ABSTRACT
The molecular identification of Fritillaria taibaiensis and its relatives was studied by real-time PCR with a TaqMan-MGB probe. DNA was extracted from F. taibaiensis and its relatives. According to the sequence of ITS1 region, the mutation sites of F. taipaiensis and its related species were identified by MEGA7.0 software. The specific primers (a pair) and a TaqMan-MGB probe were designed by Primer Premier 6.0 software. In the Roche LightCycler 96 system, the lowest limit of detection for F. taipaiensis DNA template was 0.002 39 ng·μL-1, and the optimal Tm value range was 60 and 61 ℃. Specificity identification showed that the method had good specificity for F. taipaiensis, as it could be distinguished from other 13 different Fritillaria species including F. unibracteata. Since this method could accurately identify F. taipaiensis and its related species, it provides technical support for rational development of F. taipaiensis resources, management of Chinese medicinal market and supervision of raw materials in Chinese medicine manufacturing enterprises.
ABSTRACT
Objective: To study the variation patterns of rhizospheric soil nutrient and the content of nutrient elements for Fritillaria taipaiensis,in order to provide the theoretical base for the soil improvement and balanced fertilization during the artificial cultivation. Method: Totally 14 samples of rhizospheric soil of Fritillaria taipaiensis from different origins and years were taken as the research objects. Total N,available N,total P,available P,total K,available K,organic matter,pH and 7 nutrient element contents (Ca,Mg,Na,Mn,Zn,Cu and Ni) were analyzed by the soil agrochemical analysis method combined with the atomic absorption spectrophotometry. SPSS 22.0 software was applied for data multiple comparison and correlation analysis. Result: The all results showed significant differences (PF. taipaiensis. The content of total N,available N,total P,available P and organic matter of rhizospheric soil collected from cultivated varieties decreased with the increase of years,and the content of total K,available K and pH decreased first and then increased. However,the soil physical and chemical properties of wild varieties had no obvious change with the increase of years. The content of Ca,Mg,Na and Cu of soil from cultivated varieties decreased with the increase of years,while the content of Mn decreased first and then increased. And Zn and Ni showed no significant change with the increase of years. Compared with cultivated varieties,the content of Ca,Mg,Na,Mn and Cu increased first and then decreased. The content of Zn and Ni showed no obvious change. In general,the rhizospheric soil nutrient and the content of nutrient elements for wild F. taipaiensis were superior to those of cultivated varieties. Conclusion: The third year is the turning point of F. taipaiensis growth. The rhizospheric soil nutrient and the content of nutrient elements decreased obviously after three years. Attention shall be given to the balanced fertilization,the improvement of soil quality and the prevention of the cropping during cultivation of F. taipaiensis.
ABSTRACT
In order to reveal the relationship between the amount of soil microorganisms and the quality of Fritillaria taipaiensis, both cultivated and wild F. taipaiensis were collected from Chongqing, Wuxi at different stages of their growth as objects of the research. The mycorrhizal infection rate and colonization intensity, peimisine and total alkaloid content in bulbs, the amount of microorganisms and biomass carbon content in rhizospheric soil were all determined using common methods. The results showed that the typical arbuscular-vesicle roots were formed after the AM fungi infected the F. taipaiensis roots which were collected from different origins. The mycorrhizal infection rates were ranged from 78.74% to 98.68% and the colonization intensities were ranged from 13.29% to 37.06%. The rhizospheric microorganisms of F. taipaiensis showed abundant resources. The distribution rule of them in the rhizospheric soil was as follows: the amount of bacteria>the amount of actinomycetes>the amount of fungi. The rhizospheric bacteria, decomposition inorganic phosphorus bacteria, decomposition organic phosphorus bacteria, actinomycetes amount and the total number of microbes increased first and then decreased with the increase of years, while decomposition potassium bacteria showed decreasing trend and fungi showed gradual increasing trend. The soil microbial flora content in the soil changed from "bacterial type" with a high fertility to "fungal type" with a low fertility. The mass fraction of peimisine and total alkaloid content increased first and then decreased with the increase of over the years, the same trend of culturable rhizosphere soil bacteria and actinomycetes indicated that the growth years affected the quality of soil and medicinal materials on different levels. Therefore, the diversity of microbial communities in rhizosphere soil reduced with the increase of years leading to the continuous cropping obstacles and the destruction of medicinal quality of F. taipaiensis.
Subject(s)
Alkaloids , Fritillaria , Chemistry , Microbiology , Mycorrhizae , Plant Roots , Rhizosphere , Soil MicrobiologyABSTRACT
Objective To develop a sensitive and reliable RP-HPLC method for the simultaneous determination of nine nucleosides including uracil, cytidine, guanine, uridine, adenine, guanosine, thymidine, adenosine and 2'-deoxyadenosine from Fritillaria taipaiensis P. Y. Li that had been cultivated in different producing areas; To compare the contents of these nucleosides from different producing areas. Methods The analysis was performed on a Venusil MP C18 (2) column (4.6 mm×250 mm, 5 μm) with a gradient of methanol-water at a flow rate of 1.0 mL/min;the detective wavelength was set at 260 nm; the column temperature was set at 35 ℃. Results Uracil, cytidine, guanine, uridine, adenine, guanosine, thymidine, adenosine and 2'-deoxyadenosine were obtained in the good linear range of 0.269 5–16.17 μg/mL, 0.132 1–7.927 5 μg/mL, 0.095 5–5.73 μg/mL, 1.16–69.6 μg/mL, 0.48–28.8 μg/mL, 0.571 5–57.15 μg/mL, 0.526–52.6 μg/mL, 3.307 5–198.45 μg/mL, 0.530 5–31.83 μg/mL, respectively (r≥0.999 5);the recovery was in the range of 96.49%–101.65%(RSD≤2.92%). Conclusion The contents of the nine nucleosides from different producing areas have differences. Fritillaria taipaiensis P. Y. Li from Xianyi Village, Chengkou County, Chongqing City, whether the cultivated ones or wild ones, contain the highest level of nucleosides. The established method can provide references for the perfection of quality standard for Fritillaria taipaiensis P. Y. Li.
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Objective To investigate the history, current situation and ecological environment of Chongqing wild and cultivated Fritillaria taipaiensis resources; To provide references for protection and industrial development of Fritillaria taipaiensis resources. Methods Data query, on-the-spot investigation and follow-up survey were used in the study. Results Chongqing Fritillaria taipaiensis was in the medical records since 1843. Now 5 counties have wild Fritillaria taipaiensis resources and 6 districts have cultivated Fritillaria taipaiensis resources, with 6 typical ecological types. Conclusion Chongqing wild Fritillaria taipaiensis resources are wide distributed and with long history of medical usage. Chongqing is the original producing area of Fritillaria taipaiensis, which has been destroyed by human factors. Fritillaria taipaiensis resources in several districts have become extinct, however, the cultivated Fritillaria taipaiensis resources is developing rapidly.