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      Autotoxic Ginsenosides in the Rhizosphere Contribute to the Replant Failure of Panax notoginseng

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          Abstract

          Background and Aims

          Sanqi ginseng ( Panax notoginseng) growth is often hampered by replant failure. In this study, we aimed to examine the role of autotoxicity in Sanqi replant failures and assess the role of ginsenosides in autotoxicity.

          Methods

          The autotoxicities were measured using seedling emergence bioassays and root cell vigor staining. The ginsenosides in the roots, soils, and root exudates were identified with HPLC-MS.

          Results

          The seedling emergence and survival rate decreased significantly with the continuous number of planting years from one to three years. The root exudates, root extracts, and extracts from consecutively cultivated soils also showed significant autotoxicity against seedling emergence and growth. Ginsenosides, including R 1, Rg 1, Re, Rb 1, Rb 3, Rg 2, and Rd, were identified in the roots and consecutively cultivated soil. The ginsenosides, Rg 1, Re, Rg 2, and Rd, were identified in the root exudates. Furthermore, the ginsenosides, R 1, Rg 1, Re, Rg 2, and Rd, caused autotoxicity against seedling emergence and growth and root cell vigor at a concentration of 1.0 µg/mL.

          Conclusion

          Our results demonstrated that autotoxicity results in replant failure of Sanqi ginseng. While Sanqi ginseng consecutively cultivated, some ginsenosides can accumulate in rhizosphere soils through root exudates or root decomposition, which impedes seedling emergence and growth.

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          Most cited references7

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          Pharmacological activity of sanchi ginseng (Panax notoginseng).

          T B Ng (2006)
          The pharmacological activity and constituents of the sanchi ginseng Panax notoginseng have been reviewed. The bulk of pharmacological findings have been based on the saponins or steryl glycosides, although polysaccharides with immunopotentiating activity, proteins with antifungal, ribonuclease and xylanase activity, and a triacylglycerol (trilinolein) with antioxidant activity have been reported. Protective actions against cerebral ischaemia, beneficial effects on the cardiovascular system, and haemostatic, antioxidant, hypolipidaemic, hepatoprotective, renoprotective and estrogen-like activities have been described. Various methods for authentication of P. notoginseng are available.
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            Critical Evaluation of Techniques to Detect and Measure Cell Death – Study in a Model of UV Radiation of the Leukaemic Cell Line HL60

            The reliability of eight distinct methods (Giemsa staining, trypan blue exclusion, acridine orange/ethidium bromide (AO/EB) double staining for fluorescence microscopy and flow cytometry, propidium iodide (PI) staining, annexin V assay, TUNEL assay and DNA ladder) for detection and quantification of cell death (apoptosis and necrosis) was evaluated and compared. Each of these methods detects different morphological or biochemical features of these two processes. The comparative analysis of the 8 techniques revealed that AO/EB (read in fluorescence microscopy) provides a reliable method to measure cells in different compartments (or pathways) of cell death though it is very time consuming. PI staining and TUNEL assay were also sensitive in detecting very early signs of apoptosis, but do not allow precise quantification of apoptotic cells. These three methods were concordant in relation to induction of apoptosis and necrosis in HL60 cells with the various UV irradiation time periods tested. Both AO/EB (read by flow cytometry) and annexin V‐FITC/PI failed to detect the same number of early apoptotic cells as the other three methods. Trypan blue is valueless for this purpose. Giemsa and DNA ladder might be useful as confirmatory tests in some situations.
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              Ginsenosides stimulate the growth of soilborne pathogens of American ginseng.

              Ginseng saponins (ginsenosides) were isolated from soil associated with the roots of commercially grown American ginseng (Panax quinquefolius L.), identified via LC-MS and quantified via analytical HPLC. The ginsenosides, including F(11), Rb(1), Rb(2), Rc, Rd, Re and Rg(1), represented between 0.02 and 0.098% (average 0.06%) of the mass of the soil collected from roots annually between 1999 and 2002. The same ginsenosides were also isolated from run-off of undisturbed plants grown in pots in a greenhouse using a root exudate trapping system. To investigate (1) whether these saponins could influence the growth of pythiaceous fungi pathogenic to ginseng, and (2) whether soil levels of ginsenosides were sufficient to account for any effects, bioassays were completed using a crude saponin extract and an ecologically relevant concentration of purified ginsenosides. Thus, when cultured on media containing crude saponins, the colony weight of both Phytophthora cactorum and Pythium irregulare was significantly greater than that of control, indicating a strong growth stimulation by ginsenosides. The growth of Pythium irregulare was also significantly stimulated after addition of an ecologically relevant, low concentration (i.e. 0.06%) of purified ginsenosides to culture medium. By contrast, growth of the saprotrophic fungus Trichoderma hamatum was slightly (but not significantly) inhibited under the same conditions. These results imply that ginsenosides can act as allelopathic stimulators of the growth of pythiaceous fungi in the rhizosphere, and this may contribute to the disease(s) of this crop.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                19 February 2015
                2015
                : 10
                : 2
                : e0118555
                Affiliations
                [001]Key Laboratory of Agro-Biodiversity and Pest Management of Education Ministry of China, Yunnan Agricultural University, Kunming, China
                Northwest A&F University, CHINA
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: SZ XH YZ. Performed the experiments: MY XZ YX XM LF. Analyzed the data: BJ JL ZY. Contributed reagents/materials/analysis tools: JZ ZZ. Wrote the paper: MY SZ.

                Article
                PONE-D-14-46021
                10.1371/journal.pone.0118555
                4335038
                25695831
                b18597a5-8046-4753-b945-a026df7c06de
                Copyright @ 2015

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 14 October 2014
                : 20 January 2015
                Page count
                Figures: 6, Tables: 4, Pages: 17
                Funding
                This study was partially supported by Yunnan Development and Reform Commission, the Natural Science Foundation of China (30800731 and 31260447) and the “973” program from the Ministry of Science and Technology of China (2011CB100400). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

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