ABSTRACT
The accumulation of secondary metabolites of traditional Chinese medicine (TCM) is closely related to its origins. The identification of origins and multi-components quantitative evaluation are of great significance to ensure the quality of medicinal materials. In this study, the identification of Gentiana rigescens from different geographical origins was conducted by data fusion of Fourier transform infrared (FTIR) spectroscopy and high performance liquid chromatography (HPLC) in combination of partial least squares discriminant analysis; meanwhile quantitative analysis of index components was conducted to provide an accurate and comprehensive identification and quality evaluation strategy for selecting the best production areas of G. rigescens. In this study, the FTIR and HPLC information of 169 G. rigescens samples from Yunnan, Sichuan, Guangxi and Guizhou Provinces were collected. The raw infrared spectra were pre-treated by multiplicative scatter correction, standard normal variate (SNV) and Savitzky-Golay (SG) derivative. Then the performances of FTIR, HPLC, and low-level data fusion and mid-level data fusion for identification were compared, and the contents of gentiopicroside, swertiamarin, loganic acid and sweroside were determined by HPLC. The results showed that the FTIR spectra of G. rigescens from different geographical origins were different, and the best pre-treatment method was SNV+SG-derivative (second derivative, 15 as the window parameter, and 2 as the polynomial order). The results showed that the accuracy rate of low- and mid-level data fusion (96.43%) in prediction set was higher than that of FTIR and HPLC (94.64%) in prediction set. In addition, the accuracy of low-level data fusion (100%) in the training set was higher than that of mid-level data fusion (99.12%) in training set. The contents of the iridoid glycosides in Yunnan were the highest among different provinces. The average content of gentiopicroside, as a bioactive marker in Chinese pharmacopoeia, was 47.40 mg·g⁻¹, and the maximum was 79.83 mg·g⁻¹. The contents of loganic acid, sweroside and gentiopicroside in Yunnan were significantly different from other provinces (<0.05). In comparison of total content of iridoid glycosides in G. rigescens with different geographical origins in Yunnan, it was found that the amount of iridoid glycosides was higher in Eryuan Dali (68.59 mg·g⁻¹) and Yulong Lijiang (66.68 mg·g⁻¹), significantly higher than that in Wuding Chuxiong (52.99 mg·g⁻¹), Chengjiang Yuxi (52.29 mg·g⁻¹) and Xundian Kunming (46.71 mg·g⁻¹) (<0.05), so these two places can be used as a reference region for screening cultivation and excellent germplasm resources of G. rigescens. A comprehensive and accurate method was established by data fusion of HPLC-FTIR and quantitative analysis of HPLC for identification and quality evaluation of G. rigescens, which could provide a support for the development and utilization of G. rigescens.
ABSTRACT
Objective: To establish and optimize the technique of tissue culture and rapid propagation, and compare the progeny regeneration capacity among different mating patterns in Swertia mileensis, meanwhile, to explore the reproductive assurance of the fragment populations. Methods: Sterilized seeds from different mating patterns were cultured on MS media for germination, which included self-breed, inbreeding, outbreeding, and nature pollinating, and counted the germination rate 60 d later. The study used the different source explants maintained on MS medium supplemented with different types and concentration of plant growth regulator combinations to explore its capacity of callus induction and adventitious shoots differentiation with orthogonal design. Results: There is no significant difference in the germination rates among all kinds of seed as touched above. The optimal medium for callus induction is MS + ZT 0.5 mg/L + 2,4-D 0.05 mg/L, the callus induction rates were 100%, 96.67%, 96.55%, and 96.29%; The optimal medium for adventitious shoots differentiation was MS + BA 2.0 mg/L + KT 0.1 mg/L + NAA 1.0 mg/L, and the differentiation rates all reached up to 100%; The optimal medium for rooting was 1/2 MS + NAA 1.0 mg/L, the rooting rate both were as high as 100% and there was no significant difference in the growth of plantlets. Conclusion: This study optimizes the cultivation conditions and provides an effective solution for protecting the wild resources and sprouts multiplication of S. mileensis. Meanwhile, from the plant physiological basis, it could be preliminarily predicted that the advantage of inbreeding would make up the fitness cost associated with it and the progeny regeneration capacity shows no significant difference between inbreeding and outbreeding.
ABSTRACT
In this study, the embryological characters of Pterocypsela formosana (Asteraceae) were investigated with the traditional paraffin section methods. The anther has 4 sporangiates, the anther wall development follows the dicotyledonous type and comprises of an epidermis, endothelium, a middle layer and a single-layered tapetum, the tapetum belongs to glandular type. Meiosis of the microspore mother cells is of the simultaneous type, for the formation of mostly tetrahedral tetrad, the mature pollen grains are 2 celled. The ovary is bicarpellate, unilocular, one ovule and basal placenta, the ovule is unitegmic, tenuinucellate, inverted campylotropous and with developed endothelium, archesporial cell of megaspore differentiates immediately below the nucellar epidermis and functions as megasporocyte after development and belongs to tenuinucellate ovule type. The megasporocyte undergoes meitotic to form a liner tetrad, only one chalazal megaspore becomes the functional megaspore which forms female gametophyte including 7-celled and 8-nucleated after three successive mitosis, the female gametophyte is of the Polygonum type. Two polar nuclei melt into a secondary nuclei before fertilization, the chalazal antipodal cells are ephemeral and degenerate shortly after forming. Fertilization is porogamous and belongs to premitotic type of syngamy. The division of the primary endosperm nucleus is earlier than the zygote, the endosperm is of the nuclear type with the presence of haustoria, and the embryogeny belongs to asterad type chicory variant. The developed suspensor on early stage has important significance to the embryo development.
Subject(s)
Asteraceae , Embryology , Meiosis , ReproductionABSTRACT
Objective: An efficient and stable regeneration system of Ajugae nipponensis was developed by establishing the callus induction system and selecting the best protocol for higher plant regeneration. Methods: Stem tip, stem with buds, and inflorescence were all used as explants maintained on Murashige and Skoog (MS) medium supplemented with different types and concentration of plant growth regulator combinations. Results: Inflorescence was the optimal explant for callus induction and was cultured on MS medium containing 0.1 mg/L 6-BA and 1.5 mg/L 2, 4-D in 1 week; The callus divided into green multiple shoot 2 weeks later; The optimal medium for multiple shoot proliferation was MS + 2.0 mg/L 6-BA + 0.5 mg/L NAA, the regeneration rate of callus clustered buds was as high as 100%, the regeneration coefficient was 4.10, and the proliferation increased by more than five times after 4 weeks; The optimal medium for rooting was 1/2 MS + 1.0 mg/L NAA, the plant regeneration occurred 3 weeks later and the rooting rated up to 100%; Plantlet(100%) survived after transplanting rooted plantlets into sand. Conclusion: The callus can be induced by all kinds of explants and inflorescence is the best explant for higher callus induction rate; The current study lays a solid foundation for protecting the wild resources and advancing the cultivated of A. nipponensis, meanwhile providing scientific evidence for the research of genetic transformation.