time, the specificity primers for ITS sequence of Valeriana were designed. The lengths of total ITS sequence are from 642bp to 644bp; ITS1 are from 233bp to 234 bp; 5.8S are 160 bp; ITS2 are 249 bp to 250 bp.And all the 5.8S of MMPV have the same sequence. The identity percents of ITS of MMPV are from 95.8% to 99.5%, and the 3 minimum divergences of all specimens are the divergences of V.officinalis L., V. pseudofficinalis C. Y. Cheng et H. B. Chen and V.officinalis L. var. latifolia Miq.. That results support the results of (3). Since the ITS sequences of the species of Valeriana are very stabilize, they can be used into disposaling the taxonomy of intermediate forms of Valeriana. Meanwhile, the herbs of Valeriana were clearly classfied into 3 groups by their similar ITS sequences: valeriana radix (V. pseudofficinalis C. Y. Cheng et H. B. Chen, V.officinalis L. var. latifolia Miq., V. amurensis Smir. ex Komarov and V.officinalis L.), jatamans valeriana rhizome, hardwickii valeriana herb. That is same with medicinal facts, and it means that they have particular medical efficacy themselves. The characters of ITS sequences also can be used into identification of the Valeriana herbs.